White mold, (Sclerotinia sclerotiorum), is an aggressive pathogen of beans and is capable of inflicting devastating damage on yield. Finding resistance is a major concern to bean breeders. The scarlet runner bean (Phaseolus coccineus) is generally known to have greater resistance to white mold than does the common bean, (P. vulgaris). Since it is possible to cross these two species, we have started to examine the NPGS core collection of P. coccineus for resistance to this pathogen. A straw test was used to measure physiological resistance of bean stems to white mold. A rating of one equates to a small lesion, resulting from contact with inoculum, and a rating of nine describes total plant collapse. Controls that were used were two common beans, 91G, a commercially produced, blue lake type snap bean and ExRico, a small, white dry bean. The bean 91G received a straw test rating of 8.3, which correlates to a field test rating of 8.5. ExRico rated 7.4 with the straw test and had a field test score of 6.5. Within the P. coccineus collection we found very strong resistance, with straw test values of 1 and 2 in several individual plants and in some accessions. Accessions that had individuals that displayed the strongest resistance of all the plants tested were: PI201299, PI361302, PI406938, and PI535278. These accessions appeared to be segregating for white mold resistance. Accessions showing the best average resistance were: PI313221, PI361372, PI361539, and PI583553. Because P. coccineus is outcrossed, we expected to find variation within accessions for white mold resistance. Some accessions had uniformly high levels of resistance, while other accessions showed variability.
Barbara Gilmore and James R. Myers
Melike Cirak and James R. Myers
The persistent color (pc) trait in snap bean (Phaseolus vulgaris L.) is a member of the stay-green gene family and falls into the cosmetic subclass. Cosmetic stay-green variants remain green but lose photosynthetic competence during senescence. It is an economically useful trait in snap bean as a result of its effects on pod quality. The trait produces a dark-green, uniform appearance of fresh pods, but has other pleiotropic effects, including a light-green seed color, bleached-white cotyledons on emergence, and foliage and pods that remain green even while senescing. One additional pleiotropic effect is reduced field germination and emergence compared with white- and colored-seeded genotypes. Nevertheless, with the aid of seed-applied fungicides, pc types occupy ≈40% of commercial snap bean acreage in the United States. This research project was aimed at understanding why and how germination and emergence is affected in pc beans. The effect is thought to be related to soil-borne pathogens because fungicide treatment of pc seeds increases germination and emergence rates to levels comparable to treated white- and colored-seeded genotypes. For our experiments, we increased seeds of 45 experimental lines and commercial cultivars (25 of which were pc) under uniform growing conditions. Initial experiments documented that, in the laboratory, all seeds analyzed in a tetrazolium test had high viability. Furthermore, untreated seeds of pc and non-pc types germinated in the laboratory showed no difference in germinability, whereas in the field, germination of pc types was reduced significantly. In addition, pc types showed substantially greater infection rates of seeds and seedlings, with the main pathogen being Fusarium oxysporum Schl. f. sp. phaseoli Kendrick & Snyder. Water uptake by green pc seeds was significantly more rapid than white and colored seeds. Measurements of electrical conductivity revealed that pc types had greater solute leakage than other seed types. When seed anatomic structure was examined, pc types had a significantly thinner testa, especially the osteosclereid layer. The reduction in germination and emergence appears to begin with a thinner, more fragile testa showing increased cracking that may happen during seed harvest and conditioning (but certainly does happen during imbibition), allowing more rapid water uptake during germination that leads to testa rupture. Increased and rapid solute diffusion into the surrounding spermosphere stimulates and attracts pathogens to colonize the seeds before seedlings can become established. Seed handling and conditioning processes before planting could be modified to improve field emergence and stand establishment. Selection for thicker testa may also mitigate some of the damage observed during germination of pc cultivars.
