Cytoplasmic-genic male sterility (CMS) is used to produce hybrid onion (Allium cepa L.) seed. For the most widely used source of onion CMS, male sterility is conditioned by the interaction of the male-sterile (S) cytoplasm and the homozygous recessive genotype at a nuclear male-fertility restoration locus (Ms). Maintainer lines are used to seed propagate male-sterile lines, possess normal (N) male-fertile cytoplasm, and are homozyous recessive at the Ms locus. Due to the biennial nature of onion, it takes 4 to 8 years of crossing and scoring of progeny phenotypes to establish if maintainer lines can be extracted from an uncharacterized population or family. Identification of nuclear markers tightly linked to the Ms locus would allow for molecular-facilitated selection of maintainer lines. We evaluated testcross progenies from a segregating family for nuclear restoration of male fertility over at least three environments. Although segregations in the F2 family fit the expected 1:2:1 ratio (P = 0.973), the proportion of male-sterile testcross progenies showed significant (P < 0.01) year effects and it is therefore imperative to score male-fertility restoration over environments. Too many male-sterile testcross progenies were often observed, indicating that the dominant allele conditioning male-fertility restoration for S cytoplasm may not show complete penetrance. Segregations of amplified fragment length polymorphisms and restriction fragment length polymorphisms (RFLPs) revealed RFLPs flanking the Ms locus at 0.9 and 8.6 cM. An onion cDNA showing highly significant homology to the aldehyde dehydrogenase conditioned by the rf2 locus of maize was identified and mapped to linkage group I, independent of the Ms locus. A sample of commercial onion germplasm was evaluated for putative allelic diversity at the RFLP loci linked to Ms. The genomic region corresponding to the cDNA (AOB272) revealing the closest RFLP to Ms was sequenced to reveal numerous single nucleotide polymorphisms. Single-stranded conformational polymorphisms and single nucleotide extensions were developed that revealed genomic variation at AOB272-EcoRI. The use of these molecular markers to select maintainer lines in onion is discussed.
Ali Fuat Gökçe, John McCallum, Yutaka Sato, and Michael J. Havey
Alstroemeria, also known as Lily-of-the-Incas, Inca Lily, or Peruvian Lily, has been bred at the Univ. of Connecticut since 1985. In vitro procedures have been integrated with traditional breeding techniques to create new and exciting cultivars. Embryo culture has been used to generate interspecific, intraspecific, and intergeneric hybrids that would not have been possible with traditional breeding. Somaclonal variation has been used to create new plants from spontaneous and induced mutations, but, in most cases, the plants have not been acceptable commercially. Chromosome doubling with colchicine has been used for fertility restoration of sterile diploids. Somatic embryogenesis has also been studied quite extensively; somatic embryos are easily obtained from zygotic embryos of Alstroemeria. In vitro fertilization procedures are currently being studied in order to hasten embryo development after hybridization has occurred. Because Alstroemeria plants are slow to propagate by traditional rhizome division, micropropagation is used to multiply new cultivars rapidly. Because the production of pathogen-free plants is one of the goals of our breeding and new plant introduction programs, meristem culture and thermotherapy are also being studied. All of these techniques will be described during the workshop.
Ali Fuat Gökçe and Michael J. Havey
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.
Yutaka Sato and Michael J. Havey
The production of hybrid-onion seed depends on cytoplasmic male sterility (CMS) systems. The male-sterile line is seed propagated using a normal (N) cytoplasmic maintainer line homozygous recessive at the nuclear male-fertility restoration locus (MS). Because of onion's biennial generation time, 4 to 8 years are required to establish the genotype at the MS locus. The development of maintainer lines would benefit greatly from a genetic marker linked to the MS locus. Such a marker would allow breeders to establish the nuclear genotype in seedlings and flower only those plants that are maintainers (N msms) or plants that can be used to develop maintainers (N MSms), reducing the number of plants to be testcrossed or backcrossed to a sterile line. We evaluated restriction fragment length polymorphisms (RFLPs), random amplification of polymorphic DNA (RAPDs), and amplified fragment length polymorphisms (AFLPs) to tag the chromosome region carrying the MS locus. No RAPDs or RFLPs cosegregated with MS. AFLP markers were identified that phenotypically correlated with restoration of male fertility.
Rina Kamenetsky, Idit London Shafir, Hanita Zemah, Amalia Barzilay, and H.D. Rabinowitch
An understanding of temperature and photoperiod effect on garlic (A. sativum L.) growth and florogenesis might solve the enigma of garlic sterility and provide environmental tools for flowering regulation and fertility restoration. The effect of storage temperature and growth conditions on the interactive relationships between the developing vegetative and reproductive organs was studied. A long photoperiod for more than 2 weeks was required for both dormancy induction of the axillary buds and clove formation. In contrast, combination of low temperatures with short photoperiod resulted in sprouting of the axillary buds. Four phases were recognized in the florogenesis of garlic, including: transition of the apical meristem, scape elongation, inflorescence differentiation, and completion of floral development. In garlic accession #2091, meristem transition is autonomous and occurs in growing plants under a variety of storage and growth conditions. A long photoperiod triggers the initial elongation of the scape in post-transitional plants. The temperature effect was quantitative: low storage and growth temperatures combined with long photoperiod promoted scape elongation, whereas warm temperatures and long photoperiod promoted the translocation of reserves to the cloves, and the degeneration of the developing inflorescence. Differentiation of topsets followed flower formation and was dominated by and required lengthy exposure to long photoperiod. Hence, under short photoperiod with only short interruption of long photoperiod, normal development of fertile flowers occurred. We conclude that in bolting garlic genotypes, manipulation of the environment, both before and after planting, can regulate the development of flowers and regain fertility. Normal flowering cannot be achieved if any of the four developmental stages of florogenesis mentioned above is inhibited.
Michael J. Havey and Christopher von Kohn
predict male fertility restoration for random S-cytoplasmic plants from Indian populations. This inconsistency could be due to rare recombination event(s) between AcPms1 and Ms , or possibly different male fertility restoration locus (loci) for S
Michael J. Havey and Borut Bohanec
Japanese island of Hoikkaido and has relatively high frequencies of both S cytoplasm and the dominant allele at the male-fertility restoration locus ( Ms ) ( Havey, 1995 ). This synthetic population combines the early maturity of ‘Sapporo-Ki’ with
Michael J. Havey
the female parent of the hybrid ‘Sweet Sandwich’ ( Goldman et al., 2001 ). These two chartreuse plants were intercrossed [breeding plot (BP) 23873]. The cytoplasm and genotype at the nuclear male-fertility restoration ( Ms) locus ( Jones and Clarke
Md. Mizanur Rahim Khan, Mst. Hasnunnahar, and S. Isshiki
backcrossing germinated normally. Moreover, after selfing and backcrossing, fruiting with viable seeds confirmed that the pollen and seed fertility were restored in the identified amphidiploids. Similar results of fertility restoration of sterile hybrids
Katrina J.M. Hodgson-Kratky and David J. Wolyn
Taraxacum species collected in France, the germplasm did not include russian dandelion. Genetic and molecular analyses are necessary to determine the relationship between the mechanisms of CMS and fertility restoration in the different species. Some plants