辣椒细菌性果实条斑病菌生物学特性及抑菌药剂筛选

研究辣椒细菌性果实条斑病菌的生物学特性,对病原菌进行室内药剂筛选,旨在为田间病害规律研究和病害防治提供依据。采用紫外分光光度计测定病原菌在不同温度、pH、NaCl浓度下的生长速率。采用滤纸片抑菌圈法对病原菌进行室内药剂筛选试验。结果表明,辣椒细菌性果实条斑病菌最适生长温度为28℃,最适pH 7.0,病原菌的耐盐性为12%。18种供试药剂在药剂推荐使用剂量范围内,四霉素、乙蒜素、77%志信、氯溴异氰尿酸4种药剂对病原菌有抑菌效果,对这4种药剂进行毒力测定,结果表明四霉素的毒力较强,EC₅₀值为11.75 mg/L,毒力强于其他3种试剂,四霉素可作为田间防治药剂的首选。     

马尾松叶表微生物多样性

为发掘对林木生长发育有利的优良微生物资源,并筛选适合叶表微生物的收集方法,以马尾松针叶为试验材料,分别用悬摇法和超声波法收集马尾松叶表微生物,用扩增子高通量测序技术、MUSCLE和Qiime软件研究马尾松叶表微生物的多样性。结果表明:扫描电镜观测结果显示,马尾松针叶表面定殖有大量微生物,包括真菌(菌丝及孢子)和细菌。扩增子高通量测序结果表明,马尾松叶表微生物物种丰富,包含细菌运算分类单位(OTUs)490个,真菌OTUs 1273个。马尾松叶表细菌以未分类的蓝细菌属(unidentified_Cyanobacteria)(36.53%)、未分类的拜叶林克氏菌属(unidentified_Beijerinckia)(28.60%)、鞘氨醇单胞菌属(Sphingomonas)(2.35%)为优势属;叶表真菌以枝孢属(Cladosporium)(2.45%)、拟盘多毛孢属(Pestalotiopsis)(0.92%)、无头孢菌属(Capnobotryella)(0.91%)为优势属。针对叶表细菌多样性的研究表明,悬摇法和超声波法均有较高的物种检出度;在叶表真菌多样性的研究中超声波法优于悬摇法,但超声波法样品间数据变异性较大,测定结果不稳定。     

巴音温都尔沙漠典型植物群落土壤物理性质研究

巴音温都尔沙漠是我国北方干旱气候带典型的环境敏感区和生态脆弱区,生态建设和植被恢复对巴音温都尔沙漠生态系统服务功能极为重要。本研究以巴音温都尔沙漠梭梭、唐古特白刺、球果白刺、盐爪爪、膜果麻黄群落土壤为试验对象,对不同植物群落类型土壤物理性状和持水能力进行了研究。结果表明:5种群落中土壤孔隙度、土壤毛管孔隙度与土壤非毛管孔隙度最大值均出现在盐爪爪群落,其土壤通透性最好,具备良好的通气透水能力;膜果麻黄群落土壤最为紧实,通气透水能力最差。5种群落中,盐爪爪群落土壤自然含水率最高,膜果麻黄群落最低。盐爪爪群落土壤持水量最高,麻黄群落土壤持水量最低。盐爪爪群落土壤具有良好的保水能力,膜果麻黄群落土壤保水能力处于偏低水平。     

Dynamics of Cotton Rhizosphere Bacterial Community Structure in Cotton Continuous Cropping Field Soil

