【摘要】：近年來(lái),隨著轉(zhuǎn)基因玉米的推廣和種植面積的不斷擴(kuò)大,其生物安全性也已引起人們的廣泛重視。土壤生態(tài)系統(tǒng)在物質(zhì)循環(huán)和能量轉(zhuǎn)化中占有重要的地位,轉(zhuǎn)基因玉米的外源基因可能通過(guò)根系的分泌物或作物殘茬或者其他途徑流入土壤生態(tài)系統(tǒng)中。土壤中生存的特異微生物類(lèi)群的功能性質(zhì)以及微生物群落結(jié)構(gòu)的多樣性都可能受到轉(zhuǎn)基因植株的影響。傳統(tǒng)的微生物研究方法具有一定的局限性,而一些分子生物學(xué)技術(shù)的應(yīng)用為土壤微生物的研究提供了新的有力工具,RAPD技術(shù)則是其中一種重要的分子生物學(xué)技術(shù)。本實(shí)驗(yàn)采用傳統(tǒng)平板分離法研究了不同生長(zhǎng)時(shí)期轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米和非轉(zhuǎn)基因玉米之間土壤微生物數(shù)量的變化,以及采用RAPD(Randora Amplified Polymorphie DNA)分析技術(shù)研究了不同時(shí)期可培養(yǎng)細(xì)菌與可培養(yǎng)真菌的群落結(jié)構(gòu)的變化。平板分離計(jì)數(shù)結(jié)果顯示,轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米與非轉(zhuǎn)基因玉米的根區(qū)土壤中主要微生物群落數(shù)量與其發(fā)育時(shí)期有關(guān)。根區(qū)微生物數(shù)量一般是由幼苗期開(kāi)始升高,到生長(zhǎng)期達(dá)到高峰,成熟期開(kāi)始下降。轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米與非轉(zhuǎn)基因玉米相比,在可培養(yǎng)細(xì)菌數(shù)量上,根際土壤中細(xì)菌數(shù)量在幼苗期、生長(zhǎng)期沒(méi)有差異,而在成熟期則產(chǎn)生了差異,且達(dá)到了顯著水平(P0.05)；在可培養(yǎng)真菌數(shù)量上,根際土中真菌數(shù)量在生長(zhǎng)期和成熟期均產(chǎn)生了差異,且達(dá)到了顯著水平(P_0.05)；在可培養(yǎng)放線(xiàn)菌數(shù)量上,根際土中放線(xiàn)菌數(shù)量在各發(fā)育時(shí)期均未產(chǎn)生顯著差異(P0.05)。轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米與非轉(zhuǎn)基因玉米相比,在根區(qū)外周土壤中,可培養(yǎng)細(xì)菌、真菌和放線(xiàn)菌的數(shù)量均未產(chǎn)生顯著差異(I)0.05)。由此可知,轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米與非轉(zhuǎn)基因玉米相比,在根際土中,對(duì)土壤可培養(yǎng)細(xì)菌的影響在成熟期最大,在生長(zhǎng)期、成熟期對(duì)土壤可培養(yǎng)真菌均有顯著影響,而對(duì)于可培養(yǎng)放線(xiàn)菌,各生長(zhǎng)時(shí)期的數(shù)量均無(wú)顯著變化；而在根區(qū)外周土壤中,轉(zhuǎn)基因抗病玉米對(duì)各生長(zhǎng)時(shí)期土壤中的細(xì)菌、真菌與放線(xiàn)菌的數(shù)量均無(wú)顯著影響。利用RAPD技術(shù)分析根區(qū)可培養(yǎng)細(xì)菌與可培養(yǎng)真菌群落結(jié)構(gòu)的變化。結(jié)果顯示：轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米與非轉(zhuǎn)基因玉米相比,在對(duì)可培養(yǎng)細(xì)菌群落結(jié)構(gòu)的影響上,根際土壤細(xì)菌群落結(jié)構(gòu)的相似性在不同生長(zhǎng)時(shí)期變化較大,其中,在幼苗期最大為83.33%,在生長(zhǎng)期最小為57.14%；對(duì)于根區(qū)外周土壤,細(xì)菌群落結(jié)構(gòu)相似性在收割后最小為70.80%,幼苗期最大為89.17%,不同生長(zhǎng)時(shí)期的細(xì)菌群落結(jié)構(gòu)差異變化較小。說(shuō)明轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米對(duì)根區(qū)土壤細(xì)菌的群落結(jié)構(gòu)影響較小。轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米與非轉(zhuǎn)基因玉米相比,在對(duì)可培養(yǎng)真菌群落結(jié)構(gòu)的影響上,根際土壤真菌在生長(zhǎng)期與成熟期的群落結(jié)構(gòu)的相似性較小,其中,在成熟期最小為54.17%, 幼苗期最大為75.00%；對(duì)于根區(qū)外周土壤,不同生長(zhǎng)時(shí)期的真菌群落結(jié)構(gòu)相似度則相對(duì)較高,真菌群落結(jié)構(gòu)相似度在幼苗期最大為77.08%,成熟期最小為56.25%。說(shuō)明轉(zhuǎn)幾丁質(zhì)酶基因抗病玉米對(duì)根區(qū)土壤真菌的群落結(jié)構(gòu)產(chǎn)生了一定的影響作用。
[Abstract]:In recent years, with the popularization of transgenic maize and the continuous expansion of planting area, its biological safety has attracted widespread attention. Soil ecosystem plays an important role in the material cycle and energy transformation. The exogenous genes of transgenic maize may flow into the soil through root exudates or crop residues or other ways. In soil ecosystems, the functional properties of specific microbial communities and the diversity of microbial community structure in soils may be affected by transgenic plants. Traditional methods of microbial research have some limitations, and the application of some molecular