生物技术通报2024，Vol.40Issue(9)：92-103,12.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0528
马铃薯野生种烯酰水合酶超家族基因ScDHNS的克隆与功能分析
Cloning and Function Analysis of the ScDHNS Gene of Crotonase/Enoyl-CoA Superfamily from a Wild Potato Species
摘要
Abstract
[Objective]The DHNS(1,4-dihydroxy-2-naphthoyl-CoA synthase)gene in potatoes is a potentially important in the biosynthesis of glycoalkaloid metabolism in Solanaceae plants.Researching and verifying of the function of the potato DHNS gene can provide a source of genes and materials for the selection of low-glycoalkaloid potato varieties(lines).[Method]The full-length sequence of ScDHNS cDNA was cloned from the wild potato species Solanum chacoense using the RACE technique.This sequence was analyzed for bioinformatics and subcellular localization,and its function was verified by constructing an overexpression vector pBWA(V)HS-DHNS to transform cultivated potato.[Result]The read open frame of ScDHNS cDNA sequence is 1023 bp,encodes 340 amino acids,has a molecular weight of 37.34 kD and an isoelectric point of 8.592,and contains a typical ECH domain.It belongs to the enoyl hydratase/acetyl-CoA superfamily and is present in the genomes of plants such as Brachypodium distachyon and Medicago truncatula with homologous genes,showing gene expansion and contraction events.After the overexpression of the ScDHNS gene,it was found that the expressions of the transformed plants ScDHNS and SGT1 were significantly upregulated,and the expressions were notably higher than those in wild-type(WT)potato plants.Additionally,the total glycoalkaloid content in the corresponding transformed plants was significantly higher than that in WT potato plants,reaching a maximum of 364.3 mg/kg,which is 2.4 times that of the control.Subcellular localization results indicates that ScDHNS is localized in the peroxisome.[Conclusion]The ScDHNS gene in potato may regulate the expression of the key gene SGT1,which is involved in glycoalkaloid synthesis.It influences glycoalkaloid synthesis through the β-oxidation and mevalonate pathways.This gene is significantly associated with the subcellular compartmentalization of glycoalkaloids and has considerable application value in the cultivation of potato varieties with reduced glycoalkaloid content.关键词
马铃薯野生种/恰柯薯/ScDHNS/基因克隆/亚细胞定位Key words
wild potato/Solanum chacoense/ScDHNS/gene cloning/subcellular localization引用本文复制引用
乔岩,杨芳,任盼荣,祁伟亮,安沛沛,李茜,李丹,肖俊飞..马铃薯野生种烯酰水合酶超家族基因ScDHNS的克隆与功能分析[J].生物技术通报,2024,40(9):92-103,12.基金项目
国家自然科学基金地区科学基金项目(31960441),甘肃省自然科学基金(18JR3RM236),庆阳市重点研发计划(QY-STK-2023A-077),庆阳市科技人才专项计划(QY-STK-2022A-006) （31960441）
