WOODY-LANDSCAPE

PLANTS FROM 11 ACCESSIONS OF 9 ALNUS AND BETULA TAXA PROPAGATED FROM SEED (1999), STEM CUTTINGS (1999), OR SHOOT CULTURE (2000) WERE SUBJECTED TO THREE CYCLES OF DROUGHT TO EVALUATE THEM FOR DROUGHT TOLERANCE. PLANTS WERE GROWN SINGLY IN 700 CUBIC CM OF SOIL: SPHAGNUM PEAT MOSS: PERLITE (1:3:1) IN PLASTIC CONTAINERS (TOP DIAMETER 15 CM). ONE ACCESSION PER TAXON WAS USED, EXCEPT FOR A. JAPONICA AND A. MARITIMA, WHERE TWO ACCESSIONS OF EACH WERE BULKED. ROOT ZONES OF CONTROL PLANTS WERE SATURATED DAILY WITH TAP WATER; DROUGHT WAS IMPOSED BY WITHHOLDING IRRIGATION, BASED ON DIFFERENTIAL EVAPOTRANSPIRATION RATES AMONG THE TAXA. WATER USE WAS ESTIMATED BY TRACKING THE TOTAL WEIGHT OF THE SEVEN REPLICATES OF EACH TAXON IN THE DROUGHT TREATMENT WITHIN ALL DROUGHT CYCLES. DECREASES IN WEIGHT WERE ASSUMED TO BE DUE TO EVAPOTRANSPIRATION. TREATMENT CYCLES WERE SEPARATED BY A 7-DAY RECOVERY PERIOD BETWEEN CYCLES 1 AND 2, BUT CYCLE 3 BEGAN THE DAY AFTER CYCLE 2 ENDED. DROUGHT CYCLES WERE ENDED THE FIRST DAY WHEN LEAVES OF AT LEAST FIVE OF THE NINE TAXA WERE WILTED BEFORE A DAILY IRRIGATION. DROUGHT CYCLES 1, 2, AND 3 EACH LASTED 7 DAYS. TREATMENTS WERE APPLIED SIMULTANEOUSLY. PHOTOSYNTHETIC RATE OF A STANDARD LEAF ON EACH PLANT WAS MEASURED WITH A LI-COR LI-6400 BEFORE IRRIGATION ON THE DAYS CYCLES 1 AND 3 ENDED. MOISTURE CONTENT OF UPPER 6 CM OF THE ROOT MEDIUM OF EACH PLANT WAS MEASURED (MODEL HH1 THETA PROBE/ML1 SENSOR). HEALTH OF SHOOT SYSTEMS WAS SUBJECTIVELY RATED BY TWO EVALUATORS. A RANGE OF RATINGS (1 = DEAD PLANTS THROUGH 5 = EXCELLENT HEALTH) WAS USED. TOTAL LEAF SURFACE AREA OF EACH PLANT WAS DETERMINED (LI-COR MODEL 3100 AREA METER). DATA ON PHOTOSYNTHESIS AND LEAF AREA WERE TRANSFORMED FOR GRIN BY PRESENTING THEM AS PERCENTAGES OF THE MEAN VALUES FOR THE CONTROL TREATMENTS.