biology techniques provides a new powerful tool for soil microbial research. RAPD technique is one of the most important molecular biology techniques. In this experiment, the number of soil microorganisms between chitinase-resistant and non-transgenic Maize at different growth stages was studied by traditional plate separation method, and RAPD (Randora Amplified Polymorphie DNA) analysis technique was used to study the variation of soil microorganisms at different times. The results of plate separation and counting showed that the number of major microbial communities in the root zone of transgenic and non-transgenic maize was related to their development stages. Compared with non-transgenic maize, the number of bacteria in rhizosphere soil had no difference at seedling stage and growth stage, but had a significant difference at maturity stage (P 0.05). The number of culturable actinomycetes in rhizosphere soil had no significant difference at all development stages (P 0.05). Compared with non-transgenic maize, the number of culturable bacteria, fungi and actinomycetes in rhizosphere soil had no significant difference (P 0.05). There was no significant difference in the number of soil fungi (I) between transgenic maize and non-transgenic maize (I) 0.05). Therefore, compared with transgenic maize, transgenic maize with Chitinase gene had the greatest influence on soil culturable bacteria in the rhizosphere soil at the mature stage, and had significant influence on soil culturable fungi during the growth and maturity stage, while for culturable actinomycetes, the number of each growth stage was significantly affected. There was no significant change in the amount of bacteria, fungi and actinomycetes in the peripheral soil of the root zone. RAPD technique was used to analyze the community structure of culturable bacteria and fungi in the root zone. Compared with maize, the similarity of bacterial community structure in rhizosphere soil varied greatly in different growth stages, with the maximum being 83.33% in seedling stage and the minimum being 57.14% in growth stage; the minimum similarity of bacterial community structure was 70.80% in peripheral soil of root zone after harvest and the maximum in seedling stage. The difference of bacterial community structure in different growth stages was small, indicating that transgenic chitinase-resistant maize had little effect on soil bacterial community structure in root zone. The similarity of community structure in the mature stage was 54.17% at the minimum and 75.00% at the seedling stage; the similarity of Fungi Community Structure in different growth stages was relatively high in the soil around the root zone, and the similarity of fungi community structure was 77.08% at the seedling stage and 56.25% at the mature stage. Resistant maize had a certain effect on the community structure of soil fungi in root zone.
【學(xué)位授予單位】：山西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：S154.3;S513

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 陸雅海;張福鎖;;根際微生物研究進(jìn)展[J];土壤;2006年02期

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本文編號(hào)：2178066