羽衣甘蓝自交不亲和系的选育及S_(13b)单倍型的鉴定  

Selection of self-incompatible line and identification of the S_(13b) haplotype from Brassica oleracea var. acephala

作　　者：蓝兴国[1] 解莉楠[1] 于晓敏[1] 李玉花[1] 

机构地区：[1]东北林业大学花卉生物工程研究所

出　　处：《北京林业大学学报》2006年第S2期31-39,共9页Journal of Beijing Forestry University

基　　金：国家自然科学基金项目(30371189);"863"国家高新技术项目(2001AA246102)

摘　　要：该文采用亲和指数法(种子数/自交授粉花朵数)分别在羽衣甘蓝‘赤兔’和‘白波’栽培品种的自交后代中,经过3年的自交选育获得1个自交不亲和系4·1·1和一个自交亲和系3·2·3·由于在芸苔属植物F1代杂交种的生产和自交不亲和分子机制的研究方面S单倍型鉴定是必要的,因此在该文中我们采用了PCR方法利用引物PK1和PK4获得了BoSRKx的基因组序列,此序列的大小为919bp.经过数据库检索比对后发现BoSRKx的序列与BoSRK13b的序列完全一致.通过反转录PCR的方法获得了BoSRKx的cDNA序列,结果发现BoSRKx和BoSRK13b的序列在转录水平上也是完全一致的;另一方面,我们采用SCR保守信号肽编码区和NotⅠ-oligo(dT)设计的引物获得了BoSCRx的cDNA序列,经过数据库的检索比对后发现BoSCRx的序列比BoSCR13的序列只多出3个连续的碱基.根据上述结果我们推断出Sx单倍型就是S13b单倍型;最后,通过遗传分析结合PCR-RFLP方法和DNAblot方法对自交不亲和系进行了S13b纯合体的鉴定.A strong self-incompatible line 4.1.1 and a self-compatible line 3.2.3 were selected based on self-incompatibility index (SII=(number of seeds)/(number of self-pollinated flowers)) through three self-generations from offspring of cultivars of Brassica oleracea var. acephala of ‘Red Rabbit' and ‘White Wave', respectively. It was required to identify S haplotypes from self-incompatible line for the production of F_1 hybrid seeds and molecular genetic study of SII in Brassica. Here, the BoSRK_x genomic sequences of 919 bp were obtained by PCR using a pair of primers PK1 and PK4, and database searching revealed that BoSRK_x was consist with BoSRK_ 13b completely. BoSRK_x cDNA sequences were also obtained by reverse transcription-polymerase chain reaction(RT-PCR) using a pair of primers PK1 and PK4, and then we found that there were no differences between BoSRK_x and BoSRK_ 13b in the transcriptional level. On the other hand, we amplified the cDNA sequence of BoSCR_x by RT-PCR using a primer designed based on the conserved signal peptide coding region and an NotⅠ-oligo(dT) primer. Database searching revealed that BoSCR_x was most similar to BoSCR_ 13 and 3 bp more than BoSCR_ 13 . So, we conclude that S_x is possibly to be S_ 13b . Finally, S_ 13b haplotype homozygous was confirmed by genetic analysis combining with PCR-RFLP analysis and DNA blot analysis.

关 键 词：羽衣甘蓝 自交不亲和 S位点受体蛋白激酶 S位点富含半胱氨酸蛋白 S13b单倍型 

分 类 号：S681.9[农业科学—观赏园艺]