本文關(guān)鍵詞： 膠孢炭疽菌(Colletotrichum gloeosporioides) 核桃(Juglans regia) 分生孢子萌發(fā) 附著胞形成 侵染過(guò)程　出處：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：核桃(Juglans regia L.)是重要的“木本糧油”生態(tài)樹種,為二倍體植物(2n=32),全基因組667 Mb。核桃作為“21世紀(jì)的超級(jí)食品”,果仁營(yíng)養(yǎng)成分極其適宜于人體健康,能有效預(yù)防心血管疾病,是Ⅱ型糖尿病人的首選食品。近年來(lái),隨著核桃集約栽培面積的擴(kuò)大和病害日漸嚴(yán)重。由膠孢炭疽菌(Colletotrichum gloeosporioides)引起的核桃炭疽病(walnut anthracnose)可致果實(shí)壞疽、葉片焦枯,同時(shí)還危害嫩梢,發(fā)病嚴(yán)重時(shí)可使30%~50%的青果脫落,導(dǎo)致產(chǎn)量損失,是目前核桃生產(chǎn)中的災(zāi)難性病害。本論文探究了影響膠胞炭疽菌分生孢子(conidia)萌發(fā)的條件,觀察膠胞炭疽菌在適宜條件下的生活過(guò)程;以保存于山東農(nóng)業(yè)大學(xué)林學(xué)試驗(yàn)站的核桃抗病品種‘瑞嘉’和無(wú)性系4-23、B26及B29為材料,解剖學(xué)觀察了膠胞炭疽菌侵染核桃組織的過(guò)程及核桃組織被侵染后的形態(tài)學(xué)變化;并對(duì)影響分生孢子(conidia)萌發(fā)的不同的環(huán)境因素進(jìn)行探究,以及觀察分生孢子在適宜條件下的生活過(guò)程。主要結(jié)果如下:1.影響膠孢炭疽菌分生孢子萌發(fā)的條件。孢子萌發(fā)最適萌發(fā)溫度為28-30℃、最適pH值為6.0、最適光照條件為全黑暗培養(yǎng)條件、最適濕度為100%濕度。以單糖為碳源均能促進(jìn)孢子萌發(fā),但最適合孢子萌發(fā)的碳源為葡萄糖和麥芽糖。以L-天冬氨酸、酵母提取物、牛肉膏和蛋白胨為氮源時(shí),均能促進(jìn)孢子萌發(fā),但最適合孢子萌發(fā)的氮源為酵母提取物。2.供試材料對(duì)膠胞炭疽菌抗性的鑒定。室內(nèi)接種核桃葉片和果實(shí)的發(fā)病率均高于田間接種。4-23果實(shí)平均發(fā)病率室內(nèi)和田間分別為84.44%和84%,顯著高于其他,表現(xiàn)為果實(shí)易感病,而葉片室內(nèi)和田間平均發(fā)病率分別為2.00%和1.33%,顯著低于其他品系,表現(xiàn)為葉片相對(duì)抗病�！鸺巍麑�(shí)發(fā)病率最低,室內(nèi)和田間平均發(fā)病率分別為51.11%和50%,顯著低于其他,表現(xiàn)為果實(shí)相對(duì)抗病。3.膠孢炭疽菌孢子的萌發(fā)過(guò)程。分別添加了葡萄糖為碳源及酵母提取物為氮源的培養(yǎng)基上,培養(yǎng)10h后,部分分生孢子萌發(fā)并產(chǎn)生芽管,培養(yǎng)18h后,絕大部分分生孢子萌發(fā)。培養(yǎng)24h后,分生孢子芽管的頂端膨大形成附著胞(appressorium)。培養(yǎng)36h后,附著胞的中心位置有侵染釘(penetration peg)的形成。培養(yǎng)48h后,侵染釘膨大形成初生菌絲。培養(yǎng)60h后,初生菌絲(primary hyphae)繼續(xù)生長(zhǎng),形成膨大的初生菌絲,培養(yǎng)72h后,菌絲繼續(xù)生長(zhǎng),形成次生菌絲(secondary hyphae)。4.膠孢炭疽菌對(duì)核桃的侵染過(guò)程。用分生孢子懸濁液對(duì)核桃無(wú)性系B29葉片侵染,結(jié)果發(fā)現(xiàn),接種24h,分生孢子在葉片表面萌發(fā),并形成附著胞附著于葉片表面;解剖學(xué)觀察結(jié)果顯示,葉片侵染部位表皮細(xì)胞出現(xiàn)略微凹陷。接種48h,附著胞中心部位出現(xiàn)侵染釘,同時(shí),分生孢子的形態(tài)發(fā)生略微變化;解剖學(xué)觀察結(jié)果顯示,葉片侵染部位表皮細(xì)胞出現(xiàn)明顯凹陷。接種60h,附著胞上產(chǎn)生侵染絲伸入到葉片氣孔腔內(nèi),氣孔腔周圍外層的組織中出現(xiàn)少量的初侵染絲;解剖學(xué)觀察結(jié)果顯示,葉片侵染部位表皮細(xì)胞破損,葉片內(nèi)部組織細(xì)胞凹陷明顯。接種72h,初生菌絲生長(zhǎng)并產(chǎn)生次生菌絲,細(xì)胞出現(xiàn)程序性死亡;解剖學(xué)觀察結(jié)果顯示,葉片侵染部位表皮細(xì)胞破損,內(nèi)部組織細(xì)胞出現(xiàn)部分消解。接種96h,侵染部位葉片出現(xiàn)褐色病斑,發(fā)病部位長(zhǎng)出分生孢子梗,分生孢子梗產(chǎn)生新的分生孢子,至此完成一個(gè)侵染循環(huán);解剖學(xué)觀察結(jié)果顯示,葉片侵染部位表皮細(xì)胞破損,內(nèi)部組織細(xì)胞大量消解。
[Abstract]:Walnut (Juglans regia L.) is an important "woody grain and oil" ecological species, diploid plants (2n=32), whole genome 667 Mb. walnuts as a "super food" in twenty-first Century, very suitable for nuts nutrition to human health, can effectively prevent cardiovascular disease, is the food of choice for people with type II diabetes. In recent years, with the expand the area of intensive cultivation and walnut disease. By Colletotrichum gloeosporioides (Colletotrichum gloeosporioides) walnut anthracnose caused by (Walnut anthracnose) can cause fruit gangrene, leaf scorch, also harm shoots, serious incidence of the 30%~50% fruit shedding, lead to yield loss, is currently in the production of walnut disaster disease. This paper explores the effects of Colletotrichum gloeosporioides conidia germination conditions (conidia), observe the life of Colletotrichum gloeosporioides in suitable conditions of the process; to preserve in Shandong agriculture University Forestry Experimental Station of walnut resistant cultivar 'Regal' and clones 4-23, B26 and B29 as materials, anatomical observation process of Colletotrichum gloeosporioides infection of walnut tissues and walnut tissues was morphological changes after infection; and the effects of conidia (conidia) germination of different environmental factors to explore, and