Rebecca Nelson Brown and James R. Myers
Marker-based selection for resistance to zucchini yellow mosaic virus in squash (Cucurbita spp.) would allow breeders to screen individual plants for resistance to multiple viruses. The C. moschata landrace Nigerian Local is widely used as a source of resistance in C. pepo breeding programs. We used RAPDs and bulk-segregant analysis to screen two BC1 populations for a marker linked to the dominant major gene for resistance from Nigerian Local. The initial cross was Waltham Butternut × Nigerian Local; the test populations were created from reciprocal backcrosses to Waltham Butternut. Both populations segregated 1:1 for resistance when hand-inoculated with ZYMV. RAPD primers were screened on a resistant bulk and a susceptible bulk from each population, and Waltham Butternut and Nigerian Local. Primers that gave bands linked to resistance were further screened using DNA from individual plants in each population. The potential markers will be tested on several populations derived from crosses between summer squash (C. pepo) and Nigerian Local to determine if they would be useful for selection in a C. pepo background.
Carl M. Jones and James R. Myers
Continued and mounting evidence of the health benefits provided by carotenoid and anthocyanin pigments has increased public interest in dietary sources of these important phytonutrients. Tomatoes (Lycopersicon esculentum) are the primary dietary contributor of lycopene and an important source of beta-carotene. A collection of tomatoes containing the genes hp-1, dg, ogc, Ip, B and Af that are known to affect carotenoid and anthocyanin levels have been analyzed using HPLC. Levels of lycopene, beta-carotene, phytoene, and phytofluene have been determined in these accessions. Accession LA 3005, containing the dg gene, had the highest lycopene levels of the accessions analyzed (14 mg/100 g fresh wt.). A rapid HPLC method for quantitation of carotenoid levels from tomato fruit has been developed. “Heirloom” black and purple tomatoes have also been included in the accessions analyzed and have carotenoid levels comparable to cultivated red tomatoes. Anthocyanin presence has been confirmed only in the accessions LA 1996 (Af) and in some fruit of segregating plants from LA 3668 (Abg). Total monomeric anthocyanin content of LA 1996 as measured by the pH differential method is estimated to be 5.6 mg/100 g in the outer pericarp tissues and 18.6 mg/100 g in the skin tissue.
Peter Boches, Brooke Peterschmidt, and James R. Myers
One hundred S. l. var. cerasiforme (Dunal) accessions from the ‘Tanksley’ designated core collection were evaluated for horticultural quality under greenhouse conditions. Fourteen selected accessions were grown under field conditions in a replicated trial to evaluate the fruit for phenolic content. Total fruit phenolics ranged from 44 to 82 mg/100 g gallic acid equivalents (GAE) fresh weight (FW) as measured by Folin-Ciocalteau assay (F-C), and 12 to 108 mg/100 g FW as measured by high-performance liquid chromatography (HPLC). Five accessions (LA1712, LA1455, LA2633, LA1668, and LA2632) had significantly higher total phenolics (F-C) than cultivars (P ≤ 0.05). These five accessions also possessed interesting phenolics profiles, including high levels of caffeoylquinic acids (LA1620, LA1455, and LA2633) and rutin (LA2633).
J. Erron Haggard and James R. Myers
White mold, caused by Sclerotinia sclerotiorum (Lib.) de Bary, causes major losses in dry and snap bean (Phaseolus vulgaris) production. With little genetic variation for white mold resistance in common bean, other potential sources for resistance must be investigated. Accessions of scarlet runner bean (P. coccineus) have been shown to have partial resistance exceeding any to be found in common bean. Resistance is quantitative with at least six QTL found in a P. coccineus intraspecific resistant × susceptible cross. Our goal is to transfer high levels of resistance from P. coccineus into commercially acceptable common bean lines. We developed interspecific advanced backcross populations for mapping and transfer of resistance QTL. 111 BC2F5 lines from a cross between OR91G and PI255956 have been tested in straw tests and oxalate tests, as well as in a field trial. The data show that the OR91G × PI255956 population carries a high level of resistance, but because of the quantitative nature of resistance, it may be necessary to intercross individuals to achieve higher levels. SSR, RAPD, and AFLP markers are being tested in the population to construct a linkage map for placement of QTL. QTL identified from each type of test (straw, oxalate, and field) may provide additional information about the genetic architecture of white mold resistance. Three other populations are from advanced backcrosses of the recurrent parents G122, OR91G, and MO162, with PI433251B as the donor parent in each. Analyses and advance of these populations will follow, the results of which should confirm QTL identified in the OR91G × PI255956 population, as well as possible additional resistance QTL from PI433251B.
