棉花枯、黄萎病菌营养亲和性研究进展

介绍了棉花枯、黄萎病菌营养亲和性研究方法和营养体亲和群研究,并着重总结了利用营养亲和群结合DNA多态性技术,研究尖孢镰刀菌(FusariumoxysporumSchlechtend:Fr)专化型、生理小种、亲和群之间的关系,以及棉花黄萎病菌(VerticilliumdahliaeKleb.)亲和群与致病型关系的研究进展。     

Effect of plant roots on the germination of microsclerotia ofVerticillium dahliae

Induction of germination of microsclerotia by exudates from plant roots may be important for the control ofV. dahliae. Laboratory experiments with root observation boxes were carried out to assess the influence of root tips of seven crop species and cultivars on the germination of microsclerotia ofVerticillium dahliae in soil under controlled conditions. The root density of crops was measured in a field experiment. The results of the laboratory experiments and the field experiment were combined to estimate the total effect of crops on the population of microsclerotia in the field. Germination of microsclerotia was stimulated by all crops compared to a control without a crop. Among crops, roots of potato cvs Element and Astarte had a larger stimulation effect on microsclerotia than that of potato Ostara, pea, flax, sugar beet or onion. The number of hyphae per microsclerotium decreased with distance from the root surface regardless of the crop species or cultivar. Differences in root densities, in the affected root zones and in the stimulation effect on germination of microsclerotia caused large differences among crops in the effect on the population of microsclerotia in the soil. However, growing a rop with the special purpose to reduce the level ofV. dahliae inoculum in the soil is an inefficient control measure, because only a small part of the total soil volume is affected by roots and the number of hyphae per microscleroium affected is too low.Abbreviations MS microsclerotia, microsclerotium     

Dynamics ofRhizoctonia solani (black scurf) in successive potato crops

The position of plants withRhizoctonia solani sclerotia (black scurf) on progeny tubers was mapped for an experimental field at Haren where potatoes were grown continuously and in rotation with other crops for five successive years, and for another field at Borgercompagnie with a 12 frequency of potatoes during three potato crops. Initially, the distribution of plants with black scurf on both fields was rather dense and homogeneous. In the following years the distribution became heterogeneous and patchy. The local decline ofR. solani AG 3 (the common potato pathogen) in Haren was apparently caused by an unknown factor selectively suppressingR. solani AG 3, while simultaneouslyR. solani AG 5 increased in mass. This AG 5 type proved to be an inferior competitor of AG 3 on the potato plant in a laboratory experiment. The specificR. solani antagonistVerticillium biguttatum did not play a role. A similar factor could have reduced the formation of black scurf in the experimental field at Borgercompagnie, whereV. biguttatum was also too infrequent to account for the decline.R. solani AG 5 was not present here and could not indicate the presence of a selective factor against AG 3.     

Contribution to the taxonomy and pathogenicity of fungicolous Verticillium species. II. Pathogenicity

In recent years in the Netherlands a second mushroom species,Agaricus bitorquis, which prefers higher temperatures thanA. bisporus and is less susceptible to certain diseases, is often commercially grown.Verticillium fungicola var.fungicola, the causal agent of dry bubble, is responsible for considerable damage in crops ofA. bisporus. InA. bitorquis, however, dry bubble has hardly been noticed, but brown spots due toV. fungicola var.aleophilum resulted in inferior mushroom quality. The latter variety also caused brown spots ina. bisporus, but to a minor degree. In variety Les Miz 60 ofA. bisporus, however, it also induced fruit-body deformation in a way different from dry bubble. Verticillium psalliotae, isolated fromA. bitorquis in England, induced more confluent brown spots inA. bitorquis. In the netherlands, where moreA. bitorquis is grown than in other countries,V. psalliotae has not yet been encountered in crops ofA. bitorquis. V. psalliotae, which has a high temperature optimum for mycelial growth, likeV. fungicola var.aleophilum andA. bitorquis, did not infectA. bisporus in our trials.Artificial infection ofA. bisporus orA. bitorquis could not be accomplished with the following related and/or fungicolous fungi:Verticillium lamellicola, V. fungicola var.flavidum, V. biguttatum, Nectriopsis tubariicola, Acremonium crotocinigenum andAphanocladium album.Samenvatting Vooral in Nederland wordt sinds een aantal jaren naastAgaricus bisporus ook de warmteminnende champignonsoortAgaricus bitorquis geteeld, die minder vatbaar is voor bepaalde ziekten. TerwijlVerticillium fungicola varfungicola in de teelt vanA. bisporus droge mollen en daardoor veel schade veroorzaakt, komen in de teelt vanA. bitorquis geen droge mollen voor maar wel bruine vlekken, die tot kwaliteitsverlies en dus schade leiden. De vlekken bleken veroorzaakt te worden doorV. fungicola var.aleophilum. Deze schimmel veroorzaakte ook inA. bisporus bruine vlekken, hoewel in minder ernstige mate, maar in het ras Les Miz 60 vanA. bisporus bovendien misvormde champignons, die wel op droge mollen leken, maar daaraan niet gelijk waren.OokV. psalliotae, in Engeland geïsoleerd vanA. bitorquis met vlekken, veroorzaakte wat meer samenvloeiende, bruine vlekken inA. bitorquis. In Nederland, waar meerA. bitorquis geteeld wordt dan in andere landen, isV. psalliotae nog niet aangetroffen in teelten vanA. bitorquis. InA. bisporus kon geen kunstmatige infectie worden verkregen metV. psalliotae, die net alsV. fungicola var.aleophilum enA. bitorquis warmteminnend genoemd zou kunnen worden.Met de volgendeVerticillium-achtige of van paddestoelen geïsoleerde schimmels kon evenmin op kunstmatige wijze een infectie worden opgeroepen inA. bisporus ofA. bitorquis: Verticillium lamellicola, V. fungicola var.flavidum, V. biguttatum, Nectriopsis tubariicola, Acremonium crotocinigenum enAphanocladium album.     

