【摘要】 外源染色体片段或基因的准确鉴定对作物遗传改良具有重要的作用。尽管分子标记已被广泛应用到外源染色体片段或基因的检测上,但是对于大群体中少数几个阳性植株的鉴定,逐个检测的效率极低。本研究以陆地棉(Gossypium hirsutum L.)新陆早48号与海岛棉(G.barbadense)染色体片段置换系CSSL-122的BC2F1群体147单株DNA样本为材料,通过构建二维矩阵排列DNA样本材料,分别对待测样本按行、列进行混合,获得m个行的混合样本和n个列的混合样本;以m=n=8矩阵为例,对待检m×n=64个样本行列混合获得16个混合样本并进行检测,根据检测结果在矩阵中推断确定含有外源染色体片段的阳性样本。结果表明,利用基于PCR-SSR技术构建矩阵混合池的检测方法,可以快速检测陆地棉背景中外源海岛棉染色体片段,逐个样本检测验证了其方法的准确性;统计学分析表明,阳性样本出现频率低于7.81%时,检测样本的节本率可保持在50%以上。基于PCR-SSR技术构建矩阵混合池检测方法的建立,为棉花外源染色体片段快速高效鉴定提供了新方法。更多还原

【Abstract】 To accurately identify exogenous chromosome fragments or genes plays an important role in crop genetic improvement.Although the molecular markers have been widely used in detection of exogenous chromosome fragments or genes,the efficiency of identificating a few positive plants in large group is very low.In this study,DNA samples of 147 plants in BC2F1population which was established by Gossypium hirsutum Xinluzao 48 and G.barbadense chromosome segment substitution lines CSSL- 122 through hybridization and backcross were used as materials,which were arrayed by constructing 2 dimensional matrix.The test samples were mixed according to the row and column,respectively,to get m rows of mixed samples and n columns of mixed samples,taking m=n=8 matrices as example:The m×n=64 mixed samples were obtained to test.The positive samples containing exogenous chromosome fragments in the matrix could be identified accordingly to the detection results.Results showed that exogenous Gossypium barbadense chromosome fragments in G.hirsutum could be detected quickly by using the test method of establishing a matrix mixing bank based on PCR-SSR.In addition,the accuracy of the method was verified by means of sample detection one by one furtherly.Statistical analysis showed that frequency of positive sample detection was less than 7.81%,cost saving ratio of detecting the samples could be maintained at more than 50%.Detection method of constructing the matrix mixed pool based on PCR-SSR will provide a new method for the rapid identification of exogenous Gossypium barbadense chromosome fragment in Gossypium hirsutum.更多还原