Effect of resistance variation in a natural plant host–pathogen metapopulation on disease dynamics

Existing theory suggests that increasing the diversity of resistance and virulence types in host–pathogen interactions will result in qualitative shifts in spatial and temporal dynamics, and greater among-population asynchrony in disease dynamics and prevalence. Here, data are presented from a biologically realistic metapopulation model of gene-for-gene interactions that indicate that population level variation in resistance diversity will be negatively associated with disease prevalence (fraction of individuals infected). The model also predicts that disease incidence (presence/absence) will be positively related to total resistance diversity across the metapopulation, because high resistance diversity also selects for more virulent pathogens. These results are then contrasted with empirical data from a natural host–pathogen system. While the argument that high resistance diversity should generally lead to lower disease levels has been applied extensively in agricultural situations, the connection between genetic diversity, resistance and disease dynamics has never been demonstrated in natural systems. Here, through analysis of multiyear data on disease prevalence in the context of knowledge of resistance variation among host populations in a natural plant host–pathogen metapopulation, the first evidence is provided that observed levels of asynchrony in disease dynamics may indeed be related to resistance structure.     

Variation in growth rates and aggressiveness of naturally occurring self‐fertile and self‐sterile isolates of the wilt pathogen Ceratocystis albifundus

Ceratocystis albifundus is the most important fungal pathogen of black wattle (Acacia mearnsii) grown in plantations in southern and eastern Africa. It is a homothallic fungus but also undergoes unidirectional mating type switching. As a result, the ascospore progeny can be either self‐fertile or self‐sterile. The only apparent difference between these mating types is the deletion of the MAT1‐2‐1 gene in self‐sterile isolates. There is some evidence suggesting that self‐sterile isolates grow more slowly than self‐fertile isolates, but this has not been tested rigorously. The aim of this study was to determine whether self‐sterile isolates are less fit by examining growth rate, relative germination rate and pathogenicity. Five self‐sterile isolates were generated from each of five self‐fertile isolates of C. albifundus and these 30 isolates were compared. The results showed that the self‐sterile isolates grew consistently slower and were less pathogenic than the self‐fertile isolates. The germination ratio of self‐fertile to self‐sterile isolates from single ascospores collected from the ascomata of five self‐fertile isolates was on average 7:3. This could be a consequence of the self‐sterile isolates having a lower germination rate. This observation, and the lower growth and pathogenicity levels, suggests that self‐sterile isolates are not likely to compete effectively in nature, raising intriguing questions regarding their role and value to C. albifundus and other fungi having a similar mating system.     

Combating plant diseases—the Darwin connection

Although Darwin knew of plant diseases, he did not study them as part of his analysis of natural selection. Effective plant disease control has only been developed after his death. This article explores the relevance of Darwin's ideas to three problem areas with respect to diseases caused by fungi: emergence of new diseases, loss of disease resistance bred into plants and development of fungicide resistance. Darwin's concept of change through natural or artificial selection relied on selection of many small changes, but subsequent genetic research has shown that change can also occur through large steps. Appearance of new diseases can involve gene duplication, transfer or recombination, but all evidence points to both host plant resistance and fungicide susceptibility being overcome through point mutations. Because the population size of diseases such as rusts and powdery and downy mildews is so large, all possible point mutations are likely to occur daily, even during moderate epidemics. Overcoming control measures therefore reflects the overall fitness of these mutants, and much resource effort is being directed towards assessment of their fitness, both in the presence and in the absence of selection. While recent developments in comparative genomics have caused some revision of Darwin's ideas, experience in managing plant disease control measures clearly demonstrates the relevance of concepts he introduced 150 years ago. It also reveals the remarkable speed and the practical impact of adaptation in wild microorganism populations to changes in their environment, and the difficulty of stopping or delaying such adaptation. Copyright © 2009 Society of Chemical Industry     