Phytotoxicity on cotton ex-plants of an 18.5 kDa protein from culture filtrates of Verticillium dahliae

A phytotoxic protein that evokes the typical symptoms of Verticillium wilt disease in seedlings of Gossypium hirsutum L. (Upland cotton) was isolated from culture filtrates of Verticillium dahliae. The protein was purified by ammonium sulfate precipitation, Sephadex-G100 fractionation, and native PAGE. The 18.5 kDa protein, designated VD18.5, appears to be a single subunit protein with an isoelectric point between 3 and 5. VD18.5 induces symptoms of leaf dehydration, chlorosis, necrosis and stem discoloration in seedlings of the disease susceptible cotton cultivar Siokra 1–4. The LD₅₀ of VD18.5 on protoplasts of Siokra 1–4 was 18 μg mL⁻¹. VD18.5 had no noticeable effect on Pima S-7, which is a disease resistant cultivar. Phytotoxic activity was partially destroyed at high temperature and was abolished by digestion with proteinase K. Mass spectrometry fingerprinting and protein sequence data from VD18.5 yielded no significant matches when submitted to the Mascot search engine and NCBI non-redundant protein databases, respectively. These results suggest that VD18.5 is a novel protein that may be involved in the development of some of the symptoms associated with Verticillium wilt disease in the cotton plant.     

Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

某些阳离子在大豆胞囊线虫生防系统中的作用

室内测定了不同浓度的 6种阳离子 :钼、硼、锌、铜、锰和铁对大豆胞囊线虫卵的孵化、大豆种子发芽和根系生长 ,以及对线虫生防菌———厚垣轮枝菌菌丝生长的影响。结果发现 ,在供试所有浓度下 ,铜、锰和铁明显抑制大豆胞囊线虫的卵孵化 ,锌具明显刺激作用 ;在供试浓度水平较低时 ,铜、锰和铁对大豆种子发芽及根系生长有轻微抑制 ,锰的抑制较为明显 ,但均不影响进一步发育 ;在供试浓度较低时 ,铜、锰和铁对厚垣轮枝菌菌丝生长无任何影响。铜可作为线虫生防制剂的添加剂。     

棉花黄萎病菌致病型的AFLP分析

 选用41个棉花黄萎病菌(Verticillium dahliae)代表菌系,在温室条件下,对4个棉花品种鄂荆1号(感)、中棉所12(耐)、文-5(抗)和唐棉2号(抗)进行致病性测定,结果可将供试菌系分为落叶型与非落叶型2类。选取8对AFLP引物PCR扩增的结果中,统计带型稳定、清晰且有多态性的条带,共169条作系统聚类分析,将上述菌系分为2大类,第一类为非落叶型菌系,包括10个非落叶型菌系和1个过渡菌系;第二类为30个落叶型菌系。根据聚类分析建立树状图,发现菌系与地理来源存在一定的相关性,而依据菌系致病力强弱分类则相关关系不大。选用25对EcoRⅠ和MseⅠ引物组合,对供试的41个V.dahliae进行AFLP扩增,筛选到2对引物E64(GACTGCGTACCAATTCGAC)、M53(GATGAGTCCTGAGTAACCG)和E49(GACTGCGTACCAATTCCAG)、M65(GAT-GAGTCCTGAGTAAGAG),能分别扩增出433bp和110bp2条仅为V.dahliae非落叶型菌系独有的特异片段,可将落叶型与非落叶型菌系分开,这2条特异片段被命名为EM₄₃₃和EM₁₁₀。     

应用聚合酶链式反应鉴定新疆棉花落叶型黄萎病菌

用一对棉花落叶型黄萎病菌的特异性引物D1和D2进行PCR扩增,对于落叶型黄萎病菌,该对引物可特异性地扩增产生一段550bp的产物,而非落叶型黄萎病菌则不能被扩增.供试的35个新疆黄萎菌系中,有3个菌系扩增出550bp大小的落叶型黄萎病菌特异性片段,表明目前新疆已存在落叶型黄萎病菌,用此技术可快速、准确地检疫和鉴定落叶型黄萎病菌.     

Evaluation of Olive Cultivars for Resistance to Verticillium dahliae

Resistance of 23 important olive cultivars to Verticillium dahliae has been evaluated in four experiments under controlled conditions. Nine-month-old nursery olive plants were inoculated with a cotton non-defoliating (ND) (V4) or a cotton defoliating (D) (V117) isolate of V. dahliae. Resistance was evaluated by assessing symptom severity using a 0–4 rating scale and estimating the area under disease progress curves. The percentage of plants killed and of those which recovered from the disease were used as additional parameters for including a particular cultivar into a defined category. Most of the evaluated cultivars were susceptible, although at different levels, to both isolates of V. dahliae. All cultivars were more susceptible to the D pathotype than to the ND one. A group of 11 cultivars, including several important Spanish cultivars, were susceptible or extremely susceptible to both pathotypes of V. dahliae. A second group showed differences of resistance depending on the pathotype used. They were susceptible or extremely susceptible to the D pathotype but resistant or moderately susceptible to the ND one. Finally, 'Frantoio', 'Oblonga' and 'Empeltre' were moderately susceptible to the D isolate of V. dahliae and resistant to the ND one. The resistance of 'Empeltre' was evident by the plant ability to recover from infection with either isolates. 'Empeltre' is considered to be a valuable cultivar for inclusion in breeding programmes for resistance to Verticillium wilt.