甘蓝型油菜BnEOD3基因克隆与比较测序分析  

Cloning and Comparative Analysis of BnEOD3 in Brassica napus L.

作　　者：俎峰[1] 李霞[2] 何晓莹[1] 张国建 张建昆 董云松[1] 王敬乔[1] 束正齐[1] 陈苇[1] ZU Feng;LI Xia;HE Xiao-ying;ZHANG Guo-jian;ZHANG Jian-kun;DONG Yun-song;WANG Jing-qiao;SHU Zheng-qi;CHEN Wei(Industrial Crops Institute, Yunnan Academy of Agricultural Sciences, Yunnan Kunming 650225, China;Biotechnology and Genetic Resources Institute, Yunnan Academy of Agricultural Sciences, Yunnan Kunming 650205, China;Yunnan Vocational and Technical College of Agriculture, Yunnan Kunming 650031, China)

机构地区：[1]云南省农业科学院经济作物研究所,云南昆明650225 [2]云南省农业科学院生物技术与种质资源研究所,云南昆明650205 [3]云南省农业职业技术学院,云南昆明650031

出　　处：《西南农业学报》2019年第1期20-24,共5页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金(31760394);云南省农业科学院食用油料作物育种与应用研究省创新团队(2017HC021);农业部云贵高原马铃薯与油菜科学观测试验站

摘　　要：【目的】克隆拟南芥籽粒大小发育母体效应基因EOD3在甘蓝型油菜籽粒中表达的同源基因拷贝,并在大、小籽粒材料间开展序列比较分析。【方法】以1份大籽粒与1份小籽粒种质发育中的籽粒为研究材料,利用RT-PCR克隆发育中籽粒表达的BnEOD3基因拷贝,结合生物信息学手段在大、小籽粒材料间进行差异比较分析。【结果】生物信息学分析发现BnEOD3基因在甘蓝型油菜中有4个同源拷贝(BnaC04g00760D,BnaA05g01200D,BnaC04g50960D,BnaA04g27100D)。其中BnaA04g27100D在大、小籽粒材料发育籽粒中均有表达,且在第114碱基处存在SNP变异(大籽粒为C,小籽粒为G),BnaC04g50960D则仅在小籽粒材料中检测到表达,BnaC04g00760D与BnaA05g01200D基因则在大、小籽粒中均未检测到表达。【结论】BnEOD3基因仅有2个拷贝(BnaA04g27100D和BnaC04g50960D)在发育籽粒中检测到表达,其中BnaA04g27100D在第114碱基处的SNP变异导致氨基酸序列甘氨酸(G)与丙氨酸(A)的氨基酸的差异,可能与大、小籽粒性状有关。【Objective】BnEOD3,the homologous gene of EOD3 controlling the seed development in arabidopsis,were cloned and comparative analyzed.【Method】The developing seeds obtained from 1 lager rape seed germplasm and 1 normal rape seed germplasm were used as research materials to clone BnEOD3 with RT-PCR technique.The expressed copy of BnEOD3 and the copy sequences were investigated and used for comparative analysis between the extra lager rape seed germplasm and small rape seed germplasm.【Result】There were 4 copies of BnEOD3,BnaC04g00760D,BnaA05g01200D,BnaC04g50960D and BnaA 04g27100D,existing in rape genomes,and only 2 copy,BnaA 04g27100D and BnaC04g50960D,were detected expressing in developing seed.Furthermore,the differences of the number of expressed copy and 1 SNP site were also observed between the larger and normal seed materials.【Conclusion】There was one SNP difference at the 114 bp of BnaA 04g27100D between the larger and normal seed materials,which might be related with difference of seed size.

关 键 词：油菜 基因克隆 大籽粒 比较测序 EOD3基因 

分 类 号：S565[农业科学—作物学]