植物保护学报2025，Vol.52Issue(2)：390-399,10.DOI:10.13802/j.cnki.zwbhxb.2025.2024104
麦红吸浆虫滞育不同阶段幼虫ATP含量、ATP酶活力和SmATP5A基因表达分析
ATP content,ATPase activity and SmATP5A gene expression in red wheat blossom midge Sitodiplosis mosellana larvae at different diapause stages
摘要
Abstract
To clarify the relationship between ATP content,ATPase activity and gene expression of ATPase α-subunit(ATP5A)in red wheat blossom midge Sitodiplosis mosellana and diapause develop-ment,ATP content and activities of ATP synthase and hydrolase in S.mosellana larvae at different dia-pause stages were respectively measured using an ATP fluorescence assay kit and enzyme continuous reaction colorimetry.The cDNA sequence of SmATP5A was cloned using reverse transcription PCR(RT-PCR)technique,and analyzed by bioinformatics.Quantitative real-time PCR(qPCR)and Western-blot analysis were used to detect the mRNA and protein expression patterns of the SmATP5A gene in dia-pause process of S.mosellana.Results showed that ATP content and activities of ATP synthase and hydrolyase in S.mosellana larvae significantly decreased after entering diapause,remained at low levels throughout diapause,but increased after entering post-diapause quiescent period,and increased again during post-diapause development,with levels significantly higher in the later stage compared to those during diapause and post-diapause quiescence.Open reading frame(ORF)of SmATP5A obtained is 1 659 bp(GenBank accession number:PQ412453),encoding a protein of 552 amino acids with a pre-dicted molecular weight of 59.6 kD.The amino acid sequence analysis suggested that SmATP5A con-tained the classical nucleotide-binding domain and Walker A and Walker B motifs of ATPase catalytic core.SmATP5A exhibited the highest amino acid identity and closest phylogenetic relationship to ATP5A from Contarinia nasturtii.qPCR and Western-blot analysis demonstrated that the mRNA and protein expression levels of SmATP5A showed significant differences in the course of diapause.Their expression levels were the highest at pre-diapause and post-diapause development stages,followed by post-diapause quiescence stage,and the lowest in diapause stage,which are highly consistent with change pattern of ATP content and ATPase activity.These results suggest that the decreases in ATP con-tent,ATPase activity and SmATP5A expression during diapause in S.mosellana are closely associated with diapause initiation and reduced metabolism level in diapause stage,while their increases are closely linked to the elevated energy demands required for diapause termination and post-diapause development.关键词
麦红吸浆虫/滞育/ATP含量/ATP酶活力/基因表达Key words
Sitodiplosis mosellana/diapause/ATP content/ATPase activity/gene expression引用本文复制引用
马倩,黄启通,任家欣,朱克岩,成卫宁..麦红吸浆虫滞育不同阶段幼虫ATP含量、ATP酶活力和SmATP5A基因表达分析[J].植物保护学报,2025,52(2):390-399,10.基金项目
国家自然科学基金(31371933),陕西省重点研发计划(2020NY-059),山东省自然科学基金(ZR2023QC051) （31371933）
