Methods
100 seed weight at the Western Regional Plant Introduction Station, Pullman Washington, USA
Collection pictures from the Richard C. Johnson, Harold Bockelman, Korzhenevsky, V., Boguslavsky, R. collection in 1999 from Krym, Ukraine.
Collection pictures taken in 2000 from the Kazakstan trip made by Rich Hannan, Stephanie Greene, A. Khusainov, A. Afonin, and N. Dzyubenko.
Collection pictures taken in 2006 from the Tajikistan trip by Barbara Hellier, Kenneth Street, Zebuniso Muminshoeva, Farkhod Kosimov, Shakhlo Safarzoda, John Sheppard, Natalya Rukhkyan and Sergey Shuvalov.
Tajikistan collection trip map.
Three hundred and fourteen accessions representing the NPGS M. truncatula collection were evaluated for length of growth stage, measured in days. Seeds were scarified and sown on May 5, 2003, into containers containing a soiless mix, and seedlings kept in the greenhouse for 4 weeks before being transplanted into the field, at Prosser, WA. Field trial was set up as a randomized complete block design with three replications. Each accession was represented by 30 plants (10 plants/rep). Eleven growth stages were identified, and days from sowing to each growth stage was recorded. Growth stages included 1) Full Emergence of Cotyledons, 2) Emergence of Unifoliate Leaf, 3) Emergence of 1st Trifoliate Leaf, 4) Emergence of 2nd Trifoliate Leaf, 5) Start of Vegetative Growth, 6) Start of Flowering, 7) End of Flowering, 8) First Full Size Green Pod, 9) First Brown Pod, 10) Start of Senescence, 11) End of Senescence.
Excel spread sheet of the Early growth stages.
Excel spread sheet of the Mid growth stages.
Excel spread sheet of the Late growth stages.
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Images/pictures from Prosser, Washington
Medicago truncatula seeds were surface sterilized, imbibed, and then cold treated by placing on moist filter paper in a Petri dish and incubating for at least 48 h at 4? C. Seeds were germinated in the dark at 24?C for 24 h. Seedlings were transferred to hydroponics and grown under an 18-h-light/6-h-dark regime at a constant temperature of 23?C. The light intensity was 260 uE?m-2?s-1. Twenty seedlings of each accession were used for each treatment: a modified Blaydes medium (Blaydes, 1966) plus Al, and a modified Blaydes medium minus Al. Media were aerated with a small aquarium air pump for 15 min, once per h. The Blaydes medium was modified to contain 0.5 mM CaCl2, 1.8 mM KNO3, 1.8 mM NH4NO3, 30.4 uM MgSO4 ?7 H2O, 22 uM KH2PO4, 13 uM FeSO4, 31.7 uM Ca(NO3)2, 66 uM KCl, 26 uM NaCl, 26 uM H3BO3, 26 uM MnSO4?H20, 5.2 uM ZnSO4?7 H2O, and 4.8 uM KI. Media was maintained at pH 4.3. After five d of growth in hydroponics, seedlings were removed and measured from the base of the cotyledon to the root tip. The experiment was repeated three times, once with the Al concentration at 25 uM as AlCl3 and twice at 50uM AlCl3. Relative root growth, the ratio of seedling root length grown without Al to seedling root length grown with Al, was used as a measure of Al tolerance.
Click for more information on Materials, Methods and data in a .DOC file. Click for an Excel file with root length of the control, stressed and relative along with P-value.
from 1986-1987 increase crop
from 1988-1989 increase crop
from 1989-1990 increase crop
Biomass among species done in 1991 by Craig Sheaffer and Don Barnes in Minnesota
Biomass among species done in 1991 by Noa Diwan in Beltsville, Maryland.
Biomass among species done in 1991 by Mel Rumbaugh in Utah.
Biomass among species done in 1991 by Steve Smith at Tucson, Arizona
Biomass among species done in 1992 by Joe Bouton in Georgia
Biomass among species done in 1992 by Noa Diwan in Beltsville
Biomass among species done in 1992 by Mel Rumbaugh in Utah
Biomass among species done in 1992 by Don Viands in New York
from 1992-1993 increase crop
from 1993-1994 increase crop
from 1994-1995 increase crop
from 1995-1996 increase crop
Biomass within species done in 1991 by Craig Sheaffer and Don Barnes, St. Paul, Minnesota.
