我国部分省域甘蓝主要产区黑腐病菌鉴定及致病力分析

 为了明确我国甘蓝主栽区黑腐病菌株种类与致病力水平,为优化抗病性鉴定方法、筛选抗病品种及抗病品种的合理布局提供依据,分别采集了10个省域的黑腐病病叶,经过分离纯化与分子鉴定,获得了26个典型甘蓝黑腐病菌株;选用5个甘蓝抗、感品种作为鉴别寄主,以甘蓝黑腐病菌株的悬浮液为接种体,采用喷雾接种法测定致病力。结果表明:来源于不同省域的黑腐病菌株致病力存在显著差异,致病力最强的是YU,病情指数平均值为38.31;致病力最弱的是SH,病情指数平均值为17.03。来源于不同海拔高度的甘蓝黑腐病菌株致病力最强的是G1500,病情指数平均值为37.93;最弱的是G800,病情指数平均值为19.80。根据鉴别寄主抗感反应将26个黑腐病菌株划分为12类致病型,发现来自不同地域的甘蓝黑腐病菌株间存在着显著的致病性分化现象,对甘蓝品种的致病力存在着显著性差异,其中菌株YU致病力最强,菌株YU可作为甘蓝抗病性鉴定的主要致病菌株,指导抗病品种的选育。     

我国部分省域甘蓝主要产区黑腐病菌鉴定及致病力分析

 为了明确我国甘蓝主栽区黑腐病菌株种类与致病力水平,为优化抗病性鉴定方法、筛选抗病品种及抗病品种的合理布局提供依据,分别采集了10个省域的黑腐病病叶,经过分离纯化与分子鉴定,获得了26个典型甘蓝黑腐病菌株;选用5个甘蓝抗、感品种作为鉴别寄主,以甘蓝黑腐病菌株的悬浮液为接种体,采用喷雾接种法测定致病力。结果表明:来源于不同省域的黑腐病菌株致病力存在显著差异,致病力最强的是YU,病情指数平均值为38.31;致病力最弱的是SH,病情指数平均值为17.03。来源于不同海拔高度的甘蓝黑腐病菌株致病力最强的是G1500,病情指数平均值为37.93;最弱的是G800,病情指数平均值为19.80。根据鉴别寄主抗感反应将26个黑腐病菌株划分为12类致病型,发现来自不同地域的甘蓝黑腐病菌株间存在着显著的致病性分化现象,对甘蓝品种的致病力存在着显著性差异,其中菌株YU致病力最强,菌株YU可作为甘蓝抗病性鉴定的主要致病菌株,指导抗病品种的选育。     

花生网斑病不同病斑类型及其病原菌致病力差异

本文描述了河南省发生日趋严重的花生网斑病在田间出现的一种新的病斑类型,并结合已报道的两种病斑类型,对3种病斑类型的病原菌开展致病力差异研究。研究结果表明:田间新发现的花生网斑病病斑特征明显,综合分析后将花生网斑病病斑类型分别定为网纹型、类褐斑型和污斑型;同一花生品种上3种病斑类型病原菌菌株间存在明显的致病力差异,同一病斑类型菌株对不同花生品种的致病力也表现出显著差异。     

水稻基腐病细菌和玉米茎腐病细菌的比较研究

 对水稻基腐病细菌(18个菌株)、玉米茎腐病细菌和菊欧氏杆菌的6个致病变种的标准菌株及胡萝卜软腐欧氏杆菌的两个致病变种(Erwinia carotovora pv.carotovora和E.c.pv.atroseptica)进行了58项生化试验的比较。结果水稻菌株在生化性状上与玉米菌株十分相似,最接近菊欧氏杆菌玉米致病变种(E.chrysanthemi pv.zeae),但水稻基腐病细菌在兰色素产生,丙二酸、酒石酸钠利用,产氨和过氧化氢酶活性等项上与玉米致病变种不同。在致病性比较中,水稻菌株比玉米菌株致病力强,寄主范围广。可能是一个独特的致病变种。全细胞蛋白聚丙烯酰胺圆盘电泳的研究表明水稻菌株的蛋白电泳带数与玉米菌株相同,均为16条,但电泳带的迁移率互不相同。血清学研究表明水稻基腐病细菌和玉米致病变种没有共同抗原。     

