生防菌Hhs.015对苹果枝条皮层内生细菌区系的影响  

Effect of biocontrol strain Hhs.015 on endophytic bacterial flora of apple trees

作　　者：颜霞[1,2] 张亚男[1,2] 刘聪[1,2] 赵玲云[1,2] 郭红梅[1,2] 李燕芳[1,2] 黄丽丽[3,2] 

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100 [2]旱区作物逆境生物学国家重点实验室,陕西杨凌712100 [3]西北农林科技大学植物保护学院,陕西杨凌712100

出　　处：《果树学报》2017年第9期1170-1177,共8页Journal of Fruit Science

基　　金：国家自然科学基金(31101476;31171796);陕西省科学技术研究发展计划项目(2013K01-45);杨凌示范区科技计划项目(2014NY-41)

摘　　要：【目的】用生防菌Hhs.015发酵液处理苹果发病枝条,研究其对树体微生态的影响。【方法】采用改良CTAB法提取样本中的总DNA,对细菌群落的16S r DNA的V4+V5区进行高通量测序,研究发病枝条涂抹Hhs.015菌悬液30 d后树皮内细菌区系组成变化,分析其相对丰度以及多样性指标。【结果】在属水平上,生防菌处理样本与对照样本中的细菌种类显现出明显差异,生防菌处理组Gluconobacter丰度明显升高,放线菌门菌株丰度高于对照组,Burkholderia和Luteibacter等可能与致病相关的菌属丰度大幅下降。刮去外皮后取样,糖丝菌属(Saccharothrix)丰度仍处于较高水平。【结论】Hhs.015能够在苹果树皮内定殖,并影响苹果枝条皮层细菌区系。[Objective]Apple Valsa canker is caused by Valsa mall Miyabe et Yamada which decomposes the apple tree bark and even the xylem, it is a serious fungal disease which brings significant damage to apple productivity in China. Biological control can be used on the plant surface or in vivo, regulating the balance and proportion of plant indigenous microflora and promoting plant growth to achieve the goal of prevention and stimulation. Microorganisms are an important part of the plant microflora, which are involved in the happenings, developing and declining of some life activities of the host in the level of physiology, pathology, pharmacology and many other life activities. Normal microbial flora is an important part of healthy plants. Saccharothrix yanglingensis Hhs.015 is effective in controlling apple Valsa canker both indoors and in field trials. This experiment aimed to explore the effects of Hhs.015 on the micro-ecology of apple tree twigs, which is helpful to evaluate its biosecurity. [Methods] A field trial was carried out in a ＇ Fuji＇ apple orchard in Yangling, Shaanxi. 10 new sick spots on 10 randomly selected apple tree trunks were divided into two groups. The control was treated with the biocontrol agent Saccharothrix yanglingensis Hhs.015 broth, the contrast group was treated with water. Every 5 d the scars were painted for a total of 4 times. Then 30 d after the last time, the tested samples at the boundary between diseased and healthy areas of twigs were collected. DNA was extracted using the modified CTAB method. The V4＋V5 region of 16S rDNA was amplified with a specific primer and sequenced by high throughput sequencing. From raw pyrosequencing reads, primer and tag sequences were removed, the original tags were jointed by FLASH. After filtration treatment and mosaic deletion, the final valid data were obtained. These sequences were clustered at 97% similarity using Uparse. To study the phylogenetic relationship between OTUs, these sequences were multiple sequence aligned and a tree built to pr

关 键 词：苹果 生防菌Hhs.015 苹果树腐烂病 细菌区系 高通量分析 

分 类 号：S661.1[农业科学—果树学]