瓜菜枯萎病及生防根际微生物组研究进展

枯萎病是顽固性土传病害,称为瓜菜中的“癌症”,已成为制约我国瓜菜产业可持续健康发展的瓶颈问题。本文简要介绍瓜菜枯萎病危害,并从细胞壁降解酶、毒素、信号传导和致病基因等方面综述瓜菜枯萎病灾变机制,然后从根际微生物组自身与病原菌、土壤层面和植物层面等重点阐述了根际微生物组防治和抵御瓜菜枯萎病的机理,最后对枯萎病发生和抑制关键因子挖掘、核心微生物组构建及根际微生物组分子机制等进行了展望,期望生防微生物防治病害发生进入一个崭新且高效的阶段,为加快提升作物抗逆性机理研究提供一定思路。     

微生物源挥发性物质防治采后果蔬病害的研究进展

随着化学杀菌剂弊端的日益凸显,生物防治已逐渐成为采后果蔬病害控制的研究和开发热点。其中,很多微生物产生的多种挥发性物质(volatile organic compounds,VOCs),能显著抑制多种病原菌的生长繁殖,有效控制采后果蔬病害。由于微生物源VOCs具有有效、安全、环保、易降解和无残留等优点,越来越受到各国研究者的重视和青睐。本文综述了产生VOCs的微生物的多样性、微生物源VOCs的多样性、微生物源VOCs的抑菌活性、生防效果及其主要作用机制等方面的研究进展,以期为病原菌的绿色安全防治提供基础资料。     

亚洲柑橘木虱与柑橘黄龙病菌互作的研究进展

亚洲柑橘木虱Diaphorina citri Kuwayama作为柑橘产业重要病害柑橘黄龙病的主要传播媒介,已经成为重点防治对象。该害虫与黄龙病之间的互作一直是相关研究的热点,本文就近年来该领域的研究进展做了一个总结,从亚洲柑橘木虱的获菌与传病机制、病原菌与柑橘木虱之间的互作以及病原菌感染寄主植物后对木虱的影响等方面进行了综述。期望为深入开展黄龙病相关研究、寻找防控新途径提供依据。     

湘西地区猕猴桃溃疡病病原菌分离及其全基因组分析

通过了解湘西地区猕猴桃溃疡病致病菌分类地位和基因类型,初步探讨其致病的分子机理。采用纯培养法分离猕猴桃溃疡病菌;基于16S～23S rRNA基因内转录间隔序列进行病原菌的系统发育分析;通过基因组测序和生物信息学分析解析其致病的分子机理。从“米良1号”和“红阳”猕猴桃感病枝条中分离获得5株溃疡病菌,编号为L211、L212、L321、L322、L323;通过形态特征和16S～23S rRNA基因内转录间隔序列分析,鉴定5株细菌均为丁香假单胞菌猕猴桃致病变种(Pseudomonas syringae pv.actinidae,Psa)。以菌株L211为代表进行体外猕猴桃枝条接种实验表明能引起典型溃疡病症状。通过菌株L211的全基因组测序和生物信息学分析,获得5 741条基因数目,长5 412 072 bp;基因功能注释发现菌株L211携带121种毒力因子、71个植物互作因子和77个耐药基因;同时,基因组单核苷酸多态性分析发现病原菌L211为基因Ⅲ型Psa。引起湘西地区猕猴桃溃疡病的病原菌是丁香假单胞菌猕猴桃致病变种基因Ⅲ型,与国内外报道的引起猕猴桃溃疡病大流行的致病菌一致。猕猴桃溃疡病发病...     

