A single gene in Fusarium oxysporum limits host range

Fusarium oxysoporum f. sp. radicis-cucumerinum (Forc) is able to cause disease in cucumber, melon, and watermelon, while F. oxysporum f. sp. melonis (Fom) can only infect melon plants. Earlier research showed that mobile chromosomes in Forc and Fom determine the difference in host range between Forc and Fom. By closely comparing these pathogenicity chromosomes combined with RNA-sequencing data, we selected 11 candidate genes that we tested for involvement in the difference in host range between Forc and Fom. One of these candidates is a putative effector gene on the Fom pathogenicity chromosome that has nonidentical homologs on the Forc pathogenicity chromosome. Four independent Forc transformants with this gene from Fom showed strongly reduced or no pathogenicity towards cucumber, while retaining pathogenicity towards melon and watermelon. This suggests that the protein encoded by this gene is recognized by an immune receptor in cucumber plants. This is the first time that a single gene has been demonstrated to determine a difference in host specificity between formae speciales of F. oxysporum.     

几种杀菌剂对黄瓜疫霉菌的毒力测定

采用菌丝生长速率法检测60%嘧菌酯悬浮剂、80%代森锰锌可湿性粉剂、75%百菌清可湿性粉剂、60%唑醚·代森联水分散粒剂、2%蛇床子素微乳剂、64%杀毒矾可湿性粉剂等6种杀菌剂对黄瓜疫霉菌的室内毒力。结果表明,唑醚·代森联和蛇床子素对黄瓜疫霉菌的毒力最高,EC50值分别为3.4143 mg·L-1和10.4641 mg·L-1。将两种原药混配后,表现出增效作用,在两者混配比为1∶2时增效最明显,EC50值为5.8178 mg·L-1。     

华南地区瓜类疫霉对甲霜灵的田间抗药性

【目的】了解华南地区瓜类疫霉(Phytophthora melonis)对甲霜灵的田间抗药性。【方法】2007-2010年从广西、广东两省(区)9个市冬瓜和黄瓜产区采集疫病样品,分离纯化瓜类疫霉,分别采用菌落生长速率法和叶盘漂浮法测定瓜类疫霉对甲霜灵的敏感性,并用药剂驯化方法从敏感性菌株诱导瓜类疫霉抗甲霜灵突变体。【结果】从9个市24个样点共分离纯化获得193株瓜类疫霉,抗药性检测结果表明,敏感菌株、中等抗性菌株和抗性菌株分别占测试菌株的29.0%、18.1%和52.8%;不同地区、不同寄主分离的菌株的抗性频率和抗性水平差异较大,来源于广东的菌株抗性频率和抗性水平一般高于来源广西的菌株,分离自黄瓜的菌株高于分离自冬瓜的菌株,大部分样点抗性菌株占据优势群体,个别菌株的抗性指数高达4226.9,叶盘漂浮法测定结果和菌落生长速率法相似;在含药平板上对敏感菌株进行甲霜灵抗性诱导结果表明,从60%的敏感菌株中成功诱导出对甲霜灵抗性稳定的突变体,突变体的抗性水平为敏感性亲本的189-407倍;9株来源于未施用过甲霜灵等苯基酰胺类杀菌剂样点的菌株均为敏感性菌株,其EC50值为0.0429-0.5461μg/mL,将它们EC50的平均值0.3200±0.1617μg/mL确定为华南地区瓜类疫霉对甲霜灵的敏感性基线;对两个样点的监测结果表明,瓜类疫霉抗甲霜灵菌株的频率及抗性指数有逐年增高趋势。【结论】华南广西和广东两省(区)瓜类疫霉对甲霜灵抗性普遍发生,瓜类疫霉对甲霜灵抗药性产生与其和药剂的接触密切相关。瓜类疫霉敏感性基线的建立,可为今后瓜类疫霉抗甲霜灵的评价和进一步监测提供科学依据。     

Purification and characterization of an extracellular endo-1,4-β-xylanase from Fusarium oxysporum f. sp. melonis

