Molecular characterization,comparison of screening methods,and evaluation of cross‐pathogenicity of black rot (Xanthomonas campestris pv. campestris) strains from cabbage,choy sum,leafy mustard and pak choi from Taiwan

Choy sum (Brassica rapa var. parachinensis), leafy mustard (Brassica juncea) and pak choi (B. rapa var. chinensis) are highly nutritious components of diets in Taiwan and other Asian countries, and bacterial black rot caused by Xanthomonas campestris pv. campestris (Xcc) is a major biotic constraint in these crops. As very little was known about the Xcc strains from these crops in these regions, including their cross‐pathogenicity and aggressiveness on different hosts, Xcc strains were obtained from cabbage (Brassica oleracea var. capitata), choy sum, leafy mustard and pak choi crops in Taiwan. Two previously published PCR‐based assays reliably distinguished the Xcc strains from other Xanthomonas species and subspecies. Phylogenetic analysis based on repetitive sequence‐based PCR assays placed the Xcc strains in a clade distinct from other Xanthomonas species, and also showed host specificity. Although all of the Xcc strains from the different host species were pathogenic on all five Brassica test species in both a detached leaf assay and an intact plant assay, in the intact plant assay they showed differences in virulence or aggression on the different test hosts. The Xcc strains from leafy mustard and pak choi were consistently highly aggressive on all the test host genotypes, but the strains from choy sum and cabbage were less aggressive on leafy mustard and choy sum. The intact plant assay proved more discriminating and reliable than the detached leaf assay for comparing the aggressiveness of Xcc strains on different host genotypes, and so, with the new Xcc strains isolated in this study, will be useful for screening leafy brassica germplasm accessions for resistance to black rot.     

Identification of Rhizoctonia solani AG 1-IB in Lettuce, AG 4 HG-I in Tomato and Melon, and AG 4 HG-III in Broccoli and Spinach, in Brazil

Fungi isolated in Brazil, from lettuce, broccoli, spinach, melon and tomato, were identified as Rhizoctonia solani. All lettuce isolates anastomosed with both AG 1-IA and IB subgroups and all isolates from broccoli, spinach, melon and tomato anastomosed with AG 4 subgroup HG-I, as well as with subgroups HG-II and HG-III. DNA sequence analyses of ribosomal internal transcribed spacers showed that isolates from lettuce were AG 1-IB, isolates from tomato and melon were AG 4 HG-I, and isolates from broccoli and spinach were AG 4 HG-III. The tomato isolates caused stem rot symptoms, the spinach, broccoli and melon isolates caused hypocotyl and root rot symptoms on the respective host plants and the lettuce isolates caused bottom rot. This is the first report on the occurrence in Brazil of R. solani AG 4 HG-I in tomato and melon, of AG 4 HG-III in broccoli and spinach and of AG 1-IB in lettuce.     

Characterisation of fungal pathogens causing basal rot of lettuce in Belgian greenhouses

Basal rot is a common disease in lettuce greenhouses. A 3-year study on the diversity of pathogens associated with basal rot in Belgium was carried out. A total of 150 isolates were collected originating from 56 greenhouses. Four pathogens appeared to be involved. Rhizoctonia solani was found to be the causal agent at 23 locations, Sclerotinia spp. at 14, Botrytis cinerea at 17 and Pythium spp. at seven. The isolates of R. solani were further characterised to anastomosis groups and subgroups using morphological characteristics, pectic zymogram and PCR-RFLP. Five anastomosis groups could be distinguished: AG1-1B, AG4 HGI, AG10, AG2-1, AG2-1 Nt and AG3, with isolates of AG4 HGI and AG1-1B being the most prevalent and the most aggressive. Sclerotinia sclerotiorum was found at 13 locations, while S. minor was found at only one location. Based on ITS-sequencing Pythium isolates were assigned to three different species. At 20°C, isolates of all pathogens were able to cause lesions on detached lettuce leaves, except isolates of R. solani AG3 and AG2-1 Nt. A correlation could be found between the occurrence of the pathogens and the growing season. Botrytis cinerea was the most common pathogen in winter, whereas R. solani was most frequently isolated in summer. Sclerotinia spp. and Pythium spp. were isolated in spring, summer and autumn. The information obtained in this study will be most useful in the development of an alternative control strategy for causal agents of basal rot.     

Induced resistance against Fusarium diseases of <Emphasis Type="Italic">Cymbidium</Emphasis> species by weakly virulent strain HPF-1 (<Emphasis Type="Italic">Fusarium</Emphasis> sp.)

