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鸡腿菇炭角菌病害发生机理及其防治研究

Occurrence Mechanism and Control Technology of Xylaria Pedunculata Pathogene of Coprinus Comatus

【作者】 杜爱玲

【导师】 曹支敏;

【作者基本信息】 西北农林科技大学 , 植物病理学, 2006, 博士

【摘要】 鸡腿菇是近些年迅速发展的重要食用菌之一,在我国已广泛栽培,总状炭角菌是伴随鸡腿菇栽培而出现的一种新病害,这种病害目前只在鸡腿菇生产中有报道。该菌一旦发生对鸡腿菇造成的危害极大,轻者造成鸡腿菇减产,重者绝收,且危害越来越严重。本文首次对总状炭角菌的生物学特性及其与鸡腿菇的互作关系进行了研究,并在此基础上提出了鸡腿菇炭角菌病害的综合控制方法。研究结果主要如下:1.首次系统研究了总状炭角菌的生物学特性。结果显示该菌在不同的培养基上菌落特征差异很大,菌落菌丝从匍匐形菌丝、绒状菌丝、索状菌丝到子座状菌丝团和革质状菌丝结构都有可能形成。在综合PDA斜面培养基上,一般15~20d左右长满斜面,在平板培养基上20~25d布满平面;在综合PD液体培养基上,250mL三角瓶,静止培养,20~25d菌丝布满液体表面。在菌丝顶端和菌丝中部都易形成分生孢子,菌丝形成的分生孢子与子座表面组织形成的分生孢子形态相似。菌丝在常规培养料中不能生长,在PDA培养基上仅有无性阶段,只有与鸡腿菇共培养时才能形成完整的子座及其有性阶段。该炭角菌菌丝生长温度为10~37℃,最适温度25~30℃。分生孢子萌发温度为12~40℃,最适温度25~35℃,空气相对湿度要求在90%以上。菌丝生长pH为2~11,最适5~7;分生孢子萌发pH为3~11,最适5~8。最有利于菌丝生长的碳源为麦芽糖和葡萄糖,氮源为酵母膏、蛋白胨、牛肉膏。仅含有20 g/L葡萄糖、20 g/L蔗糖或者纯水的基质不利于分生孢子萌发。固态基质和良好通气条件有利于菌丝生长和孢子萌发。光照不利于菌丝生长,但对分生孢子萌发影响不大。菌丝致死温度47~50℃,分生孢子致死温度43~45℃。分生孢子以单极萌发为主,在子座上一般不萌发。2.从生物量、电导率、胞外酶活性等生理生化角度研究了总状炭角菌对鸡腿菇菌丝生长发育的影响,结果证明总状炭角菌影响鸡腿菇生长的主要机理是产生毒素活性物质。这类物质破坏细胞,导致鸡腿菇菌丝扭曲、相互粘连,细胞膜透性增加,菌丝相对电导率升高,菌丝生长期和子实体生长期胞外羧甲基纤维素酶和蛋白酶活性降低等。3.通过总状炭角菌与平菇、香菇、黄伞和鸡腿菇等几种食用菌的平板对峙培养和出菇试验证明,有鸡腿菇生长的地方才能生长总状炭角菌,说明只有鸡腿菇具有诱导总状炭角菌发生的作用。拮抗性观察发现鸡腿菇对总状炭角菌没有拮抗作用,反而具有独特的促进作用。显微观察二者共培养交接处菌丝和鸡腿菇生产上感染总状炭角菌的菌丝,结果表明总状炭角菌对鸡腿菇的寄生现象不明显。4.通过进一步鸡腿菇代谢物对总状炭角菌菌丝生长和胞外酶活性的影响试验,初步明确了总状炭角菌病害发生机理和致病机理。即鸡腿菇含有对总状炭角菌生长和胞外酶活性提高具促进作用的特殊代谢物质,该物质的存在诱导了总状炭角菌的发生;总状炭角菌发生以后一方面与鸡腿菇产生抑制竞争作用,另一方面又产生对鸡腿菇生长不利的毒素活性物质,毒素物质破坏鸡腿菇生长细胞;而对于总状炭角菌的发生鸡腿菇又不具备抵抗力和拮抗性,不能抑制其蔓延,致使该菌成为鸡腿菇栽培中的一大病害。5.从鸡腿菇栽培季节、菇房温度、菇房环境和覆土湿度等方面研究了总状炭角菌病害的发生发展规律,在此基础上初步建立了总状炭角菌综合防治模式,提出做好鸡腿菇栽培期间的消毒工作,合理调控生长环境,改变栽培方式是控制总状炭角菌菌病害的最有效手段。

