文献类型：期刊文献

中文题名：毛白杨GM107再生体系和遗传转化体系的建立

英文题名：Establishment of Regeneration and Genetic Transformation System of Elite Clone GM107 of Populus tomentosa

作者：黄赛[1] 高凯[2] 苗得雨[1] 安新民[1]

第一作者：黄赛

机构：[1]北京林业大学生物科学与技术学院,林木育种国家工程实验室,林木、花卉遗传育种教育部重点实验室,林木花卉育种生物工程国家林业和草原局重点实验室,北京100083;[2]中国林业科学研究院亚热带林业研究所,杭州311400

年份：2025

卷号：23

期号：6

起止页码：1888-1896

中文期刊名：分子植物育种

外文期刊名：Molecular Plant Breeding

收录：;北大核心:【北大核心2023】；

基金：“十四五”国家重点研发项目“林木基因编辑技术”(2021YFD2200101-3);国家自然科学基金项目(No.31570661);国家科技重大专项(No.2018ZX08021002-002-004)共同资助。

语种：中文

中文关键词：毛白杨;再生体系;遗传转化;根瘤农杆菌

外文关键词：Populus tomentosa;Regeneration system;Genetic transformation;A grobacterium tumefaciens

分类号：S792.11

摘要：本研究以毛白杨优良无性系GM107为试验材料,通过带芽茎段外植体消毒、接种生根获得无菌组培苗,利用无菌苗叶片建立了高频率的再生体系,筛选出最佳叶片分化生芽培养基为MS+TDZ 0.1 mg/L+NAA 0.1 mg/L+BA 0.5 mg/L,分化生芽率约为90%,每小圆片生芽数可达20个。利用农杆菌介导的叶盘转化法,建立了高效的遗传转化体系,并确定了毛白杨叶片不定芽诱导卡那霉素筛选浓度为100 mg/L,不定芽生根筛选浓度为50 mg/L。Cef 250 mg/L+Tim 250 mg/L+Car 250 mg/L组合可作为筛选培养基的最佳抑菌抗生素组合。在农杆菌侵染前,对GM107小圆片外植体进行7 d预培养为宜。利用已经构建的pCAMBIA2300-VvMYBA1载体,以最佳组合进行遗传转化,获得102个转化株系,通过PCR检测,获得33个阳性株系,阳性植株达到32.35%。本研究为毛白杨基因功能研究提供了高效的遗传转化平台。
In this study,using the elite clone GM107 of Populus tomentos a as material,sterile tissue culture plantlets were obtained by disinfection of cutting with buds,inoculation and rooting.A high-frequency in vitro regeneration system was established by using the leaves of tissue culture plantlet above described.The optimum medium for leaf differentiation and bud formation was MS+TDZ 0.1 mg/L+NAA 0.1 mg/L+BA 0.5 mg/L.The rate of differentiation and bud formation was about 90%,and the number of buds per small disc could reach 20.An efficient genetic transformation system was established by using Agrobacterium mediated leaf disk transformation method.The screening concentration of kanamycin for adventitious bud induction and adventitious bud rooting of P.tomentosa was 100 mg/L and 50 mg/L.Cef 250 mg/L+Tim 250 mg/L+Car 250 mg/L were selected as the optimal antibiotics for the screening medium.Before Agrobacterium-mediated genetic transformation,it is appropriate to pre culture GM107 small disc explants for 7 days.Using the constructed pCAMBIA2300-VvMYBA1 vector,102 transformed lines were obtained by genetic transformation with the best combination.Through PCR detection,33 lines were positive,and the rate of positive plants reached 32.35%.This study provides an efficient genetic transformation platform for the study of gene function of P.tomentosa.