玉米大斑病菌黑色素缺失突变体的获得及其生物学特性

DHN黑色素是许多植物病原真菌的致病相关因子,为明确黑色素在玉米大斑病菌致病过程中的作用,采用三环唑和紫外线复合诱变的方法获得了黑色素缺失的6个突变菌株,对突变菌株的生长量、产孢量、HT-毒素活性、致病力进行测定。结果显示,突变菌株产毒能力变化不大,但生长量下降,产孢量和致病力显著下降或完全丧失,野生型菌株的产孢量和侵染频率分别是突变菌株的30～50倍和5倍。突变菌株与黑色素共培养后,菌株M01-23a和M01-23b恢复了致病力,但病斑面积和侵染频率较小。玉米大斑病菌黑色素与致病力具有一定的相关性。     

东北地区烟草靶斑病菌(Rhizoctonia solani)融合群、致病力分化及品种抗病性研究

2013-2014年吉林省、黑龙江省烟区首次大面积发生烟草靶斑病,损失严重。采集吉林省柳河县、黑龙江省林口县和宾县的烟草靶斑病病叶进行分离纯化,得到了12个菌株。将所获菌株分别与立枯丝核菌标准融合群菌株AG-2、AG-3、AG-4进行对峙培养,结果表明:此12个菌株均与立枯丝核菌AG-3标准菌株发生融合反应,不与其他融合群发生融合;随机选取3个县各1个代表性菌株,提取菌丝基因组DNA并对其ITS序列进行分析与比对,菌株HLJ-2、JL-1、HLJ-7的ITS序列与辽宁省烟草靶斑病菌菌株LF-2、LJT-8(AG-3融合群)的同源性达99%~100%,因此推断,吉林省、黑龙江省烟草靶斑病菌与辽宁省烟草靶斑病菌应属于同一个融合群,即立枯丝核菌AG-3融合群(Rhizoctonia solani AG-3)。根据各菌株在鉴别寄主上表现的抗性不同,可将吉林省、黑龙江省所有菌株分为3个致病类型,即致病类型Ⅰ、致病类型Ⅱ和致病类型Ⅲ。同一省份不同菌株间致病力有差异,黑龙江省菌株3种致病类型均有,以致病类型Ⅱ为主;吉林省菌株只有致病类型Ⅰ和致病类型Ⅱ。分别选取3个省的强致病力菌株,用离体叶片接种法鉴定了我国21个主栽烟草品种对靶斑病的室内抗病性,结果表明:没有免疫或抗病品种,‘龙江981’、‘NC297’对供试的3个强致病力菌株均表现中抗,‘中烟202’、‘云烟97’、‘NC55’、‘KRK26’和‘G28’分别对不同菌株表现中抗。     

玉米茎腐病菌毒素致病力初报

通过提取玉米茎腐病菌毒素粗提液,证实玉米茎腐病原菌(镰刀菌和腐霉菌为主)在镰刀菌毒素培养液、查氏培养液和普通培养液中,能产生和玉米小斑病菌毒素(简称HM毒素,下同)类似的致病毒素。该物质能抑制种子根的生长。接种在玉米叶片上能产生典型的萎蔫枯死斑;用上述3种培养液培养病原菌,产生的毒素致病力差异不显著;不同类型病原菌产生的毒素致病力和同一类型病原菌不同菌株产生的毒素致病力差异都极显著。玉米品系对茎腐病原菌产生的毒素抗病性差异也极显著。     

