SOYBEAN

2007 greenhouse maintenance increase, Urbana, Illinois

2008 greenhouse maintenance increase, Urbana, Illinois

G. soja accessions were planted in 15-cm diameter plastic pots filled with a commercial soil mix in the greenhouse. The temperature was maintained at 25 C with a 12-h photoperiod. Twelve seeds were planted in each pot, and plants were thinned to eight per pot for inoculation. Plants were inoculated with the K-Ho1 isolate of BPMV at the unifoliolate stage. For each entry, three pots of plants were inoculated with leaf sap from healthy plants as a control and three pots of plants were inoculated with each virus isolate. Plants were monitored for symptom expression for 3 weeks, and presence of virus was determined by enzyme-linked immunosorbent assay (ELISA) 3 weeks after inoculation. The middle leaflet from young, fully expanded, uppermost trifoliolate leaves above the inoculated leaves was collected from each plant in each pot and mixed as one sample for the ELISA assay. Reactions to BPMV were classified as mild, moderate, or severe based on symptom expression.

The soybean core collection was established using the 50K SNP dataset

Study Name: Evaluation of the USDA Glycine soja Germplasm Collection: MG 000 to I (PI 65549 to PI 603146). Field Study in screened cages. 1998-1999. Urbana, IL Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters

Study Name: Evaluation of the USDA Glycine soja Germplasm Collection: MG IV to VIII (PI 65549 to PI 603146). Field Study. 1998-1999. Stoneville, MS (Lat 36 degrees 26 minutes N)

2013 Urbana Cage Misc comparing Soja seed sources

Accessions were screened to identify accessions with low or no P34 expression. Seeds were weighed to 0.5 g for the assay for each accession was used as starting material to extract total protein as the allergen of interest is expressed highly in seeds. because of their smaller size.Pulverized seed from each accession was added to 1mL TBS-T buffer [0.05MTris, 0.5MNaCl,pH7.4 with 0.05%(v/v)Tween 20 (polyoxyethylene sorbitan monolaurate)] and incubated for 30 min on a shaker (200 rpm) at room temperature. Immuno-blot assay of the seed extracts was done by a rapid and large scale method as follows: A 10-mL aliquot of each sample extract was dot-blotted on a nitrocellulose membrane (0.45 mm; Fisher Scientific) with a P34 positive control sample (cv. Kunitz). To check for equal loading of the samples on the blots, membranes were stained with Ponceau S solution (Sigma, St. Louis, MO) for 1 min and rinsed in double distilled water. The membrane was incubated in TBS buffer (0.05 M Tris and 0.5 M NaCl, pH 7.4) for 5 min and blocked (10% nonfat dry milk powder in TBS buffer) for 1 h followed by rinsing twice in TBS buffer for 5 min each. The membranes were incubated in P34 protein specific monoclonal antibody P4B5 (diluted 1: 5000 in TBS-T buffer with 0.1% milk powder) for 1 h. The membranes were washed extensively in TBS-T buffer and incubated in the secondary antibody [antimouse IgG (Fab specific) alkaline phosphatase conjugate (Sigma)] diluted 1: 10 000 in TBS-T buffer with 0.1% milk powder for 1 h and rinsed in TBS-T buffer and TBS buffer once each. The P34 protein was detected by a color substrate system [BCIP/NBT: final concentrations 0.02% (w/v) 5-bromo-4-chloro-3-indolyl phosphate and 0.03% (w/v) nitro blue tetrazolium in 70% (v/v) dimethylformamide] (Sigma).

G. soja accessions were tested with P. sojae isolate race 3 using the plug-inoculation method. Seeds were planted in 10-cm-deep flats containing a 1:1 mix of sterilized sand and soil. The seedlings were allowed to grow for 10 days in a greenhouse with a 16-h photoperiod, a 26/21?C day/night temperature, and daily watering. Race 3 P. sojae isolate was grown for 14 days on V8 juice agar. A 3-mm cork-borer then was used to cut 3-mm plugs out of the colonies. A single plug, mycelial-side down, was placed on a cotyledon immediately adjacent to the stem of each seedling. Following inoculation, all seedlings were lightly misted with water using a hand-atomizer to increase humidity and then covered with a plastic dome that fit over individual flats. The dome-covered flats were placed about 1 m under black mesh shade cloth (80% light reduction) to prevent heat buildup inside the domes. After 5 days, the domes were removed. Two days later the shade cloth was removed and the number of seedlings that survived or died was counted. If a plug fell from the cotyledon before infection, the seedling was counted as an escape. The soybean differentials Mack and Harosoy 63 were used as controls.

Accessions were bioassayed in 2000 for SCN Race 1. Peking, Pickett, PI 88788, PI 90763, and PI 437654 were used as host differentials, with Hutcheson as the susceptible control. Bioassays were repeated with the lines initially found to be resistant and all were found to be mostly susceptible.

Expression of soybean aphid resistance was characterized in choice and nonchoice tests. Choice tests were arranged in a randomized complete block design, 2 replications of 3 plants each, where aphids were allowed to move to preferred host plants, were conducted in an air-conditioned, insecticide-free greenhouse (22-25C, continuous illumination). Leaves from Williams 82 plants infested with the Urbana aphid clone were placed on top of V1-stage seedlings. The aphids moved from the infested leaf used to transfer aphids to the test seedlings within the first day after transfer, and after about 7 to 14 d, they distributed themselves on preferred hosts and developed colonies. An aphid index was calculated by multiplying the estimate of aphid population (0, no live aphids, to 3, high aphid density) by the rating of aphid damage (0, no damage, to 3, severe damage) 21 d after transfer of aphid infested leaves. The type of resistance in resistant germplasm was studied in nonchoice tests. Total aphid populations on each plant were counted

On June 26, 2002 at the Agronomy Farm of Michigan State University (MSU), 5 seeds/accession were planted in non-replicated plots 0.3 m long with a row spacing of 0.3 m. in a 12.2 x 18.3 m polypropylene cage with a 0.49 mm mesh size that is aphid- and predator- proof. At the V1 stage, 2 plants/accession were inoculated on the partially expanded trifoliate with 2 wingless aphids each, obtained from naturally infested fields on the Agronomy Farm, MSU. The aphids were left to multiply and move among plants. Plants were visually rated 4 weeks after inoculation, using the following rating scale: 0 = No aphids, plant appears normal and healthy; 1 = Less than 100 aphids per plant, plant appears normal and healthy; 2 = 101 - 300 aphids per plant, mostly on the young leaves and the tender stem at top of plant, plant appears normal and healthy; 3 = 301 - 800 aphids per plant, leaves slightly curly and shiny, young leaves and stems covered with aphids; 4 = More than 800 aphids per plant, plants stunted, leaves severely curled, yellow, covered with sooty mold and cast skins. A damage index (DI) for each accession was calculated using the following formula: DI =sum of (Scale value x No. of plants in the category)/ (4 x Total no. of plants evaluated) ? 100. The DI ranges between 0% for no infestation and 100% for the most severe damage. Three seeds/accession were also planted in 22 cm x 23 cm plastic pots on Nov. 21, 2002 in greenhouses (22-25C) at the Horticulture Research Farm at MSU. Pots were randomly arranged. The soil used was Baccto High Porosity Professional Planting mix (Michigan Peat Company. Houston, Texas). Two of the 3 plants were inoculated at the V1 stage with 2 wingless aphids each on the partially expanded trifoliate. Four weeks after inoculation, each accession was visually rated as described above for field evaluation. The DIs obtained in the field cage evaluation and in the greenhouse evaluation for the same accession were averaged as the DI for the accession.

The accessions were evaluated 2003 at the Michigan State University (MSU) Agronomy Farm in two 12.2 x 18.3 m polypropylene cages with a 0.49 mm mesh size that are aphid- and predator- proof. In each cage, a complete set of all accessions were planted as a randomized complete block. The accessions were sown on May 30 in one cage and on June 6 in the second cage. Five seeds/accession were planted in plots 0.3 m long with a row spacing of 0.3 m. At the V1 stage, 2 plants/accession were inoculated on the partially expanded trifoliate with 2 wingless aphids each, obtained from naturally infested fields on the MSU Agronomy Farm. The aphids were left to multiply and move among plants. Plants were visually rated 4 weeks after inoculation, using the following rating scale: 0 = No aphids, plant appears normal and healthy; 1 = Less than 100 aphids per plant, plant appears normal and healthy; 2 = 101 - 300 aphids per plant, mostly on the young leaves and the tender stem at top of plant, plant appears normal and healthy; 3 = 301 - 800 aphids per plant, leaves slightly curly and shiny, young leaves and stems covered with aphids; 4 = More than 800 aphids per plant, plants stunted, leaves severely curled, yellow, covered with sooty mold and cast skins. A damage index (DI) for each accession was calculated using the following formula: DI =sum of (Scale value x No. of plants in the category)/ (4 x Total no. of plants evaluated) ? 100. The DI ranges between 0% for no infestation and 100% for the most severe damage. The DIs obtained in both field cages for the same accession were averaged as the DI for the accession.

Methods: Expression of soybean aphid (SA) resistance was characterized among 496 soybean lines in a twice-replicated field-plot test at the Eastern South Dakota Soil and Water Research Farm near Brookings, SD, in 2009. Natural infestations of SA occurred but were supplemented by placing individual stems of infested colony plants into the upper canopy of experimental plants on July 28 when plants ranged mainly from the V5 to R1 growth stages. Experimental plants were infested with SA obtained from a virus-free, multiclonal stock colony reared on the aphid-susceptible soybean line Pioneer 91B91 at NCARL. Stems used to infest field plots averaged 456 and 603 SA respectively for each replicate. Soybean infestations were allowed to progress through the summer. Test lines were evaluated using a rating scale that ranged from 1 to 5, where 1 = 0 to 25 aphids per plant; 2 = 26 to 100 aphids per plant; 3 = 101 to 500 aphids per plant; 4 = 501 to 3,000 aphids per plant; and 5 = more than 3,000 aphids per plant. Plants in replicate 1 were rated on 25 Aug and 31 Aug, and those in replicate 2 were rated on 26 Aug and Sep 2. Two persons rated each line on each date. Ratings were averaged across raters and dates. A line was considered resistant if its average rating was less than 3.0; moderately resistant if its rating was from 3.0 to 3.9; and susceptible if its rating was greater than 3.9. Six lines were resistant to SA, and these lines were aphid-resistant checks. Twelve lines were moderately resistant, including seven that have not been previously reported as aphid-resistant and five others that were also previously reported as aphid-resistant. The remaining lines were susceptible, including PI 230977, which was a resistant check .

Expression of soybean aphid (SA) resistance was characterized among 746 soybean accessions in 56 growth chamber tests at the North Central Agricultural Research Laboratory, Brookings, SD, from 2012 through 2015. Test plants were prepared by placing two seeds of a particular soybean accession into an 8.5-cm square plastic pot filled with a 2:1:1 mixture of soil, vermiculite, and peat moss. Ten to 12 days after planting, pots were thinned to one plant by selecting for similarly sized seedlings among test accessions, and the soil surface was then covered with a thin (ca. 2 cm) layer of sand. Two wks after planting, test accessions were placed into a plastic tray that held 18 square pots. Fifty-four of the 56 tests used a RCBD in which each tray comprised a replicate consisting of 14 test accessions, a susceptible check, a resistant check, and two SA-source plants. The other two tests used a CRD: one in which two paired trays comprised a replicate of 30 test accessions, and the other with two replicates of 6 test accessions per tray. Tests had from six to eight replications. Each source plant was a 4-wk-old PI 667735 soybean plant with about 250 SA that were derived from a polyclonal stock colony established in 2009, were reared on PI 667735 soybean, and had shown no virulence to known SA-resistant lines. Source plants were placed at one of two foci in each tray equidistant from immediately surrounding test accessions for even dispersion of SA. The source plants were clipped at soil level at the start of each test to induce wilting and subsequent dispersal of SA to test plants, and clipped plants were supported by attaching them via paper clip to a small wooden stake in the soil. Tests were run in environmental chambers at 16:8 L:D photoregime and temperatures from about 18oC (nighttime) to 22oC (daytime). Two weeks later when test plants were in the second trifoliolate leaf stage (V2 stage), infestations were rated by two researchers on a 1-to-6 scale, with 1 = 0 to 50, 2 = 51 to 100, 3 = 101 to 150, 4 = 151 to 200, 5 = 201 to 250, and 6 with >250 SA per test plant, respectively. For a small number of accessions that were included in more than one test, individual ratings were averaged across tests. Medians of infestation ratings are reported for each accession. Accessions were pooled across tests and ranked by median infestation rating. An accession was classified as resistant if its median rating was less than 3.0; moderately resistant if median rating was from 3.0 to 4.9; and susceptible if rating exceeded 4.9. Results show that the vast majority of accessions were susceptible to SA. However, 10 accessions were resistant to SA, and 23 were moderately resistant.

A total of 334 soybean genotypes including resistant and susceptible checks were screened for resistance to the soybean aphid (SA) in two greenhouse tests in 2011 and spring 2012. Genotypes included 33 from maturity group (MG) 00 and 293 from MG 0, and six resistant and two susceptible checks of later maturity. Each test was replicated three times. All 334 genotypes were evaluated in a greenhouse at South Dakota State University (SDSU) using a caged test and a non-caged test. Greenhouse test plants were inoculated with two, wingless, adult SA from a laboratory colony at SDSU. In the caged test, total SA per plant were counted at 1 wk and at 2 wks after inoculation; in the non-cage test, SA populations were scored by using a 1-to-5 scale at 2 weeks after inoculation (1, <=25 aphids per plant, plant appears normal and healthy; 2 = 26 to 100 aphids per plant, plant appears normal and healthy; 3 = 101 to 200 aphids per plant, older leaves may be slightly yellowing; 4 = 201 to 500 aphids per plant, plant appears slightly stunted with yellowing of older leaves and minor curling of young leaves, and slight appearance of sooty mold on stem and leaves; 5, > 500 aphids per plant, often with obvious symptoms on foliage such as leaf curling, yellowing and sooty mold development). Four genotypes (PI 430491, PI 464911, PI 603432B, and PI 603712) were resistant, and three genotypes (PI 612759B, PI 200595, and PI 603426D) were moderately resistant. Five of six resistant control lines (PI 71506, PI 243540, PI 567541B, PI 567543C, and PI 567597C) were resistant, but PI 548663 was susceptible. All other lines were susceptible.

A total of 334 soybean genotypes including resistant and susceptible checks were screened for resistance to the soybean aphid (SA) in three field tests in summer 2011. Genotypes included 33 from maturity group (MG) 00 and 293 from MG 0, and six resistant and two susceptible checks of later maturity. Each test was replicated three times. One of the field evaluations was conducted at the Eastern South Dakota Research Farm near Brookings, and the other two field tests were conducted at the Aurora Research Farm, South Dakota State University. At Brookings, a subset of 330 accessions was tested. SA infestations were scored on 9 Aug by using a 1-to-6 scale (1, =<25 aphids per plant, plant appears normal and healthy; 2 = 26 to 100 aphids per plant, plant appears normal and healthy; 3 = 101 to 200 aphids per plant, older leaves may be slightly yellowing; 4 = 201 to 500 aphids per plant, plant appears slightly stunted with yellowing of older leaves and minor curling of young leaves, and slight appearance of sooty mold on stem and leaves; 5, > 500 aphids per plant, often with obvious symptoms on foliage such as leaf curling, yellowing and sooty mold development; 6, > 500 aphids per plant, with extensive plant damage (e.g. curling, yellowing and sooty mold development). Natural infestations of SA in each replicate plot were supplemented by placing an aphid-infested stem across the plant canopy on 19 July; supplemental aphids were obtained from a colony housed at the USDA-ARS laboratory near Brookings. Most of the test plants were at R4 to R6 developmental stages when scored. A subset of 323 soybean genotypes was evaluated in one of two tests at Aurora when most of the genotypes were at R4 to R5 developmental stages. SA infestations were scored by using a 1-to-5 scale (similar as above). In the second test at Aurora, 64 selected genotypes plus 6 resistant and 2 susceptible checks. Test plants were inoculated by putting a leaflet or a part of a leaflet with 20 to 25 mixed-aged SA on the central plant of each plot at the V2 to V3 stage on 3 July. SA-infested leaves were obtained from colony plants at SDSU. Two weeks later, total number of aphids per plant was counted on five plants within each plot. Genotypes with <200 aphids per plant at the first count (and also resistant and susceptible checks) were counted again on the same five plants at four weeks after inoculation. Six weeks after inoculation, by which time most of the genotypes were at R5-R6 stage, all experimental plots were scored using the 1-to-6 scale PI 603712 was the only test accession rated as resistant, and PI 430491 was moderately resistant. Control lines PI 71506, PI 567541B, and PI 567543C were moderately resistant; however, control lines PI 243540, 543663, and 567597C were susceptible. All other lines were susceptible to soybean aphid.

A colony of soybean aphid (Aphis glycines Matsumura) was established in a growth chamber at Ohio Agricultural Research and Development Center (OARDC), Wooster, OH, during the summer of 2005 by collecting aphids from nearby soybean fields. The colony was maintained on seedlings of cultivar Williams 82 placed inside the growth chamber at temperatures between 22 and 24C with a photosynthetically active radiation of 330 umol m/2 s/1 for 15 h daily and 60 to 70% relative humidity. The colony was restarted on a fresh batch of seedlings every 3 to 4 wk by transferring aphids from the old seedlings to new ones. The experiment was conducted during September and October of 2005. Plants were grown in 10-cm-deep by 25-cm-wide by 50-cm-long plastic flats with a 8-cm space between plants and a 10-cm space between rows under 15 h light at approximately 24?C days and 9 h dark at 20?C nights. Each experimental unit was a row of three seedlings and five rows were accommodated in a flat. The experiment was arranged in a randomized complete block design (RCBD) with two replicates. At the V1-stage, seedlings were infested with 20 to 30 aphids of all developmental stages except the winged aphids by placing an infested leaf section between the petiole of the youngest expanding leaf and the stem. Aphids migrated to the leaves and stems of the seedlings within several hours after putting the infested leaf sections on the seedlings. The aphids were allowed to multiply and move freely among plants in the experiment. The plants were bottom watered to avoid disturbing the aphids. Fourteen days after infestation (DAI), each experimental unit was assigned an aphid score between 1 and 5, where 1 = <25 aphids per plant, 2 = 25 to 100 aphids per plant, 3 = 101 to 200 aphids per plant, 4 = 201 to 400 aphids per plant, and 5 = >400 aphids per plant. the LSD 0.05=0.6. Any score over 3.0 was considered highly susceptible.

A colony of soybean aphid (SA, Aphis glycines Matsumura) was established in a growth chamber at Ohio Agricultural Research and Development Center (OARDC), Wooster, OH, during the summer of 2005 by collecting aphids from nearby soybean fields. The colony was maintained on seedlings of cultivar Williams 82 placed inside the growth chamber at temperatures between 22 and 24C with a photosynthetically active radiation of 330 umol m/2 s/1 for 15 h daily and 60 to 70% relative humidity. The colony was restarted on a fresh batch of seedlings every 3 to 4 wk by transferring aphids from the old seedlings to new ones. Seeds were planted in 15 cm deep and 4 cm in diameter plastic conetainers and thinned to one seedling per conetainer at the unifoliolate-stage. Each conetainer was an experimental unit, and the experiment was arranged in a RCBD with two replicates. At the V1-stage, seedlings were infested with 20 to 30 aphids of all developmental stages except the winged aphids by placing an infested leaf section between the petiole of the youngest expanding leaf and the stem. Aphids migrated to the leaves and stems of the seedlings within several hours after putting the infested leaf sections on the seedlings. The aphids were allowed to multiply and move freely among plants in the experiment. The plants were bottom watered to avoid disturbing the aphids. Fourteen days after infestation (DAI), each experimental unit was assigned an aphid score between 1 and 5, where 1 = <25 aphids per plant, 2 = 25 to 100 aphids per plant, 3 = 101 to 200 aphids per plant, 4 = 201 to 400 aphids per plant, and 5 = >400 aphids per plant. On 28 DAI, the plants were evaluated for susceptibility to SA by using a plant damage score of 1 = <25 aphids and plant appears normal and healthy; 2 = 25 to 100 aphids per plant and plant appears normal and healthy; 3 = 101 to 300 aphids per plant and plant appears slightly stunted with slight yellowing of older leaves; 4 = 301 to 600 aphids per plant and plant appears moderately stunted with yellowing of older leaves and slight curling of young leaves; and 5 = >600 aphids per plant and plant appears severely stunted with severely curled and yellow leaves and most of the stem and leaf surfaces are covered with sooty mold and cast skins. The aphid number per plant was included in the plant damage scores because this was particularly helpful in differentiating plants with scores of 1 and 2. In both cases, the plants were fully healthy and normal looking but a plant with the score of 1 had very few if any aphids (<25) on it while a plant with a score of 2 had up to 100 aphids in some instances. This approach of separating these two types of plants allowed a conservative classification of plants as SA resistant. The plants with relatively higher numbers of aphids were classified as moderately resistant. Also, the aphid numbers at 14 DAI were not always in synchrony with the aphid numbers at 28 DAI. Some moderately resistant plants had the same number of aphids at 14 DAI as the resistant plants, but over the next 2 wk the aphid numbers on these plants increased slowly while the aphid numbers on the resistant plants did not increase at all or declined. The aphid susceptibility index (ASI) was calculated by multiplying the aphid score at 14 DAI by the plant damage score at 28 DAI with a possible ASI between 1 and 25. Resistant lines were tested again with choice and no choice tests.

G. max accessions were planted in 15-cm diameter plastic pots filled with a commercial soil mix in the greenhouse. The temperature was maintained at 25 C with a 12-h photoperiod. Twelve seeds were planted in each pot, and plants were thinned to eight per pot for inoculation. Plants were inoculated with the K-Ho1 isolate of BPMV at the unifoliolate stage. For each entry, three pots of plants were inoculated with leaf sap from healthy plants as a control and three pots of plants were inoculated with each virus isolate. Plants were monitored for symptom expression for 3 weeks, and presence of virus was determined by enzyme-linked immunosorbent assay (ELISA) 3 weeks after inoculation. The middle leaflet from young, fully expanded, uppermost trifoliolate leaves above the inoculated leaves was collected from each plant in each pot and mixed as one sample for the ELISA assay. Reactions to BPMV were classified as mild, moderate, or severe based on symptom expression.

