本文選題：小麥禾谷孢囊線蟲病 + 生物防治��； 參考：《中國農(nóng)業(yè)大學(xué)》2015年博士論文

【摘要】：禾谷孢囊線蟲又名燕麥孢囊線蟲(Heterodera avenae Wollen-weber1924, Cereal Cyst Nematodes,簡稱CCN),為世界性禾谷類作物病原線蟲,現(xiàn)已廣泛分布于我國16個省市(自治區(qū)、直轄市)小麥產(chǎn)區(qū)。近幾年土壤蟲量逐年增加,病害逐年加重,造成小麥嚴(yán)重減產(chǎn),對小麥生產(chǎn)構(gòu)成威脅,現(xiàn)在已經(jīng)成為小麥生產(chǎn)上的主要防治對象。該病害發(fā)生在小麥根部,導(dǎo)致防治難度加大。由于化學(xué)劇毒農(nóng)藥的限制使用和抗原缺乏,目前還未找到對該病害有效的防治方法,對該病害芽胞桿菌的生物防治也未見報道。本項研究主要從生物防治角度出發(fā),通過篩選對小麥禾谷孢囊線蟲病具有防控效果的有益芽胞桿菌,評價其室內(nèi)、盆栽和田間病害防治效果,并對菌株進(jìn)行了鑒定,探索了生防芽胞桿菌小麥禾谷孢囊線蟲病的生防機(jī)制,結(jié)果表明： 本研究從小麥根內(nèi)和根際土壤中共分離得到293株芽胞桿菌,通過室內(nèi)快速篩選獲得8株對小麥禾谷孢囊線蟲J2具有穩(wěn)定殺線活性的芽胞桿菌,分別是菌株BMT19、B202、BMT20、BMT8、 YLT24、YCG3、YCG4和BMT11。通過溫室盆栽復(fù)篩和3年田間防效驗證,篩選出菌株B202具有穩(wěn)定防治小麥禾谷孢囊線蟲病效果,田間最高防效達(dá)到63.6%。菌株B202可以促進(jìn)小麥植株和根系生長,增加了小麥植株根系對土壤中全氮、速效P和速效K的吸收。通過Biolog檢測碳源利用和16S rDNA基因序列分析,鑒定菌株B202為短小芽胞桿菌(Bacillus pumilus),命名為為短小芽胞桿菌(Bacillus pumilus) B202。 菌株B202可以形成生物膜,能夠產(chǎn)生蛋白酶、嗜鐵素等多種生防相關(guān)物質(zhì)。通過雙抗標(biāo)記法檢測菌株B202在小麥根際定殖定殖情況,結(jié)果表明菌株B202可以在小麥根際土壤定殖65天以上,定殖量維持在107CFU/g；菌劑B202應(yīng)用到田間小麥根部后,對根際土壤線蟲群落組成成分無明顯改變作用,對種群優(yōu)勢度有調(diào)節(jié)作用,調(diào)節(jié)優(yōu)勢屬雙胃屬Panagrolaimus為常見屬,稀有屬短體屬Pratylenchus上升為次優(yōu)勢屬。施用菌劑B202后增加了根際土壤中線蟲總豐度,提高了食細(xì)菌類群線蟲、食真菌類群線蟲結(jié)構(gòu)比例,對植物寄生類群線蟲和雜食/捕食類群線蟲有鎮(zhèn)壓作用。施用菌劑B202后,線蟲多樣性指數(shù)中香農(nóng)一威納指數(shù)(H’)無明顯變化,營養(yǎng)類群指數(shù)(TD)下降,優(yōu)勢度指數(shù)(Ig)呈升高趨勢,植物寄生線蟲指數(shù)(PPI)呈下降趨勢；采用DGGE-PCR技術(shù)對田間應(yīng)用菌株B202后小麥根際土壤中細(xì)菌真菌多樣性進(jìn)行了檢測,結(jié)果表明施用菌株B202后提高了土壤細(xì)菌類群中綠彎菌門(Chloroflexi)、芽單胞菌門(Gemmatimonadetes)、擬桿菌門(Bacteroidetes)、變形菌門(Proteobacteria)和酸桿菌門(Acidobacteria)的菌群數(shù)量,增加了土壤真菌優(yōu)勢菌群中新麗鞭毛菌門(Mastigomycota)和子囊菌門(Ascomycotina)的菌群數(shù)量,降低了接合菌門(Zygomycota)的菌群數(shù)量；通過柱層析和HPLC制備技術(shù)分離純化獲得主要存在于菌株B202發(fā)酵上清液殺線活性物質(zhì)1-16和3-11,結(jié)合HPLC-MS和NMR測定,鑒定1-16為：鄰苯二甲酸二丁酯,分子式為：C16H2204,鑒定3-11為：4,4'-二苯甲烷二氨基甲酸甲酯,分子式為：C17H18N2O4。兩種化合物為首次在短小芽胞桿菌中發(fā)現(xiàn)。
[Abstract]:Cereal cyst nematode, also known as Heterodera avenae Wollen-weber1924 (Cereal Cyst Nematodes, CCN), is a worldwide cereal crop nematode, and is widely distributed in wheat producing areas in 16 provinces and municipalities (autonomous regions and municipalities) in China. In recent years, the amount of soil insects increased year by year, and the disease was aggravated year by year, causing serious reduction of wheat. Production, which poses a threat to the production of wheat, has now become the main control object in wheat production. This disease occurs at the root of wheat, which leads to more difficulty in prevention and control. Due to the limited use of chemical toxic pesticides and the lack of antigen, the effective prevention and control method for the disease has not been found, and the biological control of the bacillus is also not found. This study mainly from the biological control point of view, through screening the beneficial bacillus that has the control effect on the wheat cereal cyst nematode disease, evaluating the effect of the prevention and control of the indoor, potted and field diseases, and the identification of the strains, and exploring the biocontrol mechanism of the Bacillus spore bacillus cereus cyst nematode disease.
In this study, 293 strains of Bacillus were isolated from Magene and rhizosphere soil, and 8 strains of Bacillus with stable nematode activity to wheat cereal cyst nematode J2 were screened through indoor rapid screening. The strain BMT19, B202, BMT20, BMT8, YLT24, YCG3, YCG4 and BMT11. were screened by greenhouse potted rescreening and 3 year field effectiveness verification. The strain B202 has a stable effect on the control of wheat cereal cyst nematode disease. The highest efficiency of 63.6%. strain B202 in the field can promote the growth of wheat plant and root, and increase the absorption of total nitrogen in the soil, the available P and the available K by the root of the wheat plant. The strain B202 is identified by the use of Biolog detection carbon source and the sequence analysis of 16S rDNA gene. Bacillus cereus (Bacillus pumilus) is named Bacillus pumilus (B202.).
Strain B202 can form a biofilm to produce a variety of biocontrol related substances, such as protease, ferric, and so on. The colonization of strain B202 in wheat rhizosphere was detected by double resistance labeling. The results showed that the strain B202 could be colonized for more than 65 days in the rhizosphere soil of wheat and the colonization of the strain was maintained at 107CFU/g. After the inoculant B202 was applied to the root of wheat in the field, the strain B202 was applied to the root of wheat in the field. There was no obvious change in the composition of the rhizosphere soil nematode community, which had a regulating effect on the population dominance, and the regulatory dominance of the genus bis Panagrolaimus was the common genus, and the rare genus Pratylenchus increased to the secondary dominant genus. The application of microbial inoculant B202 increased the total abundance of line worms in the rhizosphere soil and improved the bacteria eating nematodes and the edible fungi. The structure ratio of the group nematode has a repression effect on the plant parasitic nematode and the omnivorous / predatory nematode. After the use of B202, the Shannon one Wiener index (H ") of the nematode diversity index has no obvious change, the nutrient group index (TD) decreases, the dominance index (Ig) shows a rising trend, the plant parasitic nematode index (PPI) shows a downward trend, and uses DG. The bacterial and fungal diversity in the rhizosphere soil of wheat after field application of strain B202 was detected by GE-PCR technology. The results showed that after the application of strain B202, the phylum green gate (Chloroflexi), butoomonas (Gemmatimonadetes), Pseudomonas (Bacteroidetes), deformable bacteria gate (Proteobacteria) and acid bacilli (Acidobac) in soil bacteria groups were improved. The number of teria bacteria increased the number of Mastigomycota and Ascomycotina in the dominant fungi group of the soil fungi, and reduced the number of bacteria groups of the conjugative bacteria gate (Zygomycota). The isolation and purification by column chromatography and HPLC preparation were mainly found in the nematoactive substance 1- of the strain B202 fermentation supernatant. 16 and 3-11, in combination with HPLC-MS and NMR, 1-16 were identified as: dibutyl phthalate, the molecular formula was C16H2204, and 3-11 was identified as 4,4'- two methanate two carbamate, and the molecular formula was found for the first time in Bacillus cereus, C17H18N2O4. two compounds.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：S476.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王愛玲;侯生英;彭德良;張貴;黃文坤;姚強(qiáng);;小麥禾谷孢囊線蟲病在青海發(fā)生分布狀況調(diào)查[J];安徽農(nóng)業(yè)科學(xué);2010年24期

