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为了解甘蓝型油菜材料乙酰辅酶A羧化酶(ACCase)基因单核甘酸多态性及其与含油量的关系,该文采用高、低含油量的自交纯合系材料各6份,克隆了其同质型ACCase基因,经双向测序后用DNASTAR软件进行CLUSTAL分析,并用DNASP软件进行SNP多态性计算.结果表明:在供试材料共88 824bp的基因组核苷酸序列中共检测到317个SNPs,出现频率为1/279;其中发生在编码区的SNP有179个,非编码区138个;ACCase基因编码区的变异小于非编码区;非同义突变平均数(Ka)与同义突变平均数(Ks)比值小于1,表明该基因比较保守.在高含油量材料组中发现了一个明显高于低含油量材料的突变热点区,位于5 000~6 500bp(即全基因序列的第10 088到11 588bp)之间,该突变热点区包含了4个外显子和3个内含子,编码同质型ACCase的第1 279到1 648位共368个氨基酸的序列,这段序列位于生物素羧基载体蛋白(BCCP)与羧基转移酶(CT)功能域之间.此变异区域与ACCase活性及含油量的关系值得进一步研究.
In order to understand the single nucleotide polymorphism of acetyl CoA carboxylase (ACCase) gene and its relationship with oil content in Brassica napus, six high and low oil-content inbred homozygous materials were cloned The homozygous ACCase gene was cloned by DNASTAR software after two-way sequencing and the SNP polymorphism was calculated using DNASP software.The results showed that there were 317 genomic nucleotide sequences in 88,824bp of tested materials The frequency of occurrence of SNPs was 1/279. Among them, there were 179 SNPs in the coding region and 138 non-coding regions. The variation of coding region of ACCase gene was smaller than that of non-coding region. The average number of non-synonymous mutations (Ka) The mean (Ks) ratio was less than 1, indicating that the gene was conserved.A mutant hot spot region was found in the high oil content material group that was significantly higher than the low oil content material, located in the range of 5 000-6 500 bp (ie, 10 088 to 11 588 bp), this mutant hotspot contains 4 exons and 3 introns, encoding a sequence of 368 amino acids at positions 1 279 to 1 648 of the homologous ACCase, Located in the biotin carboxyl-carrier protein (BCCP) and carboxyl transferase (CT) domains The relationship between this area and the variability ACCase activity and oil content worthy of further study.