Tolerance to a non-host isolate of Verticillium dahliae in tomato

Tolerance to Verticillium spp. is a condition in which a host plant develops few symptoms despite substantial colonization by the pathogen. In the present paper we have shown that Craigella tomatoes are tolerant to a non-host isolate of V. dahliae, Dvd E6. Symptom expression was used to quantify disease and quantitative PCR to assess the amount of fungus in the stems. The classical incompatible and compatible interactions between Craigella resistant or Craigella susceptible near isolines and V. dahliae, race 1 were used for comparative purposes. Additional experiments using cytological assessment and quantitative PCR showed that in the tolerant interactions one plant defence response, vascular coating, was deployed as effectively as in resistant plants, limiting pathogen distribution. However, a second defence response, which causes the cyclical elimination of fungus from the stem in the classical interactions either does not occur or is substantially delayed in tolerant plants. Thus, the Verticillium population remains stable and substantial throughout the studied time course.     

Compatible Biological and Chemical Control Systems for Rhizoctonia solani in Potato

A series of chemical and biological control agents were tested for compatibility with the Rhizoctonia-specific biocontrol fungus Verticillium biguttatum aimed at designing novel control strategies for black scurf (Rhizoctonia solani) and other tuber diseases in potato. The efficacy of chemicals, alone and in combination with V. biguttatum was tested in in vitro assays on nutrient agar plates, in bio-assays with minitubers and in the field. Generally, there were both antagonistic, neutral and additive interactions with V. biguttatum among the combinations tested; there were no indications for synergistic interactions. Broad-spectrum fungicides (azoxystrobin, chlorothalonil, thiabendazole) were fungitoxic to V. biguttatum as shown in in vitro assays, and hampered black scurf control by V. biguttatum in bio-assays. Oomycete-specific chemicals (cymoxanil and propamocarb) and various biocontrol strains (Gliocladium spp., Pseudomonas spp. and Trichoderma spp.) did not interfere with the growth of V. biguttatum on agar nutrient plates and did not affect black scurf control by V. biguttatum in co-applied treatments in the minituber bio-assay. Rhizoctonia-specific (pencycuron, flutalonil) fungicides co-applied with V. biguttatum showed additive effects on black scurf control. When combinations of V. biguttatum and cymoxanil or propamocarb were applied to immature potato tubers at green crop lifting, a reduction of both black scurf and Pythium- or Phytophthora-incited tuber rot was observed at harvest. In conclusion, the biocontrol fungus V. biguttatum is compatible with selected chemical control systems and may improve control efficacy in combination with Rhizoctonia-specific fungicides or may extend control spectrum in combination with Oomycete-specific fungicides.     

Plant Phenology Influences the Effect of Mycorrhizal Fungi on the Development of Verticillium-induced Wilt in Pepper

Verticillium dahliae alters water status and consequently, growth and production of pepper plants. On the other hand, arbuscular mycorrhizal fungi (AMF) can reduce damage caused by specific soil-borne plant pathogens and improve drought resistance of pepper. Therefore, one objective of this research was to assess if AMF can modify the development of Verticillium-wilt in pepper plants. A second objective was to study the influence of plant phenology at the moment when V. dahliae was inoculated on the possible biocontrol of the disease by AMF. Results suggested that AMF reduce the deleterious effect of V. dahliae on pepper growth and yield. However, bioprotection against Verticillium-wilt was conditioned by plant phenology at the moment of pathogen attack. The highest efficacy of AMF occurred when V. dahliae was inoculated during the vegetative stage of plants. AMF allowed leaf relative water content to be maintained for longer and delayed both the appearance of disease symptoms and the decrease of photosynthesis in Verticillium-inoculated plants. These benefits on plant physiology increased pepper yield.     

蜡蚧轮枝菌固体发酵基质优化组合正交筛选

蜡蚧轮枝菌菌株VLFNL95经 5种单一固体基质和 7种一般组合固体基质的发酵 ,分析比较其产孢量和分生孢子活力 ,初筛出麦麸单一基质和玉米碎粒 +黄豆饼粉 +磷酸盐组合基质为最佳发酵基质 ,两者的产孢量很接近 ,孢子活力较高。选取黄豆饼粉、玉米碎粒、磷酸盐和谷壳为 4个组分因子 ,每个因子分别设有不同质量比例的 3个水平进行正交设计。通过对 9种正交配方发酵试验 ,筛选出 3号为最佳组合配方 ,其分生孢子产出量最高为6.90×10¹⁰个/g ,比初筛出的前 2个最佳基质提高约 1倍 ,孢子萌发率为 95.04%。     