Biomass within species done in 1991 by Noa Diwan, Beltsville, Maryland.
Biomass within species done in 1991 by Mel Rumbaugh, Logan, Utah.
Biomass within species done in 1991 by Steve Smith, Tucsno, Arizona.
Biomass within species done in 1992 by Joe Bouton, Athens, Georgia.
Biomass within species done in 1992 by Noa Diwan, Beltsville, Maryland.
Biomass within species done in 1992 by Mel Rumbaugh, Logan, Utah.
Biomass within species done in 1992 by Don Viands, Ithaca, New York.
Chromosome number taken from a literature article
Average number of coils per pod done by Robin Groose
Annual Medicago Core developed by Gary L. Bauchan, USDA-ARS, Beltsville, Maryland in 1991. See Crop Sci. 34:279-285(1994). 16 agronomic & morphologic triats on 1240 accessions were grouped by cluster analysis utilizing an unweighed pair group method with arithmetic averages. One accessions per cluster for each species made up the core.
Days to flower done by Craig Sheaffer and Don Barnes in Minnesota
Days to flower done by Noa Diwan in Beltsville, Maryland.
Days to flower done by Mel Rumbaugh in Utah.
Days to flower done by Steve Smith at Tucson, Arizona
Days to flower done by Joe Bouton in Georgia
Days to flower done by Noa Diwan in Beltsville
Days to flower done by Mel Rumbaugh in Utah in 1992
Days to flower donw by Don Viands in New York
Days to full pod production done in 1991 by Craig Sheaffer and Don Barnes, St. Paul, Minnesota.
Days to full pod production done in 1991 by Noa Diwan, Beltsville, Maryland.
Days to full pod production done in 1991 by Mel Rumbaugh, Logan, Utah.
Days to full pod production done in 1991 bt Steve Smith, Tucson, Arizona.
Days to full pod production done in 1992 by Noa Diwan, Beltsville, Maryland.
Days to full pod production done in 1992 by Mel Rumbaugh, Logan, Utah.
Plant growth habit done in 1991 by Craig Sheaffer and Don Barnes, St. Paul, Minnesota.
Plant growth habit done in 1991 by Noa Diwan, Beltsville, Maryland.
Plant growth habit done in 1991 by Mel Rumbaugh, Logan, Utah.
Plant growth habit done in 1991 by Steve Smith, Tucson, Arizona.
Plant growth habit done in 1992 by Joe Bouton, Athens, Georgia.
Plant growth habit done in 1992 by Noa Diwan, Beltsville, Maryland.
Plant growth habit done in 1992 by Mel Rumbaugh, Logan, Utah.
Plant growth habit done in 1992 by Don Viands, Ithaca, New York.
Plant height in centimeters done in 1991 by Craig Sheaffer, St. Paul, Minnesota.
Plant height in centimeters done in 1991 by Noa Diwan, Beltsville, Maryland.
Plant height in centimeters done in 1991 by Mel Rumbaugh, Logan, Utah.
Plant height in centimeters done in 1991 by Steve Smith, Tucson, Arizona.
Plant height in centimeters done in 1992 by Joe Bouton, Athens, Georgia.
Plant height in centimeters in 1992 by Noa Diwan, Beltsville, Maryland.
Plant height in centimeters in 1992 by Mel Rumbaugh, Logan, Utah.
Plant height in centimeters in 1992 by Don Viands, Ithaca, New York.
Medicgo truncatula seeds were scarified with sandpaper and sterilized in 50% (v/v) commercial bleach for 15 min, then they were rinsed (4?20 min) with sterile water. Seeds were imbibed in sterile water at 4 degrees C overnight and then germinated on 1% agar/water in Petri dishes in growth chamber conditions (25/19 degrees C, 16/8 h) for 48 h in darkness. Seedlings were transferred to the so-called Fahraeus slide hydroponic system, as described previously (Heidstra et al., 1994; Ortega-Villasante et al., 2005) with some modifications. A modified Hoagland nutrient solution was used. The relative root growth (RRG) of the seedlings was used as an indicator of metal tolerance, measuring the root length of the seedlings before and after treatment, and expressing the increase in length relative to that of control seedlings grown in the absence of Hg.
See the publication: 'Environmental and Experimental Botany 91 (2013) 90-96' for more detailed information on how the experiment was preformed.
