摘要
为了探索植物生长素极性运输载体蛋白编码基因BoPINs家族参与甘蓝自交不亲和性的成员数目与参与方式,本文通过转录组分析获得BoPINs家族在甘蓝自花和异花授粉后的表达情况,利用分子生物学技术和生物信息学方法对该家族的基因结构、蛋白进化亲缘关系和表达模式等特征进行分析。结果表明,甘蓝BoPINs基因家族包含8个成员,含有5~9个外显子和4~8个内含子;其编码的蛋白质的氨基酸残基数在350~650之间,相对分子质量为38~70kD;除了BoPIN5和BoPIN8不含中间亲水区以外,其余6个BoPINs家族成员都含有位于两端的疏水区和中间亲水环,它们可能以膜锚定蛋白的形式发挥作用;甘蓝BoPINs与芜菁BrPINs、拟南芥AtPINs基因家族亲缘关系较近;染色体定位分析表明,BoPIN1-1、BoPIN3-1、BoPIN3-2和BoPIN6与S位点之间发生不同程度的连锁;启动子活性分析表明,BoPINs家族蛋白参与甘蓝SI反应,可能受IAA、ABA等激素相互交叉影响;BoPIN1-1、BoPIN1-2、BoPIN2、BoPIN3-1、BoPIN3-2、BoPIN4、BoPIN6、BoPIN7-1和BoPIN7-2在柱头中表达量均较高;数据表达谱和荧光定量分析表明,8个家族成员中的6个BoPINs基因在自花授粉后下调表达;自花授粉后柱头生长素含量降低,与SI反应呈负相关。因此,在甘蓝BoPINs家族的8个成员中有6个BoPINs基因家族成员可能在膜上以负调节方式调控自交不亲和反应。
In order to explore the number and expression of the BoPINs gene family participating in self-incompatibilty of Brassica, their expression after self-pollination and cross-pollination were detected by transcriptome analysis, and the corresponding gene structure, phylogenetic tree and expression patterns of the family were further analyzed by bioinformatics. This gene family contained 5?9 exons and 4?8 introns. The amino acid of the encoding protein residues were between 350 and 650 and had molecular weights ranging from 38 kD to 70 kD. Except that BoPIN5 and BoPIN8 did not contain internal hydrophilic cytoplasmic regions, the remaining six BoPINs proteins contained a hydrophobic region at both ends and an internal hydrophilic ring, showing they located on membrane. The evolutionary analysis indicated that BoPINs were closely related to the BrPINs and the AtPINs gene family. Chromosome localization analysis indicated that BoPIN1-1, BoPIN3-1, BoPIN3-2, and BoPIN6 members of the family were linked to S-loucs to different degrees. Tissue-specific expression analysis indicated that BoPIN1-1, BoPIN1-2, BoPIN2, BoPIN3-1, BoPIN3-2, BoPIN4, BoPIN6, BoPIN7-1, and BoPIN7-2 had higher expression levels in the stigma. Data expression profiling and fluorescence quantitative analysis indicated that six of the eight BoPINs genes were down-regulated after self-pollination. All these results indicate that six members of the eight BoPIN gene family members on the membrane may participate in the self-incompatibility response of Brassica oleracea in a negative regulatory manner.
作者
王玉奎
张贺翠
白晓璟
廉小平
施松梅
刘倩莹
左同鸿
朱利泉
WANG Yu-Kui;ZHANG He-Cui;BAI Xiao-Jing;LIAN Xiao-Ping;SHI Song-Mei;LIU Qian-Ying;ZUO Tong-Hong;ZHU Li-Quan(College of Agriculture and Biotechnology,Southwest University,Chongqing 400715,China;College of Horticulture and Gardening,Southwest University,Chongqing 400100,China)
出处
《作物学报》
CAS
CSCD
北大核心
2019年第8期1270-1278,共9页
Acta Agronomica Sinica
基金
国家自然科学基金项目(31572127)
中央高校基本科研业务费(XDJK2017C023)项目资助~~