PLANTS FROM 11 ACCESSIONS OF 9 ALNUS AND BETULA TAXA PROPAGATED FROM SEED (1999), STEM CUTTINGS (1999), OR SHOOT CULTURE (2000) WERE SUBJECTED TO THREE CYCLES OF FLOODING TO EVALUATE THEM FOR FLOODING TOLERANCE. PLANTS WERE GROWN SINGLY IN 700 CUBIC CM OF SOIL: SPHAGNUM PEAT MOSS: PERLITE (1:3:1) IN PLASTIC CONTAINERS (TOP DIAMETER 15 CM). ONE ACCESSION PER TAXON WAS USED, EXCEPT FOR A. JAPONICA AND A. MARITIMA, WHERE TWO ACCESSIONS OF EACH WERE BULKED. ROOT ZONES OF CONTROL PLANTS WERE SATURATED DAILY WITH TAP WATER; ROOTS OF FLOODED PLANTS WERE IMMERSED CONTINUOUSLY. TREATMENT CYCLES WERE SEPARATED BY A 7-DAY RECOVERY PERIOD BETWEEN CYCLES 1 AND 2, BUT CYCLE 3 BEGAN THE DAY AFTER CYCLE 2 ENDED. TREATMENTS WERE APPLIED SIMULTANEOUSLY. FLOODED CONTAINERS WERE DRAINED AND FERTILIZED BETWEEN CYCLES. PHOTOSYNTHETIC RATE OF A STANDARD LEAF ON EACH PLANT WAS MEASURED WITH A LI-COR LI-6400 BEFORE IRRIGATION ON THE DAYS CYCLES 1 AND 3 ENDED. MOISTURE CONTENT OF UPPER 6 CM OF THE ROOT MEDIUM OF EACH PLANT WAS MEASURED (MODEL HH1 THETA PROBE/ML1 SENSOR). HEALTH OF SHOOT SYSTEMS WAS SUBJECTIVELY RATED BY TWO EVALUATORS. A RANGE OF RATINGS (1 = DEAD PLANTS THROUGH 5 = EXCELLENT HEALTH) WAS USED. TOTAL LEAF SURFACE AREA OF EACH PLANT WAS DETERMINED (LI-COR MODEL 3100 AREA METER). DATA ON PHOTOSYNTHESIS AND LEAF SURFACE AREA WERE TRANSFORMED FOR GRIN BY PRESENTING THEM AS PERCENTAGES OF THE MEAN VALUES FOR THE CONTROL TREATMENTS.

Fruit diameter measurements obtained using Adobe Photoshop CS4 with Silver Fast 6.0 (Plustek OpticPro A320) set at 300 DPI. Measurements in millimeters.

Average refractometer readings from 3 randomoly selected plants using 10 fruits from each plant. Readings obtained using Atago Pocket Refractometer (PAL-1) at time of peak harvest.

Weight of single fruit. Measurements in grams.

Ploidy level determined through University of Connecticut (Dr. Mark H. Brand). Base number is n=17; 2n=34; 4n=68. Fresh leaves were harvested and nuclei suspensions were prepared by chopping approximately 50 mg of young leaf tissue with a fresh razor blade in 55 mm plastic Petri dishes containing extraction buffer prepared according to Arumuganathan and Earle (1991). The procedure was modified in accordance with Meng and Finn (1999) by adding 2 g of PVP-10 per 50 ml of extraction buffer and fluorescently staining released nuclei with propidium iodide after filtering, rather than during the chopping process. Relative fluorescence of total DNA (FL2) for each stained nucleus was determined with a Becton-Dickinson FACS Calibur Dual Laser Flow Cytometer (Becton, Dickinson and Co., Franklin Lakes, NJ) located at the Flow Cytometry and Confocal Imaging Facility at the University of Connecticut in Storrs, CT. The cytometer was equipped with an argon ion laser emitting radiation at 488 nm. For each sample 10,000-20,000 particles were measured. Fluorescent emission data was collected and displayed by BD CellQuestTM software (Becton, Dickinson and Co., Franklin Lakes, NJ) in histograms of nuclei number according to fluorescence intensity, which was proportional to DNA content. The peaks of test samples were compared to peaks derived from control plants containing 2N DNA to determine relative ploidy levels as either diploid, triploid or tetraploid.

Seed images obtained using Adobe Photoshop CS4 with Silver Fast 6.0 (Plustek OpticPro A320) set at 300 DPI with diameter measurements captured using SmartGrain v.1.1 (Tanabata et al. 2012). Random sample of 10, intact seeds. Measurements in millimeters. Diameter defined as the widest distance through the medial portion of the seed, which is typically from the hilum to the distal portion. In general, Gymnocladus dioicus seeds are near perfectly round, but often slightly longer in length by usually 1 mm or less. Reference: Tanabata T, Yamada T, Shimizu Y, Kanekatsu M, Takano M. 2012. SmartGrain: High-Throughput Phenotyping Software for Measuring Seed Shape through Image Analysis.

