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  • Author or Editor: Michael J. Havey x
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Development of two-way onion (Allium cepa L.) hybrids is difficult due to poor seed yields on inbred female parents. Seed yield of onion is affected by inbreeding depression and the seed-production environment. A standard diallel was used to estimate combining abilities for seed yield among seven inbred onion lines. Males and hybrids differed significantly (P < 0.05) for seed yields. Combinations of relatively high-by-high seed-yielding inbred parents were not always the best combinations; combinations of medium-by-medium or medium-by-high seed yielders also produced good F1 seed yielders. For the seven inbred lines, significant correlations (P < 0.05) were observed between mean seed yield per bulb and scape height. Parent-offspring regressions revealed no significant relationship between seed yields of randomly selected, open-pollinated bulbs and their S1 families. Results indicate that relative seed yields of individual bulbs after self-pollination cannot be used to predict seed yields of progeny families. However, the seed yield of inbred lines of onion may reflect the potential seed yield of F1 male-sterile lines.

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Cytoplasmic male sterility (CMS) in onion (Allium cepa L.) is conditioned by the interaction of the male-sterile (S) cytoplasm with recessive alleles at a single nuclear male-fertility restoration locus (Ms). In order to seed propagate male-sterile plants (S msms), onion breeders must identify maintainer lines possessing normal (N) male-fertile cytoplasm and homozygous recessive at the Ms locus (N msms). Molecular markers have been identified distinguishing N and S cytoplasms and closely linked to the nuclear Ms locus. In this study, we evaluated testcross progenies from randomly selected N-cytoplasmic plants from three open-pollinated populations for nuclear restoration of male fertility over at least three environments. The Ms locus and linked restriction fragment length polymorphisms (0.9 and 1.7 cM) were at linkage equilibrium in all three open-pollinated onion populations, indicating that these linked markers cannot be used to identify maintaining genotypes in open-pollinated onion populations. However, cytoplasmic evaluations were effective in reducing the number of testcrosses required to identify CMS-maintaining genotypes.

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Hybrid leek is more uniform and higher yielding than open-pollinated cultivars and is presently produced by asexual propagation of a genic male-sterile plant. A cheaper method to produce hybrid leek seed would be a system of cytoplasmic-genic male sterility (CMS). Restriction fragment length polymorphisms (RFLPs) in the organellar genomes have correlated with CMS in many crops. We undertook gel-blot analyses of the chloroplast and mitochondrial DNAs to assess cytoplasmic diversity among 62 accessions of the major cultivated forms of Allium ampeloprasum L. (leek, kurrat, and great-headed garlic). No polymorphisms were detected in the chloroplast genome of leek and kurrat. Three accessions of leek and one of kurrat possessed one or two of seven polymorphic mitochondrial probe-enzyme combinations. Great-headed garlic differed from leek and kurrat for six polymorphisms in the chloroplast genome and for many mitochondrial probe-enzyme combinations. Our analyses revealed few organellar polymorphisms among accessions of leek and kurrat, reducing the probability that selection of polymorphic cytoplasms will reveal CMS in leek.

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A factorial mating design, using three male-sterile F1 lines in testcrosses with a sample of open-pollinated (OP) onion populations, was used to estimate combining abilities and heterosis for bulb yield, size, storage ability, pungency, soluble solids content (SSC), and water loss after 3 months in storage. Samples of testcross bulbs were flowered and scored for fertility to estimate frequencies of the nuclear allele maintaining cytoplasmic male sterility. General combining ability (GCA) estimates for OP populations (males) were significant (P < 0.05) for yield, SSC, and proportion of bulbs with diameters >7.5 cm. GCA estimates for female testers were significant for storage ability and proportion of bulbs with diameters <5.0 cm. Male × female interactions (specific combining ability estimates) were significant for SSC and storage ability. Our analyses did not reveal any storage population from which inbreds would likely yield significantly better with the male-sterile tester lines. Spanish OP populations tended to produce testcrosses with larger bulbs, lower pungency and SSC, and poorer storage ability. Heterosis estimates were most often significant for yield and SSC; less often for pungency, storage ability, and bulb size; and not significant for water loss in storage. Overall, significant GCA estimates indicate that superior onion inbreds and populations may be developed using recurrent-selection strategies that increase the frequency of desirable alleles with additive effects.

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The most commonly used source of cytoplasmic male sterility in onion (Allium cepa) is controlled by the interaction of the cytoplasm [male-sterile (S) or normal male-fertile (N)] and one nuclear male-fertility-restoration locus (Ms). Scoring of genotypes at Ms is generally done by testcrossing male-fertile to male-sterile (S msms) plants, followed by scoring of testcross progenies for male-fertility restoration. We identified two N-cytoplasmic families, one that was homozygous dominant and the other segregating at Ms. Plants from each of these two families were individually testcrossed to male-sterile onion. Nuclear restoration of male fertility in testcross progenies was evaluated in the field over 4 years. For male plants homozygous dominant at Ms, we expected testcross families to show 100% male-fertility restoration, but observed mean values between 46% and 100%. For plants segregating at Ms, we again observed lower than expected frequencies of male-fertility restoration. These results demonstrate that the dominant Ms allele shows reduced penetrance, requiring that male-fertility restoration be scored over years to more confidently assign genotypes at Ms.