野生二粒小麦叶枯病病原菌鉴定及抗病基因遗传特性的初步研究

叶枯病是小麦常见病害,由多种病菌引起。本文通过野生二粒小麦叶片的叶枯病病斑表型,菌落和孢子形态,真菌ITS1、5.8SrDNA和ITS2区域序列的进化分析以及科赫氏法则等试验表明,链格孢真菌Alternaria alternata是本研究供试野生二粒小麦叶枯病的致病菌。为进一步研究野生二粒小麦中抗病基因的遗传特性,配置了抗感杂交并获得了遗传群体。对F1、F2以及F3株系病情的遗传分析表明,在抗病亲本‘JIC1060003’中存在一对显性抗病基因,同时双亲中可能还存在微效的数量性状抗病位点。本研究为进一步通过遗传作图获得与抗病基因连锁的分子标记和抗病基因的进一步利用奠定了基础。     

The role of aggressiveness and competition in the selection of Phytophthora infestans populations

Selection within populations of Phytophthora infestans was investigated by comparing the aggressiveness of single‐lesion isolates on detached leaflets of four potato cultivars with differing levels of race‐nonspecific resistance to P. infestans. The isolates included 23 representative of Northern Ireland genotypes from the early 2000s, used to inoculate previously reported field trials on competitive selection (2003–2005), plus 12 isolates recovered from the 2003 trial. The cultivars were those planted in the previous trials: Atlantic (blight‐susceptible) and Santé, Milagro and Stirling (partially resistant). Very highly significant variation for latent period, infection frequency and lesion area was found between genotypes and cultivars; differences between genotypes were more marked on the more resistant cultivars, but no one genotype was the most aggressive across all. Detached leaflets were also inoculated with mixtures of isolates from each genotype group at three sporangial concentrations: differences in aggressiveness between genotypes were more apparent at lower concentrations and on the more resistant cultivars. Genotype groups that were the most aggressive on the more resistant cultivars tended to be those selected by the same cultivars in the field. A mixture of all isolates of all genotypes was used to inoculate detached leaflets of the same cultivars. With one exception, single spore isolates recovered from any one leaflet belonged to a single genotype, but different genotypes were recovered from different cultivars. Phytophthora infestans isolates from Northern Ireland showed significant variation for foliar aggressiveness, and pathogen genotypes exhibited differential aggressiveness to partially resistant cultivars and interacted competitively in genotype selection.     

Measurement of infection efficiency of a major wheat pathogen using time-resolved imaging of disease progress

Infection efficiency is a key epidemiological parameter that determines the proportion of pathogen spores able to infect and cause lesions once they have landed on a susceptible plant tissue. In this study, an improved method to measure infection efficiency of Zymoseptoria tritici using a replicated greenhouse experiment is presented. Zymoseptoria tritici is a fungal pathogen that infects wheat leaves and causes septoria tritici blotch (STB), a major disease of wheat worldwide. A novel experimental setup was devised, where living wheat leaves were attached to metal plates, allowing for time-resolved imaging of disease progress in planta. Because lesions were continuously appearing, expanding and merging during the period of up to 3 weeks, daily measurements were necessary for accurate counting of lesions. Reference membranes were also used to characterize the density and spatial distribution of spores inoculated onto leaf surfaces. In this way, the relationship between the number of lesions and the number of viable spores deposited on the leaves was captured and an infection efficiency of about 4% was estimated from the slope of this relationship. This study provides a proof of principle for accurate and reliable measurement of infection efficiency of Z. tritici. The method opens opportunities for determining the genetic basis of the component of quantitative resistance that suppresses infection efficiency. This knowledge would improve breeding for quantitative resistance against STB, a control measure considered more durable than deployment of major resistance genes.     

广东马铃薯黑胫病的病原鉴定

 通过对分离自马铃薯黑胫病的病原菌在选择性CVP培养基上菌落形态观察、致病性测定及16S rDNA序列比较,鉴定出引起广东省冬种马铃薯黑胫病的病原为γ-变形菌门肠杆菌科果胶杆菌属黑腐果胶杆菌(Pectobacterium atrosepticum)。生理生化试验表明,该病原菌菌株不能在37℃生长,能使明胶液化,具有运动性、耐盐性(5％NaCl),对红霉素不敏感;过氧化氢酶和蔗糖还原试验反应均为阳性,氧化酶、吲哚产生和磷酸酶活性试验均为阴性;可利用乳糖、麦芽糖、棉籽糖、蜜二糖、纤维二糖、柠檬酸盐和α-甲基酮葡萄糖苷,不能利用山梨醇、d-阿拉伯糖和丙二酸盐。研究结果可为广东冬种马铃薯黑胫病的综合防治提供科学依据。     