Vegetative-compatibility groups in Verticillium dahliae from Israel

Nitrate-nonutilizing (nit) mutants were used to determine vegetative compatibility among 34 isolates of Verticillium dahliae from cotton, potato, olive, eggplant, chrysanthemum and tomato from 12 sites in Israel. Based on the formation of complementary heterokaryons, 33 isolates were assigned to two vegetative- compatibility groups (VCGs): one VCG contained 15 isolates from cotton, eggplant, chrysanthemum and olive; and the other VCG contained 18 isolates from potato, olive and cotton. The status of an additional isolate from tomato, which was compatible with both VCGs, remained unclear. In a limited pathogenicity test with 10 isolates, two (from tomato and eggplant) were pathogenic on tomato, eggplant and cotton; most isolates from cotton were pathogenic on cotton and eggplant only; and one from cotton was non-pathogenic. Fewer isolates were pathogenic on tomato than on cotton or eggplant. The diversity of vegetative compatibility found in our V. dahliae collection is comparable to that found in studies of American populations.     

茄子黄萎病的发生及病菌生长影响因子

江苏扬州等地棚室栽培条件下,茄子黄萎病一般始见于现蕾期,5月上、中旬进入座果期开始大量发生,5月中下旬至6月上旬达发病高峰;病田土壤2年未栽作物仍能造成茄植株较高发病率.病叶混入土壤引起茄植株发病率最高,而病根混入土壤未见发病.分离自茄子、辣椒和棉花的黄萎病菌菌株在供试的玉米粉、燕麦、理查、马铃薯等培养基上均生长良好,茄子、辣椒黄萎病菌在理查培养基上出现明显的紫红色,棉花黄萎病菌则未出现;三种病菌在pH4～9的培养基上均能生长;棉叶、茄叶、辣椒叶的汁液能刺激茄黄萎病菌分生孢子的萌发,其中以茄叶汁液最明显;病菌经55℃水浴处理20 min,各菌株均未见生长.     

一种新的棉花黄萎病快速接种技术及其在病原菌致病力和寄主抗病性鉴定上的应用

 棉花品种抗黄萎病鉴定一般在田间病圃中进行,其结果受病圃中病原菌分布均匀程度、气象等因素影响极大,往往导致鉴定结果不准确。为了使鉴定方法简单、科学、可靠,我们在温室条件下比较了3种苗期接种棉花黄萎病的方法,即切根蘸孢子法(接种浓度为10⁶分生孢子/mL);菌培养物土壤接种法(0.5%、1%、2%,w/w);微菌核土壤接种法(10³个微菌核/g土)。结果表明,切根蘸孢子法导致棉苗发病均匀、严重、迅速,播种35~45 d后即可得到均匀一致的发病结果。而其它2种接种方法在播种75 d后才得到相对稳定的发病结果。同时,研究还表明接种浓度为10⁴分生孢子/mL所导致的黄萎病显著比10⁵或10⁶的轻。利用切根蘸孢子法在室内鉴定12个棉花品种或品系的抗黄萎病能力,证明该方法是抗黄萎病快速鉴定的有效方法。此外,该鉴定方法还可快速鉴定黄萎病菌不同菌株的致病性,并可应用于作物对其它土传病害的抗病性鉴定上。     

分离土壤棉花黄萎病菌选择性培养基的筛选

 采用土壤检测稀释法、水筛法和不同土样接种量,对5种土壤浸提液改良培养基进行了比较研究。结果表明,土壤水筛法对土壤棉花黄萎病菌的检测效果优于稀释法,其菌落检测数增加4~10倍。培养基每皿0.25 g土样接种量的检测效果高于0.5 g和1g。5种测试培养基中,培养基D对分离土壤棉花黄萎病菌的选择性表现最好,其土壤接种体检测回收率达63.5%~83.3%.培养基D的组份为:土壤浸提液25 ml,KH₂PO₄ 1.5 g,K₂HPO₄ 4 g,尿酸钠0.2 g,半乳糖醛酸钠(果胶)1g,山梨糖1g,Dox盐2 ml,Tergitol NP-101ml,PCNB0.1g,琼脂17g,蒸馏水1000 ml,pH6.4~6.7。每1000ml融化培养基倒皿前加抗菌素液50 ml (含氯霉素0.05 g,链霉素0.05 g,青霉素-G盐0.05 g)。     

