红松胚性愈伤组织瞬时转化体系的建立
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中图分类号：S722 文献标志码：A文章编号：1000-2006（2026）01-0012-11
Abstract：【Objective】This studyaims tooptimize atransient transformation system for Pinus koraiensisembryogenic callus mediated by Agrobacterium tumefaciens.【Method】 Using the ten-day cultured embryonic callus of P .koraiensis as the receptor material，transient transformationwascariedoutwiththe pCambial3OO vectorcarrying RUBY.Theoptimal concentrationof infectionsolution，infection time，co-cultivationtime，andrecoveryculturetime were screenedbasedon the expression level of RUBY. The vectors promoterPkDof5.6 -EGFP，promoter PkSHR -RUBY，35S-PkDof5.6-EGFP，35SPkSHR-EGFP，and 35S-PkSHR-RUBY were respectively transfected into the embryonic callus of P .koraiensistoverify the conditions of the transient transformationsystem.【Result】After transient transformation，RUBY expresion was significant. When the concentration of the inoculum（ A600 ）reached O.4，with aninfection time of ten minutes and cocultivation forone day withoutrecovery culture period，thecopy numberof the RUBYfragment was atits highest.Genes andpromotersspecificvalidationofthequantitativeandsemi-quantitativeresultsof transienttransformationindicates that this condition can achieve efficient transformation of mature callus from P koraiensis.【Conclusion】Thisstudy successfully establishedasimple，rapid，eficientand low-cost Agrobacterium-mediated transienttransformationsystem for embryogenic callus of P .koraiensis，and optimized the main factors of this system.The best transformation conditions were found to be：an infection bacterial suspension concentration （ A600 ）of 0.4，infection for ten minutes，co-cultivation for oneday，with no recoveryculture period.Thissystem isof great significance forlarge-scale genefunctional analysis of the somatic embryogenesis mechanism in P koraiensis and accelerates the breedingand large-scale propagation of high-quality P .koraiensisvarieties.
Keywords：Pinus koraiensis；transient transformation；somatic embryogenesis；molecular breeding
红松（Pinuskoraiensis）为裸子植物门松科松属常绿乔木，主要分布于中国东北的小兴安岭到长白山一带，也有少数生长在俄罗斯南部、日本以及朝鲜等地。（剩余20243字）