[Objective] The aim of this study is to understand the dynamics of cotton rhizosphere bacterial community structure in cotton continuous cropping field soil. [Method] 16S rDNA genes were sequenced by high-throughput sequencing technology to determine the community structure of cotton rhizosphere bacteria in different developmental stages using an upland cotton cultivar (Gossypium hirsutum cv. TM-1). [Result] Four dominant phyla were found in the cotton rhizosphere bacterial community including Proteobacteria, Acidobacteria, Planctomycetes, and Bacteroidetes. The four dominant phyla and Firmicutes were largely influenced by cotton root. The relative abundances of Acidobacteria, Planctomycetes, Bacteroidetes were promoted, and Proteobacteria and Firmicutes were inhibited, by cotton root. There were significant differences in community structure, but not species richness or α-diversity among different developmental stages of the cotton rhizosphere bacterial community; the differences between the flowering stage and the budding stage were greater than the differences between the budding stage and the seedling stage. The diversity of the rhizosphere bacterial community in cotton continuous cropping field soil was significantly higher than that of the bulk soil; the β-diversity values of both the rhizosphere and bulk soil bacterial communities were highest in the flowering stage. [Conclusion] The structure and dynamics of the cotton rhizosphere bacterial community in cotton continuous cropping field soil was defined by high-throughput sequencing. The effect of cotton on the rhizosphere bacterial community structure was most significant in the flowering stage.     

星油藤青枯病病原菌鉴定

 星油藤(Plukenetia volubilis L.) 是一种重要的藤本油料植物,在我国华南地区广泛种植。青枯病是近两年在海南星油藤种植区发生的新病害,为探究星油藤青枯病菌的基本特性及种下分化情况,本研究对分离的6株代表菌株进行了相关分析。细菌学鉴定及致病性测定结果表明,该病害是由类茄科雷尔氏菌(Ralstonia pseudosolanacearum)侵染引起。同时,从传统分类及分子生物学不同层面分析了星油藤青枯病菌的遗传分化情况。生理小种及生化变种的测试结果表明,星油藤青枯病菌属于1号生理小种和生化变种Ⅲ;16S rDNA和egl基因部分序列聚类分析显示,星油藤青枯病菌属类茄科雷尔氏菌演化型Ⅰ即亚洲分支菌株,序列变种34。     

High-throughput screening reveals small molecule modulators inhibitory to Acidovorax citrulli

Acidovorax citrulli is a seed-transmitted gram-negative bacterium that can cause substantial economic yield loss in watermelon and melon production worldwide. Four small-molecule libraries containing 4,952 compounds were selected for high-throughput screening against A. citrulli wild-type strain Xu3-14 by evaluation of growth inhibition. One hundred and twenty-seven molecules (2.5% hit rate) were identified as bactericidal or bacteriostatic against A. citrulli at 100 μM. Secondary screens indicated that 27 candidate compounds were more effective against A. citrulli Group II strains than Group I strains (classified using repetitive element PCR). Several compounds were inhibitory to other pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Xanthomonas campestris pv. campestris, but did not affect the growth of plant beneficial bacteria Pseudomonas fluorescens and Bacillus subtilis. More than half of the compounds did not inhibit germination of Arabidopsis or watermelon seeds. The effect of small molecules on A. citrulli seed-to-seedling transmission was evaluated by applying each compound to inoculated watermelon seeds and assessing seedling infection. Nine compounds were chosen for further investigation based on their reduction of percentage seedling infection and compiling scores on their specificity, sensitivity, and phytotoxicity obtained in the secondary screens. The five best compounds were selected (thiamphenicol, nadifloxacin, pipemidic acid, ciclopirox, and zinc pyrithione) for greenhouse tests and were found to effectively reduce the seed-to-seedling transmission of A. citrulli in both artificially and naturally infested seeds. These top five compounds provide a basis for future development of an A. citrulli-specific bactericide.     