Note many of the National Plant Germplams System (NPGS) accessions used in this experiment now have 'PI' numbers instead of 'W6' numbers. See the comment column in the GRIN data to find the W6 number that corresponds to the PI number.
For a pdf file of the article.
For a pdf file of the data.
Reaction to Erysiphe pisi was evaluated in the annual Medicago core germplasm collection of 199 USDA Plant Introduction accessions from 33 species. Seedlings at the first trifoliolate leaf stage were inoculated with conidia and maintained in growth chambers for 21 days at 200C, 70-90% relative humidity, and a 12-h photoperiod. Beginning 4 days after inoculation, plants with the first sign of powdery mildew were recorded and removed daily. Results for each accession were presented as the percentage of plants mildew-free 21 days after inoculation and also as Area Under the Disease Progress Curve (AUDPC) values expressed as standardized proportion/days. AUDPC values were useful to account for length of latent period (number of days from inoculation until first sign of mildew). Thirty-two accessions from 14 species included plants mildew-free 21 days after inoculation. Mildew-free plants in those 32 accessions ranged from 100% in M. heyniana PI 537136 and M. truncatula PI 292436 down to 1% in six accessions. All plants in 87 accessions, which included all accessions of 11 species, had powdery mildew 4 days after inoculation and thus expressed no resistance. The remaining 80 accessions included slow-mildewing plants with latent periods of 5 to14 days. This slow-mildewing trait may be a useful source of resistance to E. pisi.
Peronospora trifoliorum causes downy mildew of alfalfa throughout the temperate regions of the world, but little is known about its host range within the annual Medicago species (medics). Several medics have characteristics potentially beneficial to sustainable agriculture programs and to the genetic improvement of alfalfa. Therefore, we evaluated resistance in 199 accessions representing 33 species of the annual Medicago core germplasm collection to P. trifoliorum isolates I7 and I8 from alfalfa from Kansas and southern California, respectively. Seedlings at the cotyledonary growth stage were inoculated with conidia suspended in water. One week later, the plants were rated resistant (no conidium production) or susceptible (conidium production). High levels of resistance existed in all species. All plants in 142 (71%) of the accessions were resistant to both isolates, but plants in only 24 of those resistant accessions were free of chlorotic to necrotic hypersensitive reactions on the inoculated cotyledons. All but one of the medic accessions had a higher percentage of plants resistant to both isolates than did cv. Saranac alfalfa, the resistant control. Conidium production generally was much less intense on the susceptible medic plants than on the alfalfa controls. A greater percentage of plants in four medic accessions was more resistant to I8 and I7, and 22 were more resistant to I7 than to I8.
Reaction to Erysiphe pisi was evaluated in the annual Medicago core germplasm collection of 199 USDA Plant Introduction accessions from 33 species. Seedlings at the first trifoliolate leaf stage were inoculated with conidia and maintained in growth chambers for 21 days at 200C, 70-90% relative humidity, and a 12-h photoperiod. Beginning 4 days after inoculation, plants with the first sign of powdery mildew were recorded and removed daily. Results for each accession were presented as the percentage of plants mildew-free 21 days after inoculation and also as Area Under the Disease Progress Curve (AUDPC) values expressed as standardized proportion/days. AUDPC values were useful to account for length of latent period (number of days from inoculation until first sign of mildew). Thirty-two accessions from 14 species included plants mildew-free 21 days after inoculation. Mildew-free plants in those 32 accessions ranged from 100% in M. heyniana PI 537136 and M. truncatula PI 292436 down to 1% in six accessions. All plants in 87 accessions, which included all accessions of 11 species, had powdery mildew 4 days after inoculation and thus expressed no resistance. The remaining 80 accessions included slow-mildewing plants with latent periods of 5 to14 days. This slow-mildewing trait may be a useful source of resistance to E. pisi.
