水稻OsPIN1d基因克隆与转化及其在水稻根负向光性中的作用

发布时间：2018-02-05 03:14

本文关键词： 水稻生长素极性运输 OsPINd 负向光性种子根　出处：《西北植物学报》2017年10期 　论文类型：期刊论文

【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。
[Abstract]:The auxin polarity transport vector OsPIN1 gene family may be involved in the regulation of rice root negative light. In order to study the relationship between the OsPIN1d gene and the formation of the negative light of rice root, OsPIN1a and OsPIN1b have been proved to be involved in the negative light of rice root. According to the nucleotide sequence of OsPIN1d(Accession number: BR000830 in GenBank database, a specific primer was designed. The complete OsPIN1d gene was amplified by RT-PCR from cDNA of rice root tip. Bioinformatics analysis showed that the length of OsPIN1d was 1497 BP. The GC content of 554 amino acids was 64.08; Multiple amino acid sequence alignment and phylogenetic tree construction showed that OsPIN1d was associated with OsPIN1b in rice, OsPIN4 in maize and OsPIN1c in Arabidopsis thaliana AtPIN2. The genetic distance of genes such as OsPIN1a and AtPIN1 was close. The fusion superexpression vector pCAMBIA-1301-OsPIN1d: GFP was constructed. The transgenic rice was transformed and obtained. The results of RT-PCR detection and GUS staining showed that the exogenous fragments were successfully integrated into the rice genome and highly expressed in the roots. The negative light intensity of the transgenic rice seeds was significantly higher than that of the wild type, and the fluorescence density of OsPIN1d-GFP on the optically side was weaker than that on the backlight side. OsPIN1d was involved in the transport of IAA and NAA in rice roots, which promoted the formation of negative light in rice roots.
【作者单位】：江苏理工学院化学与环境工程学院;河南科技大学农学院;绍兴文理学院生命科学学院;
【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)
【分类号】：S511
【正文快照】： 水稻(Oryza sativa L.)是重要的粮食作物,良好的根系发育对水分和养分吸收有着重要作用。根系生长除了受重力作用外,光照亦可直接或间接地影响根系生长、侧根和根毛形成,以及根的向重性与向光性[1-2],相对于对植物向光性研究而言[3],前人对根负向光性的研究比较少[4-5]。研究

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