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Study on Microbial Community Structure in the High-Yield Apple Orchard of Weibei Loess Plateau

Author: ZhouZuo
Tutor: LiBingZhi
School: Northwest University of Science and Technology
Course: Pomology
Keywords: Apple orchard DNA extraction DGGE Nitrogen levels
CLC: S661.1
Type: Master's thesis
Year: 2010
Downloads: 119
Quote: 0
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Abstract


The soil microbial participate in biochemical cycle, and is rich in resources. This experiment the use of Microbial Molecular Ecology does not depend culture, to overcome the limitations of traditional culture methods, directly extracted soil microbial DNA genome diversity to reflect the diversity of the species, which can comprehensively and objectively found that the composition of the soil microbial and diversity. The main content of the test from the environment, to accurately extract high-quality, representative of the total DNA. The Weibei area of ??high yield and quality of the apple orchard soil for the study, four typical soil optimization the Weibei area of ??soil microbial DNA extraction method. Compare 9 manual extraction of soil microbial DNA methods, including direct extraction method, the indirect method of extraction method and kit. In high salt SDS method, the pH of the extract and the lyase to be optimized. The use of improved methods of soil microbial community structure characteristics of different spatial levels, the last simple discussion of the relationship between the different nitrogen levels apple leaf characteristics and soil microbial community structure changes. The 16SrRNA V3 region fragment amplified using PCR detection accuracy of the method. Results is the use of the level of detection of the PCR amplified DNA specific fragment DGGE DGGE system eventually identified as follows: 60 ° C under the conditions of use of 8% acrylamide / methylene (37.5:1) denaturing concentration of 30% -70% of plastic, 150V electrophoresis 7h. 9 kinds of extracts obtained by the method of DNA fragments is greater than 23kb, most important is that each method has a high diversity of strip. Among them, the indirect extraction methods of soil microbial genomic DNA of higher quality than direct extraction, but the yield is low. Little humic acid content (less organic matter content) of the soil, indirect extraction method can be directly used in the subsequent PCR without purification. DGGE is a very effective tool in the evaluation of soil microbial community structure Extract pH and off-rot buffer the pH of the quality of soil microbial DNA. The Weibei area alkaline soil extract pH value off the rot at 9:00 of the highest quality, highest purity buffer pH value of 10. The cracking soil microbial, proteinase K fragment of the population structure. Compared the distribution of the orchard soil microbes space tree plate, around the crown and between the lines, from the trunk in Shupan (about 50cm) at the surface layer of 0 ~ 10cm depth of soil microbial richest changes with depth, the total microbial reduced populations of species and diversity differences. Nitrogen levels affect the soil microbial community structure changes, and changes consistent with the apple leaf total nitrogen content. With nitrogen, the most abundant type of soil microbial populations, microbial species and the number of changes and growth of fruit trees have a positive relationship. Nitrogen levels above a certain concentration will inhibit the growth of certain bacteria. These results suggest that nitrogen management growth has a significant impact on soil microbial and fruit trees, and in some periods of the similarities.

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CLC: > Agricultural Sciences > Gardening > Fruit trees gardening > Pome > Apple
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