發(fā)布時(shí)間：2018-03-04 02:26

  本文選題：人參　切入點(diǎn)：根際微生物　出處：《延邊大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：人參Panax ginseng C.A.Meyer),是我國名貴藥用植物,也是吉林省主要特產(chǎn)之一。長白山區(qū)以其獨(dú)特的地理位置和氣候條件,成為吉林省人參主產(chǎn)區(qū)。人參屬于忌地性極強(qiáng)的植物,存在連作障礙,長期采用伐林栽參,影響可持續(xù)發(fā)展。因此,人參產(chǎn)業(yè)的發(fā)展受到了阻礙,解決人參忌連作問題迫在眉睫。大多數(shù)學(xué)者認(rèn)為連作障礙與土壤理化性狀變劣、微生物體系的變化存在密切關(guān)系。但從土壤改良方面只能從一定程度上緩解連作的癥狀,病害發(fā)生、產(chǎn)量低的問題仍然存在。本實(shí)驗(yàn)從根際微生物的角度進(jìn)行分析,利用微生物純培養(yǎng)、PCR-DGGE技術(shù)、基因克隆方法開展人參忌連作問題研究,重點(diǎn)研究真菌對植物病害的影響,以期為人參病害預(yù)防、病理研究以及開發(fā)防治人參病害殺菌劑提供基礎(chǔ)。研究的主要結(jié)論如下：1、確立了人參根際土壤真菌ITS區(qū)PCR反應(yīng)體系：25μL反應(yīng)體系DNA濃度52.5 ng、dNTPs濃度0.20 mM、Mg2+濃度1.50 mM、引物濃度4.80μmol搭配效果最佳。2、篩選出適合真菌DGGE指紋圖譜分析的引物ITS1/ITS4、ITS1-GC/ITS2,并確定真菌DGGE凝膠單體濃度為12%-6%,變性劑濃度為60%-40%。細(xì)菌分析采用引物314F-GC/518R,DGGE凝膠單體濃度為8%,變性劑濃度為60%-40%。3、研究結(jié)果表明根際真菌與非根際真菌種類存在明顯差異,并且真菌種類隨著種植年限的增加而變少。隨著種植年限的增加同種真菌在數(shù)量上也發(fā)生了變化,表現(xiàn)出變少的趨勢,而且有少量新的真菌滋生。人參根際細(xì)菌與非根際細(xì)菌多樣性指數(shù)變化不明顯,但有參地細(xì)菌量明顯多于無參地,后期滋生的細(xì)菌有可能存在致病菌。4、通過基因克隆測序得到68條測序結(jié)果,將測序結(jié)果系統(tǒng)分析,BLAST對比得出人參根際土壤中分別有菌根真菌(Mycorrhizal fungus)、擔(dān)子菌屬(Basidiomycota sp.)、真子囊菌門(Euascomycete sp.)、枝孢屬(Euascomycete sp.)、被孢霉屬(Mortierella alpina)、子囊菌屬(Ascomycete sp.)地絲霉屬(Geomyces sp.)、類球囊霉屬(Paraglomus laccatum)、隱球菌(Cryptococcus podzolicus) 、子囊菌(Ascomycota sp.) 。
[Abstract]:Ginseng Panax ginseng C.A. Meyerian is one of the most valuable medicinal plants in China and one of the main specialties of Jilin Province. Changbai Mountain area, with its unique geographical location and climatic conditions, has become the main producing area of Panax ginseng in Jilin Province. Therefore, the development of ginseng industry has been hindered, so it is urgent to solve the problem of ginseng non-continuous cropping. Most scholars think that continuous cropping obstacle and soil physical and chemical properties become worse. The changes of microbial system are closely related. But in the aspect of soil improvement, the symptoms of continuous cropping can only be alleviated to a certain extent, the problems of disease occurrence and low yield still exist. This experiment is analyzed from the point of view of rhizosphere microorganism. In order to prevent ginseng disease, PCR-DGGE technique and gene cloning method were used to study the effects of fungi on plant diseases. The main conclusions of the study are as follows: 1. The PCR reaction system of ginseng rhizosphere fungi in ITS region was established. The reaction system of PCR was established. The concentration of DNA was 52.5 渭 L and the concentration of DNA was 0.20 mg / mg _ 2 and the concentration of primer was 1.50 mm. The best concentration of 4.80 渭 mol was 4.80 渭 mol. The primers ITS1 / ITS4 / ITS1-GC / ITS2 were selected, and the concentration of fungal DGGE gel monomer was 12-6, denaturant concentration was 60-40.The bacterial analysis was conducted with primer 314F-GCr518RDGGE gel monomer concentration 8 and denaturant concentration 8. The results showed that there were significant differences between the species of rhizosphere fungi and non-rhizosphere fungi. With the increase of planting years, the number of fungi of the same species also changed, showing a trend of decreasing. The diversity index of ginseng rhizosphere bacteria and non-rhizosphere bacteria was not obvious, but the number of ginseng rhizosphere bacteria was more than that of non-ginseng soil. It is possible that the bacteria bred in the later period may have pathogenic bacteria. 68 sequences were obtained by gene cloning and sequencing. The systematic analysis of sequencing results showed that Mycorrhizal fungus, Basidiomycota sp., Euascomycete sp., Euascomycete sp., Mortierella alpina., ascomycete sp., Mycorrhizal Fungi, Ascomycete sp., Euascomycecete sp., Mortierella alpinaanus, Ascomycete sp., respectively, were found in the rhizosphere soil of Panax ginseng, and that the genus Geomyces sp., Basidiomycota sp., Euascomycete sp., Euascomycete sp., Mortierella alpinaanus, Ascomycete sp.) were found in the rhizosphere soil of Panax ginseng. Of the genus Paraglomus laccatumus, Cryptococcus podzolicus, Ascomycota sp.)
【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S567.51;S154.3

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