甜瓜细菌性果斑病生防菌株BW-6的筛选和鉴定

甜瓜细菌性果斑病是由西瓜嗜酸菌（Acidovorax citrulliSchaad et al.）引起的一种毁灭性病害。本研究以西瓜嗜酸菌为指示菌,采集97个土样,采用土壤稀释平板法和对峙培养进行拮抗菌的分离和筛选,获得19株细菌可以抑制西瓜嗜酸菌生长。通过离体叶片、盆栽试验进行生测,BW-6菌株对甜瓜果斑病的生防效果最好,且比较稳定,防效达80.3%。在甜瓜叶面进行室外定殖试验表明,BW-6菌株可以在甜瓜叶面上大量定殖,但随着时间的推移菌数逐渐减少。根据菌株BW-6的形态特征、生理生化特性以及16S rDNA序列分析综合考察,鉴定其为枯草芽胞杆菌(Bacillus subtilis Cohn)。     

瓜类细菌性果斑病研究进展

瓜类细菌性果斑病是发生在甜瓜、西瓜等葫芦科植物上的一种严重的世界性病害,此病是典型的种传细菌性病害,病原为嗜酸菌属西瓜种(Acidovorax citrulli)。本文围绕瓜类细菌性果斑病菌的分离检测、致病机理、遗传多样性及防治等方面的研究进展作一概述,阐明了瓜类细菌性果斑病的研究现状。     

Diversity of cultivable endophytic bacteria associated with halophytes in Xinjiang of China and their plant beneficial traits

Endophytic bacteria from halophytes have a wide range of application prospects in various fields, such as plant growth-promoting, biocontrol activity and stress resistance. The current study aimed to identify cultivable endophytic bacteria associated with halophytes grown in the salt-affected soil in Xinjiang Uygur Autonomous Region, China and to evaluate their plant beneficial traits and enzyme-producing activity. Endophytic bacteria were isolated from Reaumuria soongorica (PalL Maxim.), Artemisia carvifolia (Buch.-Ham. ex Roxb. Hort. Beng.), Peganum harmala L. and Suaeda dendroides (C. A. Mey. Moq.) by using the cultural-dependent method. Then we classified these bacteria based on the difference between their sequences of 16S rRNA (16S ribosomal RNA) gene. Results showed that the isolated bacteria from R. soongorica belonged to the genera Brucella, Bacillus and Variovorax. The bacteria from A. carvifolia belonged to the genera Micromonospora and Brucella. The bacteria from P. harmala belonged to the genera Paramesorhizobium, Bacillus and Peribacillus. The bacteria from S. dendroides belonged to the genus Bacillus. Notably, the genus Bacillus was detected in the three above plants, indicating that Bacillus is a common taxon of endophytic bacteria in halophytes. And, our results found that about 37.50% of the tested strains showed strong protease-producing activity, 6.25% of the tested strains showed strong cellulase-producing activity and 12.50% of the tested strains showed moderate lipase-producing activity. Besides, all isolated strains were positive for IAA (3-Indoleacetic acid) production, 31.25% of isolated strains exhibited a moderate phosphate solubilization activity and 50.00% of isolated strains exhibited a weak siderophore production activity. Our findings suggest that halophytes are valuable resources for identifying microbes with the ability to increase host plant growth and health in salt-affected soils.     

13种药剂种子处理对瓜类细菌性果斑病的防治效果

瓜类细菌性果斑病是西瓜、甜瓜等瓜类作物的重要细菌性病害。为筛选防控果斑病的有效药剂, 本研究比较了13种药剂浸种带菌西瓜种子不同时间对果斑病的防治效果及对种子发芽的影响;并通过“先喷药后接种”和“先接种后喷药”, 比较了13种药剂对西瓜苗期果斑病的预防和治疗效果, 同时测定了助剂8.6%聚乙二醇对13种药剂的增效作用。药剂浸种试验结果表明, 带菌种子在10%阿苯达唑750倍液、2%春雷霉素300倍液中浸种12 h, 对果斑病的防效均达到100%, 发芽率比清水浸种分别高16.20百分点和16.34百分点;杀菌剂1号200倍液处理12 h的防效较好, 为85.58%, 且促进种子萌发。苗期喷施药剂的试验结果表明, 13种药剂对果斑病的预防效果均好于治疗效果, 预防效果和治疗效果都超过60%的药剂有7种, 以10%阿苯达唑600倍液对果斑病的防效最好, 预防效果和治疗效果分别为87.58%和75.70%, 与8.6%聚乙二醇混配后分别提高至94.12%和81.31%, 增效作用显著。本研究结果为后续田间试验和大面积推广应用提供了技术支撑。     

