果树学报2025，Vol.42Issue(4)：732-751,20.DOI:10.13925/j.cnki.gsxb.20240629
葡萄绿枝嫁接愈合过程中内源激素含量变化及转录组解析
Changes of endogenous hormone concentrations and transcriptome anal-ysis during the healing process of greenwood grafting in grape
摘要
Abstract
[Objective]To study the changes of hormone concentrations in the graft union during the healing process of greenwood grafting,the experiment was carried out,combined with transcriptomic data analysis,so as to identify the key hormone changes in the healing process of greenwood grafting in grape,and provide references for revealing the molecular mechanism of the healing process.[Meth-ods]The stion combinations of Tiangong Liren/Beta,Tiangong Moyu/Beta and Shine Muscat/Beta were used as materials and were collected at 1,4,7,10 and 13 days after grafting(DAG),respectively.The concentrations of auxin(IAA),isopentenyladenine(IP),abscisic acid(ABA)and jasmonates(JAs)in the graft union were determined by high performance liquid chromatograph-triple quadrupole tandem mass spectrometry(HPLC-QqQ-MS/MS).Simultaniously,transcriptome sequencing was performed on the graft union of Tiangong Liren/Beta.[Results]From the perspective of morphology,Tiangong Moyu/Beta buds burst first,and the bud swelling was observed at 4 DAG.Tiangong Liren/Beta and Shine Muscat/Beta were observed to have swollen buds at 7 DAG.Different stion combinations com-pleted the healing process within 13 DAG.From the analysis of plant hormone levels,the concentration of IAA increased,while the concentrations of IP,ABA and JAs decreased during healing.From tran-scriptome level analysis,the transcriptome data of Tiangong Liren/Beta at different stages were pair-wise compared,and 6319 differentially expressed genes were screened.1 DAG vs 13 DAG had the larg-est number of differentially expressed genes,with 3846,the second was 1 DAG vs 10 DAG,with 3301 and the third was 1 DAG vs 7 DAG,with 2968.The results showed that the gene transcriptional regula-tion of 7 DAG,10 DAG and 13 DAG were significantly changed compared with 1 DAG.The 10 DAG vs 13 DAG comparison combination had the smallest number of differentially expressed genes,indicating that there was little difference in the gene expression pattern between 10 DAG and 13 DAG.6319 dif-ferentially expressed genes were divided into 6 clusters by time series analysis.Cluster 5 contained the most differentially expressed genes,which were significantly enriched in xyloglucan,including xyloglu-cosyl transferase activity,xyloglucan metabolic process and other pathways,its expression showed a downward trend in the whole healing process,and the trend was obvious at 1-4 DAG.The differentially expressed genes contained in Cluster 3 were enriched in auxin-activated signaling pathway,phloem or xylem histogenesis and other pathways,and their expression levels increased at first week after grafting and were higher at 4-10 DAG.In the process of grapevine greenwood grafting healing,27 differentially expressed genes were screened to participate in the synthesis and metabolism of plant hormones,and 3,2,12 and 10 differentially expressed genes were involved in the synthesis and metabolism of auxin,cy-tokinin,abscisic acid and jasmonic acid,respectively.In the tryptophan-dependent auxin synthesis path-way,L-tryptophan decarboxylase TDC(Vitvi07g00696)and aldehyde dehydrogenase ALDH(Vit-vi04g01402)had the highest expression levels at 7 DAG.Isopentenyltransferase IPT(Vitvi07g00154)was highly expressed at first week after grafting,and then showed a downward trend.The expression of cytokinin dehydrogenase CKX(Vitvi04g00161)decreased significantly at 1-4 DAG and 7-10 DAG by 0.66 and 0.60 times,respectively.The expression level of 9-cis-epoxycarotenoid dioxygenase NCED(Vitvil9g01356)was higher at 1-4 DAG and decreased at 4-10 DAG,which was similar to the chang-ing trend of abscisic acid concentration.The expression levels of LOX(Vitvi06g00158),AOS(Vit-vi18g00886),OPR(Vitvi18g03162 and Vitvi18g04622)and OPCL1(Vitvi18g00124)generally de-creased at 1-10 DAG,which were consistent with the changing trend of jasmonic acid concentration.49 differentially expressed genes were selected to participate in plant hormone signal transduction path-way.There were 17 differentially expressed genes involved in auxin signal transduction.They included auxin influx carrier(AUX1),transport inhibitor response protein 1(TIR1),auxin/indole-3-acetic acid(Aux/IAA),auxin response factor(ARF),acetic acid-amido synthetase(GH3),small auxin-up RNA(SAUR),auxin binding protein 1(ABP1),plasma membrane H+-transporting ATPase(AHA)and mito-gen activated protein kinase 3(TMK3).Two-component response regulators(A-ARR)(Vitvi13g01433 and Vitvi17g00732)involved in cytokinin signal transduction were highly expressed at first week after grafting.There were 8 differentially expressed genes were involved in abscisic acid signal transduction,including abscisic acid receptor PYR/PYL,protein phosphatase 2C(PP2C),sucrose non-fermenting 1-related protein kinase 2(SnRK2)and ABA-responsive element binding factor(ABF).There were 5 dif-ferentially expressed genes were involved in jasmonic acid signal transduction,including jasmonic acid-amino acid synthetase(JAR1),jasmonate ZIM domain-containing protein(JAZ)and transcription fac-tor MYC2.There were 8 differentially expressed genes were involved in brassinosteroid signal transduc-tion,including brassinosteroid resistant 1/2(BZR1/2),cyclin D3(CYCD3)and xyloglucan endotrans-glucosylase protein(TCH4).There were 3 differentially expressed genes were involved in gibberellin signal transduction,including gibberellin receptor 1(GID1)and DELLA protein.There were 3 differen-tially expressed genes were involved in ethylene signal transduction,including ethylene receptor(ETR),EIN3-binding F-box protein(EBF)and ethylene-responsive transcription factor(ERF).These three dif-ferentially expressed genes involved in salicylic acid signal transduction were pathogenesis-related pro-tein 1(PR1).By WGCNA analysis of transcriptome data,13 co-expression modules were obtained.The correlation analysis between the module and plant hormone showed that GH3(Vitvi03g00586),LOX(Vitvi06g00158),AOS(Vitvi18g00886),OPCL1(Vitvi18g00124),ABF(Vitvi12g01667)and TCH4(Vitvi11g01682)were positively correlated with the concentrations of JA and IP.AHA(Vitvi14g01888)and TIR1(Vitvi07g00248)were negatively correlated with the concentrations of ABA and IP.[Conclu-sion]The changes of plant hormone concentrations in the process of greenwood grafting healing were stage-specific.IAA played a key role in the process of grafting healing.IP,ABA and JAs regulated the healing process by interacting with IAA signal transduction.关键词
葡萄/绿枝嫁接/内源激素/转录组/差异表达基因/愈合过程Key words
Grape/Greenwood grafting/Endogenous hormone/Transcriptome/Differentially ex-pressed gene/Healing process分类
农业科技引用本文复制引用
邓嘉漪,魏嘉彦,陈爱赛,何非,于可济,王军..葡萄绿枝嫁接愈合过程中内源激素含量变化及转录组解析[J].果树学报,2025,42(4):732-751,20.基金项目
现代农业产业技术体系专项资金(CARS-29) （CARS-29）
宁夏回族自治区重点研发计划重大项目(2024BBF01002-03) （2024BBF01002-03）
