野蔷薇组培快繁和高效瞬时表达体系的建立  

Tissue Culture,Rapid Propagation and Efficient Transient Expression Systems of Rosa multiflora

作　　者：曹雪敏 包颖 张悦新 李瑞杰 苏健馨 张蔚[1,3] Xuemin Cao;Ying Bao;Yuexin Zhang;Ruijie Li;Jianxin Su;Wei Zhang(National Key Laboratory for Germplasm Innovation&Utilization of Horticultural Crops,College of Horticulture&Forestry Sciences of Huazhong Agricultural University,Wuhan 430070,China;Hebei Key Laboratory of Plant Biotechnology Research and Application,Department of Life Science,Tangshan Normal University,Tangshan 063000,China;Institute of Flowers Research,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学园艺林学学院,果蔬作物种质创新与利用全国重点实验室,武汉430070 [2]唐山师范学院生命科学系,河北省植物生物技术研究与应用重点实验室,唐山063000 [3]华中农业大学花卉研究所,武汉430070

出　　处：《植物学报》2025年第2期235-245,共11页Chinese Bulletin of Botany

基　　金：国家自然科学基金(No.32072617);河北省自然科学基金(No.C2021105002);河北省现代农业产业技术体系建设专项(No.HBCT2024200405)。

摘　　要：以野蔷薇(Rosa multiflora)当年生带芽茎段为试材,建立了其组培快繁体系。结果表明,最佳外植体是带腋芽茎段,外植体最佳消毒方法是先用75%乙醇浸泡30秒,再用10%次氯酸钠溶液浸泡20分钟,成活率可达96%;带芽茎段萌芽最佳诱导培养基为MS+1.0 mg·L^(-1)6-BA+0.01 mg·L^(-1)NAA+0.1 mg·L^(-1)GA_(3),培养30天,萌芽率可达98%;无菌再生苗增殖最佳基础培养基为WPM,增殖系数为2.87;无菌再生苗生根最佳培养基为1/2MS+1.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA,生根率可达93%;无菌再生苗移栽成活率达98%。在此基础上,以野蔷薇无菌再生苗为受体,建立了野蔷薇瞬时表达体系。结果表明,瞬时表达最佳转化条件是菌液OD600为0.8,负压为–0.10 MPa,真空抽吸2次,每次15分钟,瞬时表达效率可达96%。研究结果为建立野蔷薇再生及遗传转化体系奠定了基础,并为蔷薇属植物基因功能研究提供技术支持。A rapid propagation system via tissue culture for Rosa multiflora was established using the stem segments with buds of the current-year as the experimental material.The results showed that the best explants were stem segments with axillary buds.The best disinfection method was to soak the explants in 75%ethanol for 30 seconds,and then soak them in 10%sodium hypochlorite solution for 20 minutes.The survival rate can reach 96%.The optimal bud-induction medium was MS+1.0 mg·L^(-1)6-BA+0.01 mg·L^(-1)NAA+0.1 mg·L^(-1)GA_(3).The budding rate can reach 98%after 30 days of cultivation.WPM was the best basal medium for the proliferation of sterile regenerated plantlets,and the proliferation coefficient was 2.87.The best medium for rooting was 1/2MS+1.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA,and the rooting rate can reach 93%.The transplanting survival rate of sterile regenerated plantlets was 98%.On this basis,the transient expression system of R.multiflora was established.The results showed that the optimal transformation conditions for transient expression were OD600 of 0.8 for the bacterium culture medium,vacuum negative pressure of–0.10 MPa and vacuum suction twice for 15 minutes each time.The transient expression efficiency can reach 96%.The results of this study laid a foundation for the establishment of regeneration and genetic transformation system of R.multiflora,and also provided technical support for studying on the gene function of Rosa plants.

关 键 词：野蔷薇 快繁 无菌再生苗 真空渗透 瞬时表达 

分 类 号：S685.11[农业科学—观赏园艺]