Gymnocladus accessions held at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS) located in Ames, IA were shipped to the USDA-ARS National Center for Agricultural Utilization Research Station (Bio-Oils Research Unit) located in Peoria, IL to be analyzed for their total-oil and fatty-acid methyl-ester (FAME) profiles. Approximately 15 seeds from each accession were used for analysis. Total Oil Content was analyzed using magnetic resonance (NMR; Bruker MQ Minispec) with a 0.47 T permanent magnet maintained at 40C and provided hydrogen nuclei with a resonance of 20 MHz. The spectrophotometer was calibrated with known soybean oil standards due to the similarity in fatty acids contents. Seed samples for the analysis were randomly selected and run in 8-10 replications (1 seed/replication). The seeds were placed into an 18mm test tube and preheated to 40C before analysis. The pulsed NMR analytical method was set for 16 scans which reports grams of total oil. Oil content was calculated on a dry weight basis using the average moisture content using AOCS Official Method Ca 2c-25. The fatty acid profile analysis was carried out in three replications (10 seeds/replication) using gas chromatography (GC). The GC of fatty acid methyl esters (FAMEs) was done with a Hewlett-Packard 6890 gas chromatograph (Palo Alto, CA), equipped with a flame-ionization detector (FID) and an auto sampler/injector. Analyses were conducted on a SP 2380 30 m x 0.25 mm i.d. with a 0.2 micro M film thickness (Supelco, Bellefonte, PA). Saturated C8-C30 FAEEs standards (Nu-Check Prep Inc., Elysian, MN) were used to make FAME assignments. The SP 2380 analysis were carried out as follows: column flow 0.7 ml/min with helium head pressure of 15 psi; split ratio 100:1; programmed ramp 165C to 265C at 15C/min with a hold of 5 min at 265C ; injector and detector temperatures set at 265C. Fatty acid methyl esters were made by placing a portion of the seeds into a 4 dram vial. Sodium hydroxide/methanol solution 5 mL (0.25 M) was added to the vial and a 0.2g portion of the seed ground for 20 s with a Modular Homogenizer System (Cole-Parmer Instrument Company, Vernon Hills, IL) fitted with a 10 mm diameter shaft. The vial was then sealed with an aluminum lined cap and placed in a heating block maintained at 65C. After one half hour, the vials were removed, allowed to cool and 5ml of hexane and 5ml of saturated sodium chloride solution were added to the vials. The contents of the vials were mixed thoroughly and after the layers separated, a 0.25 ml aliquot from the top hexane layer containing the methyl esters was removed with a Pasteur pipette and diluted up to 2 ml with hexane in a GC vial. One (1) microliter of the sample was injected on the GC using the parameters described above.

Gymnocladus accessions held at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS) located in Ames, IA were shipped to the USDA-ARS National Center for Agricultural Utilization Research Station (Bio-Oils Research Unit) located in Peoria, IL to be analyzed for their total-oil and fatty-acid methyl-ester (FAME) profiles. Approximately 15 seeds from each accession were used for analysis. Total Oil Content was analyzed using magnetic resonance (NMR; Bruker MQ Minispec) with a 0.47 T permanent magnet maintained at 40C and provided hydrogen nuclei with a resonance of 20 MHz. The spectrophotometer was calibrated with known soybean oil standards due to the similarity in fatty acids contents. Seed samples for the analysis were randomly selected and run in 8-10 replications (1 seed/replication). The seeds were placed into an 18mm test tube and preheated to 40C before analysis. The pulsed NMR analytical method was set for 16 scans which reports grams of total oil. Oil content was calculated on a dry weight basis using the average moisture content using AOCS Official Method Ca 2c-25. The fatty acid profile analysis was carried out in three replications (10 seeds/replication) using gas chromatography (GC). The GC of fatty acid methyl esters (FAMEs) was done with a Hewlett-Packard 6890 gas chromatograph (Palo Alto, CA), equipped with a flame-ionization detector (FID) and an auto sampler/injector. Analyses were conducted on a SP 2380 30 m x 0.25 mm i.d. with a 0.2 micro M film thickness (Supelco, Bellefonte, PA). Saturated C8-C30 FAEEs standards (Nu-Check Prep Inc., Elysian, MN) were used to make FAME assignments. The SP 2380 analysis were carried out as follows: column flow 0.7 ml/min with helium head pressure of 15 psi; split ratio 100:1; programmed ramp 165C to 265C at 15C/min with a hold of 5 min at 265C ; injector and detector temperatures set at 265C. Fatty acid methyl esters were made by placing a portion of the seeds into a 4 dram vial. Sodium hydroxide/methanol solution 5 mL (0.25 M) was added to the vial and a 0.2g portion of the seed ground for 20 s with a Modular Homogenizer System (Cole-Parmer Instrument Company, Vernon Hills, IL) fitted with a 10 mm diameter shaft. The vial was then sealed with an aluminum lined cap and placed in a heating block maintained at 65C. After one half hour, the vials were removed, allowed to cool and 5ml of hexane and 5ml of saturated sodium chloride solution were added to the vials. The contents of the vials were mixed thoroughly and after the layers separated, a 0.25 ml aliquot from the top hexane layer containing the methyl esters was removed with a Pasteur pipette and diluted up to 2 ml with hexane in a GC vial. One (1) microliter of the sample was injected on the GC using the parameters described above.

