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  • Author or Editor: Mark W. Farnham x
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Broccoli (Brassica oleracea L. Italica group) breeders are increasingly using anther or microspore culture to produce dihaploid (diploid), homozygous lines for use in making hybrids. During the process of anther culture and subsequent plant regeneration, wherein embryos develop from microspores and plants are regenerated from the embryos, polyploidization occurs and diploid regenerants can result. However, polyploidization may not occur at all, or it may involve a tripling or quadrupling of the chromosome complement, instead of a doubling. Thus, populations may contain haploids, triploids, or tetraploids, in addition to diploids. In two cycles (1994-95 and 1995-96) of anther culture, regenerated populations from different broccoli hybrid sources were evaluated using flow cytometry to facilitate efficient identification of diploids vs. haploids, tetraploids, or others and to determine if anther donor genotype has an effect on the frequency of different ploidy levels among regenerants. In the first cycle, five broccoli hybrids had anther-derived populations in which ≈33% were haploid, 55% diploid, 37% tetraploid, and 5% aneuploid or abherent types. The hybrid, `Marathon', was different; it's regenerants were 78% diploid and only 15% tetraploid. In the second cycle, anther-derived populations had a significantly different makeup with a most hybrids giving 30% to 40% diploids and 50% to 60% tetraploids. However, consistent with the previous cycle, `Marathon' gave significantly more diploids (68%) and fewer tetraploids (25%) than other hybrids. These results indicate that anther donor genotype affects ploidy frequency among regenerants. Genotypes producing a high frequency (>60%) of diploids may be relatively uncommon.

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Collard (Brassica oleracea L. var. acephala) is an important vegetable the southeastern U. S. There are few (about 10) commercial cultivars, half being open-pollinating (OP) lines, the remainder more recent F1 hybrids. There is a potential untapped B. oleracea germplasm pool in the form of collard landraces perpetuated by southeastern gardeners and farmers. To determine the amount of genetic variation among cultivars and also whether landraces represent unique genotypes, ten cultivars and eight lines or landraces were evaluated using RAPD analysis. Decamer primers were used to amplify total genomic DNA and to differentiate collard lines and other B. oleracea crop cultivars. Additionally, individuals of an OP collard cultivar and a land-race were analyzed to evaluate intra-line variation. Virtually all primers detected polymorphic bands among lines although some identified considerably more variants. Intra-line analysis indicated that OP lines are genetically broad-based populations. Many unique RAPD markers were identified in landraces indicating that the lines represent unique genotypes and that further line collection is warranted.

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For many decades plant breeders have worked to improve vegetable crops for numerous economically important traits, like host plant resistance to disease, yield, and vegetable quality. Most improvements have been made with little knowledge as to how, or if, nutritional or phytonutrient concentrations might also be indirectly altered in the process. There have been some reports suggesting that concentrations of nutrients in vegetables have been reduced over time, possibly related to introductions of new cultivars. However, for most vegetables, current evidence indicating changes in nutrient concentrations, and specifically mineral concentrations, is circumstantial at best. To effectively test whether changes may have occurred over time as new cultivars replace older ones, appropriate field studies must be conducted wherein harvested produce from “old” vs. “new” crop cultivars is analyzed by appropriate methods and compared directly. Numerous considerations and issues such as 1) the set of cultivars to be used in field tests; 2) how nutritional concentration will be expressed; and 3) the evolution, history, and consumption changes of the crop under study must be addressed in making such direct comparisons and interpreting results.

