胞质雄性不育白菜叶绿体ndhJ-trnF区域序列发生变异  

cDNA-AFLP Reveals Variation of Sequence Region trnF-ndhJ in Cytoplasmic Male Sterile Chinese Cabbage-pak-choi

作　　者：蒋明[1] 曹家树[1] 

机构地区：[1]浙江大学蔬菜研究所细胞与分子生物学实验室,杭州310029

出　　处：《生物化学与生物物理进展》2007年第5期497-502,共6页Progress In Biochemistry and Biophysics

基　　金：浙江省重大科技项目(2005C12019-02);国家自然科学基金资助项目(30370975)~~

摘　　要：以来自Polima胞质甘蓝型油菜雄性不育源转育获得的不育白菜'Bpol97-05A'和其回交亲本即保持系'Bajh97-01B'为材料,利用cDNA扩增片段长度多态性(cDNA-AFLP)技术获得一条长约330bp的特异片段P1708,RT-PCR验证确认该序列为不育白菜材料所特有,经测序和BLAST比对,发现该片段除54bp的插入序列外,其余部分与大白菜和甘蓝叶绿体ndhJ-trnF基因之间的一段序列完全一致.根据基因区域两端的保守部位设计引物,以Polima不育白菜DNA和可育甘蓝型油菜的DNA为模板,分别获得了长约1900bp的序列,比较序列发现:不育白菜与可育白菜、甘蓝型油菜的DNA序列存在一定差异,'Bpol97-05A'中除多个位点发生变异外,另有108bp的插入序列,该插入由2个长度为54bp的重复序列组成,重复序列中除5′端3个碱基CTT外,其余部分均与trnF基因3′端51bp完全相同.By means of eDNA amplified fragment length polymorphism （cDNA-AFLP） technique, a fragment P1708 was amplified from Polima cytoplasmic male sterile Chinese cabbage-pak-choi （Brassica campestris L. ssp. chinenesis Makino, syn, B. rapa L. ssp. chinenesis） ‘Bpo197-OSA ＇ . RT-PCR showed that this fragment was specifically expressed in male sterile material. Sequencing and BLAST search in GenBank database indicated that P1708 had 100% homolog with chloroplast ndhJ-trnF gene region except a 54 bp insertion. Gene specific primer pairs were synthesized according to ndhJ-trnF gene region and two fragments about 1 900 bp were amplified respectively using genomic DNA templates of Polima cabbage and male fertile oilseed rape. The sequencing results showed that the gene region ndhJ-trnF of Polima cabbage contained two 54 bp repeats and some variation sites. The repeat part shared the same sequence as trnF gene except three bases at 5＇ ends. For the insertion of 108 bp sequence, a new open reading frame was created.

关 键 词：白菜 cDNA扩增片段长度多态性(cDNA-AFLP) ndhJ-trnF区域 序列变异 细胞质雄性不育 叶绿体基因 

分 类 号：Q78[生物学—分子生物学]