山黧豆LsSULTR基因的克隆与表达分析  

Cloning and Expression Analysis of LsSULTR Gene in Lathyrus sativus

作　　者：陈红 曾鹏 徐全乐[1] CHEN Hong;ZENG Peng;XU Quanle(College of Life Sciences, Northwest A&F University, Yangling, Shaanxi 712100, China)

机构地区：[1]西北农林科技大学生命科学学院,陕西杨陵712100

出　　处：《西北植物学报》2021年第11期1954-1961,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：陕西省自然科学基金(2020JM-167);中央高校基本科研业务费专项资金重点研究基地建设项目(lzujbky-2021-kb05)。

摘　　要：为研究山黧豆(Lathyrus sativus)硫酸盐转运蛋白(sulfate transporter,SULTR)与内源活性物质β-N-草酰-L-α,β-二氨基丙酸(β-N-oxalyl-L-α,β-diaminopropionic acid,β-ODAP)含量之间的关系,该研究采用序列比对的方法,从山黧豆转录组数据(SRP145030)中查找SULTR基因序列,并进行生物信息学分析;采用PCR方法克隆基因全长序列并进行组织特异性表达分析,以筛选与β-ODAP含量相关的SULTR基因。结果表明:(1)经序列比对共查找到13条SULTR基因序列,其编码蛋白分别隶属于SULTRⅠ-Ⅳ。其中LsSULTR3;3和LsSULTR3;5具有SULTR蛋白家族保守的STAS结构域(PF01740)和Sulfate_transp结构域(PF00916),且二者活性受到转录因子MYB和激素响应等多个顺式作用元件的共同调节,并受蛋白水平互作及磷酸化等翻译后修饰调控。(2)成功获得LsSULTR3;3和LsSULTR3;5基因全长cDNA分别为1962 bp和1923 bp,分别编码653和640个氨基酸。(3)半定量RT-PCR分析显示,LsSULTR3;3在山黧豆主根、成熟叶、茎、花、盛荚初期(S2)种子和鼓粒满期(S6)种子中表达,并在茎中的表达量较高;LsSULTR3;5在山黧豆主根、侧根、花、S2时期种子中均有表达,且花中的表达量最为显著。该研究结果为进一步研究山黧豆中硫酸盐的吸收、转运及同化、β-ODAP的生物合成途径等奠定了基础。(1)To investigate the relationship between sulfate transporter genes(SULTR)andβ-N-oxalyl-L-α,β-diaminopropionicacid(β-ODAP)content in Lathyrus sativus,we first identified 13 LsSULTR genes from transcriptomic database(SRP145030),which coding the four major groups of SULTRⅠ-Ⅳseparately.Therein,LsSULTR3;3 and LsSULTR3;5 were characterized by the conversed STAS(PF01740)and Sulfate_transp(PF00916)domain of SULTR,and gene expression levels of LsSULTR3;3 and LsSULTR3;5 strongly suggested relation withβ-ODAP biosynthesis.Bioinformatics analysis suggested that LsSULTR3;3 and LsSULTR3;5 can be regulated by various cis-acting elements such as MYB and hormone response factors.In addition,protein-protein interactions and protein phosphorylation also play an important role.(2)cDNAs of LsSULTR3;3 and LsSULTR3;5 were cloned with 1962 bp encoding a peptide of 653 amino acid residues,and 1923 bp encoding a peptide of 640 amino acid residues,respectively.(3)Semi-quantity RT-PCR analysis showed that LsSULTR3;3 gene was expressed highest in stems and followed by main roots,old leaves,flowers,seeds in early flourishing podding stage(S2)and seedfilling-flourishing stage(S6);while the LsSULTR3;5 gene was expressed highest in flowers and followed by main roots,lateral roots and seeds in S2 stage.These results will help to reveal the mechanism of sulfur transport and assimilation,β-ODAP biosynthesis in L.sativus.

关 键 词：山黧豆 硫酸盐转运蛋白 β-ODAP 基因克隆 蛋白活性调控 

分 类 号：Q785[生物学—分子生物学] Q786