Mechanisms of resistance in Brassica carinata,B. napus and B. juncea to Pseudocercosporella capsellae

Studies were undertaken to compare susceptible and resistant host responses to Pseudocercosporella capsellae in cotyledons of Brassica carinata, B. juncea and B. napus in order to define the mechanisms of resistance in these three species. On both resistant and susceptible hosts, hyphal penetration was always through stomatal openings and without infection pegs or appressoria. On resistant B. carinata ATC94129P, up to 72% of spores disintegrated and, generally, germination (<22%) and germ tube lengths (<25 μm) were comparatively low. Resistant B. napus Hyola 42 had the lowest germination (8%) and susceptible B. carinata UWA#012 had the highest (51%). On resistant B. carinata ATC94129P, germ tube extension was impeded across 24–60 h post‐inoculation (hpi) and percentage stomatal penetration lower (4%) at 60 hpi compared with susceptible B. carinata UWA#012 (26%). Stomatal densities (stomata/14 757 μm²) on resistant B. juncea Dune (2·12) and B. napus Hyola 42 (1·62) were lower than for susceptible B. juncea Vardan (2·40) and B. napus Trilogy (2·03). Resistant B. carinata ATC94129P had greater stomatal density (1·89) than susceptible B. carinata UWA#012 (1·58). Overall, B. juncea had greater stomatal density (2·26) compared with B. napus (1·83) and B. carinata (1·74). In resistant B. carinata ATC94129P, P. capsellae induced 28% stomata to close, while in susceptible B. carinata UWA#012 no such closure was induced. Epicuticular wax crystalloids were present only on resistant B. carinata ATC94129P and probably also contribute towards resistance.     

Unique infection structures produced by Pseudocercosporella capsellae on oilseed crops Brassica carinata,B. juncea and B. napus in Western Australia

White leaf spot disease (Pseudocercosporella capsellae) is widespread across oilseed, vegetable and forage brassicas. Light (LM) and scanning electron (SEM) microscope studies were undertaken to investigate host–pathogen interactions on cotyledons of resistant and susceptible Brassica carinata, B. juncea and B. napus. Under LM, unique brown structures were present, particularly on susceptible genotypes, in two morphologically distinct forms: first, as thread‐like structures within cortical tissue by 24 h post‐inoculation (hpi) and secondly, as brown ropy strand structures either within cortical tissues (internal ropy strands), or extruded out through stomatal pores (ropy strand extrusions). Under LM, these brown structures were most prevalent in highly susceptible B. juncea ‘Vardan’ that had both a high incidence within cortical tissue (70%) and of ropy strand extrusions (73%), as did susceptible B. napus ‘Trilogy’ within cortical tissue (60%). Under SEM, both these genotypes showed thread‐like structures smaller than hyphae forming highly branched networks and ropy strand‐like structures. While there were fewer brown structures in susceptible B. carinata UWA #012 (35%), fine, thread‐like structures forming networks were again prominent (SEM). In contrast, for resistant genotypes, brown structures (LM) were of very low frequency or absent; only 5% in resistant B. juncea ‘Dune’ and none in resistant B. napus ‘Hyola 42’ or highly resistant B. carinata ATC94129P. Under SEM, fine, thread‐like structures were present in the resistant B. juncea ‘Dune’ and B. napus ‘Hyola 42’. Liquid chromatographic analyses of brown structures revealed that both internal ropy strands within cortical tissues and ropy strand extrusions contained the mycotoxin cercosporin.     

Pathotypes and phylogenetic variation determine downy mildew epidemics in Brassica spp. in Australia

Isolates of Hyaloperonospora brassicae inoculated onto cotyledons of 28 diverse Brassicaceae genotypes, 13 from Brassica napus, two from B. juncea, five from B. oleracea, two from Eruca vesicaria, and one each from B. nigra, B. carinata, B. rapa, Crambe abyssinica, Raphanus sativus and R. raphanistrum, showed significant effects (P ≤ 0.001) of isolate, host and their interaction. Host responses ranged from no visible symptom or a hypersensitive response, to systemic spread and abundant pathogen sporulation. Isolates were generally most virulent on their host of origin. Using an octal classification, six host genotypes were identified as suitable host differentials to characterize pathotypes of H. brassicae and distinguished eight distinct pathotypes. There were fewer, but more virulent, pathotypes in 2015–2016 isolates than 2006–2008 pathogen populations, probably explaining the increase in severity of canola downy mildew over the past decade. Phylogenetic relationships determined across 20 H. brassicae isolates collected in 2006–2008 and 88 isolates collected in 2015–2016 showed seven distinct clades, with 70% of 2006–2008 isolates distributed within clade I (bootstrap value (BVs) of 100%) and the remaining 30% in clade V (BVs 83.3%). This is the first study to define phylogenetic relationships of H. brassicae isolates in Australia, setting a benchmark for understanding current and future genetic shifts within pathogen populations; it is also the first to use octal classification to characterize pathotypes of H. brassicae, providing a novel basis for standardizing phenotypic characterization and monitoring of pathotypes on B. napus and some crucifer species in Australia.     