稻瘟菌致病力变异初探

 从保存的4个单孢分离菌后代再分离若干单胞菌株,接种子国内7个鉴别品种上,又可以分成不同的小种。从6个单孢分离菌再分离的后代单孢菌株,分别对13个抗病品种作致病力测定,各后代单孢菌株仅对少数品种的致病力是一致的。从27个品种的小病斑或个别大病斑上分离的病菌,回接原品种后,有11个分离菌对原品种不致病。这些结果说明大多数分离菌对许多品种的致病力是易变的,只对少数品种的致病力保持相对稳定性。作者认为进行抗性遗传和抗性机制研究需要的致病力稳定分离菌,只能根据其对供试品种致病力的稳定性来选择,而不能按一组鉴别品种来选择。     

陕西省小麦茎基腐病病原菌鉴定及其致病力分析

为明确陕西省小麦茎基腐病病原菌的组成及其致病力,于2019年小麦灌浆期在陕西省宝鸡市、咸阳市、西安市和渭南市共63个采样点采集具有茎基腐病典型症状的病株,对其进行组织分离,结合培养性状和形态学特征,以及利用ITS和EF-1α序列分析鉴定分离菌株所属类群,并对不同病原菌进行致病力分析。结果表明：共分离到224株菌株,基于形态学特征和分子生物学鉴定结果,其中201株菌株鉴定为假禾谷镰刀菌Fusarium pseudograminearum,3株菌株鉴定为禾谷镰刀菌F. graminearum,14株菌株鉴定为三线镰刀菌F. tricinctum,6株菌株鉴定为层出镰刀菌F. proliferatum,其中假禾谷镰刀菌占总分离菌株的89.73%,是陕西省小麦茎基腐病的优势病原菌。不同种的镰刀菌对小麦植株均有一定的致病性,但假禾谷镰刀菌的致病力最强,而且分离自不同地区或者同一地区的假禾谷镰刀菌株间的致病力也存在一定分化。     

2019—2020年河南省小麦茎基腐病病原菌鉴定及致病力测定

为监测小麦茎基腐病病原菌的变化,2019—2020年自河南省安阳、濮阳和鹤壁等17个市129个采样点采集小麦茎基腐病病株样品并分离病原菌,对所有分离物进行形态学鉴定,利用特异性引物、ITS和EF-1α基因序列进行分子生物学鉴定,并于室内对优势病原菌菌株进行致病力测定。结果表明,共分离得到892株镰孢菌Fusarium分离物,所得分离物中假禾谷镰孢F. pseudograminearum共851株,所占比例为95.41%,为优势种;禾谷镰孢F. graminearum共16株,所占比例为1.79%。除信阳市外,其他16个市优势病原菌均为假禾谷镰孢。不同假禾谷镰孢菌株存在明显的致病力分化,但多数为强致病力菌株。     

Genetic diversity and virulence of Xanthomonas campestris pv. campestris isolates from Brassica napus and six Brassica oleracea crops in Serbia

The present study provides insight into the diversity of 147 Xanthomonas campestris pv. campestris (Xcc) isolates obtained from six Brassica oleracea vegetable crops (broccoli, cabbage, cauliflower, collard greens, kale, kohlrabi) and the winter oilseed rape crop Brassica napus, collected from different regions in Serbia in 2014. The XCF/XCR pathovar-specific primer set was used for fast preliminary identification. In repetitive sequence-based PCR (BOX, ERIC and REP) of all isolates, a higher level of genetic diversity was found in winter oilseed rape isolates compared to isolates from the other hosts. ERIC and REP-PCR showed the highest heterogeneity, with 10 and nine banding patterns, respectively. The REP-PCR results showed the highest correlation (70%) with those obtained with multilocus sequence analysis (MLSA), performed with 10 housekeeping genes (fusA, gap-1, gltA, gyrB1, lacF, lepA, rpoD, dnaK, fyuA and gyrB2). Three distinct phylogenetic groups of winter oilseed rape isolates were detected using MLSA. Two genes, gltA and rpoD, showed the greatest ability to identify and discriminate winter oilseed rape Xcc isolates from isolates of the other six hosts. The lepA gene exhibited specific three-nucleotide changes in sequences of some of the isolates. Results of virulence testing of 18 representative isolates showed statistically significant host–pathogen specialization for Xcc isolates from winter oilseed rape, cauliflower, kale and kohlrabi. In conclusion, oilseed rape isolates are more genetically diverse and show greater specialization to their host in comparison to the rest of the tested isolates from other brassica hosts.     