内蒙古马铃薯黑胫病病原菌的分离和鉴定

 马铃薯是我国的第四大主粮。内蒙古是我国最大的马铃薯生产基地,马铃薯产业是当地农民收入的主要来源。2017年8月,内蒙古锡林郭勒盟部分马铃薯田块发生了一种细菌性黑胫病,发病面积约为200亩,发病率约为20%。发病初期,植株的茎基部变色、发黑,伴有臭味,严重时茎秆腐烂,植株死亡。为明确该地区马铃薯细菌性黑胫病的病原菌种类,本研究对该病原菌进行了分离,并利用生理生化测定,Biolog分析和分子生物学手段将该病原菌鉴定为胡萝卜软腐果胶杆菌巴西亚种(Pectobacterium carotovorum subsp. brasiliense)。据以往报道,我国马铃薯黑胫病主要由黑腐果胶杆菌(P. atrosepticum)和胡萝卜软腐果胶杆菌胡萝卜亚种(P. carotovorum subsp. carotovorum)两种病原菌引起,本研究是P. carotovorum subsp. brasiliense引起马铃薯黑胫病在国内的首次报道。     

Fusarium graminearum as a dry rot pathogen of potato in the USA: prevalence,comparison of host isolate aggressiveness and factors affecting aetiology

A 2004–2005 survey of potatoes from stores in the north‐central potato‐producing region of the USA showed that the predominant causes of dry rot were Fusarium graminearum and Fusarium sambucinum. Isolates of F. graminearum originally isolated from potato tubers with dry rot (n = 15), wheat kernels with scab (n = 15), and sugarbeet tap roots with decay (n = 5) were tested for aggressiveness to potato tubers. There were no significant differences in aggressiveness among isolates of F. graminearum, regardless of original host, as measured by their ability to cause dry rot. These findings may have implications for survival of F. graminearum inoculum since potatoes, wheat and sugarbeets are frequently used in crop rotation in the region. Fusarium graminearum required larger wounds for infection of potato tubers than F. sambucinum. Plug‐removal injury, simulating a stolon‐removal injury, resulted in equal incidence of dry rot caused by the two Fusarium species, whereas abrasion and bruising injury were sufficient for infection and dry rot development by F. sambucinum, but not F. graminearum. A change in harvest practices from vine‐killing prior to harvest to mechanical vine‐killing on the day of harvest may be a factor affecting the onset of dry rot caused by F. graminearum, since this process often causes large wounds at the stem end of the tubers when the stolon is forcibly removed.     

云南弥勒甘蔗褐条病病原菌的分离鉴定

2019年10月,在云南弥勒甘蔗示范基地(23.92°N,103.33°E)发现‘云瑞10-187’和‘福农11-2907’高感甘蔗褐条病,发病率为50%～80%。为明确其病原,本研究采集病样进行了病原菌的分离、鉴定及系统发育分析,以期为该病害有效防控提供科学依据。依据形态特征、核糖体RNA的内转录间隔区(internal transcribed spacer, ITS)和甘油醛-3-磷酸脱氢酶基因(glyceraldehyde-3-phosphate dehydrogenase, GAPDH)分子鉴定及致病性测定,将病原菌鉴定为狗尾草平脐蠕孢Bipolaris setariae,是云南省甘蔗褐条病病原菌新记录种,丰富了甘蔗褐条病病原菌信息,为后续其他蔗区褐条病的研究奠定了基础。     