【Abstract】 Coprinus comatus mushroom is one of rapid development and important edible fungi in recent years,and cultivated extensively in our country.Xylaria pedunculata is a new and predominant pathogenes of Coprinus comatus.This fungus only occurred on the cultivation bed of Coprinus comatus and once it occurred, the harm on Coprinus comatus would be severe, which may lead to yield drop and even failure of Coprinus comatus production.The paper discussed the biology characteristic of Xylaria pedunculata and the interaction between Coprinus comatus for the first time. Finally,according to the research, synthesis preventing and controlling pattern were established.The main results were as follows:1. The biology characters of Xylaria pedunculata were systematically studied for the first time. The results showed that the morphology of mycelia differed from creeping, to velvet, strand or coriaceous when it was incubated in different media. On the synthetical PDA slant medium, mycelia full grew with about 15-20d, on the synthetical PDA plate medium with about 20-25d and on the synthetical PD liquid medium with about 20-25d. The conidia also could form in the top or middle of mycelium on the media.The conidia which formed from tissue on the stroma surface and mycelium on the media were similar.The mycelium of Xylaria pedunculata could not grow on the normal substrates and there was only imperfect stage on the PDA,the stroma and perfect stage formed only when it was cultured together with Coprinus comatus.For the growth of mycelia, The range of temperature was 10-37℃,with the optimum of 25-30℃, The favorable pH was 2-11 with the optimum pH 5-7.For the germination of conidia, the range of temperature was 12-40℃,with the optimum of 25-35℃,the range of pH was 3-11,with the optimum pH 5-8,and the relative humidity must exceed 90%. Among the tested carbon and nitrogen sources, glucose, maltose, yeast extract, peptone, beef extract were favorable to the growth of mycelia. 20 g/L glucose or 20 g/L sucrose or pure water only in the agar media was not suitable for germination. Solid media and aeration condition were favorable to the growth of mycelia and germination of conidia. Light inhibited the growth of mycelia, but did not have significant effect on the germination of conidia. The lethal temperature for the mycelium was 47-50℃and 43-45℃for the conidia. The germination from unit pole was as predominant type and not germination on the stroma. 2. Determination on the biomass,the conductivity and the extracellular enzyme activity of Coprinus comatus revealed that there was some toxic active substance in the metabolite of Xylaria pedunculata, and toxic substance was correlation to inhabitation mechanism of Xylaria pedunculata on the growth of Coprinus comatus.This kind of matter could led mycelium of Coprinus comatus to istortion and mutual adhesion, penetrability of cell membrane and relative conductivity increasing, and CMCase and proteinases activity decreasing both in mycelium growth phase and fruiting phase.3.The experiment on the interaction between edible fungi (pleurotus ostreatus, lentinus edode, pholiota adipdsa, Coprinus comatus) and Xylaria pedunculata were studied with methods of plate dual culture and cultivation.The results showed that Xylaria pedunculata grew only in the growth area of Coprinus comatus,which was not antagonism with its growth and reversely could induce its stroma formation.Under optical microscope, further observation on the mycelium of Xylaria pedunculata and Coprinus comatus on the dual culture plate and in the infected cultivation bags was conducted,the results showed that Xylaria pedunculata was not parasitism to Coprinus comatus.4.Further experiment on the mycelium growth and the extracellular enzyme activity of Xylaria pedunculata proved that there was some special bioactive substance in the metabolite of Coprinus comatus, which had positive effects on the growth and the extracellular enzyme activity of Xylaria pedunculata.This kind of bioactive matters was of inducing action to stroma formation of Xylaria pedunculata,and when it occurred, then compete with Coprinus comatus and destructed its cell through toxic matter.Furthermore,Coprinus comatus was not resistance and antagonism to the Xylaria pedunculata.So the bioactive substance and the promotion of extracellular enzyme activity of Xylaria pedunculata might be one of occurrence mechanisms.5. The research demonstrated that the growth and development situation of Xylaria pedunculata was correlation highly to Coprinus comatus cultivation season, room temperature, relative humidity and casing layer moisture .Basing on the research, synthesis preventing and controlling pattern were established and the author consider that the most effective way to control Xylaria pedunculata is by well sterilizing in the cultivation process and regulating the ecological environment which the Coprinus comatus mushroom grows and changing cultivation methods of Coprinus comatus.

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