The mechanism by which Fusarium diseases of cymbidium plants are suppressed by a weakly virulent strain HPF-1 of Fusarium sp. was studied. Strain HPF-1 produced microscopic, necrotic local lesions on cymbidium leaves, causing minor damage to palisade tissues at the infection sites. This weakly virulent strain remained near the site of infection and did not develop further. It systemically and nonselectively suppressed some diseases of cymbidium such as yellow spot of leaves caused by Fusarium proliferatum and F. fractiflexum, bulb and root rot caused by F. oxysporum, and dry rot of bulbs and roots caused by F. solani. Because endogenous salicylic acid levels increased in cymbidium leaves inoculated with strain HPF-1, the mechanism of disease suppression is thought to be systemic acquired resistance.     

Characterisation of Rhizoctonia solani Anastomosis Groups Causing Bottom Rot in Field-Grown Lettuce in Germany

Bottom rot caused by Rhizoctonia solani is an increasing problem in field-grown lettuce in Germany. During the growing seasons of 1999 and 2000, 95 isolates of R. solani from lettuce plants with bottom rot symptoms were collected from eight locations. The isolates were characterised using hyphal anastomosis, pectic zymograms and morphological characteristics. Ninety-three isolates were identified as anastomosis group (AG) 1-IB, one as AG 1-IC and one as AG 2-1. Optimum hyphal growth was measured over a temperature range of 20–30 °C with an optimum at 25 °C. Aggressiveness of the AG 1-IB isolates varied from weak to strong when tested on detached lettuce leaves. The pathogenic potential of six AG 1-IB isolates was determined on 14 plant species in comparison with lettuce under conditions favourable for the fungus. Radish, broccoli, kohlrabi, spinach and millet seedlings were as severely infected as lettuce seedlings. The same isolates caused little symptoms on maize, tomato and onion. Knowledge about the host range of AGs of R. solani are important for planning an effective crop rotation as part of a control management system.     

A novel pathosystem to study the interactions between <Emphasis Type="Italic">Lotus japonicus</Emphasis> and <Emphasis Type="Italic">Fusarium solani</Emphasis>

A wilt disease of the model legume Lotus japonicus was observed in a greenhouse in Tokyo, Japan in May 2004. Roots of diseased plants were rotted and dark brown with lesions spreading to lower stems and leaves, resulting in rapid plant death. The causal agent was identified as Fusarium solani based on the morphology. Sequence analysis of rDNA supported the identification. Inoculation of roots of healthy plants with conidia reproduced characteristic disease symptoms, and F. solani was reisolated from lesions, satisfying Koch’s postulates. The isolate also caused chlorotic to necrotic lesions on leaves of healthy plants after wound-inoculation. Infection by F. solani of leaves of L. japonicus was confirmed histologically. Mycelia were observed in the intercellular spaces of parenchymatous tissues in the lesion area and the surrounding tissues. This is the first report of fungal disease on L. japonicus satisfying Koch’s postulates. We named it “Fusarium root rot of L. japonicus” as a new disease. The compatibility of L. japonicus and F. solani is expected to form a novel pathosystem for studying interactions between legumes and fungal pathogens. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB258993 and AB258994.     

Rhizoctonia solani Kühn AG 2-1 on kohlrabi in Italy

Rhizoctonia solani AG 2-1 was recorded in Central Italy on kohlrabi plants showing root and stem rot. After artificial inoculation the fungus caused damping-off of 7-day-old seedlings and root and stem rot of 4-month-old plants developed after 15 days of incubation. This seems to be the first record ofR. solani AG 2-1 on kohlrabi.     

新疆南疆枣树根腐病病原的分离与鉴定

<正>枣为鼠李科(Rhamnaceae)枣属(Ziziphus M ill.)植物,在我国拥有悠久的栽培历史。枣树的喜光、喜温、耐寒、耐旱等栽培特点,非常适合在新疆这种气候条件特殊的地区生长。近年来,随着新疆红枣产业的迅速发展,枣树病害问题也逐渐突显。2010和2011年,新疆南疆多地枣园出现枣树根腐病,该病害主要造成枣树实生苗和多年生枣树根部腐烂,叶片黄化,植株树势衰弱,最终导致植株     

Molecular identification and pathogenicity of Fusarium and Alternaria species associated with root rot disease of wolfberry in Gansu and Ningxia provinces,China