玉米灰斑病菌的可溶性蛋白质及同工酶多态性

利用聚丙烯酰胺凝胶电泳技术,对采自北方玉米主产区的23个玉米灰斑病菌菌株进行可溶性蛋白质和同工酶电泳图谱分析及聚类分析,从蛋白质和酶学的多态性水平上分析玉米灰斑病菌的生理分化特征.研究表明,玉米灰斑病菌在可溶性蛋白质和SOD、MDH、PPO、POD、EST、CAT等的同工酶谱存在差异,不同菌株之间某些同工酶谱带数和同一迁移率谱带的亮度和色泽差异非常显著,说明菌株间的多态性可在同工酶水平上得到反映.研究还发现,来自不同地区的菌株同工酶谱带无明显的变化规律,反映出病菌同工酶的变异与地理位置关系不密切,也表明该病菌可能具有较广泛的地域适应性.     

茭白胡麻叶斑病病原菌分离与鉴定

茭白胡麻叶斑病发生普遍且严重,已成为茭白的常见病害。为了明确病原菌,先后从茭白主产区采集胡麻叶斑病病叶,各产区分离纯化5～10株病原菌。通过对菌株DNA的内转录间隔区ITS基因、3-磷酸甘油醛脱氢酶GPDH基因、延伸因子EF-1α基因的序列进行分析。结合形态学观察、致病力测定等方法对茭白胡麻叶斑病病菌进行鉴定。结果表明,茭白胡麻叶斑病病原菌为稻平脐蠕孢Bipolaris oryzae,与水稻胡麻叶斑病病原菌一致。本文的研究成果可为茭白胡麻叶斑病的防治提供一种新的思路。     

我国北方玉米上平脐蠕孢属和弯孢属真菌及其所致叶斑病

平脐蠕孢属(Bipolaris)和弯孢属(Curvularia)真菌可引起多种玉米叶斑病。为了解当前玉米生产上此类病害的发生情况,2014年8-9月对我国玉米主产区北京、河北、河南、黑龙江和吉林5省市玉米上疑似由该两属真菌引起的叶部病斑样品进行了采集,随后进行了真菌的分离和鉴定。共采集样品42份,根据其形状特点归为4类:长条形、椭圆形、小点状和梭形病斑。经组织分离获得平脐蠕孢属和弯孢属真菌28株,基于形态学和rDNA-ITS序列的系统发育分析共鉴定出5个种:玉蜀黍平脐蠕孢(B.maydis)、玉米平脐蠕孢(B.zeae)、玉米生平脐蠕孢(B.zeicola)、新月弯孢(C.lunata)和穂状弯孢(C.spicifera)。从长条形病斑和椭圆形病斑上分离到的主要是B.maydis和B.zeicola,从小点状病斑分离到的主要是C.lunata,其次是B.zeae。分离出C.lunata的样品病斑较为稀疏、颜色略浅、呈苍白色,分离出B.zeae的样品病斑更为密集、颜色较深。从梭型病斑分离到的是C.spicifera。有少数样品可分离到上述两种菌。采用孢子悬浮液喷雾法对温室玉米苗接种,上述5种真菌均可致病。以接种B.maydis发病最快,发病最重;接种B.zeicola、C.lunata或C.spicifera发病较慢,症状明显;接种B.zeae发病最慢,仅引起小点状病斑。研究结果可为玉米叶斑病的正确诊断提供资料和依据。     

