title: Cloning and expression analysis of S-RNase homologous gene in Citrus reticulata Blanco cv. Wuzishatangju.
abstract: S-RNase-based self-incompatibility is the most widespread form of genetically controlled mate selection in plants and that S-RNase controls pollination specificity in the pistils. 'Wuzishatangju' (Citrus reticulata Blanco), a nature bud mutant from a self-compatible (SC) cultivar 'Shatangju', displays gametophytic self-incompatibility (GSI). In this study, full-length sequences of cDNA and DNA of the S-RNase homologous gene were obtained from 'Wuzishatangju' and 'Shatangju'. There was no difference in ORF sequences of the S-RNase cDNA between 'Wuzishatangju' and 'Shatangju'. However, 13, 9 and 6 consecutive bases were missing in 'Wuzishatangju' cDNA 5' UTR, 3' UTR and genomic DNA, respectively. Tissue-specific expression of the S-RNase gene was detected using semi-quantitative RT-PCR and quantitative real-time PCR. The expression level of the S-RNase gene in styles of 'Wuzishatangju' was approximately 10- and 5-fold higher than that in leaves and pollen, respectively. When 'Wuzishatangju' was self-pollinated, the expression of S-RNase in pistils peaked at 3 days, which was approximately 10-fold higher than that at 4h and 7 days, while in cross-pollination of 'Wuzishatangju' x 'Shatangju' the expression was very weak at 3 days. Results from a Southern blot showed that two copies of the S-RNase gene existed in genomic DNA of both 'Wuzishatangju' and 'Shatangju'.
authors.affiliation: College of Horticulture, South China Agricultural University, Guangzhou, Guangdong 510642, PR China.
authors.forename: Hong-xia
authors.initials: HX
authors.lastname: Miao
authors.forename: Yong-hua
authors.initials: YH
authors.lastname: Qin
authors.forename: Jaime A
authors.initials: JA
authors.lastname: Teixeira da Silva
authors.forename: Zi-xing
authors.initials: ZX
authors.lastname: Ye
authors.forename: Gui-bing
authors.initials: GB
authors.lastname: Hu
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002957:Citrus / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D035264:Flowers / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D018515:Plant Leaves / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D011058:Pollen / Q000235:genetics; D054817:Pollination / Q000502:physiology; D018749:RNA, Plant / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012260:Ribonucleases / Q000235:genetics*; D020506:Untranslated Regions / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D018744:DNA, Plant; D010940:Plant Proteins; D018749:RNA, Plant; D020506:Untranslated Regions; D012260:Ribonucleases; C022542:ribonuclease S