Peronospora trifoliorum causes downy mildew of alfalfa throughout the temperate regions of the world, but little is known about its host range within the annual Medicago species (medics). Several medics have characteristics potentially beneficial to sustainable agriculture programs and to the genetic improvement of alfalfa. Therefore, we evaluated resistance in 199 accessions representing 33 species of the annual Medicago core germplasm collection to P. trifoliorum isolates I7 and I8 from alfalfa from Kansas and southern California, respectively. Seedlings at the cotyledonary growth stage were inoculated with conidia suspended in water. One week later, the plants were rated resistant (no conidium production) or susceptible (conidium production). High levels of resistance existed in all species. All plants in 142 (71%) of the accessions were resistant to both isolates, but plants in only 24 of those resistant accessions were free of chlorotic to necrotic hypersensitive reactions on the inoculated cotyledons. All but one of the medic accessions had a higher percentage of plants resistant to both isolates than did cv. Saranac alfalfa, the resistant control. Conidium production generally was much less intense on the susceptible medic plants than on the alfalfa controls. A greater percentage of plants in four medic accessions was more resistant to I8 and I7, and 22 were more resistant to I7 than to I8.
Medicago rigiduloides accessions were planted in sterile conditions and inoculated with a common rhizobium strain. Effective nodulation after 6-10 weeks was noted. If an accession nodulates (>0%) with this rhizobium it is not Medicago rigiduloides. The Australian at the South Australian Dept. of Agriculture have determined that the Medicago species rigidula and rigiduloides can be separated on the basis of rhizobium selectivity. For example, Medicago rigidula, the Western Mediterranean species, nodulates with a common Medic rhizobium that inoculates some 90% of annual medics, including M. polymorpha and even M. doliata. However, Medicago rigiduloides is very specific in its rhizobium requirement and will not nodulate with the common rhizobium strain as it does with other medic species. The South Australians have found a specific rhizobium strain from the Middle East that nodulates M. rigiduloides.
Visual pod/seed production done in 1991 by Noa Diwan in Beltsville, Maryland.
Visual pod/seed production done in 1991 by Mel Rumbaugh in Utah.
Visual pod/seed production done in 1991 by Steve Smith at Tucson, Arizona
Visual pod/seed production done in 1992 by Joe Bouton in Georgia
Visual pod/seed production done in 1992 by Noa Diwan in Beltsville
Visual pod/seed production done in 1992 by Mel Rumbaugh in Utah
Visual pod/seed production done in 1992 by Don Viands in New York
Pod tightness on Medicago rigidula and Medicago rigiduloides
100 seed weight at the Western Regional Plant Introduction Station, Pullman Washington, USA
Curvature of the spines on the pods done by Robin Groose 1998.
Pod spines done in 1991 by Noa Diwan, Beltsville, Maryland
Pod spines done in 1991 by Mel Rumbaugh, Logan, Utah
Pod spines done in 1991 by Steve Smith, Tucson, Arizona
Pod spines done in 1991 by Noa Diwan, Beltsville, MAryland.
Pod spines done in 1992 by Mel Rumbaugh, Logan, Utah
Spread done in 1991 by Criag Sheaffer and Don Barnes St. Paul, Minnesota.
Spread done by Noa Diwan, Beltsville, Maryland.
Spread done in 1991 by Mel Rumbaugh, Logan, Utah.
Spread done in 1991 by Steve Smith, Tucson, Arizona.
Spread done in 1992 by Joe Bouton, Athens, Goergia.
Spread done in 1992 by Noa Diwan, Beltsville, MAryland.
Spread done in 1992 by Mel Rumbaugh, Logan, Utah.
Spread done in 1992 by Don Viands, Ithaca, New York.
Pollen shape done by Robin Groose to determine the taxonomy of Medicago rigidula and Medicago rigiduloides.
Medicago accessions in the HapMap project. The HapHap project can be found at:
root@medicagohapmap.org
All accessions are Medic's except seven:
PI 504540 Medicago arborea (HM319)
PI 440491 Medicago cancellata (HM320)
PI 577445 Medicago prostrata (HM339)
PI 568100 Medicago ruthenica (HM335)
PI 314275 Medicago sativa ssp. caerulea (HM336)
PI 495200 Medicago arabica (HM318)
PI 534177 Medicago constricta (HM321)
For a xlsx file of the National Plant Germplasm System HapMap accessions
Medicago truncatula core includes acccessions without an accession suffix and accessions with an acccession suffix of SSD (Single Seed Descent).
Three hundred and fourteen accessions representing the NPGS Medicago truncatula collection were evaluated for morphological characters. Seeds were scarified and sown on May 5, 2003, into containers containing a soiless mix, and seedlings kept in the greenhouse for 4 weeks before being transplanted into the field, at Prosser, WA. Field trial was set up as a randomized complete block design with three replications.
For an excel file of the Medicago truncatula 2003 morphology data.