2 連玲麗,吳祖建,段永平,謝聯(lián)輝;線蟲寄生菌巴斯德桿菌的生物多樣性研究進(jìn)展[J];福建農(nóng)業(yè)大學(xué)學(xué)報;2005年01期

3 黃金玲;劉志明;劉紀(jì)霜;陸秀紅;潘萍紅;;植物寄生線蟲生防細(xì)菌的研究進(jìn)展[J];廣西農(nóng)業(yè)生物科學(xué);2008年03期

4 王振躍,王守正,李洪連,袁紅霞;河南省小麥孢囊線蟲病的初步研究[J];華北農(nóng)學(xué)報;1993年S1期

5 王明祖，雷智峰，肖炎農(nóng);小麥禾谷孢囊線蟲（Heteroderaavenae）的主要寄主范圍研究[J];湖北植保;1995年06期

6 王泊理,高學(xué)彪;噬線蟲真菌研究進(jìn)展[J];華南農(nóng)業(yè)大學(xué)學(xué)報;1998年04期

7 王振躍;李洪連;袁虹霞;;小麥孢囊線蟲病的發(fā)生危害與防治對策[J];河南農(nóng)業(yè)科學(xué);2005年12期

8 王振躍;高書峰;李洪連;沙廣樂;余懋群;;不同小麥品種(系)對禾谷孢囊線蟲病的抗性鑒定[J];河南農(nóng)業(yè)科學(xué);2006年05期

9 吳緒金;楊衛(wèi)星;孫炳劍;邢小萍;王振躍;李洪連;袁虹霞;;不同藥劑處理對小麥禾谷胞囊線蟲的防治效果[J];河南農(nóng)業(yè)科學(xué);2007年05期

10 吳緒金;袁虹霞;張軍鋒;邢小萍;孫炳劍;李洪連;;小麥品種抗禾谷胞囊線蟲機(jī)制的初步研究[J];河南農(nóng)業(yè)科學(xué);2009年01期

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