山黧豆LsSULTR基因的克隆与表达分析(山黧豆专辑)
投稿时间:2021-06-27  修订日期:2021-08-28  点此下载全文
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作者单位邮编
陈红 西北农林科技大学生命科学学院 712100
曾鹏 西北农林科技大学生命科学学院 712100
徐全乐 西北农林科技大学生命科学学院 712100
基金项目: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)
中文摘要:为研究山黧豆硫酸盐转运蛋白(sulfate transporter,SULTR)与内源活性物质?-N-草酰-L-?,?-二氨基丙酸(?-N-oxalyl-L-?,?-diaminopropionicacid,?-ODAP)含量之间的关系,本研究从山黧豆转录组数据(SRP145030)中比对到13条SULTR基因序列,分别编码SULTR I—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),LsSULTR3;3和LsSULTR3;5基因表达水平可能和山黧豆活性物质?-ODAP的生物合成密切相关。生物信息学分析显示,LsSULTR3;3和LsSULTR3;5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,并且蛋白水平互作及磷酸化等翻译后修饰也起到重要作用。利用PCR扩增获得LsSULTR3;3和LsSULTR3;5基因全长cDNA为1962bp和1923bp,分别编码653和640个氨基酸。半定量RT-PCR基因组织表达分析显示,LsSULTR3;3在主根、成熟叶、茎、花、盛荚初期(S2)种子和鼓粒满期(S6)种子中表达,以在茎中的表达量较高;LsSULTR3;5在主根、侧根、花、S2时期种子中均有表达,以花中的表达量最为显著。上述结果为进一步研究山黧豆中硫酸盐的吸收、转运及同化、?-ODAP的生物合成途径等奠定了基础。
中文关键词:山黧豆  硫转运蛋白  β-ODAP  基因克隆  蛋白活性调控
 
Cloning and Expression Analysis of LsSULTR Gene in Lathyrus sativus
Abstract:To investigate the relationship between sulfate transporter genes (SULTR) and ?-N-oxalyl-L-?,?-diaminopropionicacid (?-ODAP) content in Lathyrus sativus, 13 LsSULTR genes were first identified from transcriptomic database (SRP145030), which coding the four major groups of SULTR Ⅰ-Ⅳ separately. Therein, LsSULTR3;3 and LsSULTR3;5 were characterized by the conversed STAS (PF01740) and Sulfate_transp (PF00916) domain of SULTR, and gene expression level of LsSULTR3;3 and LsSULTR3;5 were suggested strongly related to ?-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 plays also an important role. And then, cDNAs of LsSULTR3;3 and LsSULTR3;5 were cloned with 1962bp encoding a peptide of 653 amino acid residues, and 1923bp encoding a peptide of 640 amino acid residues separately. Semi-quantity RT-PCR analysis showed that the LsSULTR3;3 gene was expressed highest in stems and followed by mainroots, old leaves, flowers, seeds in early flourishing podding stage (S2) and seedfilling-flourishing stage (S6) stage; 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, ?-ODAP biosynthesis in L. sativus.
keywords:Lathyrus sativus  sulfate transporter  β-ODAP  gene cloning  protein activity regulation
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