新疆加工型辣椒一种新的细菌性叶斑病病原的鉴定

2014—2015年在新疆沙湾县加工型辣椒Capsicum annuumL.上发现一种细菌性叶斑病。为明确其病原种类,采用组织分离法分离病原菌,依据柯赫氏法则进行致病性测定,并对菌株进行16S rDNA和持家基因rpoD及gry B的扩增、序列测定和系统发育分析,对其进行鉴定。结果表明,获得的8个细菌菌株间致病力无明显差异。致病细菌与已报道的黄褐假单胞菌Pseudomonas fulva菌株的16S rDNA、rpoD基因、gry B基因的同源性分别达到了99.1%~99.9%、99.4%~99.5%和97.6%~100.0%。结合革兰氏染色反应、菌体形态、培养性状、生理生化反应、寄主范围等特征,将病原菌鉴定为黄褐假单胞菌。该病原菌通过人工接种还能侵染番茄、茄子、马铃薯、黄瓜、西瓜、甜瓜、四季豆、豆角、豇豆、白菜、萝卜、胡萝卜及芹菜等多种植物。     

浙江省鲜食大豆炭疽病病原分离及抗性鉴定

炭疽病是我国南方鲜食大豆最重要的病害之一.本试验通过组织分离法,在浙江省鲜食大豆主栽区炭疽病病荚中分离得到9株疑似炭疽病菌分离物.根据分离物的形态特征、基于ITS-GAPDH-CHS-1-TUB2基因序列的系统发育分析、分离物的致病性将这些分离物鉴定为平头炭疽菌(Colletotrichum truncatum).对5...     

小麦白粉病菌分生孢子田间传播的初步研究

2015年度采用定容式孢子捕捉器对田间空气中小麦白粉病菌分生孢子传播的初步研究表明,在病害发生初期病菌分生孢子捕捉量比较低,但随着菌源中心病害的逐渐加重,病菌分生孢子在距菌源中心20 m和40 m远处捕捉量随之增大。线性弧度相关分析结果发现,距菌源中心北向20 m和40 m处的孢子捕捉量与风向存在显著正相关性;距菌源中心不同距离处的孢子捕捉量之间均存在极显著正相关性,且在相同方向上,距菌源中心20 m处孢子捕捉量显著高于40 m处孢子捕捉量。     

云南省玉米白斑病病原菌鉴定与品种田间抗性评价

为明确近年来云南省严重发生的玉米白斑病病原菌及不同玉米品种对玉米白斑病的抗性差异,2021年于云南省昆明市阿子营乡和曲靖市罗平县九龙镇采集病样,进行病斑组织分离纯化,通过形态学特征和分子生物学方法对病原菌进行鉴定,并在云南省曲靖市罗平县九龙镇采用田间自然发病方法对622份玉米品种的抗性进行鉴定。从2个采样地点的病叶中各分离出5株和9株菌株,经柯赫氏法则验证,确定了菌株WS-1、WS-191为白斑病致病菌;经形态学观察以及基于ITS、rpb2、tub2和LSU基因序列构建的系统发育树分析结果确定其分类学种名为宽颈附球菌Epicoccum latusicollum。田间抗病性试验结果表明,在622份区域试验品种中对白斑病表现高抗、抗病、中抗、感病和高感的品种分别为4、184、216、186和32份,分别占品种总数的0.64%、29.58%、34.73%、29.90%和5.14%。     

Effect of cold‐induced changes in physical and chemical leaf properties on the resistance of winter triticale (×Triticosecale) to the fungal pathogen Microdochium nivale

This study showed that several mechanisms of the basal resistance of winter triticale to Microdochium nivale are cultivar‐dependent and can be induced specifically during plant hardening. Experiments and microscopic observations were conducted on triticale cvs Hewo (able to develop resistance after cold treatment) and Magnat (susceptible to infection despite hardening). In cv. Hewo, cold hardening altered the physical and chemical properties of the leaf surface and prevented both adhesion of M. nivale hyphae to the leaves and direct penetration of the epidermis. Cold‐induced submicron‐ and micron‐scale roughness on the leaf epidermis resulted in superhydrophobicity, restricting fungal adhesion and growth, while the lower permeability and altered chemical composition of the host cell wall protected against tissue digestion by the fungus. The fungal strategy to access the nutrient resources of resistant hosts is the penetration of leaf tissues through stomata, followed by biotrophic intercellular growth of individual hyphae and the formation of haustoria‐like structures within mesophyll cells. In contrast, a destructive necrotrophic fungal lifestyle occurs in susceptible seedlings, despite cold hardening of the plants, with the host epidermis, mesophyll and vascular tissues being digested and becoming disorganized as a result of the low chemical and mechanical stability of the cell wall matrix. This work indicates that specific genetically encoded physical and mechanical properties of the cell wall and leaf tissues that depend on cold hardening are factors that can determine plant resistance against fungal diseases.     

