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| 甘蓝BYPASS1 编码基因的克隆与表达分析 | |
| 引用本文： | 刘豫东,高启国,曾静,张林成,朱利泉,任雪松,王小佳.甘蓝BYPASS1 编码基因的克隆与表达分析[J].中国蔬菜,2013,1(16):22. |
| 作者姓名： | 刘豫东 高启国 曾静 张林成 朱利泉 任雪松 王小佳 |
| 作者单位： | （;1.西南大学园艺园林学院，南方山地园艺学教育部重点实验室，重庆 400716；;2. 西南大学农学与; 生物科技学院，重庆 400716） |
| 基金项目： | 国家自然科学基金项目（30900986）， 重庆市自然科学基金项目（2009BB1298）， 西南大学博士基金项目 （SWUB2008042），中央高校基本科研业务费专项（XDJK2010B010，XDJK2009C126） |
| 摘 要： | 通过双向电泳结合MALDI-TOF-TOF/MS 质谱技术在甘蓝柱头内鉴定出1 个受自花授粉诱导
上调表达的BYPASS1 蛋白（BPS1）。利用PCR 技术扩增甘蓝BPS1 基因的编码序列， 序列分析结果表明：
该基因没有内含子，开放阅读框为1 059 bp，编码352 个氨基酸，蛋白质分子量为38.7 kD。氨基酸同源
性和系统发生树分析结果表明：甘蓝BPS1 与拟南芥BPS1 亲缘最近，氨基酸同源相似性达85%；与烟草
BPS2 关系较远。RT-PCR 检测结果表明：甘蓝BPS1 基因在开花前1～2 d 的花瓣、萼片、花药、柱头和
叶片中均有表达，柱头中的表达量最高。实时荧光 定量PCR 分析结果表明：甘蓝柱头内BPS1 表达水平在
自花授粉后逐渐升高，异花授粉后柱头内BPS1 表达水平先升高后降低再升高。 |
| 关 键 词： | 甘蓝 自花授粉 异花授粉 BYPASS1 表达分析 |
Encoding Gene Cloning and Expression Analysis of BYPASS1 in Brassica oleracea L. |
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| LIU Yu-Dong,GAO Qi-Guo,ZENG Jing,ZHANG Lin-Cheng,ZHU Li-Quan,REN Xue-Song,WANG Xiao-Jia.Encoding Gene Cloning and Expression Analysis of BYPASS1 in Brassica oleracea L.[J].China Vegetables,2013,1(16):22. | |
| Authors: | LIU Yu-Dong GAO Qi-Guo ZENG Jing ZHANG Lin-Cheng ZHU Li-Quan REN Xue-Song WANG Xiao-Jia |
| Institution: | （;1.College of Horticulture and Landscape Architecture， Southwest University， Key Laboratory of Horticulture Science; for Southern Mountainous Regions， Ministry of Education， Chongqing 400716， China； ;2.College of Agronomy; and Biotechnology， Southwest University， Chongqing 400716， China） |
| Abstract: | An up-regulated protein in Brassica oleracea L. stigma induced by self-pollination was identified as BYPASS1 protein by 2-DE electrophoresis and MALDI-TOF-TOF/MS. The genomic DNA and cDNA coding sequences of BYPASS1（ BPS1）were amplified by PCR．The sequence analysis showed that BPS1 had no intron. Its ORF was 1 059 bp，encoded a polypeptide with 352 amino acids with a predicted molecular mass of 38.7 kD. Phylogenetic tree analysis using MEGA 5.1 showed that Brassica oleracea L. BPS1 was more close to Arabidopsis thaliana BPS1 rather than Nicotiana benthamiana BYPASS1，amino acid homologous similarity is 85%．RT-PCR analysis showed that 1-2 days before flowering，BPS1 expressed in petal，sepal，pollen，stigma and leaf，and the express level in the stigma was higher than the other organs．Real-time fluorescence quantitative PCR analysis showed that the relative expression level of BPS1 in stigma was continually increased after self-pollination，and after cross-pollination its expression level was increased firstly，than decreased and finally increased again. |
| Keywords: | Brassica oleracea L Self-pollination Cross-pollination BYPASS1 Expression analysis analysis |
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