Wolfberry (Lycium barbarum) ranks in the top 10 best-selling medicinal plants in China and it has been used for centuries as a medicine and a food supplement. It is suggested to have benefits on human health due to the rich content of polysaccharides, carotenoids, flavonoids, and alkaloids contained in its fruits, leaves, and root bark. Recently, severe root rot diseases have been causing plant losses in major growing areas. Here, we report fungi causing root rot disease in Chinese wolfberry plants. The analysis of nucleotide sequences of the internal transcribed spacer (ITS) region revealed a total of 92 isolates isolated from both soil and plant material samples. Fusarium spp. were the most abundant (58%), followed by Penicillium spp. (9%), and Alternaria spp. (5%). Fusarium spp. included F. oxysporum (36%), F. solani (30%), F. chlamydosporum (9%), F. nematophilum (9%), and F. tricinctum (8%). Sequences from the translation elongation factor 1-α gene (TEF-1α) were used to confirm the identity of Fusarium spp. and showed the predominance of F. oxysporum and F. solani. To confirm the pathogenicity of isolates, four isolates belonging to Fusarium spp. and one isolate belonging to Alternaria spp., isolated from wolfberry root tissues with root rot symptoms, were tested in outdoor and laboratory conditions. Results revealed that the five tested isolates were pathogenic with varying degrees of aggressiveness and ability to induce symptoms of root rot in wolfberry seedlings. The five isolates were recovered from inoculated seedlings, completing Koch's postulates. This is the first report on causative agents of root rot in Chinese wolfberry.     

First report of white leaf rot on Chinese chives caused by Rhizoctonia solani AG-2-1

Delayed sprouting and white rot of leaf tips were found on Chinese chives in a greenhouse in Hokkaido, Japan, in the spring of 2006 and 2007. The causal fungus was identified as Rhizoctonia solani anastomosis group (AG)-2-1 and discriminated from the leaf rot pathogen R. solani AG-4 HG-I in terms of pathogenicity. Symptoms and the time of year that the disease occurs also apparently differ for the two pathogens. This is the first report of white leaf rot on Chinese chive caused by R. solani AG-2-1.     

Pythium rot of Chinese cabbage (Brassica rapa L. subsp. pekinensis) caused by Pythium aphanidermatum

Severe rot was found at the base of leaves and stems of Chinese cabbage (Brassica rapa L. subsp. pekinensis) in Ibaraki Prefecture every year in early September from 2002 through 2004. The causal fungus was identified as Pythium aphanidermatum (Edson) Fitzpatrick. This is the first report of P. aphanidermatum on Chinese cabbage. A similar disease of Chinese cabbage caused by P. ultimum Trow var. ultimum is known as Pythium rot. We propose adding P. aphanidermatum as a new pathogen of this disease.     

Phylogenetic analysis of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> subgroups associated with web blight symptoms on common bean based on ITS-5.8S rDNA

Sixty-eight Rhizoctonia solani isolates (31 AG-1, 37 of AG-2-2) associated with web blight (WB) of common bean, Phaseolus vulgaris, were examined for sequence variations in the ITS-5.8S rDNA region. The isolates were collected in bean-growing lowland and mountainous regions in Central and South America. Sequences of these isolates were aligned with other known R. solani sequences from the NCBI GenBank and distance and parsimony analysis were used to obtain phylogenetic trees. WB isolates of AG-1 formed two clades separated from known AG-1 subgroups. WB isolates of AG-2-2 formed one clade separated from known AG-2-2 subgroups. Other isolates belonged to AG-1 IA and AG-1 IB. Based on phylogenetic analysis, we confirmed that at least five genetically different subgroups incite WB of common beans. Three new subgroups of R. solani have been identified and designated as AG-1 IE, AG-1 IF and AG-2-2 WB. DNA sequences of these isolates provided needed information to design taxon-specific primers that can be employed in ecological/epidemiological studies and seed health tests.     

玉米内生菌L10的分离、鉴定及拮抗活性

为获得对玉米茎腐病主要病原菌禾谷镰孢Fusarium graminearum有明显拮抗作用的玉米内生菌,采用平板对峙法从成熟健康玉米茎秆中筛选禾谷镰孢拮抗菌株,并分析其抗菌谱;通过形态特征、生理生化特性及16S rDNA序列分析进行菌种鉴定;利用盆栽生防试验检测其对玉米茎腐病的防治效果。结果表明,共分离获得了164株玉米内生细菌菌株,其中L10菌株对禾谷镰孢具有较好的抑制效果,抑菌圈半径达1.68 cm;该菌对玉米大斑病菌Setosphaeria turcica、层出镰孢F. proliferatum、禾谷镰孢F. graminearum、拟轮枝镰孢F. verticilliodes、玉米弯孢叶斑病菌Curvularia lunata、玉米小斑病菌Bipolaris maydis、立枯丝核菌Rhizoctonia solani、茄链格孢Alternaria solani共8种植物病原菌均有拮抗作用,尤其对禾谷镰孢抑制效果最佳;结合形态特征、生理生化性质及16S rDNA序列分析,将L10菌株鉴定为多粘类芽胞杆菌Paenibacillus polymyxa。L10菌株脂肽类物质对禾谷镰孢菌具有较好的抑制活性,且盆栽生防试验结果显示该菌株对玉米茎腐病具有一定的防治效果。表明菌株L10对玉米镰孢茎腐病的防治具有一定潜力。     