广东省冬枣叶枯病病原菌的分离与鉴定

2018年2月,广东省清远市冬枣种植区发生了一种叶枯病,发病率约为17％,严重影响植株生长.为明确引起广东省清远市冬枣叶枯病的病原,采用常规病组织分离法获得2株菌株,通过致病性、生理生化特征和分子生物学对2株菌株进行鉴定.致病性测定结果显示,该细菌可侵染引起冬枣叶片产生枯死症状,与田间病株的症状相同.生理生化测定结果显...     

Analysis of resistance to Xylella fastidiosa within a hybrid population of Pera sweet orange × Murcott tangor

Resistance to Xylella fastidiosa was evaluated within a population of 20 interspecific hybrids of Pera sweet orange and Murcott tangor under greenhouse conditions. Efficiency of inoculation, multiplication of bacteria within the plants, xylem vessel morphology, and symptom expression were analysed. The rate of infection ranged from 40 to 100% (average 70%) for all genotypes analysed. Xylella fastidiosa populations ranged from log 0·59 to log 2·13 cells mg⁻¹ tissue for the resistant hybrids. These values were significantly different ( P  = 0·05) from those obtained for the tolerant (no symptoms but bacteria recovered) or susceptible (symptoms and bacteria recovered) hybrids (log 3·02 to log 4·06 cells mg⁻¹). Xylella fastidiosa was recovered from all hybrids (log 2·31 to 5·03 CFU mg⁻¹ tissue) except the resistant ones. The first foliar symptoms appeared at least 90 days post-inoculation, the time varying according to genotype. No correlation between xylem vessel morphology and disease expression was observed, indicating that the resistance was the result of a genetic response of the host. According to this hypothesis, a high broad-based heritability index for resistance was obtained (0·96) at 210 days from X. fastidiosa inoculations, using bacterial quantification by real-time PCR, which indicated that the influence of the number of bacteria was the result of genetic rather than environmental variations.     

Development of Oculimacula yallundae and O. acuformis (eyespot) on leaf sheaths of winter wheat in the UK in relation to thermal time

In a controlled environment (15/10°C) (day/night) container experiment on winter wheat (cv. Avalon), eyespot incidence (percentage of plants affected) and number of leaf sheaths penetrated after 6 weeks increased with inoculum concentration (10²−10⁶ conidia mL⁻¹) of Oculimacula yallundae (OY) or Oculimacula acuformis (OA), but there was no difference between the two species. In an outdoor container experiment, seedlings inoculated with OY 2 weeks after sowing had a greater incidence of eyespot than those inoculated with OA, when assessed 7 weeks after inoculation. Seedlings inoculated with OA at 10 or 20 weeks after sowing developed more severe eyespot by maturity than those inoculated with OY. In an experiment at 15/10°C with seedlings inoculated with OY + OA 2 weeks after sowing, more leaf sheaths were penetrated by OY (3·0 per plant) than OA (2·3 per plant) 6 weeks after inoculation. Field experiments with winter wheat consistently showed leaf sheath production, leaf sheath death, and number of leaf sheaths infected or penetrated by OA or OY were related linearly to thermal time (°C days) after sowing. Depending on cultivar, season and sample, a new leaf sheath was produced in 116–216°C days; a leaf sheath died in 221–350°C days; and infection of a new leaf sheath occurred in 129–389°C days. The mean number of living leaf sheaths infected differed between samples, cultivars and seasons for both OY and OA. Regression analysis of the 1985/86 data suggested that OY progressed more rapidly than OA through the leaf sheaths, and that both the pathogens progressed more rapidly than the rate of leaf sheath death, but more slowly than the rate at which leaf sheaths were produced. It also suggested that OA progressed more slowly than the rate at which leaf sheaths died in 1987/88, but OY did not.