引起我国甜樱桃叶斑病的间座壳菌种类鉴定

为明确引起我国甜樱桃叶斑病的病原菌种类,于2019年在北京市、四川省、山东省和辽宁省对与已报道的其他叶斑病症状明显不同的一种甜樱桃叶斑病进行调查并采集病叶,通过组织分离法进行病原菌分离纯化,结合形态学特征和多基因(ITS、EF1-α、TUB2和CAL)系统发育分析对病原菌的种类进行鉴定,并采用离体叶片接种法进行致病性测定。结果显示,新发现的这种甜樱桃叶斑病通常从叶边缘产生圆形或不规则灰褐色病斑,病斑周围具黄色晕圈,有时形成穿孔。从病样中共分离纯化得到10株单孢系菌株,结合其菌落形态、分生孢子形态和大小等,以及多基因系统发育分析结果,最终将10株菌株鉴定为甜樱间座壳Diaporthe eres(5株)和大豆间座壳D. sojae(5株),致病性测定结果显示甜樱间座壳和大豆间座壳对甜樱桃叶片均具有致病性,完成柯赫氏法则验证。     

山东省生姜细菌性黄叶斑病病原鉴定

Ginger (Zingiber officinale Roscoe) is an important economic crop in China. Bacterial leaf spot disease is the major limiting factor for the ginger crop production in Shandong. In recent years, a bacterial leaf spot disease was observed on ginger in Laiwu and Anqiu, that became a major limiting factor in ginger production. Yellowish bacterial colonies that were obtained after incubation at 28°C for 2 days from infected leaves were purified by streaking on nutrient agar. The purified isolates were initially identified as Pantoea ananatis by the phenotypic and biochemical characterization. Further molecular identification based on the 16S rDNA sequence analysis and multilocus sequence analysis with the gyrB, rpoB, and atpD sequences confirmed that the isolates were P. ananatis. To our knowledge, this is the first report of bacterial leaf spot of ginger caused by P. ananatis in Shandong. Our study will provide basic references for disease management of this pathogen.     

燕麦叶斑病病原菌鉴定及其生物学特性

为明确内蒙古、河北地区燕麦叶斑病病原菌种类及其生物学特性,采用组织分离法对病原菌进行分离,通过形态学特征及18SrDNA序列分析进行分类鉴定,以离体和活体叶片接种进行致病性测定,并采用十字交叉法对病原菌的生物学特性进行研究。结果表明,共分离到3株菌株R1、H44和B8,根据形态学特征和18SrDNA序列分析将3株菌株均鉴定为燕麦内脐蠕孢菌Drechslera avenacea(M.A.Curtis ex Cooke)Shoemaker,有性态为燕麦核腔菌Pyrenophora avenae S.Ito et Kurib.。3株菌株均为致病菌,在离体和活体叶片上均能产生腐烂坏死病斑。3株菌株的菌丝在5~30℃、pH 5~11范围内均可生长,其中菌株R1和H44的最适生长温度为25℃、最适生长pH为8,菌株B8的最适生长温度为20~25℃、最适生长pH为7~8。3株菌株对碳源的利用效果中均以淀粉最好,对乳糖利用效果最差;菌株R1的最适氮源是硝酸铵,对蛋白胨和尿素利用效果最差;菌株H44和B8的最适氮源是蛋白胨,菌株H44对硫酸铵利用效果较差,3株菌株都不能利用碳酸铵,且菌株B8也不能利用尿素。     

Morphological and molecular diversity of Colletotrichum spp. causing pepper spot and anthracnose of lychee (Litchi chinensis) in Australia

Since the 1980s a new disease has been affecting Australian lychee. Pepper spot appears as small, black superficial lesions on fruit, leaves, petioles and pedicels and is caused by Colletotrichum gloeosporioides, the same fungus that causes postharvest anthracnose of lychee fruit. The aim of this study was to determine if a new genotype of C. gloeosporioides is responsible for the pepper spot symptom. Morphological assessments, arbitrarily‐primed PCR (ap‐PCR) and DNA sequencing studies did not differentiate isolates of C. gloeosporioides from anthracnose and pepper spot lesions. The ap‐PCR identified 21 different genotypes of C. gloeosporioides, three of which were predominant. A specific genotype identified using ap‐PCR was associated with the production of the teleomorph in culture. Analysis of sequence data of ITS and β‐tubulin regions of representative isolates did not group the lychee isolates into a monophyletic clade; however, given the majority of the isolates were from one of three genotypes found using ap‐PCR, the possibility of a lychee specific group of C. gloeosporioides is discussed.     