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Commercial bulb-onion (Allium cepa L.) growers often complain that hybrids they have grown successfully for a few years fail to perform at the expected level. Inbreds used to produce hybrid-onion seed rarely have been self-pollinated for more than two generations and retain a high level of heterozygosity. Over time, selection, drift, or contamination of inbreds may contribute to disappointing hybrid performance. We identified randomly amplified polymorphic DNA (RAPD) between two inbred onion lines, demonstrated their Mendelian inheritance, and tried to distinguish among and examine changes in independently maintained, publicly released inbred lines of onion. We observed poor agreement between data sets based on genetically characterized and uncharacterized RAPD markers. Our analyses used only genetically characterized RAPD markers and revealed that contamination, in addition to-drift and/or selection, likely contributed to differences among independently maintained, publicly released inbreds. However, RAPD markers were not able to distinguish confidently among four related inbreds. RAPD markers will be useful in Allium genetics and breeding, but identifying and characterizing reliable polymorphisms is critical.

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Garlic (Allium sativum) is cultivated worldwide and appreciated for its culinary uses. In spite of primarily being asexually propagated, garlic shows great morphological variation and adaptability to diverse production environments. Molecular markers and phenotypic characteristics have been used to assess the genetic diversity among garlics. In this study, we undertook transcriptome sequencing from a single garlic plant to identify molecular markers in expressed regions of the garlic genome. Garlic sequences were assembled and selected if they were similar to monomorphic sequences from a doubled haploid (DH) of onion (Allium cepa). Single nucleotide polymorphisms (SNPs) and insertion–deletion (indel) events were identified in 4355 independent garlic assemblies. A sample of the indels was verified using the original complementary DNA (cDNA) library and genomics DNAs from diverse garlics, and segregations confirmed by sexual progenies of garlic. These molecular markers from the garlic transcriptome should be useful for estimates of genetic diversity, identification and removal of duplicate accessions from germplasm collections, and the development of a detailed genetic map of this important vegetable crop.

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Cucumber (Cucumis sativus) plants regenerated from cell cultures occasionally possess mosaic (MSC) phenotypes on cotyledons and leaves. Lines MSC3 and MSC16 have distinct MSC phenotypes and originated from plants regenerated from different cell-culture experiments established using a highly inbred wild-type cucumber. Both the mitochondrial (mt) DNA and MSC phenotype of cucumber show paternal transmission, and MSC3 and MSC16 have different mt coding regions at significantly lower copy numbers relative to wild-type plants. A nuclear locus, Paternal sorting of mitochondria (Psm), conditions a high proportion of wild-type progenies, specifically when MSC16 is crossed as the male with wild-type female plants. During this research, we identified plants that produced a high proportion of wild-type progenies in crosses with MSC3 as the male parent. Plants from an F2 family were crossed with MSC3 as the male, progenies were scored for numbers of MSC vs. wild-type plants, and single-nucleotide polymorphisms (SNP) were identified for genetic mapping. A major quantitative trait locus on chromosome 3 was associated with a higher frequency of wild-type progenies from MSC3 as the male parent, and the 1.5-logarithm-of-odds interval for the most significant SNP was located 627 kb from Psm. These results reveal that separate genetic factors control sorting to the wild-type phenotype in progenies from crosses with different MSC parents. The identification of causal genes controlling mitochondrial sorting in cucumber should provide insight regarding nuclear-mitochondrial interactions affecting the prevalence of specific mitochondrial DNA in plants.

Open Access

Hybrid-onion (Allium cepa) seed is produced using systems of cytoplasmic male sterility (CMS) and two different CMS systems have been genetically characterized. S cytoplasm was the first source of onion CMS identified in the 1920s, followed by T cytoplasm that was described in the 1960s. Numerous studies have documented polymorphisms in the organellar DNAs differentiating S and T cytoplasms from the normal male-fertile cytoplasm of onion. There may be additional source(s) of onion CMS that have been described as “T-like” and appear to be more similar to N and T cytoplasms than S cytoplasm. In this study, onion breeding lines from commercial entities were evaluated for molecular markers distinguishing sources of onion CMS. Our results reveal that bona fide T cytoplasm is rarely used commercially to produce hybrid-onion seed, and both S cytoplasm and “T-like” cytoplasm are widely used. We propose that this “T-like” cytoplasm be labeled as “R” cytoplasm because it may have originated from population(s) of ‘Rijnsburger’ onion in the Netherlands. The results of this study also help to clarify inconsistent reports regarding nuclear male-fertility restoration for different sources of onion CMS.

Open Access

Restriction fragment length polymorphisms (RFLPs) in the nuclear genome were used to assess genetic diversity among cultivated open-pollinated populations (OPs) of bulb onion (Allium cepa L.). Twenty OPs of contrasting day-length responses [long (LD) and short (SD) day] were examined with 104 random cDNA probes and two to four restriction enzymes. Sixty-one probes detected polymorphisms among the OPs for at least one restriction enzyme. Parsimony and cluster analyses were completed and no distinct grouping was observed between LD and SD OPs. Parsimony analysis generated a consensus tree that grouped all but two LD OPs and their relationships to the SD OPs were not resolved. Cluster analysis grouped all but three LD OPs and the distances from that group to the others were not greater than those among LD or SD OPs. These results suggest that LD and SD onions do not represent distinct germplasm sources and LD OPs possess a more narrow genetic background. The paucity of unique fragments among OPs indicates that phenotypic variation, e.g., day-length response or bulb color or shape, does not reflect diverse sources of germplasm.

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