摘要
生长素极性运输输出载体OsPIN1基因家族可能参与调控水稻根负向光性,其中OsPIN1a和OsPIN1b参与水稻根负向光性已得到证实。为了研究OsPIN1d基因与水稻根负向光性形成的关系,依据GenBank数据库中OsPIN1d(Accession number:BR000830)的核苷酸序列,设计特异性引物,通过RT-PCR从水稻根尖的cDNA中扩增出完整的OsPIN1d基因全长。生物信息学分析表明,OsPIN1d的序列全长为1 497bp,编码554个氨基酸,GC含量为64.08%;氨基酸序列多重比对及系统发育树构建表明,OsPIN1d与水稻OsPIN1b、玉米OsPIN4以及拟南芥AtPIN2、OsPIN1c、OsPIN1a和AtPIN1等基因的遗传距离较近。通过构建融合超表达载体pCAMBIA-1301-OsPIN1d∷GFP,转化并获得其转基因水稻,经RT-PCR检测和GUS染色结果显示,外源片段已成功整合到水稻基因组内,并在根部高效表达;受单侧光照射后,转基因水稻种子的根负向光性明显大于野生型,且向光侧OsPIN1d-GFP荧光密度明显弱于背光侧。研究表明,OsPIN1d参与了水稻根负向光性的IAA和NAA的运输,从而促进了根负向光性的形成。
OsPIN1a and OsPIN1b of auxin efflux carrier OsPIN1family genes were demonstrated in regulating negative phototropism of rice roots.To study relationship between OsPIN1d and negative phototropism of rice roots,we amplified the complete open reading frame of OsPIN1d through reverse transcriptase polymerase chain reaction(RT PCR)based on sequence deposited in GenBank(Accession number:BR000830).Bioinformatics analysis showed that OsPIN1d was consisted of1497bp,coding554amino acids(AA),and GC content was64.08%.AA sequence comparison and phylogenic tree indicated that OsPIN1d gene belongs to plant PINs,which is closed to OsPIN1b in rice,OsPIN4in maize,AtPIN2,OsPIN1c,OsPIN1a and AtPIN1in Arabidopsis.Fusion expression vector pCAMBIA1301OsPIN1d∷GFP containing OsPIN1d gene was constructed to obtain T2generation transgenic rice.RT PCR molecular detection andβ-glucuronidase staining showed that the target construct was integrated into genome of rice,mainly locating in root.Irradiated with unilateral light,the negative phototropic curvatures of transgenic rice seed roots were higher than those of wild type.Moreover,OsPIN1d GFP in shaded side was significantly higher than that in irradiated side in elongation zone of transgenic rice root tip.It was indicated that OsPIN1d is involved in transportation of IAA and NAA,thus promoting formation of negative phototropism in rice roots.
作者
梁国斌
胥华伟
方群
莫亿伟
Liang Guobin;Xu Huawei;Fang Qun;Mo Yiwei(School of Chemistry and Environmental Engineering, Jiangsu University of Technology, Changzhou, Jiangsu 213001, China;College of Agriculture, Henan University of Science and Technology, Luoyang, Henan 471003, China;College of Life Science, Shaoxing University, Shaoxing, Zhejiang 312000, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2017年第10期1896-1903,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
浙江省自然科学基金(LY14C130003)
国家自然科学基金(31071353)
江苏省产学研前瞻联合性研究基金(BY2015028-04)