蓝莓叶斑病病原菌鉴定及其生物学特性

为明确山东省蓝莓叶斑病病原菌的种类及其生物学特性,采用组织分离法获得菌株B1,通过形态学特征并结合r DNA-ITS序列分析对该菌株B1进行鉴定,以离体和活体叶片接种法测定菌株B1的致病性,并对其生物学特性进行研究。结果表明,菌株B1菌落呈白色绒毛状,背面淡黄色,分生孢子纺锤形,5个细胞,中间3个细胞为褐色,具有2～4根顶端附属丝;菌株B1的ITS序列与GenBank中棒状拟盘多毛孢Pestalotiopsis clavispora的相似性达99%以上,结合形态特征与rDNAITS序列分析将病原菌鉴定为棒状拟盘多毛孢,GenBank登录号为MG009201。菌株B1接种蓝莓叶片后产生褐色病斑,后期密生黑色分生孢子盘,与蓝莓田间自然发病症状一致。该菌株在5～35℃、pH 5～11范围内均可生长,最适温度范围为25～30℃,最适pH范围为5～9;但仅在25～30℃时产生分生孢子,最适产孢温度为25℃;光照条件对菌丝生长无明显影响,但连续黑暗有利于产孢;以葡萄糖为碳源时利用率最高,对乳糖利用效果最差;最适氮源为硝酸钠、硫酸铵和蛋白胨,几乎不能利用尿素。     

湖北省黄连叶斑病病原鉴定及其生物学特性

为明确湖北省黄连叶斑病病原菌种类及其生物学特性,采用常规组织分离法获得病原菌菌株HL-1,通过形态学特征并结合基于ITS序列分析对菌株进行鉴定,并研究该病原菌的生物学特性,用生长速率法测定了6种杀菌剂对该病原菌菌丝生长的抑制作用。结果表明,黄连叶斑病病原菌分生孢子器黑褐色,大小为100.5~120.0μm×175.3~210.0μm;分生孢子无色,单孢,大小为2.0~2.5μm×4.1~5.4μm;该菌株ITS序列系统发育进化分析结果表明,其与耧斗菜茎点霉Phoma aquilegiicola(HM222537.1)的同源性为100%,通过形态学特征及分子鉴定结果确定该菌株为耧斗菜茎点霉。该病原菌在4~25℃内均能生长,最适温度为20℃;不同光照条件对病原菌生长的影响无显著差异;在供试碳(氮)源中病原菌对麦芽糖和蛋白胨的利用效果最好;菌丝的致死温度为50℃、10 min。供试6种药剂中10%苯醚甲环唑对病原菌菌丝生长的抑制效果最好,其EC_(50)最小,为0.03μg/mL。     

广西香蕉真菌性叶斑病病原菌种群结构分析

2009年2-3月对广西香蕉主产区真菌性叶斑病病原进行抽样鉴定。结果显示,广西香蕉真菌性叶斑病病原至少有10种,主要病原为Cordana musae,分布广,检出率为56.77%~92.19%;Corynespora cassiicola和Deightoniella torulosa为次,两种病原菌均主要集中在南宁地区,检出率分别为32.34%和22.61%;叶斑病类型以单一病原侵染为主,检出率为78.13%~92.19%;复合侵染的叶斑病类型以2种病原共同侵染居多。     

越南槐圆斑病病原鉴定

豆科槐属多年生植物越南槐(Sophora tonkinensis Gagnep.)是我国传统中药材山豆根的基源植物,药用其干燥根及根茎,具有清热解毒、消肿利咽的功效,在历版《中华人民共和国药典》中均有收载[1].广西是山豆根药材的道地产区[2],近年来随着山豆根市场需求的日益增大,越南槐在广西的种植面积由最初的60多公...     

Phenotypic,molecular and pathogenic characterization of Phlyctema vagabunda,causal agent of olive leprosy

Olive leprosy, caused by the fungus Phlyctema vagabunda, is a classic fruit rot disease widespread in the Mediterranean basin. From 2009 to 2013, new disease symptoms consisting of small circular necrotic leaf lesions, coin branch canker and shoot dieback were observed in Spanish and Portuguese olive orchards showing intense defoliation. Phlyctema‐like anamorphs were consistently isolated from leaves and shoots with symptoms. Representative isolates from affected leaves, shoots and fruits were characterized based on morphology of colonies and conidia, optimum growth temperature and comparison of DNA sequence data from four regions: ITS, tub2, MIT and rpb2. In addition, pathogenicity tests were performed on apple and olive fruits, and on branches and leaves of olive trees. Maximum mycelial growth rate ranged between 0.54 and 0.73 mm per day. Conidia produced on inoculated apple fruits showed slight differences in morphology among the representative fungal isolates evaluated. Phylogenetic analysis clustered all of the Phlyctema‐like isolates in the same clade, identifying them as Phlyctema vagabunda. On fruits, influence of wounding, ripening and cultivar resistance was studied, with cv. Blanqueta being the most susceptible cultivar. On branches, a mycelial‐plug inoculation method reproduced olive leprosy symptoms and caused shoot dieback. On leaves, Koch's postulates were fulfilled and the pathogen caused characteristic necrotic spots and plant defoliation. This is the first time that the pathogenicity of P. vagabunda in olive leaves has been demonstrated.     

