南京林业大学学报（自然科学版）2026，Vol.50Issue(1)：12-22,11.DOI:10.12302/j.issn.1000-2006.202501010
红松胚性愈伤组织瞬时转化体系的建立
Establishment of a transient transformation system for embryogenic callus tissue of Pinus koraiensis
摘要
Abstract
[Objective]This study aims to optimize a transient transformation system for Pinus koraiensis embryogenic callus mediated by Agrobacterium tumefaciens.[Method]Using the ten-day cultured embryonic callus of P.koraiensis as the receptor material,transient transformation was carried out with the pCambia1300 vector carrying RUBY.The optimal concentration of infection solution,infection time,co-cultivation time,and recovery culture time were screened based on the expression level of RUBY.The vectors promoterPkDof5.6-EGFP,promoterPkSHR-RUBY,35S-PkDof5.6-EGFP,35S-PkSHR-EGFP,and 35S-PkSHR-RUBY were respectively transfected into the embryonic callus of P.koraiensis to verify the conditions of the transient transformation system.[Result]After transient transformation,RUBY expression was significant.When the concentration of the inoculum(A600)reached 0.4,with an infection time of ten minutes and co-cultivation for one day without recovery culture period,the copy number of the RUBY fragment was at its highest.Genes and promoters specific validation of the quantitative and semi-quantitative results of transient transformation indicates that this condition can achieve efficient transformation of mature callus from P.koraiensis.[Conclusion]This study successfully established a simple,rapid,efficient and low-cost Agrobacterium-mediated transient transformation system 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 one day,with no recovery culture period.This system is of great significance for large-scale gene functional analysis of the somatic embryogenesis mechanism in P.koraiensis and accelerates the breeding and large-scale propagation of high-quality P.koraiensis varieties.关键词
红松/瞬时转化/体胚发生/分子育种Key words
Pinus koraiensis/transient transformation/somatic embryogenesis/molecular breeding分类
农业科技引用本文复制引用
贺欣,徐秀月,亢溢敏,杨玲..红松胚性愈伤组织瞬时转化体系的建立[J].南京林业大学学报（自然科学版）,2026,50(1):12-22,11.基金项目
国家重点研发计划(2023YFD2200103). （2023YFD2200103）
