采后ASM诱导处理对鸭梨果实黑霉病的控制

 Acibenzolar-S-methyl (ASM) 是重要的植物抗病基因诱导剂。鸭梨果实经过负压渗透ASM诱导处理后接种黑霉病(Alternaria alternata) 。结果表明, 0.5 mmol·L ^{- 1} ASM能有效抑制鸭梨果实损伤接种病害和自然侵染病害的发生。ASM处理果实具有较高的苯丙氨酸解氨酶( PAL) 和几丁质酶(CHT) 活性, 积累了较高水平的H₂O₂ 和酚类物质。离体试验表明, ASM不对病原菌生长产生抑制作用。ASM处理对果实病害的抑制作用可能与其增强了果实抗性系统有关。     

Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001     

Microscopic detection of reactive oxygen species generation in the compatible and incompatible interactions of Alternaria alternata Japanese pear pathotype and host plants

 Reactive oxygen species (ROS) generation was examined in the interaction of Alternaria alternata Japanese pear pathotype and host plants using three methods: nitro blue tetrazolium (NBT) method for microscopic detection of O₂ ⁻, diaminobenzidine (DAB) methods for microscopic detection of H₂O₂, and cerium chloride methods for ultrastructural detection of H₂O₂. ROS generation was detected by NBT and DAB methods at appressoria on leaves of susceptible cultivars and heat-shocked leaves of resistant cultivars but not in leaves of resistant cultivars. Ultrastructural detection by the cerium chloride method identified ROS generation at cell walls of appressoria and penetration pegs in susceptible, resistant leaves and heat-shocked leaves. These differences in the ultrastructural and microscopic data in resistant areas were due to the restriction of ROS generation in limited areas, the side facing the plant surface, of appressoria and penetration pegs. Therefore, ROS generation was apparently induced regardless of the resistance or susceptibility of the cultivar with the difference being in the volumes generated. After evaluating the pathological role of ROS generation in fungal structures, such generation was found to be associated with early penetration of cell walls in pear plants. Additionally, ROS generation in plants was also found in degrading pectin layers near infected hyphae and in plasma membrane modification sites in susceptible leaves but not in resistant leaves. ROS generation in susceptible leaves might be accompanied with plasma membrane damage, although the role of ROS generation in the pectin layers is not clear. ROS generation in both fungal and plant cells during their interaction was likely associated with the expression of susceptibility. Received: June 3, 2002 / Accepted: July 31, 2002     

肉牛源细极链格孢菌的分离鉴定

为鉴定肉牛皮肤病病原及其致病性并筛选其敏感药物,本研究取患皮肤病肉牛病变部位皮屑、毛发和痂皮进行病原菌的分离,并对分离菌株进行了形态学鉴定、ITS序列分析、小鼠致病性试验和药敏试验。结果显示分离到1株形态特征与细极链格孢菌极其相似的菌株(PY2-1-2);经PCR扩增、测序和BLAST序列比对,结果显示分离株(PY2-1-2)的ITS基因序列与细极链格孢菌(MG975630.1)的ITS基因序列的相似性为99%,分离株PY2-1-2的ITS基因序列长度为543 bp,该片段包括ITS1、5.8S rDNA和ITS2的全部基因序列以及18S rDNA和28S rDNA的部分基因序列,提交GenBank(MH656780.1)。根据形态学结合ITS基因序列分析结果确定该分离株为细极链格孢菌。小鼠致病性试验结果显示该菌对小鼠有致病性。药敏试验结果显示:该菌对灰黄霉素、氟康唑、伊曲康唑、酮康唑的最小抑菌浓度(MIC)值分别为0.5μg/mL、4μg/mL、1μg/mL、1μg/mL。本研究为今后进一步探究细极链格孢菌引起的皮肤病诊治提供科学有效的参考。     

The culturable leaf mycobiome of Viscum album subsp. austriacum

In Poland's pine forests, mistletoe (Viscum album subsp. austriacum) abundance has increased over the past 20 years. This mistletoe infestation has decreased the growth and vigour of Scots pine trees. In this study, we surveyed the culturable fungi from healthy and diseased V. album subsp. austriacum leaves from two stands in Poland. In total, 63 distinct species were identified, 99.8% of which belonged to the phylum Ascomycota. The community compositions of fungi in mistletoe leaves were similar in healthy and diseased leaves as Alternaria alternata, Aureobasidium pullulans, Botryosphaeria visci, Fusarium paeoniae and Microsphaeropsis olivacea were consistently found in leaves of all symptom types. The most frequently isolated fungus from asymptomatic leaves was M. olivacea, followed by A. alternata, A. pullulans and Hypoxylon rubiginosum. In comparison, the most frequently isolated fungi from leaves with rusty-brown necrotic spots were (in decreasing order) M. olivacea, B. visci, F. paeoniae and A. alternata, while the most frequently isolated fungi from leaves with black or dark brown spots were M. olivacea, A. alternata, A. pullulans, Epicoccum layuense and F. paeoniae. This study was the first comprehensive report showing that certain fungal species may be pathogens of V. album subsp. austriacum in Poland. This study was also the first report of F. paeoniae, F. juglandicola, Diaporthe vacuae and Heterotruncatella spartii from V. album, and the first report of D. vacuae and H. spartii in Poland.     

