The biological processes leading to sex expression in plants are of tremendous practical significance for fruit production of many agricultural and horticultural crops. Sex-expression studies in cucumber showed that the different sex types are determined by three major genes: M/m, F/f and A/a. The M/m gene in the dominant condition suppresses stamina development and thus leads to female flowers. The F/f gene in the dominant condition shifts the monoecious sex pattern downwards and promotes femaleness by causing a higher level of ethylene in the plant. To investigate the molecular character of the gene F/f, we used nearly isogenic gynoecious ( MMFF) and monoecious ( MMff) lines (NIL) produced by our own backcross programme. Our investigations confirmed the result of other groups that an additional genomic ACC synthase (key enzyme of ethylene biosynthesis) sequence ( CsACS1G) should exist in gynoecious genotypes. A linkage was also verified between the F/f locus and the CsACS1G sequence with our plant material. After the exploration of different Southern hybridization patterns originating from different CsACS1 probes, a restriction map of the CsACS1 locus was constructed. By using this restriction map, the duplication of the CsACS1 gene and following mutation of the CsACS1G gene could be explained. The promoter regions of the genes CsACS1G and CsACS1 were amplified in a splinkerette PCR and sequenced. An exclusive amplification of the new isolated sequence ( CsACS1G) in gynoecious ( MMFF) and sub-gynoecious ( MMFf) genotypes confirmed that the isolated gene is the dominant F allele.
The chimerical mitochondrial CMS(1)-specific sequence in chives ( Allium schoenoprasum) was used to develop a PCR-marker that distinguishes both male-sterility inducing cytoplasms, CMS-(S) and CMS-(T), from the normal cytoplasm in onion ( Allium cepa). In combination with a previously described marker for CMS-(S), which anchors in the upstream region of the mitochondrial gene cob, all of the three known cytoplasms in the onion are distinguishable. The PCR-marker system was tested in 361 onion plants, which were selected from F(1)-hybrids and different open-pollinated varieties. The latter are mainly landraces from Turkey, in which all three cytoplasm types were detected.
The complete coding and 3;-flanking region of the mitochondrial gene atp9 of chives ( Allium schoenoprasum L.) was determined in order to develop primers that allow the identification of atp9-related sequences in subsequent PCR-amplifications. One of these sequences is of a chimerical nature, consisting of atp9-homologous regions on its end, interrupted by an insertion that is composed of one atp6-homologous part and one part of unknown origin. This PCR-fragment is 762 bp in size and exclusively amplified in the sterility inducing cytoplasm of CMS(1). Thus it can be used as a PCR-marker in order to distinguish this cytoplasm type from the remaining cytoplasm types of chives. The chimerical marker sequence forms a putative open reading frame ( orfA501), from which CMS(1) might originate.
Sex determination is the most widely studied subject in cucumber. The sex of cucumber plants can be monoecious, hermaphrodite, gynoecious, androecious, or andromonoecious. Besides environmental factors, three major genes, F/f, M/m, and A/a mainly govern the sex types in cucumber. Regardless of their sex all floral buds are bisexual at the early bud stage. A stage specific arrest of either stamen or carpel leads to unisexual flower development. The possible downstream product of the interaction of the sex determining genes that may directly allow the growth or selectively arrest stamen or pistil is not yet identified. Therefore, in the current study, we performed suppression subtractive hybridization using floral buds from nearly isogenic gynoecious and hermaphrodite cucumber plants and identified for the first time a cDNA homologous to nucleotide sugar epimerase. The expression level of the isolated putative nucleotide sugar epimerase is weak in female floral buds but strong in bisexual and male flowers. The weak level of the putative nucleotide sugar epimerase may be an indication for its improper functioning, which may influence stamen development in cucumber plants.
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