Are herbicide‐resistant crops the answer to controlling Cuscuta?

BACKGROUND: Herbicide‐resistant crop technology could provide new management strategies for the control of parasitic plants. Three herbicide‐resistant oilseed rape (Brassica napus L.) genotypes were used to examine the response of attached Cuscuta campestris Yuncker to glyphosate, imazamox and glufosinate. Cuscata campestris was allowed to establish on all oilseed rape genotypes before herbicides were applied. RESULTS: Unattached seedlings of C. campestris, C. subinclusa Durand & Hilg. and C. gronovii Willd. were resistant to imazamox and glyphosate and sensitive to glufosinate, indicating that resistance initially discovered in C. campestris is universal to all Cuscuta species. Glufosinate applied to C. campestris attached to glufosinate‐resistant oilseed rape had little impact on the parasite, while imazamox completely inhibited C. campestris growth on the imidazolinone‐resistant host. The growth of C. campestris on glyphosate‐resistant host was initially inhibited by glyphosate, but the parasite recovered and resumed growth within 3–4 weeks. CONCLUSION: The ability of C. campestris to recover was related to the quality of interaction between the host and parasite and to the resistance mechanism of the host. The parasite was less likely to recover when it had low compatibility with the host, indicating that parasite‐resistant crops coupled with herbicide resistance could be highly effective in controlling Cuscuta. Published 2009 by John Wiley & Sons, Ltd.     

Detection methods for Xanithomonas campestris pv. graminis on forage grasses1

In Norway seven different grass species were found to be attacked by Xanthomonas campestris pv. graminis (bacterial wilt of forage grasses). Among these, Phleum pratense is the most susceptible and common host. Considerable damage may occur after long periods of hot and dry weather, while in cool and wet periods hardly any diseased plants are found. The pathogen may be identified by isolation and biochemical tests. Specific antibodies used in immunofluorescence and ELISA had a high degree of sensitivity and specificity against the target bacterium. The two methods were used for screening pure cultures and detecting bacteria directly in plant tissue extracts. Their application revealed the presence of low numbers of bacteria in symptomless plants and a discontinuous distribution within the plant.     

Detection and identification of the crucifer pathogen, <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> pv. <Emphasis Type="Italic">campestris</Emphasis>, by PCR amplification of the conserved Hrp/type III secretion system gene <Emphasis Type="Italic">hrcC</Emphasis>

The development of a rapid detection method for Xanthomonas campestris pv. campestris (Xcc) in crucifer seeds and plants is essential for high-throughput certification purposes. Here we describe a diagnostic protocol for the identification/detection of Xcc by PCR amplification of fragments from the pathogenicity-associated gene hrcC. Under stringent conditions of amplification, a PCR product of 519 bp from hrcC was obtained from a collection of 46 isolates of Xcc, with the exception of two isolates from radish. No amplicons were obtained from 39 pure cultures of the phytopathogenic bacteria Xanthomonas campestris pv. cerealicola, X. campestris pv. juglandis, X. campestris pv. pelargonii, X. campestris pv. vitians, X. arboricola pv. pruni, X. axonopodis pv. phaseoli, X. axonopodis pv. vesicatoria, X. vesicatoria, Pseudomonas syringae pv. phaseolicola, P. syringae pv. syringae, P. syringae pv. tomato, P. fluorescens, P. marginalis, Pectobacterium atrosepticum, P. carotovorum subsp. carotovorum. In addition, PCR reactions were negative for fifty unidentified environmental isolates purified from the surface of crucifers. The PCR fragment was obtained from four strains previously classified as X. campestris pv. aberrans, X. campestris pv. armorociae, X. campestris pv. barbarae and X. campestris pv. incanae using pathogenicity assays. Our PCR protocol specifically detected Xcc in inoculated leaves, seeds and naturally infected leaves of crucifers.     