Fifty-two soybean accessions were selected to represent the major ancestral lines of NA cultivars. Williams 82 was used as the susceptible check. Bean pod mottle virus isolate 98 was collected from infected soybean at the CSREC, Urbana, IL, and was maintained by continuous transfer and stored long term in desiccated refrigerated leaves. It belongs to subgroup I. Screening was done in 2002 and 2003. In the greenhouse, eight seeds of each line tested were planted in a pasteurized soil mixture (1:1:1 soil-sand-peat) in plastic flats (52- by 37-cm) or in 10-cm plastic pots with a soilless mix (Sunshine Mix LC1, Sun GroHorticulture, Inc., Bellevue, WA) and covered with coarse vermiculite. All entries without symptoms were retested using five to six plants. Inoculum consisted of extracts from infected leaves of Williams 82 plants maintained in the greenhouse that were prepared by homogenizing infected leaves in chilled 0.025 M KPO4 buffer, pH 7.1, plus 0.01 M sodium sulfite with sterilized pestles and mortars. Pestles were used to apply inoculum to carborundum-dusted leaf surfaces. Plants were inoculated with BPMV 7 to 10 d after planting at growth stage Vi. Two to three weeks after inoculation, trifoliolate leaves were examined for systemic virus symptoms. Resistance reactions were recorded when plants were symptomless. In cases where it was difficult to distinguish between mild and no symptoms, ELISA results were used to determine the reaction. At least three plants were tested by ELISA to verify phenotypic observations. Lines with a negative ELISA reaction and no systemic symptoms were considered virus resistant.

Accessions in maturity groups 000 to IV were evaluated between 1981 and 1984 for resistance to Brown Stem Rot (BSR) caused by Phialophora gregata. Lines were evaluated in naturally infested fields at latitudes to which they were adapted. In 1985 and 1986, putatively resistant lines were evaluated in replicated tests. The early maturity material was tested at Rosemount, MN, Hancock, WI, and Ames, IA. The late maturity material was tested at Ames, IA, and Urbana, IL. Each line was also evaluated in the greenhouse at Urbana, IL. Ratings at Hancock, WI, and in the greenhouse were based on leaf symptoms. All other ratings were based on stem symptoms.

Brown Stem Rot Resistance in Soybean Germ Plasm from Central China. Based on field results, putatively resistant lines were selected and evaluated in the greenhouse from Oct. 1994 through May 1996, using the root-dip inoculation method of Willmot and Nickell. Sixty-four accessions, 3 susceptible, and 3 resistant standards were replicated 6 times in a randomized complete block design. The resistant standards were L78-4094 (Rbs1), PI 437833 (Rbs2), and PI 437970 (Rbs3). The susceptible standards were cvs. Century 84, Resnik, and Flyer. Entries were inoculated with a monoconidial type I isolate of P. gregata, PgOh2. Plants were evaluated for BSR reaction after 8 to 9 weeks (R1 to R3 growth stages). Individual pots were rated for incidence of foliar (percentage of plants with foliar symptoms) and stem symptoms. Stem symptoms were measured by splitting each stem lengthwise and recording the proportion of total nodes exhibiting internal browning. Accessions were classified as resistant if the foliar and stem symptoms were significantly lower than the corresponding means of the susceptible standards and not significantly different from the corresponding means of the resistant standards.

Evaluation of Soybean Plant Introductions from China for Resistance to Brown Stem Rot. Modified root dip inoculation as described in: Sebastian, S.A., C.D. Nickell, and L.E. Gray, 1985. Efficient selection for brown stem rot resistance in soybeans under greenhouse conditions. Crop Sci 25:753-757. Lines were considered resistant if they were less susceptible than the most susceptible resistant check (PI 437833 with 0.02% brown nodes/plant BSR score). Any PI's that showed a resistant or unconvincing response the first time were retested. The screening was done between Sept and May 1999-2001.

Samples reported upon are from the USDA Germplasm Collection based in Urbana, IL. Soybean samples were scanned by NIR at the University of Minnesota's Soybean Breeding Project laboratory. Whole soybean samples received from USDA-NASS were first ground and then scanned on a FOSS 6500 NIR Instrument. NIR Spectra from the FOSS 6500 were predicted using ISIPredict Software version 1.10.2.4842. Calibrations, provided by FOSS North America, were used to predict soybean composition from the NIR spectra. Presented as a comment for each reported compositional value is a value for Global "H". As stated in FOSS Application Note FNA538-01. "Global H value-measures how far this sample is from the center of the calibration database (The samples selected for this spectra). The distance is normalized and a value exceeding 3 is typically considered an outlier." A number of factors can contribute to a sample being flagged as an outlier when using a given set of calibration equations. With soybeans, this could include seed coat color if the calibrations were developed using only soybeans with yellow seed coats. In addition to soybeans with yellow seed coats, this dataset includes soybeans bearing other colors of seed coat. Sample preparation and presentation to the instrument can also contribute to higher H values. In addition to the previously stated reasons, another reason for a high H value could be that a sample is truly unique.

Chorosis scores were taken on Maturity Group I soybeans planted in 2001 at Boyd, Minnesota. The chlorosis score is a composite of an early and late chlorosis score, 2 reps. The scores are least square means using neighboring plant scores as a covariate to adjust the mean. This test was repeated at Morgan, MN (see SOYBEAN.CHLOROSIS.LS.MORGAN.2001)

Chlorosis scores were taken on Maturity Group 0 soybeans planted in 2004 at Buffalo Lake, Minnesota. The chlorosis score is a late chlorosis score, 3 reps. The scores are least square means using neighboring plant scores as a covariate to adjust the mean. The minimum score was 1.0, maximum 5.0, average 3.4. Early chlorosis scores are reported in the comments for each accession. This test was repeated in Wood Lake, Morgan, and SleepyEye, MN (see SOYBEAN.CHLOROSIS.LS.WOODLAKE.2004, SOYBEAN.CHLOROSIS.LS.MORGAN1.2004, SOYBEAN.CHLOROSIS.LS.MORGAN2.2004, SOYBEAN.CHLOROSIS.LS.SLEEPYEYE.2004

Chorosis scores were taken on Maturity Group I soybeans planted in 2001 at Morgan, Minnesota. The chlorosis score is a composite of an early and late chlorosis score, 2 reps. The scores are least square means using neighboring plant scores as a covariate to adjust the mean. This test was repeated at Boyd, MN (see SOYBEAN.CHLOROSIS.LS.BOYD.2001)

Chlorosis scores were taken on Maturity Group 0 soybeans planted in 2004 at Morgan, Minnesota. The chlorosis score is a late chlorosis score, 3 reps. The scores are least square means using neighboring plant scores as a covariate to adjust the mean. The minimum score was 2.4, maximum 5.0, average 4.4. Early chlorosis scores are reported in the comments for each accession. This test was repeated at a second location in Morgan and in Buffalo lake, Wood lake, and SleepyEye, MN (see SOYBEAN.CHLOROSIS.LS.BUFFALOLAKE.2004, SOYBEAN.CHLOROSIS.LS.WOODLAKE.2004, SOYBEAN.CHLOROSIS.LS.MORGAN2.2004, SOYBEAN.CHLOROSIS.LS.SLEEPYEYE.2004)

Chlorosis scores were taken on Maturity Group 0 soybeans planted in 2004 at Morgan, Minnesota. The chlorosis score is a late chlorosis score, 3 reps. The scores are least square means using neighboring plant scores as a covariate to adjust the mean. The minimum score was 1.6, maximum 5.0, average 3.5. Early chlorosis scores are reported in the comments for each accession. This test was repeated at a second location in Morgan and in Buffalo lake, Wood lake, and SleepyEye, MN (see SOYBEAN.CHLOROSIS.LS.BUFFALOLAKE.2004, SOYBEAN.CHLOROSIS.LS.WOODLAKE.2004, SOYBEAN.CHLOROSIS.LS.MORGAN1.2004, SOYBEAN.CHLOROSIS.LS.SLEEPYEYE.2004)

Chlorosis scores were taken on Maturity Group 0 soybeans planted in 2004 at SleepyEye, Minnesota. The chlorosis score is a late chlorosis score, 3 reps. The scores are least square means using neighboring plant scores as a covariate to adjust the mean. The minimum score was 1.0, maximum 3.7, average 1.2. Early chlorosis scores are reported in the comments for each accession. This test was repeated in Buffalo lake, Wood lake, and Morgan, MN (see SOYBEAN.CHLOROSIS.LS.BUFFALOLAKE.2004, SOYBEAN.CHLOROSIS.LS.WOODLAKE.2004, SOYBEAN.CHLOROSIS.LS.MORGAN1.2004, SOYBEAN.CHLOROSIS.LS.MORGAN2.2004)

Chlorosis scores were taken on Maturity Group 0 soybeans planted in 2004 at Wood Lake, Minnesota. The chlorosis score is a late chlorosis score, 3 reps. The scores are least square means using neighboring plant scores as a covariate to adjust the mean. The minimum score was 1.1, maximum 4.5, average 2.7. Early chlorosis scores are reported in the comments for each accession. This test was repeated in Buffalo lake, Morgan, and SleepyEye, MN (see SOYBEAN.CHLOROSIS.LS.BUFFALOLAKE.2004, SOYBEAN.CHLOROSIS.LS.MORGAN1.2004, SOYBEAN.CHLOROSIS.LS.MORGAN2.2004, SOYBEAN.CHLOROSIS.LS.SLEEPYEYE.2004)

The soybean core collection was established using the multivariate proportional selection strategy

The soybean core collection was established using the 50K SNP dataset

Methods: Levels of defoliation from chewing insects were characterized among 330 soybean lines in a thrice-replicated field-plot test at the Eastern South Dakota Soil and Water Research Farm near Brookings, SD, in 2010. Soybeans were planted at 4-cm depth on 26 (replicate 1) and 27 May (replicates 2 and 3) with a two-row cone seeder at 25 seeds per replicate plot (1.1 m). Row width was 0.75 m. Weeds within plots were managed by mechanical field cultivation, herbicides, and hoeing. Soybean plants incurred defoliation mainly during mid- to late season from natural infestations of chewing insects, primarily from green cloverworms, soybean loopers, and grasshoppers, and to a minor extent from bean leaf beetles. Test lines were evaluated using a rating scale that ranged from 1 to 3, in which '1' = 0 to 25 percent defoliation of plants within a replicate plot, mostly from a few, small rounded feeding areas within interveinal area of leaves ('shot holes') and along leaf edges, with leaflets intact and virtually no yellowing of leaves. A rating of '2' = 26 to 74 percent defoliation, with numerous shot holes and large sections of leaf edges eaten, but leaflets mostly intact; and minor yellowing of leaves. A rating of '3' = greater than 75 percent defoliation, with large coalesced feeding areas, often with entire leaves consumed or only veins remaining; moderate yellowing of remaining leaf tissue. Plants in replicates 1 and 2 were rated on 18 Aug, and those in replicate 3 were rated on 19 Aug. Two persons provided a consensus rating for each replicate per line on each date. Ratings are reported as averages across replicates. Plants ranged from vegetative V6 (in late maturity checks) to full-seed R6 growth stages (in early MG) when rated. Ratings of 1.0 to 1.3 were considered resistant (R), 1.7 to 2.0 mostly resistant (MR), and 2.1 and above susceptible (S). Of the 330 soybean lines, three lines had average ratings of 1 (PI 468906, PI 468909, PI 612753A), 16 lines had average ratings of 1.3, and 67 had average ratings of 1.7. The remaining 244 lines had ratings of 2 or greater, including 13 with average ratings of 3.

