梅花花瓣瞬时转化体系构建

doi:10.16420/j.issn.0513-353x.2024-0448

摘要/Abstract

摘要： 以梅花（Prunus mume）花瓣为受体，β–葡萄糖苷酸酶基因（GUS）为报告基因，通过单因素、正交试验对影响梅花花瓣瞬时转化效率的7个主要因素进行了分析。对不同侵染方式、侵染时间、发育阶段和基因型进行单因素试验。结果表明，整花真空法、侵染时间为15 min、发育阶段为初开期和‘雪梅’基因型的GUS瞬时表达率最高。三因素水平正交试验结果表明，对梅花花瓣GUS瞬时表达的影响大小依次为共培养时间、农杆菌类型、侵染浓度（OD₆₀₀值），其中，以农杆菌EHA105、OD₆₀₀值为0.6、共培养5 d的花瓣GUS瞬时表达率最高（32.67%），并以梅花花青素合成基因（PmANS）为目的基因对筛选出的最优组合进行了成功验证。

关键词: 梅花, 花瓣, 农杆菌介导法, 瞬时转化, GUS

Abstract: This study takes the petals of Prunus mume as the receptor and GUS gene as the reporter gene. The influences of 7 main factors on the transient transformation efficiency of Prunus mume petals were conducted through an orthogonal design and single factor analysis method. The effects of infection mode，infection time，development stage and genotype on the transient transformation efficiency of Prunus mume petals were investigated by single factor experiment. The results showed that the whole flower vacuum method，infestation time of 15 min，development stage of initial blooming，and‘XueMei’ genotype had the highest transient expression rate of GUS gene，respectively. The orthogonal experiment was further designed to explore the effect of the combination of Agrobacterium strain type，OD₆₀₀ value，Coculture time on the transient transformation efficiency of Prunus mume petals. The results showed that the three factors affected GUS transient expression in the order of coculture time，Agrobacterium strains type，OD₆₀₀ value. Among them，the combination of Agrobacterium EHA105，OD₆₀₀values is 0.6 and coculture time is 5 d showed the highest GUS transient expression effect of Prunus mume petals（32.67%）. Then，the optimal combination screened was verified by applying an anthocyanin synthesis gene（PmANS）as the target gene.

Key words: Prunus mume, petal, Agrobacterium-mediated method, transient transformation, GUS

中图分类号:

S 685.17

田壮, 房晨曦, 刘雅婷, 王雨桐, 薛林蕾, 石嘉欣, 任天放, 张俊卫, 包满珠, 张杰. 梅花花瓣瞬时转化体系构建[J]. 园艺学报, 2025, 52(4): 908-920.

TIAN Zhuang, FANG Chenxi, LIU Yating, WANG Yutong, XUE Linlei, SHI Jiaxin, REN Tianfang, ZHANG Junwei, BAO Manzhu, ZHANG Jie. Construction of Transient Transformation System of Petals in Prunus mume[J]. Acta Horticulturae Sinica, 2025, 52(4): 908-920.

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