芽胞杆菌在防治水稻稻瘟病中的应用及生防机制

芽胞杆菌具有人畜安全、不污染环境、病原菌不易产生抗药性、抗逆性强和促进植物生长等优点,是稻瘟病防治上的重要生防菌。芽胞杆菌的生防机制主要包括竞争作用、拮抗作用和诱导抗病性。芽胞杆菌定殖在水稻植株上,产生抗菌活性物质抑制稻瘟病菌的生长,诱导水稻产生抗病性,对水稻植株具有促生作用,可以挽回水稻产量损失。芽胞杆菌可以制备生防制剂用来防治我国南方稻区和北方稻区的稻瘟病危害,在水稻产业的可持续发展中对稻瘟病的生物防治具有指导意义。本文主要综述芽胞杆菌在防治水稻稻瘟病中的应用研究、芽胞杆菌在防治水稻稻瘟病中的生防机制、影响稻瘟病生防芽胞杆菌防效的因素。     

柑橘木虱抗药性研究进展

柑橘木虱Diaphorina citri Kuwayama主要危害柑橘等芸香科植物,传播柑橘黄龙病病菌。柑橘树感染黄龙病后,叶片黄化,果实早熟脱落,畸形,最终死亡,给柑橘产业带来了极大的损失。柑橘木虱对各种常见的杀虫剂已产生了不同程度的抗药性。本文从柑橘木虱的抗性测定方法、抗药性现状、交互抗性、抗药性机理和抗性治理等方面进行了综述,为今后柑橘木虱的防治提供一定的参考。     

食源性致病微生物危害风险及其防控用抗菌生物活性肽研究进展

食源致病微生物是污染食品、食用农产品的主要生物危害物,严重威胁人类及动物健康;同时也给食品、食用农产品产业造成巨大经济损失,是制约产业健康可持续发展的关键危害因素。传统依赖抗生素对致病微生物的防控效果显著,但由于抗生素长期的不合理使用甚至是滥用所诱发产生的致病微生物耐药性,以及对人类和动物体内脏器、肠道组织、神经和代谢系统正常功能构成的潜在危害性等问题也同样突出。抗菌生物活性肽作为具有抗菌功能的生物多肽类蛋白质,被认为是对人类、动物以及生态环境友好的抗菌物质,有望替代抗生素用于致病微生物的防控,现已成为探索新型安全抗菌类药物研发的热点。全面梳理了食源致病微生物种类、污染特点及其主要危害风险;系统概述了抗菌生物活性肽主要类型、来源及其创制技术;着重介绍了当前抗菌生物活性肽在食源致病微生物防控上的研究状况,并深入探讨了其应用前景、存在问题及拟解决对策,旨在为食品、食用农产品中食源性致病微生物筛查以及靶向防控技术提供最新的有参考价值的文献资料和创新思路。     

群体感应及其在动物病原菌致病中的作用

摘要: 群体感应是指微生物群体某些基因的表达受到与群体密度相关的信号分子调控的现象。微生物以酰基高丝氨酸内酯化合物,某些短肽分子,呋喃酮类化合物,以及一些小分子物质为信号分子,介导不同的群体感应系统。各群体感应系统之间以平行协同或层次串连的方式组织起来调控微生物各种基因。众多病原菌致病基因的表达与群体感应密切相关,主要表现在：群体感应帮助微生物对宿主的侵袭和定殖;调控毒力因子的产生和作用于宿主;以及介导病原菌对宿主的免疫能力和药物抗性。进行群体感应对微生物致病过程调控的研究,将有利于从群体感应入手进行病原菌防控新策略的探索。     

柑橘木虱抗药性研究进展

柑橘木虱Diaphorina citri Kuwayama主要危害柑橘等芸香科植物,传播柑橘黄龙病病菌。柑橘树感染黄龙病后,叶片黄化,果实早熟脱落,畸形,最终死亡,给柑橘产业带来了极大的损失。柑橘木虱对各种常见的杀虫剂已产生了不同程度的抗药性。本文从柑橘木虱的抗性测定方法、抗药性现状、交互抗性、抗药性机理和抗性治理等方面进行了综述,为今后柑橘木虱的防治提供一定的参考。     

