水稻线粒体基因orf290功能的初步研究
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国家自然科学基金面上项目(31170296) 


Function of mitochondrial gene or f 290 in rice
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    摘要:

    从红莲型不育系粤泰A(YTA)线粒体基因组中克隆并鉴定了一个6.7 kb 的CMS相关片段HL- sp1 ,序列预测发现了2个 orf ,其中一个编码290个氨基酸,暂命名为 orf290 。构建带有线粒体信号肽的超表达载体 35S∶∶Rf1b5’ ∶∶ orf290 ,通过农杆菌介导转化受体保持系粤泰B(YTB),以水稻的单拷贝基因蔗糖磷酸合成酶基因( SPS ) 作为内参基因,以潮霉素抗性基因( hpt )作为目的检测基因,基于实时荧光定量PCR鉴定转基因T0代植株中外源基因的拷贝数,并对阳性植株进行花粉镜检和结实率统计。结果显示获得了2株单拷贝转35 S ∶∶ Rf1b5′ ∶∶ orf290 植株,并且都有 orf290 的表达。单拷贝超表达 orf290 植株的平均花粉可育度30.8%,平均套袋结实率41.6%。这些结果初步证实,线粒体基因 orf290 可能与水稻细胞质雄性不育相关。

    Abstract:

    HL- sp1 , a 6.7 kb HL-CMS-related fragment, was cloned from the mitochondrial genome of HL-CMS sterile line YTA. One of the two ORFs predicted encodes 290 amino acids and is tentatively named orf290 . One overexpression vector 35S∶∶Rf1b5′∶∶orf290 with a mitochondrial signal peptide was constructed and transformed by Agrobacterium into YTB, a HL-CMS maintain line. The copy numbers of the exogenous gene in the 35S∶∶Rf1b5′∶∶orf290 transgenic T0 plants were estimated according the ratio of hygromycin B phosphotransferase ( hpt ) gene and the single copy sucrose phosphate synthase ( SPS ) gene in rice with qPCR based on DNA. The pollen fertility and the seed set rate of the T0 positive transgenic plants were investigated. The results showed that two transgenic plants with single copy of 35S∶∶Rf1b5′ ′∶∶ orf290 were obtained and the expression level of orf290 in the T0 positive transgenic plants were detected with qPCR. The average pollen fertility was 30.8% and the average seed set rate was 41.6% in the single copy transgenic lines. It is indicated that mitochondrial gene orf290 may be related to cytoplasmic male sterility in rice.

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杨梦醒,姜 慧,田 泽,刘学群,谭艳平,王春台.水稻线粒体基因orf290功能的初步研究[J].华中农业大学学报,2018,37(06):1-6

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  • 收稿日期:2018-04-10
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  • 在线发布日期: 2018-10-31
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