文档介绍：中南民族大学
硕士学位论文
不同生态条件和耕作制度烟田土壤微生物多样性的研究
姓名:王家昕
申请学位级别:硕士
专业:生物化学与分子生物学
指导教师:李晓华
2011-05-23
中南民族大学硕士学位论文
摘要
采用 Bead beating 法提取了不同生态条件和耕作制度的烟田土壤微生物总 DNA,
以细菌 16S rDNA V3 区通用引物对进行降落 PCR 扩增,产物进行变性凝胶梯度电泳
(DGGE),经过对不同生态条件和耕作制度烟田土壤 DGGE 指纹图谱的分析,主要
结论如下:
Bead beating法提取土壤微生物总DNA,既保证了DNA不受机械损伤,又兼顾了
DNA的质量,提取的微生物DNA可直接用于后续酶反应。
单一施用有机肥对土壤微生物多样性影响最大。施用75%无机肥配25%有机肥利
于较快提高土壤中微生物多样性。单一施用无机肥降低根际土中微生物多样性,能提
高非根际土中微生物多样性。
在烟草生长过程中,连作土壤中微生物种群变化幅度最大,微生物多样性最高,
其次为间隔2年土壤样品;间隔4年土壤中微生物多样性随烟草生长降低。
海拔 746m 土壤中,非根际土微生物多样性从旺长期到采收期呈降低趋势,根际
土相反。海拔 933m 和海拔 1095m 土壤中微生物多样性从旺长期到采收期升高,其中
海拔 1095m 的土壤微生物多样性大于海拔 933m 土壤。随海拔升高,非根际土与对应
根际土的相似性降低。海拔越低,旺长期土壤与采收期该土壤的相似性越高。
黄棕壤非根际土移栽期时微生物多样性最低,根际土移栽期微生物多样性最高,
微生物多样性从烟草旺长期到采收期变化较小。紫色土微生物多样性低于黄棕壤。从
旺长期到采收期紫色土微生物多样性降低明显。黄棕壤旺长期与采收期的相似性大于
紫色土旺长期与采收期的相似性。

关键词:土壤微生物多样性;PCR-DGGE;耕作制度;生态条件;

I
不同生态条件和耕作制度烟田土壤微生物多样性的研究
Abstract
This article uses Bead beating method to extract total DNA in different ecological
condition and cultivation system, touchdown PCR was performed using 16S rDNA V3
region consensus primer set, and the products were analyzed by denaturing gradient gel
electrophoresis (DGGE). The main results are shown as follows:
The DNA-extracting method of Bead beating not only protected DNA from mechanical
damage but also ensured the DNA's quality, and the extracted DNA could be directly used
in follow enzyme reactions.
Single use anic fertilizer had the biggest effect on soil microbial diversity. Using
75% anic fertilizer with 25% organic fertilizer helped increasing soil microbial
diversity rapidly. The microbial diversity reduced in single anic fertilized rhizosphere
soils, meanwhile, increased microbial diversity of non-rhizosphere soils.
During the growth phase of o, the change of microbe population in continuous
cropping soil was substantial, and the microbial diversity of continuous cropping soil was
the highest, soil which interval year is 2 was the second. T