Bacterial fruit blotch of melon: screens for disease tolerance and role of seed transmission in pathogenicity

Bacterial fruit blotch (BFB) of cucurbits, caused by Acidovorax avenae subsp. citrulli, is a serious threat to the watermelon and melon industries. To date, there are no commercial cultivars of cucurbit crops resistant to the disease. Here we assessed the level of tolerance to bacterial fruit blotch of various commercial cultivars as well as breeding and wild lines of melon, using seed-transmission assays and seedling-inoculation experiments. Selected cultivars were also tested in a greenhouse experiment with mature plants. All tested cultivars/lines were found to be susceptible to the pathogen, and most of them showed different responses (relative tolerance vs. susceptibility) in the different assays; however, some consistent trends were found: cv. ADIR339 was relatively tolerant in all tested assays, and cv. 6407 and wild lines BLB-B and EAD-B were relatively tolerant in seed-transmission assays. We also provide evidence supporting a strong correlation between the level of susceptibility of a cultivar/line and the ability of the pathogen to adhere to or penetrate the seed. To the best of our knowledge, this is the first attempt to assess melon cultivars/lines for bacterial fruit blotch response.     

厚皮甜瓜细菌性果斑病标准化综合防控技术

为有效防控厚皮甜瓜细菌性果斑病的发生,提出一套综合防控技术,通过田间开展各单项防控技术的重要性试验,建立标准化综合防控技术体系。结果表明:各单项技术可提高对细菌性果斑病的防效。与对照相比,72%农用硫酸链霉素可溶粉剂浸种可使防效提高16%,病果率降低3%;标准化喷药能使防效提高17%以上,病果率降低3.1%;整枝打杈前后进行防控能使防效提高39%以上;厚皮甜瓜整个生育期不浇水,能够减少病果的数量;在此基础上组装而成的标准化综合防控技术体系防效达89.2%以上,病果率仅为2%。     

荧光假单胞工程菌株对甜瓜细菌性果斑病的生物防治

从植物病理学研究室保存的30余株染色体整合2,4-二乙酰基间苯三酚合成基因簇的荧光假单胞(Pesudomonas flrorescens)工程菌株中,通过离体平板对峙培养法筛选出对甜瓜细菌性果斑病菌有颉颃作用较强的菌株18株.经过进一步复筛,从中选出颉颃性最强的2P24-4、P32-40、CPF10-2等3个菌株,并将这些菌株对甜瓜果斑病进行温室防病试验,进一步验证其生防作用.结果表明,这3株生防茵的防病能力都较强,生防效果在60%以上.其中菌株P32-40的防治效果最好,达到76.1%;菌株CPF10-2的防效为71.9%;菌株2P24-4的防效为64.1%.对这3株工程菌在甜瓜叶面进行了室内、外定殖试验.研究表明,这3个菌株都可以在甜瓜叶面上大量定殖,室内和室外定殖数量上差异不明显.     