Methods: Levels of defoliation from chewing insects were characterized among 11 soybean lines in a thrice-replicated field-plot test at the Eastern South Dakota Soil and Water Research Farm near Brookings, SD, in 2010. Soybean seed was obtained from the USDA-ARS Soybean Germplasm Center, except that lines K1639-1 and KS4202 were obtained from Kansas State University. Soybeans were planted at 4-cm depth on 21 May with a two-row cone seeder at 25 seeds per replicate plot (1.1 m). Row width was 30 inches. Weeds within plots were managed by mechanical field cultivation, herbicides, and hoeing. Soybean plants incurred defoliation mainly during mid- to late season from natural infestations of chewing insects, including green cloverworms, soybean loopers, and grasshoppers, and to a minor extent from bean leaf beetles. Test lines were evaluated using a rating scale that ranged from 1 to 3, in which '1' = 0 to 25 percent defoliation of plants within a replicate plot, mostly from a few, small rounded feeding areas within interveinal area of leaves ('shot holes') and along leaf edges, with leaflets intact and virtually no yellowing of leaves. A rating of '2' = 26 to 74 percent defoliation, with numerous shot holes and large sections of leaf edges eaten, but leaflets mostly intact; and minor yellowing of leaves. A rating of '3' = greater than 75 percent defoliation, with large coalesced feeding areas, often with entire leaves consumed or only veins remaining; moderate yellowing of leaf tissue. Plants were rated on 19 Aug. Two persons produced a consensus rating for each replicate per line on each date. Ratings are reported as averages across replicates. Plants ranged from vegetative V6 (in late maturity checks) to full-seed R6 growth stages (in early MG) when rated because of the wide span in maturity of individual lines. PI 227687, PI 548654 and KS4202 had average ratings of 1, and PI 548374 had an average rating of 1.33. Other lines had ratings of 2 or greater. Ratings of 1.0 to 1.3 were considered resistant (R), 1.7 to 2.0 mostly resistant (MR), and 2.1 and above susceptible (S).

Observations supplied by the developer for the specific variety. Contact the developer for additional information.

Soybean Germplasm screened in the field during 1974, 1975, 1976 and 1977 for corn earworm resistance.

Descriptive characteristics, agronomic performance, seed composition and disease reaction data of soybean accessions FC 01.547 to PI 266.807 in maturity groups I and II was recorded. Also included were cultivars, in these same maturity groups, developed at public institutions in the United States and Canada, and released by 1966. Data were gathered from a test planted on the Agronomy South Farm, University of Illinois, Urbana (Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters Soil class: Flanagan Silt Loam). Two replications of each maturity group were grown in plots two rows wide and 2.4 m long, with 100 cm between rows. Replication 1 was planted on May 14, 1964 and replication 2 was planted on June 3, 1964. Because the plots were not end trimmed at maturity, plots were considered to be 3.1 meters long when original yield data were converted to a per acre value. All original yield per acre values have been converted to megagrams per hectare at 13 percent moisture. Tests were blocked by maturity group to minimize competition effects. A composite sample from the two replications was used to obtain seed composition data. Protein percentages were obtained using the Kjeldahl method. Oil percentages were determined by the extraction method Methionine values were obtained using Krober's modification of the McCarthy-Sullivan colorimetric method using enzymatic hydrolysis. Fatty acid composition was obtained using the spectrophotometric method of Collins and Sedgwick.

Descriptive characteristics, agronomic performance, seed composition and disease reaction data of soybean accessions FC 01.547 to PI 266.807 in maturity groups III and IV were recorded. Also included were cultivars, in these same maturity groups, developed at public institutions in the United States and Canada, and released by 1966. Data were gathered from a test planted on the Agronomy South Farm, University of Illinois, Urbana (Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters Soil class: Flanagan Silt Loam). The accessions were planted on May 20, 1965 and on May 26 & 27, 1966, with 1 replication/year. Plots were two rows wide and 2.4 m long, with 100 cm between rows. Because the plots were not end trimmed at maturity, plots were considered to be 3.1 meters long when original yield data were converted to a per acre value. All original yield per acre values have been converted to megagrams per hectare at 13 percent moisture. Since there were no border rows in any of the tests, all tests were blocked by maturity group to minimize competition effects A composite sample from the two replications was used to obtain seed composition data. Protein percentages were obtained using the Kjeldahl method. Oil percentages were determined with nuclear magnetic resonance. Methionine values were obtained using Krober's modification of the McCarthy-Sullivan colorimetric method using enzymatic hydrolysis. Gas-liquid chromatography was used to obtain fatty acid composition.

Descriptive characteristics, agronomic performance, seed composition and disease reaction data of soybean accessions FC 01.547 to PI 266.807 in maturity groups 000 through 0 was recorded. Also included are cultivars, in these same maturity groups, developed at public institutions in the United States and Canada, and released by 1966. Descriptive data were obtained from observations of plants and seeds at Urbana, and a chlorosis score obtained from single-row plots grown in a high-lime field near Lamberton, Minnesota, in 1964. Performance data were gathered from a test planted om May 24, 1963 at St. Paul, MN (45.0 N. lat.). Maturity group 000 had not yet been designated when this evaluation was conducted, so accessions were tested as Groups 00 and 0. Two replications of each maturity group were grown in plots two rows wide and 2.4 m long, with 90 cm between rows. All original yield per acre values have been converted to megagrams per hectare at 13 percent moisture. Plots were blocked by maturity group to minimize competition effects. A composite sample from the two replications was used to obtain seed composition data. Protein percentages were obtained using the Kjeldahl method. Oil percentages were determined by the extraction method. Methionine values were obtained using Krober's modification of the McCarthy-Sullivan colorimetric method using enzymatic hydrolysis. Fatty acid composition was obtained using the spectrophotometric method of Collins and Sedgwick.

Study Name: Soybean Evaluation 2 of Maturity Groups 000 to IV. Field Study. 1980-1981. Maturity Groups I through III Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters

Study Name: Soybean Evaluation 2 of Maturity Groups 000 to IV. Field Study. 1980-1981 Maturity Group early IV's Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters

Study Name: Soybean Evaluation 2 of Maturity Groups 000 to IV. Field Study. 1980-1981. Maturity Group late IV Latitude: 38 Degrees 02 Minutes N Longitude: 84 Degrees 30 Minutes W

Study Name: Soybean Evaluation 2 of Maturity Groups 000 to IV. Field Study. 1981-1982. Mat Groups 000 to 0 Latitude: 45 Degrees 00 Minutes N Longitude: 93 Degrees 07 Minutes W

Study Name: Soybean Evaluation 3 of Maturity Groups 000 to IV Experiment Type: Field Field Study Year: 1983 Exp. Design: Maturity Group I Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters Year started: //1982 Year ended: //1983 Experiment length: 2 years

Study Name: Soybean Evaluation 3 of Maturity Groups 000 to IV Experiment Type: Field Field Study Year: 1983 Exp. Design: Mat Groups III & IV Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters Year started: //1982 Year ended: //1983 Experiment length: 2 years

Study Name: Soybean Evaluation 3 of Maturity Groups 000 to IV. Field Study. 1982, 1984. Maturity Group II. Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters

Study Name: Soybean Evaluation 3 of Maturity Groups 000 to IV Experiment Type: Field Field Study Year: 1983 Exp. Design: Mat Groups 000 & 00 Latitude: 45 Degrees 00 Minutes N Longitude: 93 Degrees 07 Minutes W Year started: //1982 Year ended: //1983 Experiment length: 2 years

Study Name: Soybean Evaluation 3 of Maturity Groups 000 to IV Experiment Type: Field Field Study Year: 1983 Exp. Design: Maturity Group 0 Latitude: 45 Degrees 00 Minutes N Longitude: 93 Degrees 07 Minutes W Year started: //1982 Year ended: //1983 Experiment length: 2 years

Study Name: Soybean Evaluation 4 of Maturity Groups 000 to IV Experiment Type: Field Field Study Year: 1987 Exp. Design: Mat Groups I to IV Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters Year started: //1986 Year ended: //1987 Experiment length: 2 years Comment: Evaluated 2 years with 1 replication per year. Planted on May 6, 1986 and on May 5, 1987. Data presented is an average of these 2 replications.

Study Name: Soybean Evaluation 4 of Maturity Groups 000 to IV Experiment Type: Field Field Study Year: 1987 Exp. Design: Mat Groups 000 to 0 Latitude: 45 Degrees 00 Minutes N Longitude: 93 Degrees 07 Minutes W Year started: //1986 Year ended: //1987 Experiment length: 2 years Comment: Evaluated 2 years with 1 replication per year. Planted on May 22, 1986 and June 1, 1987. Data presented is an average of these 2 replications.

Accessions in maturity groups I to IV, PI 490765 to PI 507573 were evaluated for descriptive characteristics, agronomic performance, and seed composition on the Agronomy-Plant Pathology South Farm, University of Illinois, Urbana (40.13N lat.). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 1988. All tests were replicated once per year. Plots were four rows wide, with rows 4.7 m long and 75 cm between rows. The test was blocked by maturity group. The center two rows were end trimmed to 3.2 m one month after planting. Only the center two rows of each plot were harvested for yield. Planting dates were May 13, 1989 and May 8, 1990. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters. Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis.

Accessions in maturity groups 000 to 0, PI 490765 to PI 507573 were evaluated for descriptive characteristics, agronomic performance, and seed composition at the University of Minnesota, St. Paul (45.0N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 1987. All tests were replicated once per year. Plots were four rows wide, with rows 3.7 m long and 75 cm between rows. The test was blocked by maturity group. The center two rows were end trimmed to 2.4 m when plants reached maturity. Only the center two rows of each plot were harvested for yield. Planting dates were May 26, 1990 and May 31, 1991. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters. Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis.

Descriptive characteristics, agronomic performance, and seed composition data of soybean (Glycine max (L.) Merr.) germplasm were evaluated on accessions initially classified in maturity groups I through IV in the range from PI 578371 to PI 612761, plus some accessions that were omitted from earlier publications. Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 2001. The accessions were evaluated in 2001 and 2002 in Urbana, Illinois (Lat. 40o 00' N). Planting dates were May 4, 2001 and May 27, 2002. Plots were 4 m long with 4 rows 76 cm apart. They were trimmed to 2.4 m after maturity and the middle two rows harvested. Accessions initially classified as maturity group I were stunted by herbicide damage and were not harvested in 2002. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989, Bannon et al. 1982). Oil and protein for samples with yellow seed coats was analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Descriptive characteristics, agronomic performance, and seed composition data of soybean (Glycine max (L.) Merr.) germplasm were evaluated on accessions initially classified in maturity groups I through IV in the range from PI 612762 to PI 654045, plus some accessions that were omitted from earlier publications. Also included were cultivars in the same maturity groups, developed in the United States and Canada, and released by 2009.

