蒙古冰草GDSL酯酶/脂肪酶基因的序列及表达分析  

Sequence and expression analysis of GDSL esterase/lipase gene in Agropyron mongolicum

作　　者：闫秀秀 范菠菠 张学峰 于卓[1] 房永雨 吴婧 赵彦 马艳红[1] YAN Xiuxiu;FAN Bobo;ZHANG Xuefeng;YU Zhuo;FANG Yongyu;WU Jing;ZHAO Yan;MAYanhong(Agronomy College,Inner Mongolia Agricultural University,Hohhot 010019,China;Institute of Grassland,Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences,Hohhot 010031,China;College of Grassland and Resources and Environment,Inner Mongolia Agricultural University,Hohot 010018,China)

机构地区：[1]内蒙古农业大学农学院,呼和浩特010019 [2]内蒙古自治区农牧业科学院草原研究所,呼和浩特010031 [3]内蒙古农业大学草原与资源环境学院,呼和浩特010018

出　　处：《中国农业大学学报》2023年第4期41-56,共16页Journal of China Agricultural University

基　　金：国家自然科学基金(31860670);内蒙古自治区草原英才创新团队。

摘　　要：为挖掘蒙古冰草(Agropyron mongolicum)中的GDSL酯酶/脂肪酶基因,以干旱胁迫下蒙古冰草转录组测序得到的Unigenes为对象,利用生物信息学方法对蒙古冰草GDSL脂肪酶基因进行理化性质、亚细胞定位、同源性以及系统发育进化分析,同时对不同干旱处理条件下的GDSL脂肪酶基因进行表达分析。结果表明:蒙古冰草中52个GDSL脂肪酶基因,蛋白质分子量在6.80~44.77 kU,66~420个氨基酸,pI 4.27~9.42;52个蒙古冰草GDSL脂肪酶基因亚细胞定位预测显示细胞外基质、叶绿体、细胞核、细胞质、细胞膜、细胞壁均有分布;将其与13个已知具有抗旱功能的GDSL脂肪酶基因进行同源性对比及进化树构建,蒙古冰草GDSL脂肪酶基因与水稻、大麦、拟南芥、木豆、大豆、蜡梅、辣椒的GDSL脂肪酶基因均有较高相似性。在干旱处理0.5、1.0、2.0、3.0、5.0、7.0 d及复水1.0 d,与对照(CK)相比,14个基因呈显著下调表达,38个基因呈现为上调表达。对6个蒙古冰草GDSL脂肪酶蛋白跨膜及信号肽和磷酸化预测结果显示,DN26944_c0_g1属于跨膜蛋白;DN26944_c0_g1、DN14677_c0_g2、DN29240_c0_g2这3个蛋白均含有信号肽,推测其为外分泌蛋白。通过磷酸化位点分析发现DN26944_c0_g1、DN14677_c0_g2、DN29240_c0_g2、DN17531_c0_g6、DN29240_c0_g3、DN20405_c0_g1蛋白磷酸化程度总体不高,其中DN26944_c0_g1最高有34处磷酸位点,DN14677_c0_g2最低有7处磷酸位点。模拟干旱处理下蒙古冰草DN26944_c0_g1、DN14677_c0_g2、DN29240_c0_g2、DN17531_c0_g6、DN29240_c0_g3、DN20405_c0_g1这6个基因的表达量均显著高于对照组(CK),说明这6个酯酶/脂肪酶基因在蒙古冰草干旱胁迫中起正调控作用。In order to exploit GDSL lipase gene of Agropyron mongolicum, GDSL lipase genes were screened from the unigenes of A. mongolicum under drought stress. The physicochemical properties, subcellular localization, homology and phylogenetic evolution of the selected GDSL lipase gene were analyzed by bioinformatics methods. Meanwhile, the expression of GDSL lipase gene under different drought conditions was analyzed. The results showed that: A total of 52 GDSL lipase genes were screened from A. mongolicum. The protein size was ranged from 66 to 420 aa, the molecular weight was ranged from 6.80 to 44.77 kU, and the theoretical isoelectric point was ranged from 4.27 to 9.42. The 52 GDSL lipase genes were predicted to be distributed in extracellular, chloroplast, nucleus, cytoplasm, cell membrane and cell wall. Compared with 13 known GDSL lipase genes with drought resistance function and an evolutionary tree was constructed, the GDSL lipase gene of A. mongolicum has high homology with the GDSL lipase gene of rice, barley, arabidopsis, pigeonpea, soybean, wintersweet and pepper. Among drought treatments at 0.5, 1.0, 2.0, 3.0, 5.0, 7.0 d and rehydration 1.0 d, 14 genes were significantly down regulated and 38 genes were up-regulated compared to control(CK). The prediction results of transmembrane, signal peptide and phosphorylation of six GDSL lipase proteins of A. mongolicum showed that DN26944_c0_g1 belongs to transmembrane protein, DN26944_c0_g1, DN14677_c0_g2, DN29240_c0_g2 all contain signal peptides, and it is speculated that they are exocrine proteins. Phosphorylation site analysis found that the phosphorylation degrees of DN26944_c0_g1, DN14677_c0_g2, DN29240_c0_g2, DN17531_c0_g6, DN2 9240_c0_g3, DN20405_c0_g1 protein were not high in general, among which DN26944_c0_g1 had 34 phosphorylation sites, and DN14677_c0_g2 had 7 phosphorylation sites. The differential expression analysis of the six GDSL lipase genes of A. mongolicum under simulated drought treatment showed that the expression levels of DN26944_c0_g1, DN1467

关 键 词：蒙古冰草 GDSL酯酶/脂肪酶基因 生物信息学分析 抗旱 

分 类 号：S543.9[农业科学—作物学]