甘草根腐病病原菌鉴定

 甘草(Glycyrrhiza uralensis Fish.)别名甜草、蜜草、甜根子, 为豆科多年生草本植物, 以根与根茎入药, 具有补脾益气、清热解毒、祛痰止咳、缓急止痛、调和诸药之功效, 是我国临床常用的中药材, 也可用作食品添加剂。甘草主要分布在我国的内蒙古、甘肃和宁夏, 在青海、陕西、新疆、黑龙江、辽宁、吉林、河北、山西等地局部地区也有分布。宁夏盐池县是我国乌拉尔甘草的重要产区, 面积大、贮量多、品质好, 1995年被誉为“中国甘草之乡” ^[1]。近年来, 野生甘草遭到了大规模采挖, 甘草蕴藏量急剧减少, 目前主要通过人工种植来满足市场需求。随着甘草种植面积的不断扩大, 甘草病虫害日趋严重, 根腐病危害尤为突出, 直接影响甘草的产量和品质, 造成巨大经济损失。     

Pathogenic fungi involved in root rot of peas in the Netherlands and their physiological specialization

Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.     

The first report of tomato foot rot caused by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-3 PT and AG-2-Nt and its host range and molecular characterization

Foot rot of mature tomato plants was found in four cities of Hokkaido, Japan, from 2004 to 2007. Six of eight isolates obtained from damaged tissues were identified as Rhizoctonia solani anastomosis group (AG)-3, and the remaining two isolates belonged to AG-2-1. We compared these isolates with nine reference isolates including the different subgroups in AG-3 (PT, TB and TM) and AG-2-Nt (pathogen of tobacco leaf spot) within AG-2-1 in terms of pathogenicity to tomato, tobacco and potato. All eight isolates caused foot rot on tomato. The six AG-3 isolates caused stem rot on young potato plants. While, all reference isolates of AG-3 PT causing stem rot of young potato plants incited foot rot on tomato. The two AG-2-1 isolates and an AG-2-Nt reference isolate caused severe leaf spot on tobacco leaves. The sequences of rDNA- ITS region and rDNA-IGS1 region of the AG-3 isolates showed high similarity to that of AG-3 PT isolates. Phylogenetic tree based on ITS and IGS1 regions of rDNA indicated that the AG-2-1 isolates from tomato formed a single clade with AG-2-Nt isolates and that they were separate from Japanese AG-2-1 isolates (culture type II). Pathogenicity tests and DNA sequence evaluation of the causal fungi revealed that the present isolates of AG-3 and AG-2-1 belonged to AG-3 PT and AG-2-Nt, respectively. This is the first report of tomato foot rot caused by R. solani in Japan.     

枸杞根腐病病原菌鉴定及其拮抗菌的筛选

为了针对根腐病实施有效的生物防治,分离鉴定枸杞根腐病病原菌及其拮抗菌,采用组织分离法获得腐根上的病原菌,结合形态学与分子生物学特征鉴定病原菌;利用稀释涂布、滤纸片法和共培养法从健康根际土壤筛选拮抗细菌,通过形态学观察、生理生化鉴定和16S rDNA序列分析对拮抗效果较好的菌株进行鉴定。结果表明：从枸杞根腐病分离得到腐皮镰刀菌（Fusarium solani）和尖孢镰刀菌（Fusarium oxysporum）两种主要致病菌,从健康枸杞根际土壤中分离获得12株对两种镰刀菌具有拮抗作用的细菌,其中J7-3、J7-8、J7-9和J10-8对腐皮镰刀菌的抑菌率均在80%以上,J7-3和J10-8对尖孢镰刀菌的抑菌率均在70%以上。菌株J7-3和J10-8对根腐病病原菌的抑菌作用较好,分别鉴定为解淀粉芽孢杆菌（Bacillus amyloliquefaciens）和枯草芽孢杆菌（Bacillus subtilis）,其中J10-8的拮抗作用和溶菌效果最好,可作为研发微生物菌剂的菌种资源。     

