余婷婷，袁莲莲，李莹，焦裕冰，申莉莉，王凤龙，黄坤，王勇，李斌，章松柏，杨金广

摘  要：為挖掘具有纳米硒化能力的生防菌株，开发烟草病毒病的绿色防治材料，从烟草病毒病重病田块健株根际土壤中筛选获得到一株荧光假单胞菌Pseudomonas fluorescens KBD-1，对其进行了纳米硒化，采用Real-time PCR和Western blot测定了两者对烟草常见病毒基因表达的影响，并采用接种法测定了其对烟株相应病毒病的防治作用。结果表明，纳米硒化后的KBD-1菌体外部存在高密度电子颗粒，在X射线11.22 keV处出现硒的特征吸收峰。纳米硒化后的P. fluorescens KBD-1菌液浓度在5×105 cfu/mL（单质硒浓度0.25 mg/L）时，对烟草花叶病毒（TMV）、马铃薯Y病毒（PVY）、黄瓜花叶病毒（CMV）、番茄斑萎病毒（TSWV）4种病毒病的防治效果均在88.0%以上，对CMV防治效果最高达91.4%，该浓度处理对烟株促生效果显著。荧光假单胞菌KBD-1可成功将亚硒酸钠还原生成纳米硒，并能增强原始菌株的抗烟草病毒活性和促生效果。

关键词：纳米硒;荧光假单胞菌;烟草病毒病;病毒抑制;促生作用

Inhibition of Tobacco Virus Diseases by Nano-selenated Pseudomonas fluorescens KBD-1

YU Tingting1，2， YUAN Lianlian2， LI Ying2， JIAO Yubing2， SHEN Lili2， WANG Fenglong2， HUANG Kun3，

WANG Yong4， LI Bin5， ZHANG Songbai1*， YANG Jinguang2*

（1. College of Agronomy， Yangtze University， Jingzhou， Hubei 434025， China; 2. Tobacco Research Institute， Chinese Academy of Agricultural Sciences， Qingdao 266101， China; 3. Honghe Company of Yunnan Tobacco Company， Kunming 650000， China; 4. Liangshan Company of Sichuan Provincial Tobacco Company， Xichang， Sichuan 615000， China; 5. Sichuan Tobacco Company， Chengdu 610000， China）

Abstract： In order to explore biocontrol strains with nano-selenized ability and develop green antiviral materials to prevent tobacco virus diseases， Pseudomonas fluorescens KBD-1 was identified from the rhizosphere soil of healthy plants in the field with serious tobacco virus diseases. The nano-selenized activity of P. fluorescens KBD-1 was investigated in this study， and its inhibitions against four main tobacco viruses were determined by using the methods including Real-time PCR and Western blot under the treatments of original strain and nano-selenized strain. The results showed that there were high density electron particles outside the nano-selenized KBD-1 and the characteristic absorption peak of selenium appeared at 11.22 keV X-ray. When the concentration of P. fluorescens KBD-1 bacterial fluid after nano-selenization was 5×105 cfu/mL （0.25 mg/L selenium）， the control effects against Tobacco mosaic virus （TMV）， Potato virus Y （PVY）， Cucumber mosaic virus （CMV） and Tomato spotted wilt virus （TSWV） were all above 88.0%， and the highest control effect against CMV was 91.4%. The treatment at this concentration also had significant growth-promoting effect on tobacco plants. This study showed that P. fluorescens KBD-1 successfully reduced sodium selenite to produce nano-selenium， and enhanced the antiviral activity and growth-promoting effect of the original strain.

Keywords： nanometer selenium; Pseudomonas fluorescens; tobacco virus diseases; virus inhibition; role in the growth

病毒病是烟草普遍发生的一类侵染性病害[1]，其中造成严重经济损失的种类主要包括烟草花叶病毒病（TMV）、马铃薯Y病毒病（PVY）、黄瓜花叶病毒病（CMV）、番茄斑萎病毒病（TSWV）等[2]。研究表明TMV、CMV、PVY复合侵染现象普遍，产量和品质受损严重[3-4]，防治主要以早期预防为主，但收效甚微，迄今还没有效果很好的单一防治措施，病害发生后有效治疗药剂更是缺乏[5]。

基金项目：中国烟草总公司贵州省公司遵义市公司项目（201921）;中国烟草总公司四川省公司科技项目（SCYC202008）;中国烟草总公司项目硒是植物必需的微量元素之一[6]，然而，如果硒含量过高，会导致严重健康问题。……