亮氨酸对早期断奶湖羊瘤胃发育及细菌菌群的影响

本试验旨在研究日粮中添加亮氨酸对早期断奶湖羊瘤胃发育及瘤胃细菌菌群组成的影响。选用36只5日龄的湖羊羔羊,依据体重相近原则随机分成对照组(C,0 g/d)、亮氨酸低剂量添加组(L,0.66 g/d)和高剂量添加组(H,1.33 g/d),每组4个重复,每个重复3只羔羊。于30日龄断奶当天每组每个重复取1只羔羊屠宰以获得瘤胃组织及内容物样品(n=4)。结果表明:H组瘤胃乳头的长度较C组显著增加(线性,P<0.01);添加亮氨酸可使瘤胃挥发性脂肪酸水平显著提高(二次,P=0.05),但并未改变乙酸、丙酸和丁酸的摩尔比例(P>0.05);与C组相比,L与H组的瘤胃氨态氮浓度均显著降低(线性,P<0.05;二次,P>0.05);L组羔羊瘤胃内容物中微生物蛋白含量显著高于C组(二次,P=0.05);亮氨酸的添加并未改变瘤胃细菌物种丰富度和多样性指数,但对菌群组成有一定影响;L组瘤胃内最多的是厚壁菌门(49.5%);H组拟杆菌门(51.4%)显著高L和C组(二次,P=0.05);H组的普雷沃氏菌属最多(线性,P>0.05;二次,P>0.05),而L组的巨型球菌属最多(二次,P <0.01);与C组相比,L组的双歧杆菌属(二次,P<0.05)和光岗菌属显著增加(二次,P<0.05)。由此可见,添加亮氨酸可促进羔羊瘤胃发育,改变瘤胃细菌菌群组成。     

白叶枯病菌效应子XopN在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用

【目的】黄单胞菌细胞外泌蛋白质(Xanthomonas outer proteins, Xops)是植物病原黄单胞菌高度保守的毒性效应子或毒性辅助组分,通过细菌Ⅲ型分泌系统分泌到细菌细胞外部,然后转入植物细胞而发挥病理作用。水稻黄单胞菌水稻致病变种即水稻白叶枯病菌的标准菌株PXO99^A分泌的XopN是一种毒性效应子,通过影响寄主免疫反应而使水稻发病。但是,XopN对病菌毒性的影响是否因水稻品种的不同而异,还有待研究。【方法】利用同源双交换技术,先敲除了PXO99^A的XopN基因,获得了∆XopN突变体,又经过遗传互补,得到了回补菌株∆XopN/XopN。通过营养肉汤液体培养,测定了XopN对病菌繁殖能力的影响;根据文献选用14个水稻品种,通过接种实验,测定了PXO99^A对这些品种的毒性与XopN敲除或回补的影响;在XopN发挥毒性作用的水稻品种上,测定了隐性抗病基因OsSWEET11/xa13与显性感病基因OsSWEET11/Xa13受病菌侵染而表达的情况,分析了XopN敲除或回补的影响。【结果】在营养肉汤液体培养过程中,病菌突变体菌株∆XopN的繁殖速度明显低于野生型PXO99^A。PXO99^A、∆XopN和∆XopN/XopN接种水稻后,根据其在水稻叶片组织内的繁殖量及随后产生的白叶枯病症状的严重程度,将供试的14个水稻品种分为两种情况。一是感病程度与病菌XopN是否敲除或回补无关,这有10个水稻品种（IRBB1、IRBB3、IRBB8、IRBB10、IRBB14、IR24、IRBB203、IRBB204、IRBB205和IRBB211）,PXO99^A、∆XopN和∆XopN/XopN对它们的毒性无明显差别。二是XopN对病菌毒性发挥作用的水稻品种,包括高度感病的3个品种（IRBB208、Asominori和日本晴）和低感品种IRBB13。与PXO99^A或∆XopN/XopN相比,∆XopN对这4个水稻品种的毒性大为降低。在IRBB13上,病菌侵染对隐性抗病基因OsSWEET11/xa13在叶片内的表达发生抑制作用,这一效应与XopN的毒性功能相关。相反,日本晴显性感病基因OsSWEET11/Xa13却受病菌侵染的诱导,在叶片内的表达水平大幅度提高。IRBB208和Asominori携带OsSWEET11/Xa13同源基因,该同源基因在叶片内的表达水平也因病菌侵染而大幅提高。在这4个水稻品种上,PXO99^A和∆XopN/XopN能够诱导OsSWEET11/Xa13或其同源基因表达,但∆XopN无此作用。另外,XopN对病菌在非寄主植物烟草上诱发过敏反应有量变贡献,相比PXO99^A或∆XopN/XopN,∆XopN引起过敏反应的程度有所降低。【结论】XopN是一个有限广谱性效应子,在拥有OsSWEET11同源基因的水稻品种上发挥毒性作用。XopN也是病菌繁殖所需要的,对病菌在非寄主植物上诱导过敏反应有一定贡献。