枯草芽孢杆菌L1-21在柑橘上的定殖能力及其防治绿霉病效果  

Colonization ability of Bacillus subtilis L1-21 in Citrus and its control effect on green mold

作　　者：李健 何鹏飞[1] 李咏梅 吴毅歆[2] 何鹏搏 孔宝华[1] 李兴玉 MUNIR Shahzad 何月秋[1] LI Jian;HE Pengfei;LI Yongmei;WU Yixin;HE Pengbo;KONG Baohua;LI Xingyu;MUNIR Shahzad;HE Yueqiu(College of Plant Protection,Yunnan Agricultural University,Kunming 650201,China;College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学植物保护学院,昆明650201 [2]云南农业大学农学与生物技术学院,昆明650201

出　　处：《华中农业大学学报》2021年第5期31-36,共6页Journal of Huazhong Agricultural University

基　　金：国家重点研发计划项目(2018YFD0201508)。

摘　　要：为明确枯草芽孢杆菌L1-21在柑橘叶片、果实内的定殖能力及其防治果实绿霉病的效果,通过向柑橘叶片喷施10^(6)cfu/mL的L1-21菌株的绿色荧光标记菌(L1-21-GFP)发酵液,测定其在叶片内的定殖数量及传导能力;采用10^(8)cfu/mL的菌液喷施和浸泡处理柑橘果实,检测其在果实内的定殖情况,用注射法接种Penicillium digitatum孢子并计算发病率,测定其对绿霉病的防控效果。结果显示,处理后10 min,L1-21-GFP在叶表及叶肉中的定殖量分别达到1.36×10^(3)和8.08×10^(2)cfu/cm^(2),并呈现持续增长的趋势。处理后1 h,L1-21-GFP向下传导到叶表的菌含量为1.51×10^(2)cfu/cm^(2),并稳定存在于叶表中;处理后2 h,传导到叶肉的菌含量为5.67 cfu/cm^(2),逐渐增加到30 h时的1.15×10^(3)cfu/cm^(2)。L1-21-GFP能在果实内定殖,浸泡1 h果肉和果皮中的定殖量分别为9.51×10^(3)和3.51×10^(4)cfu/g,但在不同柑橘品种果实间定殖能力有显著性差异,椪柑中定殖量最高。浸泡30 min后,L1-21对果实绿霉病防效第3天为100%,第5天为80.36%。以上结果表明,枯草芽孢杆菌L1-21能高效定殖于柑橘叶片及果实内,可以利用该菌株防治柑橘病害。In order to determine the colonization ability of Bacillus subtilis L1-21 in Citrus leaves and fruits and its control effect on fruit green mold,the colonization dynamics and downward-conduction of B.subtilis in citrus leaves were investigated by foliar spraying with 10^(6)cfu/mL green fluorescent labelled strain(L1-21-GFP),colonization in citrus fruits was checked through spraying and soaking with 10^(8)cfu/mL L1-21-GFP,and control effect on green mold was evaluated by spore injection with Penicillium digitatum and incidence investigation.After treatment for 10 min,the colonization of B.subtilis L1-21-GFP in leaf surface and mesophyll reached 1.36×10^(3)and 8.08×10^(2)cfu/cm^(2),respectively,and displayed a continuous increasing trend.At 1 h after treatment,the GFP-tagged endophyte was stable in leaf surface with 1.51×10^(2)cfu/cm^(2).The bacterial concentration reached 5.67 cfu/cm^(2)in mesophyll at 2 h after treatment,which gradually increased to 1.15×10^(3)cfu/cm^(2)after 30 h.Endophyte L1-21-GFP could easily colonize in fruits,and bacterial concentration in pulp and peel after 1 h was 9.51×10^(3)cfu/g and 3.51×10^(4)cfu/g,respectively.However,there was significant difference in the colonization ability in fruits among different citrus varieties,and Citrus reticulata cv Ponkan had the highest colonization.In addition,after soaking for 30 min,the control effect of L1-21 on fruit green mood was 100%on the third day,and 80.36%on the fifth day.Taken together,B.subtilis L1-21 could colonize in citrus leaves and fruits effectively,providing a theoretical basis for further application of this strain to control citrus diseases.

关 键 词：枯草芽孢杆菌 内生菌 定殖 防效 绿霉病 柑橘 

分 类 号：Q939.124[生物学—微生物学] S435.661.1[农业科学—农业昆虫与害虫防治]