Accessions in maturity groups I through IV ranging from PI 507670 to PI 574486 were evaluated for descriptive characteristics, agronomic performance, and seed composition in 1994 and 1995 in Urbana, Illinois (Lat. 40o 00' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by early 1993. Tests were replicated once per year. Planting dates were May 20-21, 1994 and May 31-June 1, 1995. Plots were 4 m long with 4 rows 76 cm apart. They were trimmed to 3.2 m after maturity and the middle two rows harvested. Because of the difference in planting date, plants flowered about 5 days later in 1995 than in 1994. Accessions in maturity groups I and II also matured about 5 days later in 1995 than in 1994. Rainfall in August and early September of 1994 was much greater than in the same period of 1995, which may have contributed to a longer growing season for the later maturing accessions in 1994. A frost on September 22, 1995 cut the growing season short; therefore maturity dates from 1995 were not included if the accessions had not matured before the frost. Accessions in maturity group III matured about 10 days earlier in 1995 than in 1994 where data from both years were available. The seed composition data were collected at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. To obtain oil and protein percentages of the seeds with yellow seed coats, approximately 7 g of seeds were placed in a beaker and dried in a forced air oven for 3 hours at 130 C. The seeds were then transferred to 50 g bottles, sealed, and allowed to cool for 1 hour. Samples were then ground in a Varco electric dry food grinder and returned to the 50 g bottles. Seed samples were analyzed with an Infratec 1255 food and grain analyzer (Perstorp Analytical Company). Samples were scanned from 800 - 1100 nm. The analyzer was calibrated with at least 40 soybean samples having a protein range of 33 to 50 percent and an oil range of 12 to 24 percent. Protein and oil concentrations for samples with colored or very heavily mottled seed coats were obtained with the Kjeldahl procedure for protein and Butt extraction for oil. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters. Seeds were ground in a small food grinder and stored at ?20 C until analyzed. Approximately 200 mg of ground sample was placed in a 25 ml vial, and 5 ml of sodium methoxide added in two 2.5 ml aliquots with an automatic syringe in such a way as to ensure mixing. (The sodium methoxide solution was prepared daily by adding 1 g of sodium metal to 100 ml of reagent grade methanol.) The sodium methoxide, ground sample suspension was allowed to stand for 45 minutes, after which 1 ml of 10 percent acetic acid solution was added, followed immediately by 10 ml of heptane (in two 5 ml aliquots). The samples were completely mixed after each reagent addition. This mixture was allowed to stand for several minutes so that the layers could separate. A 2-ml aliquot of the heptane layer was extracted for analysis in a Hewlett Packard model 6890 gas chromatograph equipped with a Model 6890 auto injector and flame ionization detector. Columns were 30 m by 0.32 mm capillaries coated internally with 5 percent diphenyl dimethyl siloxane. In the HP 6890, chromatography was isothermal and flow rates for helium, hydrogen, and air were 40, 40, and 450 ml min-1 respectively. The injection volume was 1 ?l with split ratios used, dependent upon the concentration of sample. Total analysis time was approximately 5 minutes. The integration, peak identification, data storage, and report printing were all performed by the Hewlett Packard Chemstation software and a modified Excel spreadsheet.

Descriptive characteristics, agronomic performance, and seed composition data of soybean (Glycine max (L.) Merr.) germplasm were evaluated on accessions initially classified in maturity groups 000 through 0 in the range from PI 578371 to PI 612761, plus some accessions that were omitted from earlier publications. Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 2001. The accessions were evaluated in 2001 and 2002 in Rosemount, Minnesota (Lat. 45o 02' N). Planting dates were June 8, 2001 and May 28, 2002. Plots were 3.6 m long with 4 rows 76 cm apart. Plots were trimmed to 2.4 m after maturity and the middle two rows harvested. Because of the difference in planting date, plants matured an average of six days later in 2001 than in 2002. Growing conditions in 2001 were hot and dry, while 2002 was wet most of the growing season. As a result, seed size and yield in 2002 was generally greater than in 2001 Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989, Bannon et al. 1982). Oil and protein for samples with yellow seed coats was analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Descriptive characteristics, agronomic performance, and seed composition data of soybean (Glycine max (L.) Merr.) germplasm were evaluated on accessions initially classified in maturity groups 000 through I in the range from PI 613561 to PI 653905, plus some accessions that were omitted from earlier publications. Also included were cultivars in the same maturity groups, developed in the United States and Canada, and released by 2009

Accessions in maturity groups 000 through I ranging from PI 507670 to PI 574486 were evaluated for descriptive characteristics, agronomic performance, and seed composition in 1994 and 1995 in Rosemount, Minnesota (Lat. 45o 02' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by early 1993. Tests were replicated once per year. Planting dates were May 31, 1994 and May 5, 1995. Plots were 3.6 m long with 4 rows 76 cm apart. Plots were trimmed to 2.4 m after maturity and the middle two rows harvested. Because of the difference in planting date, plants flowered and matured later in 1994 than in 1995. The difference in planting was almost 4 weeks but the average difference between years in flowering and maturity dates was only about 14 days. There was more rain in 1994 and yields were generally much higher in 1994 than in 1995. The seed composition data were collected at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. To obtain oil and protein percentages of the seeds with yellow seed coats, approximately 7 g of seeds were placed in a beaker and dried in a forced air oven for 3 hours at 130 C. The seeds were then transferred to 50 g bottles, sealed, and allowed to cool for 1 hour. Samples were then ground in a Varco electric dry food grinder and returned to the 50 g bottles. Seed samples were analyzed with an Infratec 1255 food and grain analyzer (Perstorp Analytical Company). Samples were scanned from 800 - 1100 nm. The analyzer was calibrated with at least 40 soybean samples having a protein range of 33 to 50 percent and an oil range of 12 to 24 percent. Protein and oil concentrations for samples with colored or very heavily mottled seed coats were obtained with the Kjeldahl procedure for protein and Butt extraction for oil. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters. Seeds were ground in a small food grinder and stored at ?20 C until analyzed. Approximately 200 mg of ground sample was placed in a 25 ml vial, and 5 ml of sodium methoxide added in two 2.5 ml aliquots with an automatic syringe in such a way as to ensure mixing. (The sodium methoxide solution was prepared daily by adding 1 g of sodium metal to 100 ml of reagent grade methanol.) The sodium methoxide, ground sample suspension was allowed to stand for 45 minutes, after which 1 ml of 10 percent acetic acid solution was added, followed immediately by 10 ml of heptane (in two 5 ml aliquots). The samples were completely mixed after each reagent addition. This mixture was allowed to stand for several minutes so that the layers could separate. A 2-ml aliquot of the heptane layer was extracted for analysis in a Hewlett Packard model 6890 gas chromatograph equipped with a Model 6890 auto injector and flame ionization detector. Columns were 30 m by 0.32 mm capillaries coated internally with 5 percent diphenyl dimethyl siloxane. In the HP 6890, chromatography was isothermal and flow rates for helium, hydrogen, and air were 40, 40, and 450 ml min-1 respectively. The injection volume was 1 ?l with split ratios used, dependent upon the concentration of sample. Total analysis time was approximately 5 minutes. The integration, peak identification, data storage, and report printing were all performed by the Hewlett Packard Chemstation software and a modified Excel spreadsheet.

Accessions in maturity group V, FC 30265 to PI 408435 (received by 1976) were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 1998 and some accessions received earlier which had not been previously evaluated. All tests were replicated once per year. Plots were four rows wide, with rows 3.6 m long and 76 cm between rows. Only the center two rows of each plot were harvested for yield. All accessions were planted again in single rows in 2004 to verify plant descriptors. Planting dates were May 13, 1999 and April 27, 2001. Plants flowered an average of 15 days later and matured 2 days later in 1999 than in 2001. Plants were taller and yields were generally greater in 1999 than in 2001. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989; Bannon et al. 1982). Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Accessions in maturity group V, PI 416758 to PI 561398 (received between 1977 and 1991) were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 2001 and some accessions received earlier which had not been previously evaluated. All tests were replicated once per year. Plots were four rows wide, with rows 3.6 m long and 76 cm between rows. Only the center two rows of each plot were harvested for yield. All accessions were planted again in single rows in 2004 to verify plant descriptors. Planting dates were April 27, 2000 and May 14, 2002. Plants flowered an average of 25 days later and matured 7 days later in 2002 than in 2000. Plants were taller and yields were generally greater in 2002 than in 2000. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989; Bannon et al. 1982). Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Accessions in maturity group V PI 597469 to PI 612614 (received between 1996 and 1998) were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 2000 and some accessions received earlier which had not been previously evaluated. All tests were replicated once per year. Plots were four rows wide, with rows 3.6 m long and 76 cm between rows. Only the center two rows of each plot were harvested for yield. All accessions were planted again in single rows in 2004 to verify plant descriptors. Planting dates were April 27, 2001 and May 14, 2002. Plants flowered an average of 20 days later, matured 4 days later and were generally taller and higher yielding in 2002 than in 2001. A hurricane on Oct. 3, 2002 damaged many plots. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989; Bannon et al. 1982). Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Accessions in maturity groups VI through VIII, PI 597465 to PI 606436B (received between 1996 and 1998) were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 2001 and some accessions received earlier which had not been previously evaluated. All tests were replicated once per year. Plots were four rows wide, with rows 3.6 m long and 76 cm between rows. Only the center two rows of each plot were harvested for yield. All accessions were planted again in single rows in 2004 to verify plant descriptors. Planting dates were April 27, 2001 and April 14, 2003. Plants flowered an average of 9 days later and 2 days later and were generally taller and higher yielding in 2003 than in 2001. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989; Bannon et al. 1982). Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Descriptive characteristics, agronomic performance, and seed composition data of soybean (Glycine max (L.) Merr.) germplasm were evaluated on accessions initially classified in maturity groups V through VIII in the range from PI 614156 to PI 654832, plus some accessions that were omitted from earlier publications. Also included were cultivars in the same maturity groups, developed in the United States and Canada, and released by 2009.

Accessions were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Accession ranges included were FC 03.659 to PI 520.732 in maturity group VI. Also included were cultivars in the same maturity group that were developed at public institutions in the United States and Canada and released by 1991. The evaluation was planted in mid-May, 1992 and May 17, 1993. The test was replicated once per year. 1993 was much drier than 1992 and supplemental irrigation was required. Almost all entries flowered earlier in 1992 than in 1993 but matured at about the same time in both years. Plant height and yield were both significantly lower in 1993 than in 1992. Plots were four rows wide, with rows 3.6 m long and 91 cm between rows. Only the center two rows of each plot were harvested for yield. Plots were not end trimmed at maturity so yield values reported here slightly overestimate the actual yield. For the MG VI evaluation, only 1993 yield data could be used. Based on data collected in these evaluations, maturity groups for some accessions were changed from those reported in earlier publications. The seed and oil composition data were collected at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Protein and oil percentages for lines with yellow seed coats were obtained using infrared instrumentation (NIR). Composition percentages for those with colored or very heavily mottled seed coats were obtained with the Kjeldahl procedure for protein and Butt extraction for oil. Fatty acid composition was determined by gas-liquid chromatography.

Accessions were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Accession ranges included were FC 30.267 to PI 567.181B in maturity group VII and FC 31.592 to PI 567.235B in maturity group VIII. Also included were cultivars in the same maturity group that were developed at public institutions in the United States and Canada and released by 1991. The evaluation was planted on May 10, 1994 and May 23-24, 1995. In 1994, yield was much greater and seed quality much higher than in 1995. The total rainfall during each growing season was not significantly different but rainfall during the grain filling period was much greater in 1994 than in 1995. Some supplemental irrigation was required in both years. The test was replicated once per year. Plots were four rows wide, with rows 3.6 m long and 91 cm between rows. The evaluation was blocked by maturity group. Only the center two rows of each plot were harvested for yield. Plots were not end trimmed at maturity so yield values reported here slightly overestimate the actual yield. Based on data collected in these evaluations, maturity groups for some accessions were changed from those reported in earlier publications. The seed and oil composition data were collected at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Protein and oil percentages for lines with yellow seed coats were obtained using infrared instrumentation (NIR). Composition percentages for those with colored or very heavily mottled seed coats were obtained with the Kjeldahl procedure for protein and Butt extraction for oil. Fatty acid composition was determined by gas-liquid chromatography.