丁香假单胞菌猕猴桃致病变种的遗传多样性及进化关系

猕猴桃细菌性溃疡病是一种严重威胁猕猴桃生产的毁灭性病害,在世界各大猕猴桃产区发病严重。引起该病的病原菌为丁香假单胞菌猕猴桃致病变种(Pseudomonas syringae pv.Actinidiae,Psa)。综合当前国内外关于该病原菌的相关研究,就其分类地位、遗传多样性及起源进化等方面进行了系统综述。     

Modification of the PthA4 effector binding elements in Type I CsLOB1 promoter using Cas9/sgRNA to produce transgenic Duncan grapefruit alleviating XccΔpthA4:dCsLOB1.3 infection

Citrus canker caused by Xanthomonas citri subspecies citri (Xcc) is a severe disease for most commercial citrus cultivars and responsible for significant economic losses worldwide. Generating canker‐resistant citrus varieties will provide an efficient and sustainable solution to control citrus canker. Here, we report our progress in generating canker‐resistant grapefruit by modifying the PthA4 effector binding elements (EBEs) in the CsLOB1 Promoter (EBE_PthA4‐CsLOBP) of the CsLOB1 (Citrus sinensis Lateral Organ Boundaries) gene. CsLOB1 is a susceptibility gene for citrus canker and is induced by the pathogenicity factor PthA4, which binds to the EBE_PthA4‐CsLOBP to induce CsLOB1 gene expression. There are two alleles, Type I and Type II, of CsLOB1 in Duncan grapefruit. Here, a binary vector was designed to disrupt the PthA4 EBEs in Type I CsLOB1 Promoter (TI CsLOBP) via epicotyl transformation of Duncan grapefruit. Four transgenic Duncan plants with targeted modification of EBE_PthA4‐T1 CsLOBP were successfully created. As for Type I CsLOB1 promoter, the mutation rate was 15.63% (#D13), 14.29% (#D17), 54.54% (#D18) and 81.25% (#D22). In the presence of wild‐type Xcc, transgenic Duncan grapefruit developed canker symptoms similarly as wild type. An artificially designed dTALE dCsLOB1.3, which specifically recognizes Type I CsLOBP, but not the mutated Type I CsLOBP or Type II CsLOBP, was developed to infect Duncan transformants. Consequently, #D18 had weakened canker symptoms and #D22 had no visible canker symptoms in the presence of XccΔpthA4:dCsLOB1.3. Our data suggest that activation of a single allele of susceptibility gene CsLOB1 by PthA4 is sufficient to induce citrus canker disease, and mutation in the promoters of both alleles of CsLOB1 is probably required to generate citrus canker‐resistant plants. This work lays the groundwork to generate canker‐resistant citrus varieties via Cas9/sgRNA in the future.     

New genes of Xanthomonas citri subsp. citri involved in pathogenesis and adaptation revealed by a transposon-based mutant library

Background  

Citrus canker is a disease caused by the phytopathogens Xanthomonas citri subsp. citri, Xanthomonas fuscans subsp. aurantifolli and Xanthomonas alfalfae subsp. citrumelonis. The first of the three species, which causes citrus bacterial canker type A, is the most widely spread and severe, attacking all citrus species. In Brazil, this species is the most important, being found in practically all areas where citrus canker has been detected. Like most phytobacterioses, there is no efficient way to control citrus canker. Considering the importance of the disease worldwide, investigation is needed to accurately detect which genes are related to the pathogen-host adaptation process and which are associated with pathogenesis.     

