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基于BSA-seq和植物激素联合分析筛选番茄耐热基因OA北大核心CSTPCD

Screening of Heat Tolerance Genes in Tomato(Solanum lycopersicum)Based on BSA-seq and Plant Hormone Combined Analysis

中文摘要英文摘要

高温影响番茄(Solanum lycopersicum)的生理和激素代谢,番茄耐热相关基因的定位可以为后续基因克隆和功能分析提供理论依据和技术支持.本研究以电导率为鉴定指标,通过对苗期2个番茄品系进行高温耐热鉴定,以耐热品系CH与不耐热品系DH及其F1代自交构建成的F2代分离群体为材料,对亲本CH与DH进行植物激素检测,F2代分离群体采用混合分离群分析法(bulked sergeant analysis,BSA)进行重测序,通过二者联合分析筛选出与番茄耐热相关的候选基因.结果表明,通过植物激素差异代谢物筛选及KEGG注释获得了4个与植物非生物胁迫相关的代谢通路,分别为甘氨酸、丝氨酸和苏氨酸代谢途径,氨基酸的生物合成途径,色氨酸代谢途径和氨酰-tRNA生物合成途径;通过BSA-seq测序,在3号染色体上获得了1个大小为6.32 Mb的候选区间,其中包括752个候选基因,结合差异代谢通路,最终在候选区域代谢通路内筛选出5个与番茄耐热相关的候选基因,分别为D-甘油酸3-激酶(D-glycerate 3-kinase,DG3K)、羟基丙酮酸还原酶(hydroxypyruvate reductase,HYDR)、脯氨酰-tRNA合成酶(prolyl-tRNA synthetase,PARS2)和磷酸核酮糖-3-差向异构酶(ribulose phosphate 3-epimerase,RPE)、醛脱氢酶(aldehyde dehydrogenase,ALDH);通过qPCR对候选基因进行表达量分析,候选基因DG3K、PARS2和RPE在高温胁迫3h后表达量极显著增加(P<0.01),推断这3个候选基因可能在调控番茄耐热能力上起到重要作用.本研究对分子辅助选育番茄耐热品系具有重要意义.

High temperatures affect the physiology and hormone metabolism of tomatoes(Solanum lycopersicum).Localization of genes involved in heat tolerance in tomato can provide theoretical and technical support for subsequent gene cloning and functional analysis.In this study,electrical conductivity was used as the identification index,and 2 tomato strains at seedling stage were identified by high temperature and heat resistance,and the F2 generation isolated population formed by heat-resistant strain CH and heat-sensitive strain DH and their F1 generation were used as materials to detect plant hormones in parents CH and DH.The F2 generation isolated population was resequenced by bulked sergeant analysis(BSA),and candidate genes related to tomato heat resistance were screened through the combined analysis.The results showed that 4 metabolic pathways related to abiotic stress were obtained by phytohormone differential metabolite screening and KEGG annotation,namely glycine,serine and threonine metabolic pathways,amino acid biosynthetic pathway,tryptophan metabolic pathway and aminoacyl-tRNA biosynthetic pathway;6.32 Mb size candidate interval was obtained on chromosome 3 by BSA-seq sequencing.which included 752 candidate genes.Combined with the differential metabolic pathways,5 candidate genes associated with heat tolerance in tomato were finally screened within the candidate regional metabolic pathways,namely:D-glycerate 3-kinase(DG3K),Hydroxypyruvate reductase(HYDR),prolyl-tRNA synthetase(PARS2)and ribulose phosphate 3-epimerase(RPE),aldehyde dehydrogenase(ALDH).The expression of the candidate genes was analysed by qPCR,the expression of candidate genes DG3K,PARS2,and RPE increased extremely significant after 3 hours of high temperature stress(P<0.01),it was hypothesized that these 3 candidate genes might play an important role in the regulation of heat tolerance in tomato.This study has important implications for molecular assisted selection of heat-resistant tomato lines.

姜悦畅;段金虎;李毅丰;毛秀杰;王帅;张宁

河北科技师范学院园艺科技学院,秦皇岛 066004河北科技师范学院园艺科技学院,秦皇岛 066004||河北省特色园艺种质挖掘与创新利用重点实验室,秦皇岛 066004||河北省燕山农业特色产业技术研究院,秦皇岛 066004||河北科技师范学院河北省高校特色园艺植物生物育种应用技术研发中心,秦皇岛 066004河北科技师范学院园艺科技学院,秦皇岛 066004||河北省特色园艺种质挖掘与创新利用重点实验室,秦皇岛 066004||河北科技师范学院河北省高校特色园艺植物生物育种应用技术研发中心,秦皇岛 066004河北科技师范学院园艺科技学院,秦皇岛 066004||河北省特色园艺种质挖掘与创新利用重点实验室,秦皇岛 066004

园艺学与植物营养学

番茄耐热鉴定植物激素混合分离群分析(BSA)

TomatoHeat resistance identificationPlant hormoneBulked sergeant analysis(BSA)

《农业生物技术学报》 2024 (005)

995-1007 / 13

河北省科技厅现代种业科技专项(20326325D)

10.3969/j.issn.1674-7968.2024.05.002

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