Incidence,pathogenicity and diversity of Alternaria spp. associated with alternaria leaf spot of canola (Brassica napus) in Australia

Studies were undertaken to determine Alternaria spp. associated with leaf spot symptoms on canola (Brassica napus) in two cropping seasons (2015, 2016) across southern Australia. Major allergen Alt a1 and plasma membrane ATPase genes were used to identify Alternaria spp. In 2015, 112 isolates of seven Alternaria spp. were obtained, with A. metachromatica predominating. In 2016, 251 isolates of 12 Alternaria spp. were obtained, with A. infectoria predominating. Alternaria spp. isolates were morphologically and phylogenetically identified and studies to determine their pathogenicity on both B. napus (cv. Thunder TT) and B. juncea (cv. Dune) confirmed 10 species (A. alternata, A. arborescens, A. brassicae, A. ethzedia, A. hordeicola, A. infectoria, A. japonica, A. malvae, A. metachromatica and A. tenuissima) as pathogenic on both Brassica species. Alternaria ethzedia, A. hordeicola and A. malvae were recorded for the first time in Australia on any host and the record of A. arborescens was the first for New South Wales (NSW) and South Australia (SA). Other first records included A. infectoria on B. napus in NSW; A. japonica on B. napus in NSW and Western Australia (WA); A. metachromatica on any host in NSW, Victoria (VIC), WA and SA; and A. tenuissima on B. napus in NSW, SA and WA. It is evident that alternaria leaf spot on canola across southern Australia is not solely caused by A. brassicae, but that a range of other Alternaria spp. are also involved to varying degrees, depending upon the year and the geographic locality.     

引起江苏茅山地区宣木瓜叶斑病的病原鉴定

<正>原产于安徽省宣城市的地道中药材宣木瓜(Chaenomeles sinensis)隶属蔷薇科(Rosaceae),为常用中药木瓜的上品,是与藏红花、甘枸杞、川杜仲齐名的四大名贵中药材之一。除药用之外,近年也被用于化妆品和保健品领域[1]。此外,由于树冠优美、花团锦簇,宣木瓜还被栽培用于观赏;同时,因其耐旱耐瘠,对土壤要求不高,又常被作为退耕还林的引种树种。因此,宣木瓜是一种集经济、生态、社会等多种效益于一身的优良树种,被广泛种植于安徽、云     

引起江苏茅山地区宣木瓜叶斑病的病原鉴定

A new leaf disease on Chinese quince (Chaenomeles sinensis) was recently noted in Mount Maoshan, Jurong City, Jiangsu Province, China. Symptoms began with irregular lesions on tips or margins of the leaves, which gradually developed into concentric rings. The lesions eventually spread to cover one half or the entire leaf, leading to eventual desiccation and defoliation. Disease survey and responsible agent isolation was conducted in this study. Based on morphological characteristics, combined gene sequence analysis of ITS, G3PDH, HSP60 and RPB2, and Koch’s postulation, the causal agent of this new disease was identified as Botrytis cinerea.     

青海省樱桃叶斑病病原菌的分离与鉴定

为明确引起青海省樱桃叶斑病病原菌的种类,本研究从西宁市城北区、海东市乐都区和贵德县的樱桃上采集有叶斑症状的叶片,采用组织分离法分离出病原菌,通过观察病原菌的形态特征,结合rDNA-ITS、EF-1α和Alt a 1基因序列分析对病原菌的种类进行了鉴定,并用柯赫氏法则进行验证。结果表明,共获得279株病原菌菌株,分属链格孢Alternaria alternata、细极链格孢A.tenuissima和刺盘孢属Colletotrichum spp.,分离频率分别为84.95%、5.02%和10.03%。采用柯赫氏法则进行离体叶片致病性测定,接种叶片100%发病,说明引起青海省樱桃叶斑病的病原菌为链格孢、细极链格孢和刺盘孢属真菌,且链格孢为主要病原菌。该研究结果可为青海省樱桃叶斑病的综合防治提供理论依据。