Accessions in maturity groups V through VIII, PI 566960 to PI 592914 (received between 1990 and 1994) were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 1994 and some accessions received earlier which had not been previously evaluated. All tests were replicated once per year. Plots were four rows wide, with rows 3.6 m long and 76 cm between rows and blocked by maturity group. Only the center two rows of each plot were harvested for yield. All accessions were planted again in single rows in 2004 to verify plant descriptors. Planting dates were May 14, 1996 and May 22, 1997. Plants flowered an average of 8 days later and matured 2 days later in 1997 than in 1996. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989; Bannon et al. 1982). Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Accessions in maturity groups V through VIII, PI 593948 to PI 594904 (received between 1995 and 1996) were evaluated for descriptive characteristics, agronomic performance, and seed composition in Stoneville, Mississippi (Lat. 33o 26' N). Also included were cultivars in the same maturity groups, developed at public institutions in the United States and Canada, and released by 1997 and some accessions received earlier which had not been previously evaluated. All tests were replicated once per year. Plots were four rows wide, with rows 3.6 m long and 76 cm between rows and blocked by maturity group. Only the center two rows of each plot were harvested for yield. All accessions were planted again in single rows in 2004 to verify plant descriptors. Planting dates were May 18, 1998 and May 14, 1999. Plants flowered an average of 8 days later and matured 5 days later in 1998 than in 1999. Yields were generally greater in 1999 than in 1998. Seed quality, seed mottling, seed shape, and chemical analysis of maturity group VIII accessions were not recorded in 1998 due to extremely poor seed quality. Seed composition was analyzed at the USDA Northern Center for Agricultural Utilization Research in Peoria, IL. Fatty acid composition was obtained by gas liquid chromatography of the methyl esters (Christie 1989; Bannon et al. 1982). Oil and protein for samples with yellow seed coats were analyzed using the near infrared method for whole-grain analysis (AACC Method 39-21). Protein concentrations for samples with colored or heavily mottled seed coats were obtained using the improved Kjeldahl method (AACC Method 46-16) and oil by the Butt Extraction method (AOCS Official Method Ac 3-44).

Data from Expired Plant Variety Protection certificates

About 10 seed from each accession were germinated on paper towels and the 3-day-old seedlings were examined under an ultraviolet light. Remnant seed of nonfluorescent accessions were planted at the Bruner Farm near Ames, Iowa. Cross-pollinations were made between nonfluorescent accessions and the fluorescent root cultivars BSR 101 or Harosoy. Cross-pollinations were made between non fluorescent accessions and tow nonfluorescent testers, near isogenic lines Hark-fr1 and Hark-fr2.

To screen for root fluorescence, 10 seeds of each accession were germinated on paper towels. After 4 days in a growth chamber, the seedlings were observed in the dark under ultraviolet light.

Study Name: Soybean Germplasm Resistant to Cercospora sojina. Evaluation of Maturity Groups VI and VII for resistance to Frogeye leaf spot, C-32 isolate. Experiment type: Greenhouse. Greenhouse Study Year: 1999. Exp. Design: Randomized with three replications, 1 pot with 4 plants/ replicate.

Evaluation location: Asociados Don Mario Research Center in Chacabuco, a city of Buenos Aires province in Argentina. Natural infection on the fields. The data was taken on 2 m. row per line, at flowering time. From the differential evaluation, race 11 was present. Photographs of scale used

Comment: The data recorded in this study has no specific environmental information. For additional information, contact Randy Nelson at the USDA Soybean collection in Urbana, IL 61801. Phone: (217) 244-4346.

A genotyping by sequencing (GBS) haplotype method characterized accessions for their allelic status at the GmPhyA3 gene.

1988 soybean germplasm increase trial, Urbana, Illinois. Field S700, planted 5-14-1988. Sprockets 20 to 14, Series: 23 x 11.5' Harvest Length 7.5', Alley: 4'. Row width 30 inches.

1989 soybean germplasm increase trial, Urbana, Illinois. Field S600, planted 5-17-1989. Sprockets 20 to 14, Series: 23 x 11.5' Harvest Length 7.5', Alley: 4'. Row width 30 inches.

1990 soybean germplasm increase trial field R1S, Urbana, Illinois

1990 soybean germplasm increase trial, Urbana, Illinois. Field S700, planted 5-09-1989. Sprockets 20 to 22, Series: 20 x 13' Harvest Length 9.5', Alley: 3.5'. Row width 30 inches.

1997 soybean germplasm increase trial, Urbana, Illinois.

1998 soybean germplasm increase trial, Urbana, Illinois.

1999 soybean germplasm increase trial, Urbana, Illinois.

2000 soybean germplasm increase trial, Stoneville, Mississippi.

2000 soybean germplasm increase trial, Urbana, Illinois.

2001 soybean germplasm increase trial, Urbana, Illinois.

2002 soybean germplasm increase trial, Urbana, Illinois.

2003 soybean germplasm increase trial, Urbana, Illinois.

2004 soybean germplasm increase trial, Stoneville, Mississippi.

2005 soybean germplasm increase trial, Urbana, Illinois.

2006 soybean germplasm increase trial, Urbana, Illinois.

2007 soybean germplasm increase trial, Urbana, Illinois.

2008 soybean germplasm increase trial, Urbana, Illinois.

2009 soybean germplasm increase trial, Stoneville, Mississippi.

2009 soybean germplasm increase trial, Urbana, Illinois.

2010 soybean germplasm increase trial, Stoneville, Mississippi.

2010 soybean germplasm increase trial, Urbana, Illinois.

2011 soybean germplasm increase trial, Urbana, Illinois.

2012 soybean germplasm increase trial, Stoneville, Mississippi

2012 soybean germplasm increase trial, Urbana, Illinois

2013 soybean germplasm increase trial, Stoneville, Mississippi

2013 soybean germplasm increase trial, Urbana, Illinois

2014 soybean germplasm increase trial, Stoneville, Mississippi

2014 soybean germplasm increase trial, Urbana, Illinois

2015 soybean germplasm increase trial, Stoneville, Mississippi

2015 soybean germplasm increase trial, Urbana, Illinois

2016 soybean germplasm increase trial, Urbana, Illinois

2017 soybean germplasm increase trial, Urbana, Illinois

2018 soybean germplasm increase trial, Stoneville, Mississippi

2019 soybean germplasm increase trial, Stoneville, Mississippi

2020 soybean germplasm increase trial, Stoneville, Mississippi

2020 soybean germplasm increase trial, Urbana, Illinois

2006 soybean germplasm increase trial, Upala, Costa Rica

2007 soybean germplasm increase trial, Upala, Costa Rica

2013 1 or 2 row maintenance increase, Urbana, Illinois

2014 soybean germplasm increase trial, Upala, Costa Rica

2016 soybean germplasm increase trial, Upala, Costa Rica

2017 1 or 2 row maintenance increase, Stoneville, Mississippi

2018 soybean germplasm increase trial, Upala, Costa Rica

2019 1 or 2 row maintenance increase, Stoneville, Mississippi

2018 soybean germplasm increase trial, Upala, Costa Rica

2020 1 or 2 row maintenance increase, Stoneville, Mississippi

Field screen for resistance to Mexican Bean Beetle foliar feeding based on natural infestations in the field. 1971-1990. University of Maryland Poplar Hill Research Farm, Quantico, MD. Lines scored 1, 2, or 3 were retested to confirm rating.

2013 soybean germplasm misc, Urbana, Illinois. Seed increases of very low seed or accessions that have purity questions.

2017 soybean germplasm misc, Stoneville, Mississippi. Seed increases of accessions that have purity questions.

2019 Misc. soybean germplasm to be tested for RR trait

2020 soybean germplasm misc, Stoneville, Mississippi. Seed increases of accessions that have purity questions.

2020 soybean germplasm misc, Urbana, Illinois. Seed increases of very low seed or accessions that have purity questions.

Study Name: Northern Stem Canker Evaluation 1986 - 1993. Inoculation technique was similar to that described by Keeling, B.L., Phytopathology 72:807-809 (1982). Stoneville, MS 12 to 15 plants of each line were inoculated with a tooth pick infested with the Northern Stem Canker organism 25 to 30 days after emergence. Disease ratings were made 60 days after inoculation.

Soybean accessions were surveyed using a monoclonal antibody against P34

Accessions were screened to identify accessions with low or no P34 expression. Individual seed of each accession was used as starting material to extract total protein as the allergen of interest is expressed highly in seeds. Pulverized seed from each accession was added to 1mL TBS-T buffer [0.05MTris, 0.5MNaCl,pH7.4 with 0.05%(v/v)Tween 20 (polyoxyethylene sorbitan monolaurate)] and incubated for 30 min on a shaker (200 rpm) at room temperature. Immuno-blot assay of the seed extracts was done by a rapid and large scale method as follows: A 10-mL aliquot of each sample extract was dot-blotted on a nitrocellulose membrane (0.45 mm; Fisher Scientific) with a P34 positive control sample (cv. Kunitz). To check for equal loading of the samples on the blots, membranes were stained with Ponceau S solution (Sigma, St. Louis, MO) for 1 min and rinsed in double distilled water. The membrane was incubated in TBS buffer (0.05 M Tris and 0.5 M NaCl, pH 7.4) for 5 min and blocked (10% nonfat dry milk powder in TBS buffer) for 1 h followed by rinsing twice in TBS buffer for 5 min each. The membranes were incubated in P34 protein specific monoclonal antibody P4B5 (diluted 1: 5000 in TBS-T buffer with 0.1% milk powder) for 1 h. The membranes were washed extensively in TBS-T buffer and incubated in the secondary antibody [antimouse IgG (Fab specific) alkaline phosphatase conjugate (Sigma)] diluted 1: 10 000 in TBS-T buffer with 0.1% milk powder for 1 h and rinsed in TBS-T buffer and TBS buffer once each. The P34 protein was detected by a color substrate system [BCIP/NBT: final concentrations 0.02% (w/v) 5-bromo-4-chloro-3-indolyl phosphate and 0.03% (w/v) nitro blue tetrazolium in 70% (v/v) dimethylformamide] (Sigma).

Study Name: New Sources of Resistance to Phytophthora sojae in the Soybean Plant Introductions. Experiment Type: Laboratory (hypocotyl inoculation)

Accessions from Southern China were evaluated for their response to races 1, 3, 4, 5, 7, 10, 12, 17, 20, and 25 of Phytophthora sojae using the hypocotyl inoculation technique in the greenhouse at Urbana, ILlinois in 1997 and 1997.

Study Name: Origin of soybean alleles for Phytophthora resistance in China. Method: Hypocotyl inoculation in the Greenhouse

Hypocotyl inoculation

Plants were rated for their response to P. sojae 6 days after inoculation. The number of living and dead plants was recorded. Accession with greater than 70% live plants were rated as resistant; less than or equal to 30% live plants were rated as susceptible; less than 70% but greater then 30% live plants were rated as heterogeneous.

