作物学报2018，Vol.44Issue(2)：177-184,8.DOI:10.3724/SP.J.1006.2018.00177
自花授粉诱导的甘蓝功能基因BoSPI的克隆与表达分析
Cloning and Expression Analysis of BoSPI Induced by Self-pollination in Bras-sica oleracea L.var.capitata
摘要
Abstract
Self incompatibility is a complex and comprehensive genetic mechanism formed in the long-term evolution, which prevents inbreeding and promotes heterosis. Mining functional genes involving in self incompatibility has an important signifi-cance for the study of self incompatibility in Brassica oleracea L.var.capitata.In this study,we identified a gene named BoSPI which expression was up-regulated and induced by self-pollination based on the stigma transcriptome data in 0–60 min self-pollination and cross-pollination.BoSPI contains an open reading frame(ORF)with the length of 534 bp,encoding a protein of 177 amino acid residues without introns, which contains four conserved EF-hand domains without signal peptide and trans-membrane domain, the theory isoelectric point of BoSPI is 4.21. In addition, diverse cis-acting promoter elements involved in fungal elicitor response, metabolic regulation and organ formation were discovered in the upstream 2000 bp of initial codon of BoSPI.BoSPI could be expressed as a 17 kD protein in E.coli BL21(DE3).The expression level of BoSPI was the highest in stigmas and lower in petals, sepals, leaves and stamens of cabbage after self-pollination. Subcellular localization analysis revealed that BoSPI encoded a protein localized in the cell membrane and cytoplasm.The expression of BoSPI gene was significantly in-duced by self pollination after 30 minutes.These results suggest that BoSPI is involved in the molecular processes of the stigma response to self-pollen stimulation,which may be a new functional gene related to the self incompatibility of Brassica oleracea L. var.capitata.关键词
自花授粉/结球甘蓝/基因BoSPI/转录组测序Key words
self-pollination/Brassica oleracea L.var.capitata/gene(BoSPI)/transcriptome technology引用本文复制引用
蒲敏,朱利泉,罗绍兰,廉小平,张贺翠,白晓璟,王玉奎,左同鸿,高启国,任雪松..自花授粉诱导的甘蓝功能基因BoSPI的克隆与表达分析[J].作物学报,2018,44(2):177-184,8.基金项目
本研究由国家自然科学基金项目(31572127)和重庆市研究生科研创新项目(CYS16076)资助.This study was supported by the National Natural Science Foundation of China(31572127)and the Research Project of Graduate Students in Chongqing(CYS16076). （31572127）
