牡丹组织培养过程中抑菌剂的筛选及内生细菌的鉴定

Screening of Bacteriostatic Agents and Identification of Endophytic Bacteria in Tissue Culture of Paeonia suffruticosa

为研究不同抑菌剂对牡丹内生菌生长的影响,本研究以牡丹品种‘太平红’(Paeonia suffruticosa‘Taipinghong’)试管苗为材料,分离纯化出4种内生菌(PS01,PS02,PS03和PS04)并进行鉴定。结果表明,培养20 d后,培养基添加50 mg/L氯霉素对白色菌类(PS01)抑制效果较好,污染率仅为25%;添加1.5 g/L山梨酸钾对黄色菌类(PS02)抑制效果较好,未出现污染;添加300 mg/L硫酸链霉素对红色菌类(PS03)抑制效果较好,污染率仅为2.78%;但6种抑菌剂对透明开花内生菌类(PS04)抑制效果均不佳,培养20 d后,污染率均大于70%。测序结果表明,PS01、PS02、PS03和PS04分别为农杆菌属、新鞘脂菌属、甲基杆菌属和阿菲波菌属。本研究为降低牡丹组培污染系数和提高牡丹试管苗的生根率提供理论基础和技术依据。

In order to study the effects of different bacteriostatic agents on the growth of peony endophytes,this study used the test tube seedlings of Paeonia suffruticosa’Taipinghong’as materials to isolate and purify 4 kinds of endophytes(PS01,PS02,PS03 and PS04)and perform identification.The results showed that after 20 d of culture,adding 50 mg/L chloramphenicol to the medium had a better inhibitory effect on white fungi(PS01),and the pollution rate was only 25%.Adding 1.5 g/L potassium sorbate to yellow fungi(PS02),the inhibitory effect was good,and no pollution occurs.The addition of 300 mg/L streptomycin sulfate had a good inhibitory effect on red fungi(PS03),the pollution rate was only 2.78%.But the 6 kinds of bacteriostatic agents can affect transparent flowering endophytic fungi(PS04),the inhibitory effect was not good,and the pollution rate was greater than 70%after 20 d of culture.The sequencing results showed,PS01,PS02,PS03,and PS04 are Agrobacterium,Neosphingomyce,Methylobacterium and Afipia.This research aims to provide a certain theoretical basis and technical basis for reducing the pollution coefficient of peony tissue culture and increasing the rooting rate of peony test-tube plantlets.