Specific Detection of Tomato Pathotype of Verticillium dahliae by PCR Assays

Verticillium dahliae Klebahn is the causal agent of tomato wilt disease. Isolates of V. dahliae can be classified based on pathogenicity to tomato, but the pathotypes are indistinguishable in morphology. We designed PCR primers for specific detection of isolates pathogenic to tomato (tomato pathotype) from the sequences of a pathotype-specific gene, vdt1. With the primer pair Tg5/Tc3, a PCR product (approximately 3.2 kb) specific to tomato pathotype was amplified from the genomic DNA of isolates. Using the primer pair, a tomato pathotype isolate was specifically detected from hypocotyls of inoculated tomato and eggplant. On the other hand, no amplification was observed from non-tomato pathotype isolates of V. dahliae, some other wilt pathogens of tomato and a healthy host plant. Therefore, the primer pair can be useful for pathotype-specific detection of V. dahliae as well as for diagnosis of wilt disease of tomato plant. Received 7 September 2001/ Accepted in revised form 3 December 2001     

Effects of granular nematicides on growth and microbial antagonism to Rhizoctonia solani

Effects of nematicides on growth and microbial antagonism toRhizoctonia solani were investigated as part of a study on the mechanisms involved in the increased incidence of this pathogen in nematicide-treated potato crops.Ethoprophos inhibited mycelial growth ofR. solani on potato dextrose agar (PDA), Czapek Dox agar (CDA) and on water agar (WA). Aldicarb stimulated its growth on PDA up to 14% but not on CDA and WA. Oxamyl inhibited mycelial growth on CDA and WA, but not on PDA.Ethoprophos and aldicarb stimulated development of the mycoparasiteVerticillium biguttatum on cultures ofR. solani. The effect was dependent on the medium on which the host fungus was grown. ForRhizoctonia cultures on PDA, growth of the mycoparasite was highly promoted by aldicarb and to a lesser extent by ethoprophos. WhenR. solani was grown on CDA, the development of the mycoparasite was not affected by aldicarb, slightly stimulated by ethoprophos and slightly inhibited by oxamyl. On water agar, its development on the host mycelium was not affected.In field trials on sandy soil, nematicides encouragedV. biguttatum probably by increased availability of substrate (i.e.Rhizoctonia mycelium) perhaps through reduced activity of the mycophagous fauna.Soil fungistasis was increased by ethoprophos and to a lesser extent by aldicarb at very high doses. At normal field rates, no effects can be expected on fungistasis. So the increased stem and stolon infection of potatoes in nematicide-treated fields was not caused by a direct effect of the nematicides on growth ofR. solani or by suppressing the microbial antagonism.Samenvatting De invloed van granulaire nematiciden op de groei vanRhizoctonia solani en op het microbiële antagonisme tegen deze schimmel werd onderzocht in het kader van een studie over de mechanismen die een rol spelen bij de toename van de aantasting in een met nematiciden behandeld aardappelgewas.Ethoprofos remde de myceliumgroei vanR. solani op aardappeldextrose agar (PDA), Czapek Dox agar (CDA) en op wateragar (WA). Aldicarb stimuleerde op PDA de groei met maximaal 14%. Op CDA en WA werd geen effect van aldicarb waargenomen. Oxamyl veroorzaakte groeiremming op CDA en WA, maar niet op PDA.Ethoprofos en aldicarb stimuleerden de ontwikkeling van de mycoparasietVerticillium biguttatum op cultures vanR. solani. De mate van groeistimulering was afhankelijk van de voedingsbodem waarop de waard,R. solani, werd gekweekt. De groei vanV. biguttatum werd sterk gestimuleerd door aldicarb en in geringere mate door ethoprofos, wanneer de waard gekweekt werd op PDA. Aldicarb had geen effect op de mycoparasiet wanneerR. solani op CDA gekweekt werd, terwijl ethoprofos de groei wel iets stimuleerde en oxamyl een gering remmend effect had. Op WA werd geen effect van de nematiciden op het mycoparasitisme vastgesteld.In veldproeven op zandgrond stimuleerden de nematiciden het voorkomen vanV. biguttatum op de stolonen. Het effect werd waarschijnlijk veroorzaakt door een verhoogde substraat beschikbaarheid (d.w.z. mycelium vanR. solani). De verhoogde beschikbaarheid van dit mycelium kan samenhangen met een door nematiciden gereduceerde activiteit van de fungivore bodemfauna.De bodemfungistase werd verhoogd door ethoprofos en, in geringere mate, door aldicarb bij hogere doseringen. Bij de in de praktijk aanbevolen doseringen kan echter geen effect op de fungistase verwacht worden. De toename in stengel- en stolonaantasting van aardappelen, geteeld in met granulaire nematiciden behandelde percelen, kon niet worden toegeschreven aan een direct effect van de nematiciden op de groei vanR. solani of aan een vermindering van het microbiële antagonisme.