Phenotypic and Genotypic Characterization of Xanthomonas campestris pv. zinniae Strains

During 1997 and 1998, serious outbreaks of bacterial leaf spot disease were observed on zinnia plants grown in home and commercial gardens in Ohio, USA. Twenty-two strains of Xanthomonas campestris pv. zinniae, isolated from diseased zinnia plants and contaminated seeds, were identified based on morphological, physiological and biochemical tests, fatty acid methyl ester analyses and pathogenicity tests on zinnia cv. Scarlet. Host range studies indicated that all of the X. campestris pv. zinniae strains were pathogenic on zinnia and tomato, but not on cabbage, lettuce, pepper and radish. The phenotypic and genotypic relationships among the strains determined based on serological reaction pattern, fatty acid profiles, repetitive extragenic palindromic-polymerase chain reaction (rep-PCR) fingerprints and sequence analysis of the 16S–23S rDNA spacer region suggested that X. campestris pv. zinniae strains were closely related to each other, but clearly distinct from other Xanthomonas species including X. campestris pv. campestris, X. axonopodis pv. vesicatoria, X. vesicatoria and X. hortorum pv. vitians tested in this study. The results also demonstrated that rep-PCR fingerprinting is rapid, reliable and the most practical method for routine detection and identification of X. campestris pv. zinniae strains.     

Inference of the phylogenetic diversity and population structure of Xanthomonas campestris affecting Brassicaceae using a multilocus sequence typing‐based approach

Xanthomonas campestris pathovars are widely distributed throughout the globe and have a broad host range, causing severe economic losses in the food and ornamental crucifers markets. Using an approach based on multilocus sequence typing, phylogenetic diversity and population structure of a set of 75 Portuguese and other Xanthomonas campestris isolates from several cruciferous hosts were assessed. Although this population displayed a major clonal structure, neighbour‐net phylogenetic analysis highlighted the presence of recombinational events that may have driven the ecological specialization of X. campestris with different host ranges within the Brassicaceae family. A high level of genetic diversity within and among X. campestris pathovars was also revealed, through the establishment of 46 sequence types (STs). This approach provided a snapshot of the global X. campestris population structure in cruciferous host plants, correlating the existing pathovars with three distinct genetic lineages. Phylogenetic relationships between the founder genotype and remaining isolates that constitute the X. campestris pv. campestris population were further clarified using goeBURST algorithm. Identification of an intermediate link between X. campestris pv. campestris and X. campestris pv. raphani provided new insights into the mechanisms driving the differentiation of both pathovars. Wide geographic distribution of allelic variants suggests that evolution of X. campestris as a seedborne pathogen was not shaped by natural barriers. However, as Portuguese isolates encompass 26 unique STs and this country is an important centre of domestication of Brassica oleracea crops, a strong case is made for its role as a diversification reservoir, most probably through host–pathogen coevolution.     

Interactions in the Brassica napus–Pyrenopeziza brassicae pathosystem and sources of resistance to P. brassicae (light leaf spot)

Pyrenopeziza brassicae, cause of light leaf spot (LLS), is an important pathogen of oilseed rape and vegetable brassicas and has a wide geographic distribution. Exploitation of host resistance remains the most sustainable and economically viable solution for disease management. This study evaluated 18 oilseed rape cultivars or breeding lines for host resistance against P. brassicae in glasshouse experiments. Selected cultivars/lines were inoculated with eight single-spore isolates of the pathogen obtained from three different regions in England. Analysis of P. brassicae infection-related changes on host plants identified leaf deformation as a characteristic feature associated with P. brassicae infection, this showed poor correlation to LLS severity measured as the amount of pathogen sporulation on infected plants. Resistant host phenotypes were identified by limitation of P. brassicae sporulation, with or without the presence of a necrotic response (black flecking phenotype). Investigation of this pathosystem revealed significant differences between cultivars/lines, between isolates, and significant cultivar/line-by-isolate interactions. In total, 37 resistant and 16 moderately resistant interactions were identified from 144 cultivar/line-by-isolate interactions using statistical methods. Most of the resistant/moderately resistant interactions identified in this study appeared to be nonspecific towards the isolates tested. Our results suggested the presence of isolate-specific resistant interactions for some cultivars. Several sources of resistance have been identified that are valuable for oilseed rape breeding programmes.     