Gymnocladus accessions held at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS) located in Ames, IA were shipped to the USDA-ARS National Center for Agricultural Utilization Research Station (Bio-Oils Research Unit) located in Peoria, IL to be analyzed for their total-oil and fatty-acid methyl-ester (FAME) profiles. Approximately 15 seeds from each accession were used for analysis. Total Oil Content was analyzed using magnetic resonance (NMR; Bruker MQ Minispec) with a 0.47 T permanent magnet maintained at 40C and provided hydrogen nuclei with a resonance of 20 MHz. The spectrophotometer was calibrated with known soybean oil standards due to the similarity in fatty acids contents. Seed samples for the analysis were randomly selected and run in 8-10 replications (1 seed/replication). The seeds were placed into an 18mm test tube and preheated to 40C before analysis. The pulsed NMR analytical method was set for 16 scans which reports grams of total oil. Oil content was calculated on a dry weight basis using the average moisture content using AOCS Official Method Ca 2c-25. The fatty acid profile analysis was carried out in three replications (10 seeds/replication) using gas chromatography (GC). The GC of fatty acid methyl esters (FAMEs) was done with a Hewlett-Packard 6890 gas chromatograph (Palo Alto, CA), equipped with a flame-ionization detector (FID) and an auto sampler/injector. Analyses were conducted on a SP 2380 30 m x 0.25 mm i.d. with a 0.2 micro M film thickness (Supelco, Bellefonte, PA). Saturated C8-C30 FAEEs standards (Nu-Check Prep Inc., Elysian, MN) were used to make FAME assignments. The SP 2380 analysis were carried out as follows: column flow 0.7 ml/min with helium head pressure of 15 psi; split ratio 100:1; programmed ramp 165C to 265C at 15C/min with a hold of 5 min at 265C ; injector and detector temperatures set at 265C. Fatty acid methyl esters were made by placing a portion of the seeds into a 4 dram vial. Sodium hydroxide/methanol solution 5 mL (0.25 M) was added to the vial and a 0.2g portion of the seed ground for 20 s with a Modular Homogenizer System (Cole-Parmer Instrument Company, Vernon Hills, IL) fitted with a 10 mm diameter shaft. The vial was then sealed with an aluminum lined cap and placed in a heating block maintained at 65C. After one half hour, the vials were removed, allowed to cool and 5ml of hexane and 5ml of saturated sodium chloride solution were added to the vials. The contents of the vials were mixed thoroughly and after the layers separated, a 0.25 ml aliquot from the top hexane layer containing the methyl esters was removed with a Pasteur pipette and diluted up to 2 ml with hexane in a GC vial. One (1) microliter of the sample was injected on the GC using the parameters described above.

Data collected: 2008. North Central Regional PI Station in Ames, Iowa staff recorded data. For additional information contact Dr. Mark Widrlechner at (515)-294-3454.

Listing of Omernik Level III Ecoregion where germplasm was obtained. Ecoregions determined using Omernik, J.M., 1987, Ecoregions of the conterminous United States (map supplement): Annals of the Association of American Geographers, v. 77, no. 1, p. 118-125, scale 1:7,500,000.