Abstract Fusarium oxysporum f. sp. melonis produces extracellular endo-1,4-β-xylanase and β-xylosidase when grown in shaken culture at 26°C in a mineral salts medium containing oat spelt xylan and glucose as carbon sources. Endo-1,4-β-xylanase was purified 251 times from 5-day-old culture filtrates, by Sephacryl S-200, ion exchange and gel filtration HPLC. The purified sample yielded a single band in SDS polyacrylamide gels with a molecular mass of 80 kDa on electrophoretic mobility and 83 kDa by gel filtration behavior. High activity of the endo-1,4-β-xylanase against xylan was observed between 5 and 8 pH, and between 40 and 60°C, the optimum pH and temperature being 5.0 and 50°C, respectively. Kinetic properties of the enzyme are similar to those of other fungal xylanases, showing high affinity towards oat spelt xylan with a K _m of 1 mM expressed as xylose equivalent.     

Phytophthora melonis Associated with Fruit and Vine Rot Disease of Pointed Gourd in India as Revealed by RFLP and Sequencing of ITS Region

Pointed gourd is an important tropical high value vegetable crop, which is mainly affected by fruit and vine rot disease in field conditions. Causal organism of this devastating disease is Phytophthora melonis as revealed through morphological criteria as well as by molecular tools based on the restriction fragment length polymorphism (RFLP) of non‐coding Internal Transcriber Spacer (ITS) region and ITS sequencing. Sequencing of ITS region of our Ph. melonis isolate has 100% similarity with the five isolates of GenBank including a Ph. sinensis. The pathogen, Ph. melonis, is a new report from India and as regards host ranges a possible new report globally.     

Safflower Seedling a Selective Host to Discriminate Phytophthora melonis from Phytophthora drechsleri

Safflower seedlings were used to discriminate two morphologically similar species of Phytophthora namely P. melonis and P. drechsleri . All isolates of P. melonis from different sources could not infect safflower seedlings under high inoculum potential whereas all isolates of P. drechsleri from various hosts attacked safflower within a short period. Of 31 authentic Phytophthora species inoculated to safflower seedlings only seven species including Phytophthora asparagi , Phytophthora cactorum , Phytophthora cryptogea , Phytophthora drechsleri , Phytophthora erythroseptica , Phytophthora palmivora and Phytophthora quercina caused hypocotyls infection. All cucurbit isolates of Phytophthora from different parts of Iran and one from China could not infect safflower seedlings and were identified as P. melonis which had been confirmed previously by molecular analysis.     

Ultrastructural Study on the Interactions Between Three Respectively Different Disease- resistant Cucumber Cultivars and Phytophthora melonis

Ultrastructural changes during the process of interactions between three different diseaseresistant, susceptible, moderately resistant and resistant cucumber (Cucumis sativus L. ) cuhivars and Phytophthora melonis Katsura were respectively compared and the associated factors in the observed differences in colonization were determined. During the process of interaction in the susceptible cultivar, both inter- and intracellular growth of hyphae associated with extensive colonization of healthy hyphae and marked alteration of mesophyll cells, such as aggregation of cytoplasm and disintegration of cellular organelle were demonstrated. The general disorganization of host cell was observed in areas directly adjacent to the fungal hyphae and in cells several layers from the nearest visible hyphae. In the moderately resistant cultivar during interaction the host showed limited hypha growth with responses of resistance including 1 ) breakage of plasmodesmata to prevent the host cells to contact the hyphae, 2) increasing the quantity of endoplasmic reticulum and Golgi apparatus so as to increase the quantity of secretory protein, 3) a plasmalemma invagination containing granular material at the site in contact with the intercellular hyphae. The interaction in the resistant cuhivar was characterized as a hypersensitive reaction (HR) which means a rapid necrosis of the infected cells and the cells surrounding the invaded hyphae, resulting in death of the invaded hyphae. Formation of cell wall appositions right at the penetration sites of hyphae and haustorium-like structure was observed. The results indicate that the mechanism of host response to P. melonis invasion was differ- ent between the moderately resistant and the resistant cucumber cultivars.