Cloning and Function Analysis of the ScDHNS Gene of Crotonase/Enoyl-CoA Superfamily from a Wild Potato Species
[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.
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