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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。
The cytoplasmic male sterility lines used for the production of japonica hybrid rice in China are only Yunnan type Ⅰ and BT type. The pollen produced by their male sterile lines is stained and has the same restorer relationship, which is the same as that of gametophyte sterility. Therefore, based on the characteristics of pollen abortion and the relationship between the restoration and security and other conventional techniques can not identify the two sterile lines. In this study, 27 Yunnan Dian I-CMS lines and 9 BT-type CMS lines were amplified by PCR using the mitochondrial DNA-specific primer LD24. The results showed that there was no amplified fragment in Dian I-type sterile line and one 605 bp fragment. The DNA sequence of this fragment was 99% identical to a 604 bp fragment of the rice LD genomic cytoplasmic mitochondrial genome. The fragment contained a 230 bp sequence downstream of the rps1 gene and a 374 upstream of the rps1 and orf187 intergenic regions bp sequence. The results of this study indicate that the LD24-labeled PCR product can effectively identify the cytoplasm of rice Dian Ⅰ and BT genotypes.