Study Name: Response of soybean accessions from southern China to races 1, 3, 7, and 25 of Phytophthora sojae. Hypocotyl Inoculation Technique with Zoospores. Zoospore suspensions were injected into soybean plants. Each plant was injected three times at 1 cm intervals below the cotyledonary node in the center of the hypocotyl. Plants were not watered for at least 12 hours after inoculation to avoid washing off the inoculum. Plant Technique and Greenhouse Environment: Ten seed of each soybean line were planted 2.5 cm deep in 10 cm plastic pots filled with sand. Pots were watered to saturation each day prior to plant emergence and following emergence. Plants were grown and inoculated in the greenhouse under a 14-hour photoperiod with temperatures ranging between 210C and 290C. Seedlings were inoculated 8 to 14 days after planting, when the unifoliolate leaves had just opened. Plants with enlarged hypocotyls or late emerging plants were removed from the pot prior to inoculation. Plant Response Rating: Plants were rated for their response to P. sojae 4 to 5 days after inoculation. The number of living and dead plants in each pot was recorded. Pots containing greater than 70% alive plants or plants with non-killing lesions were rated as resistant; pots containing less than 30% alive plants or plants with non-killing lesions were rated as susceptible; pots containing less than 70% alive plants or plants with non-killing lesions but greater then 30% alive plants or plants with non-killing lesions were rated as heterogeneous. Resistant and susceptible checks were included in each test to evaluate the virulence of the pathogen.

Seedlings were inoculated in the field for Peanut Mottle Virus using the airbrush technque. Field-resistant Maturity Group II and III strains were retested under greenhouse conditions using the mortar and pestle inoculation.

Soybean seeds of 30 soybean cultivars obtained from the Arkansas soybean variety testing program, were germinated in vermiculite in a greenhouse. In the cotyledonary stage, individual seedlings were transplanted into 10-cm-d clay pots containing 500 cm 3 fine sandy loam (ca. 91% sand, 5% silt, 4% clay, <1% O.M.). Ten pots of each cultivar were used in 1991; in 1992, 10 fallow pots were added. One-half of the pots of each cultivar and fallow treatment were infested with ca. 1,000 vermiform reniform nematodes. Pots were arranged in a randomized complete block design with five replications per treatment. Vermiform reniform nematodes were obtained from greenhouse-grown soybean plants (cv. Braxton). Soil was washed from the roots, suspended in water, and poured through nested 841- (20 mesh) and 38 (400 mesh) um-pore sieves. The material on the 38-um-pore sieve was placed on a tissue in a Baermann funnel. All vermiform stages of R. reniformis were collected after 16 hours and injected with an autopipette into three 1-inch deep holes made in the soil with a dibble at the proper dilution. After ca. 60 days plants were harvested, roots and shoots were weighed, and nematodes were extracted from roots and soil. To determine the final nematode population , a 100-cm 3 aliquot of well-mixed soil from each pot was suspended in water and poured through nested 841- and 38-um-pore sieves to remove plant debris and extract the nematodes. The nematode suspension from the 38-um-pore-sieve was clarified by sucrose centrifugal flotation, counted, and multiplied by five to give the number per pot. The eggs and vermiform nematodes in the egg masses on roots were extracted with a 0.525% sodium hypochlorite solution and counted. The total number of R. reniformis per pot was calculated by adding the number from the soil to the number from the roots. The reproductive index was calculated as the final nematode population density/initial nematode population density.

Soybean seeds were germinated in flats of vermiculite, and single seedlings were transplanted at the cotyledon stage into 10-cm-diam. clay pots containing 500 cm 3 fine sandy loam (91% sand, 5% silt, 4% clay; < 1% O.M.). The soil in each pot was inoculated with ca. 1,000 vermiform reniform nematodes placed in three 1-inchdeep depressions. The R. reniformis population was from an Arkansas isolate obtained from a Jefferson County, Arkansas cotton field in 1986, maintained in a greenhouse on cotton cv. DP-50, and increased on soybean cv. Braxton for these studies. Greenhouse ambient air temperature ranged from 28 to 34?C. All test pots were watered twice daily (8 am and 4 pm) and fertilized once a week with soluble 20- 20-20 (N-P-K). Final nematode populations for each pot were determined by combining the soil (sieving, sucrose centrifugation extraction) and root (sodium hypochlorite shaker extraction) populations. A reproductive index (RI = final population/ initial population) was calculated for each entry.

Twenty-two lines resistant to Soybean Cyst Nematode (SCN) race 3 or race 5 were tested in a greenhouse for resistance to reniform nematode because of preivious reports linking SCN race 3 resistance to reniform resistance.

Greenhouse study. Soybean accessions were spray inoculated between the first and second trifoliate stage using a mixture of urediniospores (60,000 spores per ml) from four Phakopsora pachyrhizi isolates, incubated overnight in a dew chamber at 22-25 C, and placed a greenhouse at 20-25 C for 14 days. Disease severity was evaluated on the first trifoliate leaves for most accessions, however the unifoliate leaves were evaluated for a few accessions with slow germination. A disease severity scale, based on symptom and lesion development on 1 to 5 scale, was used where 1 = no visible symptoms, 2 = light infection with few lesions present, 3 = light to moderate infection, 4 = moderate to severe infection, and 5 = severe infection. TAN, RB, or mixed lesion types were also recorded. The TAN lesion type is a susceptible reaction, while the RB or "reddish-brown" lesion type has been associated with resistance in the known single genes. The mixed reaction was recorded when both RB and TAN lesions were observed on the same leaf.

Field Study. 1990, Stoneville, MS (Lat 36 degrees 26 minutes N). Soybean looper adults were released inside 0.17 acre field cages in which the accessions were grown.

Study Name:Soybean Plant Introductions With Resistance to Races 3, 4, or 5 of Soybean Cyst Nematode. Seeds were planted in field soil infested with 20 to 30 cysts/100 g soil, grown for 30 d in a greenhouse at 24 C. The rating scale used was: 0=no cyst; 1=1-9 cysts, Resistant; 2=10-30 cysts, Moderately resistant; 3=31-60 cysts, Moderately susceptible; and 4=61+ cysts per plant compared with 100 on Essex (susceptible cultivar), Susceptible.

Soybean accessions were screened for resistance to Soybean Cyst Nematode race 1 (HG type 2.5.7), race 2 (HG type 1.2.5.7), race 3 (HG type 0), and race 5 (HG type 2.5.7). A SCN bioassay using revised classification system, HG Type (Niblack et al.,2002) was used to evaluate soybean germplasm in the greenhouse. HG Type of classification includes seven indicator lines and a susceptible cultivar for determination of soybean reaction to SCN. The indicator lines included PIs 548402 (Peking), 88788, 90763, 437654, 209332, 89772, 548316 ( Cloud) and, a standard susceptible control cv. Lee-74 ( PI 548658). Lee-74 was replaced with cv.5601T which is also a susceptible variety. The current HG Type classification system can't distinguish Race 1 population from Race 5 but cv. Pickett was included as a differential line which is resistant to Race 1 but susceptible to race 5. Soybean germplasm accessions were screened with near-homogeneous populations of SCN races or HG Types to obtain consistent reaction. The source and method for developing near-homogeneous populations of SCN Races or HG Types used for screening soybean accessions was already described (Qui et al.,1999). The method for SCN bioassay performed in the greenhouse followed established protocols (Arelli et al., 2000) with the modifications described in Arelli and Wang (2008). Each plant was grown in a 7-cm in diameter clay pot filled with steam sterilized soil on greenhouse bench top with an evaporative cooling and under bench heating systems. The duration of lighting, heating and, cooling systems in the green house were regulated during the bioassay for proper growth of soybean seedlings and nematodes. Ten seedlings for each accession, indicator lines and a susceptible control were included in each experiment and maintained at 27+or-2 degrees C. When the seedlings were 4 or 5 days old each plant was inoculated with a given nematode Race or HG Type using eggs and juveniles. Each seedling was inoculated with 5 ml of suspension (de-ionized water) consisting approximately 2000 eggs and juveniles. The inoculum was delivered close to the roots of seedling. Approximately 30 days after inoculation, plant roots were washed individually with a strong jet of water to dislodge the females and cysts. These were counted under a stereomicroscope and, a female index (FI%) was calculated for the number of females developing on each plant (Golden et al.,1970). Female index is the number of SCN females occurring on a soybean plant expressed as the percentage of mean number of females on susceptible Lee-74 or cv.5601T. Ratings of resistant (FI=0 to 9%), moderately resistant (FI=10 to 30%), moderately susceptible (FI=31 to 60%) and, susceptible (FI>60%) used to classify the reaction of plants were based on Schmitt and Shannon (1992). Each experiment was repeated and FI% was averaged for a given race or HG type. The screening procedure with some modifications has been published in Euphytica ( 2009 ) 165: 1-4.

Accessions were bioassayed during 1997 and 1998 for SCN Race 1 and 2 in thermoregulated water baths in the greenhouse. Peking, PI 90763, PI 88788, and Picket 71 were used as host differentials, with Hutcheson as the susceptible control.

Bioassays were performed in the greenhouse for two SCN populations: Races 1 and 3 (HG Types 2.5.7 and 0, respectively), during 2011-2014. The methods used were described by Arelli et al. (2015. New soybean accessions evaluated for reaction to Heterodera glycines populations. Crop Sci 55:1236-1242). In brief, for each nematode population, seven seedlings were evaluated for each of 239 accessions, susceptible control, and indicator lines. Each seedling within a genotype represented a single replication; the test was completely randomized and repeated twice. Approximately 30 d after inoculation, plant roots were individually washed with a strong jet of water to dislodge white females and cysts. These were counted under a stereomicroscope, and a female index (FI%) was calculated for the number of females developed on each line in each replication. Data were combined for the two tests for each nematode population for ANOVA of female and means separated with Fisher’s LSD based on a significant F test. Ratings of resistant (FI=0-9%), moderately resistant (FI=10-30%), moderately susceptible (FI=31%-60%), and susceptible (FI>60%) used to classify the reaction of the accessions were based on Schmitt and Shannon (1992. Differentiating soybean responses to Heterodera glycines races. Crop Sci 32:275-277)

Accessions were bioassayed during 1995 and 1996 for SCN Race 1 and 2 in thermoregulated water baths in the greenhouse. Peking, PI 90763, PI 88788, and Picket 71 were used as host differentials, with Hutcheson as the susceptible control.

Evaluations were conducted once for each race in a greenhouse. One seed of each accessions was planted in each of three pots in soil infested with either race 3 or race 4 of soybean cyst nematode (SCN), Heterodera glycines. After the completion of the first generation in approximately one month, numbers of females on roots were determined and a resistance score was assigned based on a scale of 0 - 4 where 0 = no females, 1 = 1 - 4 females, 2 = 6 - 10 females, 3 = 11 - 30 females, and 4 = > 30 females per root system. If the resistance score is either 0 or 1, an accession is considered highly resistant and is moderately resistant if the score is 2.