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Private and public vegetable breeders are interested in using current and emerging PCR-based marker systems in their respective improvement programs. However, before new systems are employed to replace existing ones, the new systems must prove to be efficient and cost-effective alternatives. Sequence related amplified polymorphisms (SRAPs), amplified fragment length polymorphisms (AFLPs), and simple sequence repeats (SSRs) were compared for their ability to differentiate individuals of a diverse group of 24 elite broccoli (Brassica oleracea L. italica) inbreds. Genomic DNA was assayed using 24 AFLP, 24 SRAP, and 44 SSR primer pairs. In this assessment, SSRs produced an average of only two bands per primer, with 25% of these bands being monomorphic, and the remaining bands detecting very few differences among the inbreds. Although the AFLP method resulted in a lower rate (63%) of polymorphism than the SSRs, it produced about 20 bands per primer. SRAPs produced an average of 14 bands per primer, with 82% of these bands being polymorphic. Since AFLP and SRAP markers had a higher multiplex ratio and SSRs were frequently monomorphic, AFLP and SRAPs were more effective in differentiating the elite broccoli inbreds examined in this study. Similarity matrices were generated from the AFLP and SRAP data, and resulting dendographs were compared.

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Downy mildew, caused by Peronospora parasitica (Pers. ex Fr.), is one of the most economically important diseases in broccoli (Brassica oleracea L. Italica group). Previous studies reported that resistance to downy mildew in broccoli depends on plant age and that seedling resistance appears to be independent of mature-plant resistance. The objectives of our studies were to evaluate resistance and susceptibility of USDA broccoli inbreds to downy mildew and to investigate the interaction between the host and pathogen at two plant stages with single or double inoculation. Multiple screening tests at both cotyledon and three-expanded leaf stages using 38 entries, including USDA inbreds and commercial hybrids, were conducted in randomized complete-block designs. In these tests, every leaf of each plant was thoroughly sprayed with P. parasitica isolate PP1 at a concentration of 10,000 sporangia per ml at both stages. Ratings for downy mildew reaction phenotype were made at 9 days postinoculation on a 0-9 scale of increasing disease severity. We found significant phenotypic variation to infection among broccoli entries. We observed three general phenotypes: 1) resistance at both stages; 2) susceptible at cotyledon stage combined with resistance at three-expanded leaf stage; and 3) susceptibility at both stages. Additionally, inoculation at the cotyledon stage had no effect on inoculation at the three-expanded leaf stage.

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The discovery that broccoli (Brassicaoleracea L., Italica Group) sprouts contain high levels of sulforaphane, a constituent that may provide chemoprotection against certain carcinogens, has stimulated much interest in seed production of this crop. Studies were undertaken to determine the potential for producing broccoli seed using self-compatible selections from open-pollinated (OP) populations or doubled-haploid (DH) programs. In all outdoor and greenhouse trials, three OP selections and seven DH lines produced selfed seed, but seed weight per plant and number per plant varied significantly among the entries. In all environments there were individuals with relatively high (i.e., >3 g/plant) production that were significantly different from low (i.e., <2 g/plant) producers. The relative productivity of some lines varied greatly between experiments, which indicates that seed production of particular genotypes is affected differently by environmental conditions. This indicates the importance of identifying lines that are high producers of selfed seed across different environments. Two OP cultivar-derived lines (USVL102 and USVL104) and two DH lines (USVL062 and USVL093) were identified that consistently produced relatively high yields in greenhouse and screen cage trials. These lines are good candidates for evaluating seed production in field tests and as possible sources of seed for sprouting.

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Collard and kale (Brassica oleracea L. var. acephala) cultivars and several landraces obtained from southeastern growers were tested for potential winter production. Collard and kale entries were grown in four winter environments in South Carolina from 1993 to 1995. Transplants were set in the field during November or December, and leaf production and plant fresh weight were monitored through the winter. When plants reached a 22-leaf stage, a plot subsample was harvested and weighed. The date at which 50% of the plants per plot had bolted was also recorded. Essentially all entries survived the conditions of four winter environments. However, whether an entry reached harvest size depended on its date of bolting. Collard entries typically bolted earlier than kale entries, and most kale and several collard entries attained harvestable-size before bolting. The ranking of genotypes for days to 50% bolting was consistent among environments. `Blue Max' and a landrace of collard, and `Squire' and `Blue Knight' kale usually never reached 50% bolt.

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