Peter J. Mes* and James R. Myers
Tomato lines carrying the genes Aft, atv, Abg, hp-1, and an as yet undetermined gene from the introgression line LA2099 have been combined to produce fruit with elevated anthocyanin content. The antioxidant activity of juice made from anthocyanin-expressing tomatoes was compared to juices made from tomatoes with varied carotenoid content. The contribution of anthocyanin to the total antioxidant activity of the whole fruit in current material is small, but with potential for significant improvement. The increase in flavonoids in the elevated anthocyanin lines has increased water-soluble antioxidant activity of the fruit in vitro.
Rebecca Nelson Brown and James R. Myers
A molecular and morphological marker map would improve our knowledge of Cucurbita genetics, and would facilitate efforts to breed improved summer and winter squash cultivars. Random amplified polymorphic DNA (RAPD) markers were used to construct a partial map of the Cucurbita genome. The mapping population was the BC1 progeny of the Cucurbita pepo L. yellow straightneck inbred A0449 and the tropical Cucurbita moschata Duchesne ex Lam. landrace `Nigerian Local'. A0449 was the recurrent parent. This cross was chosen because of the relatively greater economic importance of summer squash, traits of value to be introgressed from the C. moschata parent, and maximized genetic variation from the interspecific cross. The map contains 148 RAPD markers in 28 linkage groups. Loci controlling five morphological traits were placed on the map. The map covers 1,954 cM, which is estimated to be 75% of the Cucurbita genome. The qualitative traits placed on the map include the B gene for fruit which turn yellow before anthesis, the M gene for silver mottling of leaves, and a locus controlling the intensity of rind color on mature fruit. Quantitative trait loci (QTL) associated with fruit shape and the depth of the indentations between primary leaf veins were identified.
Peter J. Mes*, James R. Myers, and Balz Frei
A nutritional study was initiated to determine which carotenoids found in tomato result in decreased lipid oxidation ex vivo. To compare the carotenoids in a human diet without the use of purified supplements, tomatoes expressing nonfunctional enzymes in the carotenoid pathway were used. Tomato lines carrying the genes t, B, ogc, Del, or r were grown to produce fruit containing with high levels of prolycopene, beta-carotene, lycopene, or delta-carotene respectively, or low total carotenoids in r. Juices were processed from these lines and used in a dietary intervention study. Plasma samples were drawn before and after consumption of each juice. These samples were subjected to a battery of tests to analyze the contribution of carotenoids to the total lipid antioxidant status. Results of these tests are discussed.
Peter J. Mes, Peter Boches, James R. Myers, and Robert Durst
Cultivated tomatoes (Solanum lycopersicum L.) produce anthocyanins in vegetative tissues and certain flavonols can be found in the fruit. Some related wild species do produce anthocyanins in the fruit, and this trait has been transferred into cultivated tomato. Fruit with the genes Abg, Aft, and atv exhibit varying degrees of anthocyanin production in the epidermis, but not in the fruit pericarp. Fruit with these alleles in various combinations were analyzed to characterize the anthocyanidin profile, moieties, and total anthocyanin content. In general, combining atv with either Aft or Abg substantially increased anthocyanin production in the fruit. Over 23 different anthocyanins were detected, petunidin-3-(p-coumaryl)-rutinoside-5-glucoside being predominant. The highest level of anthocyanin expression was observed in small fruit with the genotype Abg- atvatv and AftAft atvatv, well in excess of 100 mg/100 g fresh weight of epidermis and subepidermis depending on the size of the fruit. Nonanthocyanin flavonoids were also upregulated in proportion to the anthocyanin concentration. The anthocyanin genes were also combined with genes affecting carotenoid composition and content. Reduced carotenoid content conditioned by the alleles B (Beta) and r (yellow flesh) was associated with lower total anthocyanins, an unexpected observation because the carotenoid and anthocyanin pathways are thought to be independent. The level of anthocyanin did not affect carotenoid profiles or amounts.