Plant responses underlying nonhost resistance of Citrus limon against Xanthomonas campestris pv. campestris

Citrus is an economically important fruit crop that is severely afflicted by citrus canker, a disease caused by Xanthomonas citri ssp. citri (X. citri); thus, new sustainable strategies to manage this disease are needed. Although all Citrus spp. are susceptible to this pathogen, they are resistant to other Xanthomonas species, exhibiting non-host resistance (NHR), for example, to the brassica pathogen X. campestris pv. campestris (Xcc) and a gene-for-gene host defence response (HDR) to the canker-causing X. fuscans ssp. aurantifolii (Xfa) strain C. Here, we examine the plant factors associated with the NHR of C. limon to Xcc. We show that Xcc induced asymptomatic type I NHR, allowing the bacterium to survive in a stationary phase in the non-host tissue. In C. limon, this NHR shared some similarities with HDR; both defence responses interfered with biofilm formation, and were associated with callose deposition, induction of the salicylic acid (SA) signalling pathway and the repression of abscisic acid (ABA) signalling. However, greater stomatal closure was seen during NHR than during HDR, together with different patterns of accumulation of reactive oxygen species and phenolic compounds and the expression of secondary metabolites. Overall, these differences, independent of Xcc type III effector proteins, could contribute to the higher protection elicited against canker development. We propose that Xcc may have the potential to steadily activate inducible defence responses. An understanding of these plant responses (and their triggers) may allow the development of a sustained and sustainable resistance to citrus canker.     

Prediction of citrus canker epidemics generated through different inoculation methods

Citrus canker epidemics were generated with 10^8?cfu/ml of Xanthomonas axonopodis pv. citri (ex Hasse) on Citrus limonia cv. China lemon, Citrus reticulate cv. kinnow, Citrus jambhiri, Citrus reticulate cv. Feutral’s early and Citrus limettioides using four inoculation techniques. Natural inoculum was also relied upon for infection. Overall, the injection infiltration method led to maximum disease generation followed by spray, pinprick and smear inoculation methods. Citrus canker incidence along with environmental data were recorded and subjected to stepwise regression analysis. Except relative humidity, the relationship of weekly air temperature (maximum and minimum), rainfall and wind speed with citrus canker disease development in all citrus cultivars was positively correlated and best explained by linear regression. Overall, two environmental variable model containing maximum and minimum air temperature fit the data well explaining 93% variability in disease development. The observed citrus canker incidence values and those predicted by the model were close in most of citrus cultivars. This two environmental variable model can be used to issue advance warning forecasts for the timely management of the citrus canker in Pakistan.     

Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker

Citrus is a highly valued tree crop worldwide, while, at the same time, citrus production faces many biotic challenges, including bacterial canker and Huanglongbing (HLB). Breeding for disease‐resistant varieties is the most efficient and sustainable approach to control plant diseases. Traditional breeding of citrus varieties is challenging due to multiple limitations, including polyploidy, polyembryony, extended juvenility and long crossing cycles. Targeted genome editing technology has the potential to shorten varietal development for some traits, including disease resistance. Here, we used CRISPR/Cas9/sgRNA technology to modify the canker susceptibility gene CsLOB1 in Duncan grapefruit. Six independent lines, D_LOB2, D_LOB3, D_LOB9, D_LOB10, D_LOB11 and D_LOB12, were generated. Targeted next‐generation sequencing of the six lines showed the mutation rate was 31.58%, 23.80%, 89.36%, 88.79%, 46.91% and 51.12% for D_LOB2, D_LOB3, D_LOB9, D_LOB10, D_LOB11 and D_LOB12, respectively, of the cells in each line. D_LOB2 and D_LOB3 showed canker symptoms similar to wild‐type grapefruit, when inoculated with the pathogen Xanthomonas citri subsp. citri (Xcc). No canker symptoms were observed on D_LOB9, D_LOB10, D_LOB11 and D_LOB12 at 4 days postinoculation (DPI) with Xcc. Pustules caused by Xcc were observed on D_LOB9, D_LOB10, D_LOB11 and D_LOB12 in later stages, which were much reduced compared to that on wild‐type grapefruit. The pustules on D_LOB9 and D_LOB10 did not develop into typical canker symptoms. No side effects and off‐target mutations were detected in the mutated plants. This study indicates that genome editing using CRISPR technology will provide a promising pathway to generate disease‐resistant citrus varieties.