Soil samples were collected to a depth of 20 cm using a 3.0 m × 3.0 m grid in 900 m 2(30.0 m ×30.0 m) plots for analysis of spatial patterns of soil in undisturbed area of China fir(Cunninghamia lanceolata) ...Soil samples were collected to a depth of 20 cm using a 3.0 m × 3.0 m grid in 900 m 2(30.0 m ×30.0 m) plots for analysis of spatial patterns of soil in undisturbed area of China fir(Cunninghamia lanceolata) and Masson pine(Pinus massoniana) mixed forest in the Three Gorger Reservoir Areas.The investigation data showed that the average of soil DNA content was 922.71 μg·g-1 water-free soil,the maximal value was 6 761.30 μg·g-1 water-free soil,and the less value 58.52 μg·g-1 water-free soil.The geostatistical analysis indicated that the spatial distribution characteristic was not obviously reflected by the liner graphs while samples were randomly distribution and independent each other.The separation distance among samples was related to precision of soil DNA content,the effects of non-structure factors(environment factors,disturbed factors) on spatial correlation structure would disappeared due to the separation distance in excess of 18.84 m,and the spatial correlation of soil DNA faded down in plots.Spatial variability in soil DNA in the region was mostly contributed by the influencing factors including soil nitrogen,soil organic matter and content of soil water.展开更多
为了优化以土壤微生物DNA为模板的PCR反应条件,采用E.Z.N.A.soil DNA kit试剂盒提取土壤微生物总DNA,对16SrDNA V3可变区的PCR反应体系和反应条件进行优化。主要从DNA模板用量、引物浓度、退火温度、热启动方式4个方面进行筛选试验,最...为了优化以土壤微生物DNA为模板的PCR反应条件,采用E.Z.N.A.soil DNA kit试剂盒提取土壤微生物总DNA,对16SrDNA V3可变区的PCR反应体系和反应条件进行优化。主要从DNA模板用量、引物浓度、退火温度、热启动方式4个方面进行筛选试验,最后得出最适宜的土壤DNA扩增体系为:10.5ng模板DNA、5μL 10×buffer、4μL 2.5mmol/L dNTP、10μmol/L引物各1.5μL,2.5UTaq酶,加无菌ddH2O补足至50μL;PCR循环程序为:94℃预变性5min;94℃变性1min,52℃退火1min,72℃延伸70s,29个循环;72℃延伸5min。试验结果表明:选择热启动方式和合适的退火温度是获得高质量PCR产物的关键。展开更多
文摘Soil samples were collected to a depth of 20 cm using a 3.0 m × 3.0 m grid in 900 m 2(30.0 m ×30.0 m) plots for analysis of spatial patterns of soil in undisturbed area of China fir(Cunninghamia lanceolata) and Masson pine(Pinus massoniana) mixed forest in the Three Gorger Reservoir Areas.The investigation data showed that the average of soil DNA content was 922.71 μg·g-1 water-free soil,the maximal value was 6 761.30 μg·g-1 water-free soil,and the less value 58.52 μg·g-1 water-free soil.The geostatistical analysis indicated that the spatial distribution characteristic was not obviously reflected by the liner graphs while samples were randomly distribution and independent each other.The separation distance among samples was related to precision of soil DNA content,the effects of non-structure factors(environment factors,disturbed factors) on spatial correlation structure would disappeared due to the separation distance in excess of 18.84 m,and the spatial correlation of soil DNA faded down in plots.Spatial variability in soil DNA in the region was mostly contributed by the influencing factors including soil nitrogen,soil organic matter and content of soil water.