Characterization of isolates that cause black rot of crucifers in East Africa

A study was conducted in the East African countries of Kenya, Tanzania and Uganda in the months of July and August 2009 with the objectives of assessing the status of black rot and race structure of Xanthomonas campestris pv. campestris in the three countries. Samples infected with black rot were collected from farmers’ fields mainly from Brassica oleracea crops (broccoli, cabbage, cauliflower and kales). A total of 399 farms were surveyed of which 260 were from Kenya, 91 from Tanzania and 48 from Uganda. Following successful isolations, a total of 249 isolates of the causal agent, Xanthomonas campestris pv. campestris were recovered. Pathogenicity of all isolates was confirmed on B. oleracea susceptible cultivars Copenhagen Market F1 and Wirosa F1. Sixty of the 250 isolates were race-typed using a differential set Brassica spp. Only two races, 1 (Kenya and Tanzania) and 4 (Kenya, Tanzania and Uganda) were observed however, another race (5) was observed from one isolate recovered from a B. rapa sample obtained from Tanzania in 2003. Genomic fingerprinting with repetitive-PCR revealed clusters that did not depict significant correlations between isolates and geographical location, isolates and host adaptation or isolates and race. However, it did demonstrate existence of genetic differences within the East African X. campestris pv. campestris population indicating that it is not a similar clonal population of the same genetic background.     

菜豆壳球孢菌与棉花的关系

用棉花的4个品种(Stoneville 825,CABCS′-1-81,LE~2BOS-1-82和Tamcot CAMD-E)来鉴定从大豆、棉花和花生(编号为MP7,MP9,MP14)上分离到的菜豆球壳孢菌(Macrophomina phaseolina)的致病力。3个分离物对4个棉花品种均能侵染。花生分离物(MP14)致病力较强。受侵棉花幼苗较健株矮。病菌不需经伤口就能侵入。在幼苗阶段,品种TamcotCAMD-E和CABCS′-1-81对MP14更易感病,而品种Stonevill825较其他3个品种抗病。说明品种之间对M.phaseolina的抗性有差别。受害棉株有时表现为死株而不落叶,不严重时肉眼看不出根部伤痕,但主根的中柱部份以及根、主茎下部的木髓部变色,从棕褐色到黑色。从病部能分离到该菌的菌核。     

Development of a specific molecular tool for detecting Xanthomonas campestris pv. musacearum

A specific and rapid diagnostic tool has been developed to detect Xanthomonas campestris pv. musacearum, the causal agent of bacterial wilt of banana. PCR primers were developed from intergenic regions of X. campestris pv. musacearum following its partial sequence. A total of 48 primers were tested for specificity to X. campestris pv. musacearum strains collected from various regions in Uganda. These were also tested for specificity against related Xanthomonas species from the vasicola group, Xanthomonas species pathogenic to other crops, and against those existing saprophytically on banana plants. Seven primer sets (Xcm12, Xcm35, Xcm36, Xcm38, Xcm44, Xcm47 and Xcm48) were found to be very specific to X. campestris pv. musacearum. These primer sets directed the amplification of the expected product for all 52 strains of X. campestris pv. musacearum collected from different locations in Uganda. No amplification products were obtained with unrelated phytopathogenic bacteria or endophytic/epiphytic bacteria from banana. A detection limit of 10³ CFU mL^?1 corresponding to about four cells per PCR reaction was observed when X. campestris pv. musacearum cells were used for all the seven primer sets. The DNA samples from symptomless plant tissues also tested positive with primer set Xcm38. The specific PCR method described here is a valuable diagnostic tool which can be used to detect the pathogen at early stages of infection and monitor disease.