贡柑黑腐病病原菌生物学特性研究

摘要：在贡柑黑腐病病原菌分离鉴定的基础上,对其病原菌的生物学特性进行研究。结果表明,该病原菌（Alternaria citri Ellis & Everhart）在PDA培养基上生长最佳。菌丝生长和分生孢子萌发的适宜温度分别为21~30 ℃和18~30 ℃,适宜pH值分别为5~12和4~7。供试的6种碳源中,葡萄糖和可溶性淀粉较利于菌丝生长,蔗糖较利于孢子萌发;供试的6种氮源中,牛肉膏较利于菌丝生长,酵母膏较利于孢子萌发。光照条件对病原菌产孢影响不大,完全光照与光暗交替12 h较利于菌丝生长和孢子萌发。分生孢子致死温度为51 ℃水浴10 min。     

贡柑黑腐病病原菌的分子鉴定

贡柑黑腐病是近年来在贡柑上发生的一种新型病害,该病害可导致嫩梢快速枯死,大量嫩叶脱落,严重影响树势和产量。本研究从病区采集发病枝叶,对贡柑黑腐病病原菌进行分离、纯化,并采用rDNA-ITS序列分析技术对其进行分子鉴定。结果表明,贡柑黑腐病病原菌为链格孢属（Alternaria Nees）真菌,rDNA-ITS序列与GenBank中芸薹链格孢菌(Alternaria brassicae (Berk.) Sacc.)和柑桔链格孢菌(Alternaria citri Ellis & Everhart)的ITS序列的同源性为99%。结合形态学特征,推断该病原菌为柑桔链格孢菌。     

抗柑桔褐斑病菌和炭疽病菌宽皮柑桔种质的筛选

由交链格孢菌Alternaria alternata和胶孢炭疽菌Colletotrichum gloeosporioides侵染引起的柑桔褐斑病和炭疽病是重庆万州地区红桔的主要真菌病害。为筛选适合万州地区发展的柑桔品种,采用离体叶片室内人工接种和病情指数抗性评价方法探讨了16份宽皮柑桔种质对柑桔褐斑病菌和炭疽病菌的抗性。结果表明：对于柑桔褐斑病菌,不知火桔橙、W?默科特、春见桔橙和温州蜜柑为高抗种质;对于柑桔炭疽病菌,不知火桔橙、W?默科特、蔓生红桔和苏红为高抗种质。综合考虑,重庆万州地区可选择抗病性较强的鹅蛋红桔、福桔、安江红桔、不知火桔橙和W?默科特等宽皮柑桔进行品种替换和提升。     

银海枣褐斑病的病原鉴定与化学防治

对广东省新近发现的一种银海枣叶部病害进行了病原鉴定,该病病原为链格孢Alternaria alternata（Fr.）Keissl．以生长速率法和孢子萌发法对该病害进行化学防治试验的结果表明：在参试的11种杀菌剂中,叶斑净和世高的防治效果最好,对病斑扩展的抑制率分别达52．82％和33．62％．     

无机营养N,P,Ca和K对黑斑病菌侵染甘蓝幼苗的影响

将甘蓝幼苗分别培养在缺N(oN)、缺P(oP)、缺Ca(oCa)或缺K(oK)的不完全营养液中,培养56 d后分别于离体(Seg)和非离体(Int)叶片上接种黑斑病菌,以N,P,Ca和K营养素齐全的全营养液为对照,通过调查不同营养液处理后甘蓝叶片黑斑病的病斑大小,探讨N,P,Ca和K等无机营养素对黑斑病菌侵染甘蓝幼苗的影响。试验分2部分完成:第1部分缺N,缺P和缺Ca 3个处理同时进行,试验结果表明,不同营养液培养的甘蓝叶片黑斑病病斑大小有显著差异,顺序为对照>缺P>缺Ca>缺N处理;而在对照、缺P和缺Ca处理中,同一处理的非离体叶片病斑明显大于离体叶片病斑,缺N处理情况则相反。第2部分缺K处理的试验中,接种后5 d的病斑明显小于对照,非离体叶片与离体叶片接种后的病斑分别比对照小53%和14%;而缺K处理的离体叶片病斑显著比非离体叶片大(37%),并且病情随着接种后时间的延长而加重,在接种22 d后植株叶片和离体叶片的病斑大小趋于平衡。相关性分析表明,接种后5 d甘蓝幼苗活体组织和衰老组织的症状表达没有明显的相关性,但从试验2接种后22 d以及整个试验所有数据的结果分析,两者之间存在一定的相关性,相关系数达r=0.61。