我国瓜类细菌性果斑病发生现状和综合防控策略

瓜类细菌性果斑病是西甜瓜等葫芦科作物上的一种检疫性病害,本文从病害的发生历史、传播及流行特点等方面综述了瓜类细菌性果斑病在我国的发生现状,提出了政府部门、种子种苗生产单位和种植户应共同采取措施对病害进行综合防控,以防止病害进一步传播扩散的策略。     

甘肃省西瓜细菌性果斑病的诊断

对从甘肃采集到的疑似西瓜果斑病的病瓜及种子中分离获得的菌株,通过形态学特征、革兰氏染色、致病性测定以及利用美国Agdia公司的专化型免疫凝聚试剂条测定和16S rDNA分子生物学方法进行了诊断鉴定,结果表明,该菌株符合燕麦噬酸菌西瓜亚种(Acidovorax avenae subsp. citrulli)的特性,因此,确定甘肃省已发生西瓜果斑病,结果为甘肃省有效控制此病提供了依据。     

Studies on control of seed borne bacterial vegetable disease: studies on epidemiology and control of cucurbits bacterial fruit blotch

Journal of General Plant Pathology -     

西瓜品种资源抗细菌性果斑病苗期鉴定

为筛选高抗瓜类细菌性果斑病的西瓜资源,以24份西瓜品种资源为试材,采用苗期喷雾接种法分别接种分离自甜瓜上的瓜类嗜酸菌Acidovorax citrulli菌株pslb96和ZZ-1,鉴定各品种资源对瓜类细菌性果斑病的苗期抗性。结果表明,24份西瓜品种资源中未发现有对2株菌株表现免疫的材料,有7份资源对菌株pslb96表现高抗,12份资源对菌株ZZ-1表现高抗;9份资源对菌株pslb96表现中感或感病,7份资源对菌株ZZ-1表现中感和感病;对2株菌株均表现高抗的品种资源有野生型种质资源A9及商品种华欣、申蜜968、申选958和申抗988,占总品种资源的20.83%。部分品种资源A4、A13和申蜜7号对菌株pslb96和ZZ-1的抗性表现出明显的差异,表明相同寄主来源的2株不同菌株致病力存在差异。     

Hypersensitive response suppression by type III effectors of plant pathogenic bacteria

The suppressor activity of four representative avirulence (avr) genes from the Pseudomonas syringae group against elicitors of a general hypersensitive response (HR) was examined in tobacco leaves inoculated with double transformants of Pseudomonas fluorescens containing both a cosmid plasmid (pHIR11) carrying the hrp gene cluster and a plasmid carrying each avr gene. The double transformants Pf (pHIR11) containing avrB, avrRps4, or avrPto failed to induce an HR, but that carrying avrRpt2 did induce an HR as Pf (pHIR11 + empty vector) did. Thus, some Avr proteins seem to have suppressor activity against a general HR and should promote aggressiveness of the pathogens.     

药剂处理对三倍体西瓜种子发芽势的影响及对果斑病的防治效果

选用本实验室组配的1号杀菌剂和3种常用药剂,比较了不同药剂及不同浓度的1号杀菌剂处理人工接菌的西瓜种子后对种子发芽、幼苗生长的促进作用和对果斑病的防治效果。结果显示:与对照CK1(种子破壳,清水处理)相比较,72%农用硫酸链霉素可溶粉剂7.2×102μg/mL处理2h、40%甲醛1.6×103μg/mL处理1h和1号杀菌剂0.5×104μg/mL处理2h,发芽势分别高出10.5%、6.7%和4.9%。供试药剂中1号杀菌剂处理种子对果斑病有较好的防治效果,1号杀菌剂0.5×104μg/mL处理2h、1%盐酸3.7×103μg/mL处理30min、40%甲醛溶液处理1h和72%农用硫酸链霉素可溶粉剂7.2×102μg/mL处理2h防治效果分别为74.7%、73.7%、58.5%和49.3%。     

