The isolation of cDNA clones from cucumber (Cucumis sativus L.) floral buds coming from plants differing in sex.
In this study, we found flower cDNA clones which may be connected with the development of flower sex in cucumber. Two pairs of nearly-isogenic lines: gynoecious GY3 (FFMMGG) versus hermaphrodite HGY3 (FFmmGG) and monoecious B10 (ffMMGG) versus gynoecious 2gg (ffMMgg) were used for clone isolation. To obtain differentially-expressed clones, we applied the differential screening method. 454 clones from GY3 and 478 from B10 cDNA libraries were isolated. The results of RFLP analysis with 56 cDNA clones showed no clones which cosegregated with sex in cucumber. The 28 cDNA B10 and 33 cDNA GY3 clones isolated using the differential screening method were sequenced. Some of them seem to may play a role in cell differentiation or flower development. Among the 61 identified clones, 14 show high homology to plant proteins, although of unknown function. 11 show high homology to known proteins, and the possible function of some of them is discussed. For 3 clones, no significant similarity was found. The 31 clones displayed high homology to plant cDNA in EST database. The patterns of expression of five differential cDNA clones, 35GY3, 216GY3, 47GY3, 100B10 and 157B10, were analyzed in cucumber flower buds using in situ RT-PCR. The most interesting clone is 35GY3, because of its possible role in the inhibition of the development of male specific elements in the female cucumber flower.
Przybecki
Zbigniew
Z
Department of Plant Genetics, Breeding and Biotechnology, Faculty of Horticulture, Warsaw Agricultural University, Nowoursynowska 166, 02-787 Warsaw, Poland. przybecki@alpha.sggw.waw.pl
Kowalczyk
Magdalena Ewa
ME
Siedlecka
Ewa
E
Urbańczyk-Wochniak
Ewa
E
Malepszy
Stefan
S
D000595:Amino Acid Sequence; D003433:Crosses, Genetic; D018553:Cucumis sativus / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary; D035264:Flowers / Q000378:metabolism*; D020869:Gene Expression Profiling; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary