甘蓝型油菜BnCHI-1基因的克隆和表达特征分析  

Cloning and characterization of BnCHI-1 from Brassica napus

作　　者：许本波[1] 谢伶俐[1] 李加纳[2] 柴友荣[2] 曾柳[3] 

机构地区：[1]长江大学生命科学学院,湖北荆州434025 [2]西南大学农学与生物科技学院,重庆北碚400715 [3]中国农业科学院油料作物研究所,湖北武汉430062

出　　处：《中国油料作物学报》2016年第2期142-149,共8页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金(31171177)

摘　　要：为了解Bn CHI-1基因的表达与粒色差异之间的关系,采用RACE技术从甘蓝型油菜黑籽系5B中克隆了Bn CHI-1全长c DNA序列和基因组序列。Bn CHI-1的DNA序列为1 809bp。965bp的Bn CHI-1 mRNA含有一个756bp的ORF,编码包含251个氨基酸的多肽。预测发现Bn CHI-1蛋白存在查尔酮-黄烷酮异构酶结构域,底物2S-柚皮苷的结合位点以及氢键网络氨基酸残基活性位点。Southern杂交结果表明在甘蓝型油菜中有6个CHI基因成员。RT-PCR揭示,甘蓝型油菜5B的11个器官组织均有Bn CHI转录,其在茎、叶、蕾和花中的转录水平比较高,其次为种子和根,在子叶中最低。比较1对近等基因系(黑籽系L1和黄籽系L2)中Bn CHI转录水平,发现在花蕾、花、花后20d种子和花后30d种子中,二者的转录水平没有显著差异,但在开花后10d的种子中,L2中Bn CHI转录水平显著低于L1中Bn CHI转录水平。To investigate the relationship between Bn CHI- 1 gene and rapeseed color,a Bn CHI- 1 gene was cloned from B. napus stock line 5B by RACE amplification. The genomic sequences of Bn CHI- 1 was 1 809 bp and contained 3 introns. The 965 bp Bn CHI- 1 mRNA had a 756 bp ORF encoding 251 amino acids. NCBI conserved domain search indicated that Bn CHI- 1 gene contained conserved gnl ｜ CDD ｜ 8301 domain（ chalcone- flavanone isomerase）,（ 2S）- naringenin binding cleft and residues of the active site hydrogen bond network. Southern blot result indicated likely that 6 CHI genes existed in B. napus. RT- PCR indicated that the tRNA of Bn CHI genes were ubiquitously expressed in 11 detected organs of B. napus line 5B. Relatively higher expression levels were detected in stems,leaves,buds and flowers,followed by seeds and roots,and the cotyledons had the lowest expression level. No significant difference of the total transcription of Bn CHI family was detected between L1 and L2（ a pair of near- isogenic lines,black line and yellow line respectively） in buds,flowers,20 DAF seeds and 30 DAF seeds. But in 10 DAF seeds of L2,the expression was significantly lower than that of L1.

关 键 词：甘蓝型油菜 BnCHI-1基因 克隆 表达 查尔酮异构酶 

分 类 号：S565.403[农业科学—作物学]