大棚甜瓜三种主要真菌病害拮抗细菌的筛选与鉴定

从甜瓜根际土壤、牦牛粪、蚯蚓粪等样品中分离出245株细菌,用平板对峙法筛选出14株对甜瓜枯萎病菌具有良好抑制作用的菌株,分别采用管碟法和凹玻片法测定拮抗细菌发酵液对甜瓜枯萎病的抑制效果和对甜瓜白粉病菌、甜瓜霜霉病菌孢子囊萌发的抑制效果。抑菌试验结果显示,菌株FCJ2发酵液对甜瓜枯萎病菌的抑制率为64.7%,对甜瓜白粉病菌、甜瓜霜霉病菌孢子萌发抑制率为62.3%、49.2%。盆栽试验结果显示,FCJ2对甜瓜枯萎病、甜瓜白粉病和甜瓜霜霉病的防效分别为88.5%、93.6%和82.6%,与农药对照组相比均具有显著差异(P0.05)。经形态特征鉴定、生理生化特征鉴定及16S rDNA序列分析,确定FCJ2为芽孢杆菌属枯草芽孢杆菌Bacillus subtilis。     

Endophytic bacteria associated with halophyte Seidlitzia rosmarinus Ehrenb. ex Boiss. from saline soil of Uzbekistan and their plant beneficial traits

Endophytic bacteria of halophytic plants play essential roles in salt stress tolerance. Therefore, an understanding of the true nature of plant-microbe interactions under extreme conditions is essential. The current study aimed to identify cultivable endophytic bacteria associated with the roots and shoots of Seidlitzia rosmarinus Ehrenb. ex Boiss. grown in the salt-affected soil in Uzbekistan and to evaluate their plant beneficial traits related to plant growth stimulation and stress tolerance. Bacteria were isolated from the roots and the shoots of S. rosmarinus using culture-dependent techniques and identified by the 16S rRNA gene. RFLP (Restriction Fragment Length Polymorphism) analysis was conducted to eliminate similar isolates. Results showed that the isolates from the roots of S. rosmarinus belonged to the genera Rothia, Kocuria, Pseudomonas, Staphylococcus, Paenibacillus and Brevibacterium. The bacterial isolates from the shoots of S. rosmarinus belonged to the genera Staphylococcus, Rothia, Stenotrophomonas, Brevibacterium, Halomonas, Planococcus, Planomicrobium and Pseudomonas, which differed from those of the roots. Notably, Staphylococcus, Rothia and Brevibacterium were detected in both roots and shoots, indicating possible migration of some species from roots to shoots. The root-associated bacteria showed higher levels of IAA (indole-3-acetic acid) synthesis compared with those isolated from the shoots, as well as the higher production of ACC (1-aminocyclopropane-1-carboxylate) deaminase. Our findings suggest that halophytic plants are valuable sources for the selection of microbes with a potential to improve plant fitness under saline soils.     

甜瓜白粉病内生拮抗细菌的筛选鉴定及其防治效果的研究

［目的］从高抗白粉病的甜瓜品种植株内分离筛选出对甜瓜白粉病有较好防效的内生拮抗细菌菌株,为甜瓜白粉病生物防治探寻新途径。［方法］ 采用温室盆栽试验和叶盘漂浮法分离筛选有较好防效的内生拮抗菌,并对其进行形态学、生理生化和分子生物学鉴定。［结果］ 从高抗白粉病的甜瓜品种‘MR-1’植株内分离获得31株内生细菌,其中1株内生菌Mg15对甜瓜白粉病盆栽试验防效达到74.3%。在离体叶盘上,经Mg15发酵液处理12 h后白粉菌孢子萌发率仅为9.29%,14 d后产孢量与对照相比降低了60%。扫描电镜观察发现,Mg15发酵液处理后的病菌菌丝皱缩生长,甚至产生畸形。Mg15发酵原液温室防病效果达80.66%。［结论］ Mg15对甜瓜白粉病具有显著防治效果,经鉴定该菌株为枯草芽胞杆菌（Bacillus subtilis)。     

侵染猕猴桃的柑橘叶斑驳病毒外壳蛋白多克隆抗体的制备及检测应用

柑橘叶斑驳病毒(Citrus leaf blotch virus,CLBV)是β线形病毒科(Betaflexiviridae)柑橘病毒属(Citrivirus)的代表种,前期研究表明CLBV在陕西栽培猕猴桃中广泛发生.为进一步简化CLBV的检测,并探究其致病机制,通过RT-PCR扩增得到CLBV猕猴桃分离物外壳蛋白(c...