Listing of USDA Cold Hardiness Zone where germplasm was obtained. Cold Hardiness Zones determined using the USDA Plant Hardiness Zone Map, 2012. Agricultural Research Service, U.S. Department of Agriculture.

Digital image of bark characteristics.

Digital image of plant(s) at peak fall color.

Digital image of fruit(s).

Digital image of herbarium specimen.

Digital image of plant(s) under cultivation.

Digital image of flower(s).

Digital image of leaf or leaves.

Digital image of plant(s) at collection site in nature.

Scanned image of fruit(s).

Scanned image of seed.

Scanned image of one seed per parent tree.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2007.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2009.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2010.

Observational data collected from ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2011.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2012.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2013.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2014.

Observational data collected from ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2015.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2018.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2020.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2021.

Observational data collected from woody ornamentals growing at the North Central Regional Plant Introduction Station in Ames, Iowa throughout 2022.

In 2009 and 2010, Aronia accessions were established at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS), Iowa State University, Ames, IA. Seedlings were started by germinating seeds and then cultivating them in the greenhouse until seedlings were approximately 15-30 cm in height. They were then transplanted by hand in 2009 and 2010 by hand to a field at the NCRPIS farm (42 degrees 00' 40"N, 93 degrees 39' 33"W). Seedlings were grouped by accession in a single row. Plants were spaced 45-cm apart within rows and 550-cm apart between rows in a randomized block design. Observations were taken in 2012 for the following descriptors: date of peak flowering, flower diameter, number of inflorescences per 30.5 cm, fruit color, fruit diameter, date of peak fruit ripening, plant height, and fruit weight.

In 2009 and 2010, Aronia accessions were established at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS), Iowa State University, Ames, IA. Seedlings were started by germinating seeds and then cultivating them in the greenhouse until seedlings were approximately 15-30 cm in height. They were then transplanted by hand in 2009 and 2010 by hand to a field at the NCRPIS farm (42 degrees 00' 40"N, 93 degrees 39' 33"W). Seedlings were grouped by accession in a single row. Plants were spaced 45-cm apart within rows and 550-cm apart between rows in a randomized block design. Observations were taken in 2013.

In 2009 and 2010, Aronia accessions were established at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS), Iowa State University, Ames, IA. Seedlings were started by germinating seeds and then cultivating them in the greenhouse until seedlings were approximately 15-30 cm in height. They were then transplanted by hand in 2009 and 2010 by hand to a field at the NCRPIS farm (42 degrees 00' 40"N, 93 degrees 39' 33"W). Seedlings were grouped by accession in a single row. Plants were spaced 45-cm apart within rows and 550-cm apart between rows in a randomized block design. Observations were taken in 2014.

In 2009 and 2010, Aronia accessions were established at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS), Iowa State University, Ames, IA. Seedlings were started by germinating seeds and then cultivating them in the greenhouse until seedlings were approximately 15-30 cm in height. They were then transplanted by hand in 2009 and 2010 by hand to a field at the NCRPIS farm (42 degrees 00' 40"N, 93 degrees 39' 33"W). Seedlings were grouped by accession in a single row. Plants were spaced 45-cm apart within rows and 550-cm apart between rows in a randomized block design. In 2013, LAI was measured using an AccuPAR Ceptometer (Model LP-80; Decoagon Devices, Inc.; Pullman, WA). Measurements were attained at mid-section of the plot, at ground level, ~10 cm parallel from the row of plants. The AccuPAR Ceptometer probe is 19 cm x 9.5 cm, and contains 8 segmented PAR sensors. Measurements were recorded three times on each side of the plant. The AccuPAR calculated LAI based on the above and below-canopy PAR measurements along with other variables (i.e. zenith angle, fractional beam measurement value, and leaf area distribution parameter). An average LAI was calculated from a total of six measurements.

Length and width of the pistillate catkin (measured in mm) using SmartGrain v.1.1 (Tanabata et al. 2012). Random sample of 20, intact catkins. Length is defined as distance between distal to proximal ends. Length is defined as distance between distal to proximal ends. Tanabata T, Yamada T, Shimizu Y, Kanekatsu M, Takano M. 2012. SmartGrain: High-Throughput Phenotyping Software for Measuring Seed Shape through Image Analysis.