呼和浩特市苜蓿根腐病病原菌鉴定及致病性测定

为确定引起呼和浩特市苜蓿根腐病的病原菌种类,采用常规组织分离法对采集的疑似苜蓿根腐病病样进行病原菌分离与培养,利用形态学观察结合分子生物学方法对分离物代表菌株进行鉴定,并采用土壤接种法对代表菌株的致病性进行测定。结果表明,共分离获得6类形态学特征不同的分离物,各随机选择1株代表菌株进行鉴定,结合分子生物学鉴定结果确定呼和浩特市苜蓿根腐病病原菌有6种,分别是麦根腐平脐蠕孢菌Bipolaris sorokiniana、立枯丝核菌Rhizoctonia solani、木贼镰刀菌Fusarium equiseti、变红镰刀菌F. incarnatum、锐顶镰刀菌F. acuminatum和织球壳枯萎菌Plectosphaerella cucumerina,分别分离到1、7、14、26、7和14株菌株,占总分离菌株数的1.45%、10.14%、20.29%、37.68%、10.14%和20.29%。其中,立枯丝核菌的致病力最强,接种苜蓿幼苗发病的病情指数达82.67,其次为木贼镰刀菌、变红镰刀菌、锐顶镰刀菌、麦根腐平脐蠕孢菌和织球壳枯萎菌,病情指数分别为72.67、62.67、58.67、52....     

Characterization of Rhizoctonia solani Associated with Soybean in Brazil

Rhizoctonia solani causes pre- and post-emergence damping-off, root and hypocotyl rot and foliar blight in soybean. Foliar blight has resulted in yield losses of 31–60% in north and northeast Brazil. The aim of this study was to characterize isolates of R. solani associated with soybean in Brazil. Among 73 Rhizoctonia isolates examined, six were binucleate and 67 were multinucleate. The multinucleate iso1ates were characterized according to hyphal anastomosis reaction, mycelial growth rate, thiamine requirement, sclerotia production, and RAPD molecular markers. Four isolates that caused hypocotyl rot belonged to AG-4 and using RAPD analysis they grouped together with the HGI subgroup. Another isolate that caused root and hypocotyl rots was thiamine auxotrophic, grew at 35°C, and belonged to AG-2-2 IIIB. All 62 isolates that caused foliar blight belonged to AG-1 IA. RAPD analysis of R. solani AG-1 IA soybean isolates showed high genetic similarity to a tester strain of AG-1 IA, confirming their classification. The teleomorph of R. solani, Thanatephorus cucumeris was produced in vitro by one AG-1 IA isolate from soybean. The AG-4 and AG-2-2 IIIB isolates caused damping-off and root and hypocotyl rots of soybean seedlings cv. FT-Cristalina, under greenhouse conditions. The AG-2-2 IIIB isolate caused large lesions on the cortex tissue, that was distinct from the symptoms caused by AG-4 isolates. The AG-1 IA isolates caused foliar blight in adult soybean plants cv. Xingu under the greenhouse and also in a detached-leaf assay.     

枯草芽胞杆菌B1409对番茄和辣椒的防病促生作用

为探讨枯草芽胞杆菌Bacillus subtilis菌株B1409对番茄早疫病和辣椒疫霉病的防效和生防机制,采用平板对峙法和盆栽法测定了该菌株对番茄早疫病菌和辣椒疫霉病菌菌丝生长的抑制作用、对2种病害的盆栽防效以及对番茄和辣椒植株促生长效果和防御酶活性的影响。结果表明:菌株B1409能明显抑制番茄早疫病菌和辣椒疫霉病菌菌丝生长,且导致菌丝发生畸变。10~8CFU/mL菌株B1409菌液对番茄早疫病和辣椒疫霉病的预防效果分别为67.82%和61.22%,治疗效果分别为41.22%和56.43%。不同浓度B1409菌液均能促进番茄和辣椒植株生长,并能增强其体内超氧化物歧化酶、过氧化物酶和过氧化氢酶活性,且浓度越高促进效果越明显。番茄和辣椒植株的平均干重分别在10~2CFU/mL和10~4CFU/mL B1409菌液处理后显著高于对照,增长率分别为42.35%和4.87%。番茄和辣椒植株经10~2CFU/mL B1409菌液处理后,体内超氧化物歧化酶活性比对照显著增加,增长率分别为91.23%和19.58%。研究表明枯草芽胞杆菌B1409菌株可通过直接抑制菌丝生长及诱导植物体自身抗病性等方式来有效防治番茄早疫病和辣椒疫霉病。