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6.7kb的CMS相关片段HLsp1,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35S::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。
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 T 0 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 T 0 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 T 0 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.
作者
杨梦醒
姜慧
田泽
刘学群
谭艳平
王春台
YANG Mengxing;JIANG Hui;TIAN Ze;LIU Xuequn;TAN Yanping;WANG Chuntai(Key Laboratory of State Ethnic Affairs Commission for Biological Technology/Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China/College of Life Sciences,South-Central University for Nationalities,Wuhan 430074,China)
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2018年第6期1-6,共6页
Journal of Huazhong Agricultural University
基金
国家自然科学基金面上项目(31170296)
作者简介
杨梦醒,硕士研究生.研究方向:植物分子生物学.E-mail:1182867670@qq.com;通信作者:王春台,博士,教授.研究方向:水稻分子生物学.E-mail:wangchuntai@mail.scuec.edu.cn。
