核桃不定根发生过程酵母文库构建与JrAGD3上游调控因子筛选

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中图分类号：S718.46 文献标志码：A 文章编号：1673-923X（2026）03-0015-08

Construction of yeast library during walnut adventitious root formation and screening of upstream regulatory factors of JrAGD3 gene

ZHANG Pu，SONG Li， SONG Xiaobo，PEIDong

（a.ResearchInstituteofForestry;b.StateKeyLaboratoryofTreeGeneticsandBreeding;c.KeyLaboratoryofTreeBredingand CultivationoftheNationalForestryndGrassandAdminstration，CineseAcademyoforestryBeingo，ina）

Abstract：【Objective】Upstreamregulatory factorsof theJrAGD3 gene were screenedbyconstructingayeast libraryandusing yeastne-ybridtechologymingtorovideatheoreticalbasisforelucidatingtemolecularmechanismyichteJrGDgene mediatesrejuvenationand promotes adventitious rot formation.【Method】Tisues from rejuvenated walnut cutings undergoing adventitiousrootformationwereusedasmaterials.AyeastcDNAlibrarywasconstructedusingSMART（SwitchingmechanismAt 5′ endofRNAtemplate）techologyThepromotersequenceoftheJrAGD3genewasusedasabaittoscrenforupstreamregulatory factors via the yeast one-hybrid （YlH） system.【Result】The promoter region of JrAGD3 contains multiple types of cis-acting elements，cteisaoslstpod regulation.The constructedcDNAlibrary hadacapacity of 3×106CFU/mL ，witha total clone count of 1.20×107CFU anaverage insert fragment length of approximately1 ooo bp，and a recombination rate of 100% .Through yeast one-hybrid screening，19 potential upstreamregulatoryfactorsofJrAGD3wereidentifed，including twotranscriptionfactors：WOX4andWRKY11.【Concsio】his studysuccessfullyonstructedaigqualityyeastlibraryfortheejuvenatio-iducedadventitiousootfomationprocessalnut cutings，providingavaluableresourceforscreeinginteractingproteinsusingtheyeastone-hybridsystem.UsingtheJrAGD3gene promoterequeeasbitWOX4ndasitictosredefdpeliailyealingeotetialol regulatorynetworkofJrAGD3.Thesefindingsnotonlyenrichedthemolecularunderstandingofadventitiousotformationinwalnut butalso providednewisightsforoptimizingcutingpropagationsystemsandimprovingtheconalpropagationefcencyofwalutand otherimportant economic forest tree species.

Keywords：walnut;JrAGD3 gene;cDNA library;yeast one-hybrid; transcription factor

核桃Juglansregia是世界范围内重要的坚果和木本油料树种之一，具有较高的营养、经济和生态价值。（剩余14509字）