Cloning of HEMG gene from Hordeum bogdanii Wilensky and effect of endophytic fungi on its relative expression

YAN Jiahui; LIU Xiaozhen; MA Yunfei; SHIFA Tumai; YANG Hongfeng; WEI Yidan; CHEN Shuihong

doi:10.13557/j.cnki.issn1002-2813.2025.14.014

Abstract：The study aimed to investigate the influence of endophytic fungi on the heterologous enzyme for malfunctioning protoporphyrinogen oxidase(HEMG)gene in host plants.The protein-coding region(CDS)of the HEMG gene from Hordeum bogdanii Wilensky was cloned and sequenced using reverse transcription polymerase chain reaction(RT-PCR).Bioinformatics analysis was conducted on the obtained sequence,and subcellular localization confirmed the nuclear localization of the HEMG protein.Real time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the relative expression levels of HEMG genes in Bunton barley with endophytic fungi(GI,WI)and without a endophytic fungi(GF,WF)in Wensu County,Xinjiang and Linze area,Gansu,as well as in Bunton barley implanted with Wensu bacteria(HE2)and Bunton barley implanted with Wensu bacteria(HE3)in Gansu.The results showed that the cloned HEMG CDS was 927 bp in length,encoding 308 amino acid residues.HEMG protein may be located in the nucleus without transmembrane domain and signal peptide.The homology of HEMG gene between Burton barley and Uraltou wheat was the highest,reaching 98.04％.The relative expression of the HEMG gene in GI was significantly higher than that in GF(P＜0.05),the relative expression of the HEMG gene in WI was significantly lower than that in WF(P＜0.05),the relative expression of the HEMG gene in HE3 was significantly higher than that in WI(P＜0.05),and the relative expression of the HEMG gene in HE2 was significantly lower than that in GI(P＜0.05).The study shows that the strains from Linze,Gansu can increase the relative expression of the HEMG gene,while those from Wensu,Xinjiang suppress it.