observation conidia under suitable conditions of the life process. The main results are as follows: 1. the effect of Colletotrichum gloeosporioides conidia germination conditions. The spore germination of optimal germination temperature of 28-30, the optimum pH value is 6, the optimal light conditions for the dark culture condition, the optimum humidity is 100% with humidity. Monosaccharide as carbon source could promote spore germination, but the most suitable for spore germination of carbon source was glucose and maltose. L- aspartic acid, yeast extract, beef extract and peptone as nitrogen source, could promote spore germination, but the most suitable for spore germination The nitrogen source was yeast extract.2. for identification of test materials of Colletotrichum gloeosporioides resistance. Indoor inoculation walnut leaves and fruits of the incidence rate higher than the average incidence rate of.4-23 fruit field inoculation indoor and field were 84.44% and 84%, significantly higher than the other, as susceptible fruits, and leaves the average incidence of indoor and field rates were 2% and 1.33%, significantly lower than that of other strains, were relatively resistant. "Regal" leaves the lowest incidence of fruit, indoor and field average incidence rates were 51.11% and 50%, significantly lower than that of the other, for the germination process of fruit disease.3. against Colletotrichum gloeosporioides. Spores were added to glucose carbon source and yeast extract as nitrogen source on the 10h after culture, part of the conidia germination and germ tube, after 18h, the vast majority of conidia germination. After 24h culture, top spore germ tube Enlargement of appressorium formation (appressorium). After 36h culture, the center position of appressorium infection (penetration peg) nail formation. After 48h culture, the development of primary hyphae. Nail infection after 60H, primary hyphae (primary hyphae) continue to grow, to form a swollen primary hyphae, after 72h, hyphae continue to grow the formation of secondary hyphae (secondary, hyphae).4. infection process of Colletotrichum gloeosporioides on walnut. With conidia suspension of B29 infected leaves, walnut department found that inoculation with 24h clones, conidia germination on the leaf surface, and appressorium formation attached to the leaf surface; anatomical observation showed that slightly sunken leaves the infected part of epidermal cells. 48h inoculation, appressorium Center site infection nail, at the same time, conidia morphology changes slightly; anatomical observation showed that the leaf epidermal cells showed obvious infection site Sag. Inoculation of 60H, appressorium on produce infection into silk leaf stomatal cavity, stomatal cavity around the outer tissue in small primary infection silk; anatomy observation showed that the leaf epidermal cells of the infected part is damaged, the blade internal organization cell depression significantly. 72h vaccination, primary hyphae grow and produce secondary hyphae, the cells appeared program death; anatomical observation showed that the leaf epidermal cells of the infected part is damaged, the internal organization of cell part digestion. Inoculation with 96h infection leaves brown spots appeared, incidence of long conidiophores, conidia to produce new conidia, thus completing a cycle of infection; anatomical observation showed that leaf epidermal cells of the infected part is damaged, a large number of internal tissue digestion.

【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S436.64

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