Length and width of a winged nutlet from a single specimen (measured in mm) using SmartGrain v.1.1 (Tanabata et al. 2012). Random sample of 50, intact single winged nutlets. Length is defined as distance between distal to proximal ends. Tanabata T, Yamada T, Shimizu Y, Kanekatsu M, Takano M. 2012. SmartGrain: High-Throughput Phenotyping Software for Measuring Seed Shape through Image Analysis.

Length of peduncle (measured in mm) from pistillate flowers using SmartGrain v.1.1 (Tanabata et al. 2012). Random sample of 20, intact catkins. Length is defined as distance between distal to proximal ends. Length is defined as distance between distal to proximal ends. Tanabata T, Yamada T, Shimizu Y, Kanekatsu M, Takano M. 2012. SmartGrain: High-Throughput Phenotyping Software for Measuring Seed Shape through Image Analysis.

Length and width of a pericarp from a single specimen (measured in mm) using SmartGrain v.1.1 (Tanabata et al. 2012). Random sample of 20, intact single winged nutlets. Length is defined as distance between distal to proximal ends. Tanabata T, Yamada T, Shimizu Y, Kanekatsu M, Takano M. 2012. SmartGrain: High-Throughput Phenotyping Software for Measuring Seed Shape through Image Analysis.

Ploidy level determined through Woody Landscape Plant Germplasm Repository (Richard Olsen).

Seed length and width measurements obtained using Tomato Analyzer 3.0 (Department of Horticulture and Crop Science, The Ohio State University/Ohio Agricultural Research and Development Center, Wooster, Ohio 44691) with Silver Fast 6.0 (Plustek OpticPro A320) set at 300 DPI. Random sample of 20 intact seeds. Measurements in millimeters.

Nut measurements (Ercisli et al.,2012) obtained using a digital caliper (Mitutoyo Absolute Digimatic Caliper Series 500). Random sample of 50 intact nuts. Measurements in millimeters. Reference: Ercisli, S., B. Sayinci, M. Kara, C. Yildiz, and I. Ozturk. 2012. Determination of size and shape features of walnut (Juglans regia L.) cultivars using image processing. Scientia Horticulturae 133: 47-55.

Seed length and width measurements obtained using SmartGrain V. 1.1 (Tanabata et al. 2012) with Silver Fast 6.0 (Plustek OpticPro A320) set at 600 DPI. Random sample of 50, single intact seeds. Length is defined as distance between distal to proximal ends and width is defined as widest distance between distal and proximal ends. Measurements in millimeters. Tanabata T, Yamada T, Shimizu Y, Kanekatsu M, Takano M. 2012. SmartGrain: High-throughout Phenotyping Software for Measuring Seed Shape through Image Analysis.

Fruit diameter measurements obtained using Adobe Photoshop CS4 with Silver Fast 6.0 (Plustek OpticPro A320) set at 300 DPI. Measurements in millimeters.

Refractometer reading from bulk fruit harvest. Readings obtained using Atago Pocket Refractometer (PAL-1) at time of peak harvest.

Weight of single fruit. Measurements in grams.

Fruit diameter measurements obtained using Adobe Photoshop CS4 with Silver Fast 6.0 (Plustek OpticPro A320) set at 300 DPI. Measurements in millimeters.

Average refractometer readings using 9 fruits from a single specimen or population. Readings obtained using Atago Pocket Refractometer (PAL-1).

Weight of single fruit. Measurements in grams.

Fruit diameter measurements obtained using using SmartGrain v.1.1 (Tanabata et al. 2012). Adobe Photoshop CS4 with Silver Fast 6.0 (Plustek OpticPro A320) set at 300 DPI. Random sample of 25 fruits Measurements in millimeters.

Length and width (mm) of a Sapindus seeds from a single specimen measured using SmartGrain v.1.1 (Tanabata et al. 2012). Random sample of 25, Length is defined as distance between distal to proximal ends.

Ploidy level determined through USDA-ARS (Dr. Alan Whittemore). Flow cytometry was carried out on a CyFlow Space flow cytometer, following manufacturers suggested protocol. DNA was stained with propidium iodide, and Glycine max 'Williams 82' was used as an internal standard.