To evaluate resistance to Heterodera glycines, five seeds of each cultivar were planted in 420-ml styrofoam cups filled with soil naturally infested with either race 3 or race 14 (7). The soil infested with race 3 (cyst index: Peking, 0; Pickett, 1; P188788, 0; P190763, 0) was a Cecil coarse sandy loam (72% sand, 12% silt, 16% clay; a member of the clayey, kaolinitic, thermic family of Typic Hapludults) collected from the Plant Sciences Farm near Athens, Georgia. The soil infested with race 14 (cyst index: Peking, 37; Pickett, 76; PI 88788, 1; PI 90763, 11) was a Dothan sandy loam (85% sand, 12% silt, 3% clay; a member of the fine-loamy, siliceous, thermic family of Plinthic Paleudults) collected near Waynesboro, Georgia. One week after planting, seedlings were thinned to one per cup and watered as needed. After 30-40 days, when white females and cysts had developed on susceptible standard cultivars, the soil was gently shaken from each root system, white females and cysts were enumerated under a magnifying lamp, and a resistant or susceptible rating was assigned. Plants were assigned a rating of susceptible (generally >or = 11 white females or cysts per plant) or resistant (generally < or = 10 white females or cysts) on the basis of number of white females or cysts relative to the standard resistant cultivars. The resistant and susceptible standards were Centennial and Lee for race 3 and Leflore and Centennial for race 14, respectively. Each cultivar was evaluated a minimum of 2 years in a randomized complete block experimental design with four replications. Plants were grown with supplemental light provided by 400-W metal halide lamps and fertilized weekly with 6 mg N, 3 mg P, and 5 mg K.

Study Name:Soybean Germplasm Evaluated for Resistance to Races 3, 5, or 14 of Soybean Cyst Nematode. Seeds were planted in field soil infested with 20 to 50 cysts/pot, grown for 30 d in a greenhouse at 27 C. The number of SCN cysts and mature females on the roots of each plant were visually rated. 1=0 to 5, 2=6 to 10, 3=11 to 20, 4=21 to 40, 5=>40 cysts.

Soybean Germplasm Evaluated for Resistance to Races 3, 5, and 14 of Soybean Cyst Nematode. Lines with mean rating < 2 for any race in evaluation with field soil infested with 20 to 50 cyst/pot were reevaluated along with SCN race differentials Lee, Peking, Pickett 71, PI 88788 and PI 90763 in a RCBD with 10 replications for each race. Seeds of each line were planted in heat-sterilized soil infested with 1000 nematode eggs/plant. Cyst and females were counted after 30 d.

Study Name: Soybean Germplasm Resistant to Races 3, 5, or 14 of Soybean Cyst Nematode. Heat-sterilized silt loam soil was infested with 1000 eggs/pot. Plants were grown for 30 d in a greenhouse at 27 C. The number of H. glycines females was visually rated: 1=0-5, 2=6-10, 3=11-20, 4=21-40, 5=>40 females.

Soybean Germplasm Resistant to Races 3, 5, or 14 of Soybean Cyst Nematode. Accessions with a mean visual rating <2 for Races 3 or 5 or <2.2 for Race 14 in initial evaluations were tested again along with H. glycines race determination differentials Lee 68, Peking, Pickett, PI 88788, and PI 90763 in a RCBD with 5 replications for each race. Seeds of each accession were planted in heat-sterilized soil infested with 1000 eggs/pot. Females were counted 35 d after planting.

Four liters of a steam-treated soil mix (1:1 sand/soil) was placed in 50 x 35 x 10 cm flats. A template was used to make seven furrows 2 cm deep and 35 cm long, and 10 cm3 of sorghum seed infested with F. solani f. sp. glycines isolate Mont-1 was evenly distributed in each furrow. The inoculum was then covered with 2 cm of soil, and the template was reapplied to make a 0.5-cm-deep furrow directly over the inoculum. Soybean seed was added to each furrow and covered with another 2 cm of soil. Plants were then placed on a greenhouse bench and grown under a 16-h photoperiod. Plants were watered to saturation after planting and maintained at near field capacity throughout the study. Two to 4 weeks after inoculation, plants were rated for SDS foliar symptoms on 4 separate dates. Plots were rated if two or more seeds emerged. Five seeds per entry were planted in flats containing 20 entries and one check. The susceptible check was Great Lakes 3302, and the moderately resistant checks were PI 520733 and PI 567374 . The entire set was screened in nonreplicated subsets of 500 PIs. Entries that had a mean disease severity rating and an AUDPC value 1.0 standard deviation below the mean of the subset or were not significantly different from either of the MR checks were advanced to the second stage. The second stage (1,302 entries from stage one) was planted as a nonreplicated set. The entries that had a mean disease severity rating and an AUDPC value 1.0 standard deviation below the mean of the set were advanced to the third stage. For the third stage, 212 accessions from stage two were arranged in a RCBD with four replications. Of these 212 entries, 57 were repeated in the final stage. These 57 PIs had foliar symptoms that were not significantly different from the MR check PI 567374 in the third stage.

Ancestral soybean lines representing approximately 99% of the genes in modern North American cultivars and lines identified in the pedigrees of previously identified mostly resistant (MR) lines were screened for resistance to SDS. Four liters of a steam-treated soil mix (1:1 sand/soil) was placed in 50 x 35 x 10 cm flats. A template was used to make seven furrows 2 cm deep and 35 cm long, and 10 cm3 of sorghum seed infested with F. solani f. sp. glycines isolate Mont-1 was evenly distributed in each furrow. The inoculum was then covered with 2 cm of soil, and the template was reapplied to make a 0.5-cm-deep furrow directly over the inoculum. Soybean seed was added to each furrow and covered with another 2 cm of soil. Plants were then placed on a greenhouse bench and grown under a 16-h photoperiod. Plants were watered to saturation after planting and maintained at near field capacity throughout the study. 24 days after inoculation, plants were rated for SDS foliar symptoms. Plots were rated if two or more seeds emerged. Five seeds per entry were planted in flats containing 20 entries and one check. The susceptible check was Great Lakes 3302, and the moderately resistant checks were PI 520733 and PI 567374. The ancestral set was evaluated 3 times and the pedigree set was evaluated twice using a randomized complete block design with 3 replications.

An isolate of F. solani, Monticello, was increased on 200 g of sorghum seeds soaked overnight and autoclaved twice in a 1-liter flask. Each flask was inoculated with 10 plugs (4 mm diameter) of fungal mycelium and incubated for 2 weeks. Two liters of a steam-treated soil mix (1:1 sand/soil) was placed in 8 x 12-cm trays. A template was used to make three furrows that were 12 cm long, 2 cm deep, and 3 cm apart, and 5 cm3 of infested sorghum seeds was evenly distributed in each furrow. The infested seeds were covered to a depth of 2 cm. The template was reapplied to make a 2-cm-deep furrow directly over the inoculum. 12 seeds per entry were added to each furrow and covered with another 2 cm soil. After seedling emergence, the soil was maintained at near water-holding capacity by flooding the flats twice daily as needed. Three weeks after planting, each plant was evaluated for SDS leaf symptoms: 1 = no symptoms, 2 = slight symptom development, with mottling and mosaic on leaves (1 to 20% foliage affected), 3 = moderate symptom development, with interveinal chlorosis and necrosis on foliage (21 to 50% foliage affected), 4 = heavy symptom development, with interveinal chlorosis and necrosis (51 to 80%), and 5 = severe interveinal chlorosis and necrosis (81 to 100% foliage affected). In the first stage, all genotypes plus 3 standards (PI 520.733 and cvs. Ripley and Great Lakes 3202) were evaluated in the greenhouse. The entries were arranged in 8 sets. Entries within sets that had a mean disease severity of 1 standard deviation (SD) below the mean of the set were advanced. In the second stage, 133 PIs from stage one and the standards plus cvs. Jack (intermediate) and Spencer (susceptible) were inoculated in two sets. Entries with mean disease severity ratings less than 1 SD from the mean disease severity of the set were tested in a randomized complete block design with two reps. The experiment was repeated in a growth chamber at 25C and 12 hours of light per day.

Soybean were planted in clay pots containing autoclaved, composted soil. Seven pots containing 6 to7 plants were inoculated with each of the seven SMV strains and grown in the greenhouse between March and October, 1980. Tests were done at least twice. Notes on symptom development were taken for 1 month after inoculations.

Seeds were sown and plants were grown in 15-cm-diameter plastic pots (approx. eight plants per pot) in the greenhouse with temperature controlled at 25 +/-3 C and a 14-h photoperiod. Each germplasm was grown in three pots for each treatment (each inoculated SMV strain). Within each treatment, one pot was used as an uninoculated control and the plants in the other two pots were inoculated at the unifoliolate leaf stage with one of the seven SMV strains. Inoculated plants were monitored for symptom expression for 4 wk after inoculation.

Fifteen plants of each line were inoculated with a tooth pick infested with Diaporthe phaseolorum var. meridionalis cepario 7-12 days after planting and placed in 100% relative humidity, 28-30C for 72 h. Disease ratings were made 10-15 days after inoculation. Death/total was calculated as [(infected/2)+ dead]/total*100%. Resistant = 0-25% death/total, Mostly Resistant = 26-50%, Mostly Susceptible = 51-75%, Susceptible = 76-90%, Extremely susceptible = >=91% . Protocol in Spanish

The purpose of this research was to identify soybean germplasm possessing positive traits affecting high seed germinability for seed produced under high-temperature environments, such as in the early soybean production system (ESPS) of the midsouthern United States. Seed was produced in the ESPS for 486 U.S. plant introductions (PI), 25 ancestral lines of U.S. cultivars, and four U.S. cultivars at Stoneville, MS, in 2002 and 2003. Standard and accelerated aging germination percentages, hardseededness, seed coat wrinkling, and incidence of P. longicolla were estimated for each line. Accessions having a mean standard field germination of greater than or equal to 90%, less than 10% hard seed and P. longicolla infection, and less than or equal to 10% wrinkled seed coat were considered "T" (High germinability in high temperature environments); between 80 and 89% standard field germination, less than 10% hard seed and P. longicolla infection, and less than or equal to 10% wrinkled seed coat were considered "I" (Intermediate germinability in high temperature environments); and the rest "S" (Poor germinability in high temperature environments).

The first-stage screen for N2 fixation tolerance to soil-water deficit was the measurement of petiole ureide levels under well-watered conditions. The entries sampled were accessions in maturity groups V through VIII from the USDA Soybean Germplasm Collection and were grown at Stoneville, MS, on a Bosket fine-sandy loam soil (fine-loamy, mixed thermic, Mollic Hapludalf). Entries were grown in either four-row evaluation plots, four-row seed increase plots, or one-row comparison plots. Approximately 1000 plant introductions, which were blocked by maturity group, were screened each year. These introductions represented nearly 40 different countries but most of the accessions came from China (40%), Japan (21%), Indonesia (12%), S. Korea (5%), India (5%), and USA (5%). Plots were maintained under well-watered conditions and irrigated as necessary. In each of three years (1995, 1996, and 1997), petioles were harvested on one day for all cultivars. The harvest day was selected to be at approximately mid-flowering for many of the genotypes and ranged from R1 to R4. Petioles were harvested from the uppermost, fully-expanded leaves from three plants, placed in coin envelopes, and stored in ice chests for approximately 6 h. Subsequently, the petioles were oven dried at 80.C and finely chopped. Approximately 30 mg of petiole tissue was extracted in 1 mL of 0.2 M NaOH for 30 min at 100.C Ureide concentration was determined with an autoanalyzer based on the method of Van Berkum and Sloger (1983) with the modification that the alkaline hydrolysis step was omitted.

Study Name: Wild Soybean Evaluation 1 of Mat Groups 000 to IV Experiment Type: Field Field Study Year: 1985 Exp. Design: Mat groups 000 to IV Latitude: 40 Degrees 08 Minutes N Longitude: 88 Degrees 12 Minutes W Elevation: 227 meters Year started: //1984 Year ended: //1985 Experiment length: 2 years Comment: Evaluated 2 years with 1 replication per year. Planted on May 14 and 15, 1984 and May 20, 1985. Data presented is an average of these 2 replications.