水稻滇Ⅰ型和BT型细胞质雄性不育系的PCR标记鉴别

Identification of CMS Lines on Dian Type I and Boro Type Cytoplasm by a PCR Marker

中国用于杂交粳稻生产的细胞质雄性不育系仅有滇Ⅰ型和BT型,它们的不育系产生的花粉都为染败型,而且具有相同的恢保关系、同为配子体不育。因此,基于花粉败育特点和恢保关系等常规技术无法鉴别这两种不育系。本研究利用线粒体特异引物LD24对27个滇Ⅰ型不育系和9个BT型不育系进行PCR扩增,结果显示滇Ⅰ型不育系无扩增片段,BT型不育系具有1个605 bp的片段。该片段的DNA序列与水稻LD型雄性不育细胞质线粒体基因组的一个604 bp片段相似性达99%,该片段包括一个位于rps1基因下游的一段230 bp序列及rps1与orf187基因间隔区上游的一个374 bp序列。本研究的结果说明利用LD24标记的PCR产物可以有效地鉴别水稻滇Ⅰ型和BT型雄性不育细胞质。

Cytoplasmic male sterility(CMS) of Dian typeⅠ(CMS-DT) and Boro type(CMS-BT) are the only two CMS used in japonica hybrid rice production in China. Since CMS lines on the both CMS do not only produce the same abortive pollens, stained abortive type, but also share the same maintainers and restorers, and belong to gametophytic sterility, it is impossible to distinguish both CMS lines based on pollen abortive type and restorer/maintainer relationship or other regular methods. In this study, 27 CMS-DT lines and 9 CMS-BT lines were assayed by a PCR marker LD24 specific on rice mitochondrial DNA. The results showed that no a product was amplified with CMS-DT lines, but a 605 bp fragment was amplified with CMS-BT lines. Sequencing and alignment analysis revealed that the PCR fragment had 99% identity with a 604 bp fragment on mitochondrial genome in CMS-LD(lead rice). The fragment covered a 230 bp downstream sequence of rps1 plus 374 bp intergenic region between rps1 and orf187. This research demonstrated that CMS lines on CMS-DT/BT can be identified effectively by a PCR product amplified with LD24.