Research On Soil Fertility And Microbial Diversity And Physiological Activities Of The Rhizosphere Of Vitis Vinifera

The annual variations of the root zone and the rhizosphere microbial flora of grapevines(bacteria,actinomycetes and fungi)were analyzed through the analysis of microbial flora in the growing period,the dynamic changes of soil fertility and TDS contents in grapevines rhizosphere soil were determined at the same time,combined with the analysis of microbial diversity in rhizosphere,to separate the endogenous rhizosphere microorganisms of grapevine and to screen plant growth promoting rhizobacteria,the main research areas were as follows:1.The dynamic changes of grapevine root rhizosphere soil microbial community and soil elements during the whole growth cycle at Jiayuguan Zixuan vineyards were studied.The results showed that:(1)Grapevine variety was one of the factors affecting the quantity of microbial community in rhizosphere.There existed a close relationship between the actinomycetes and fungi in the rhizosphere soil and the growth of grapevines;the number of rhizosphere bacteria of the cultivated varities of grapes(H07 Bei Binghong)were relatively higher,the number of rhizosphere actinomycetes of the cultivated varities of grapes(J02 Vidal,A03 Amur grape and Q02 Zuo Youhong)were relatively higher,and the number of fungi in the rhizosphere soil of grape(Q02 Zuo Youhong)was relatively higher.(2)The variety of grapevine was one of the factors affecting the contents of the elements in the rhizosphere.There were obvious differences in TDS contents in rhizosphere soil and the absorption and utilization of soil elements conformed to a certain change trend in the whole growth cycle.The contents of organic matter and Ca were relatively lower in the period of expansion and coloration.The N content was relatively lower in the expansion,coloration and maturity stage.In the coloring and maturity period,the contents of P,Zn and Fe were relatively lower.The elements of K,Mg,Mn and Cu were lower in maturity period.The values of TDS of these grapes(J02 Vidal,F01 Chardonnay,N08 Merlot,F06 Vidal and E05 Cabernet Sauvignon)in the rhizosphere soil were relatively higher,and the numbers of soil microorganisms with high TDS were relatively lower.2.Using Illumina MiSeq sequencing technology to make diversity analysis about microbial communities in rhizosphere soil of grapes in mature stage at Kefeng located in the southeast of China and Zixuan vineyard located in the northwest of China,respectively.The results showed that:(1)Analysis of bacterial diversities in rhizosphere soils of wine grape:the difference between diversitied of rhizosphere bacterial communities and varieties of grape were minor,and the composition and proportion of the bacterial communities in the rhizosphere soil of the same species were different in the different planting years.The longer the years of plant,the higher the richness and diversity of bacterial communities of the correspondingthe plants.According to the OTU cluster analysis,the bacterial communities in the rhizosphere soil of wine grape were more abundant with a higher potential for resource utilization.According to the analysis of community structure and cluster,Actinomycetes,Bacteroidetes and Firmicutes were more abundant,including Nocardioides,Cellulomonas,Flavobacterium,promicromonospora,Streptomyces,Bacillus,Bacteroides,Lactobacillus,Lactococcus and other species of the genus.The longer planting years of different varieties of grapevine,the higher proportions of Gemmatimonadetes and Acidobacteria in the corresponding root soils of the plants.Based on the function of gene prediction,bacterial communities in rhizosphere soil of grape in the function(Membrane Transport,Carbohydrate Metabolism,Amino Acid Metabolism,Material Degradation and Detabolism,Nucleotide Detabolism,Cofactor and Vitamin Metabolism,Lipid Metabolism,Energy Metabolism and so on)showed the strongest correlation,indicating that the rhizosphere soil bacterial community resource of grapes at Zixuan vineyard was abundance with a higher potential of resource utilization.(2)Analysis of fungal diversity in rhizosphere soils of wine grape:There were significant differences in the fungal communities in the rhizosphere soil of different varieties of wine grape.The composition and proportion of the fungal communities in the rhizosphere soil of the same cultivars were different.The longer planting years,the lower richness and diversity of fungal community in the corresponding root soils of the plants.Further analysis of the fungal community structure in the rhizosphere soil of wine grape found that the longer the planting year,the higher the number of Pezizomycetes in the rhizosphere soil.And the rhizosphere soil fungal communities of the varieties of grape(Q02 Zuo Youhong,N08 Merlot and F06 Vidal)had more potential of resource utilization.(3)Analysis of microbial diversity in rhizosphere soil of wine grape in different regions:There were obvious differences between different regions in bacterial community composition in rhizosphere soil of grape(diversity of soil bacterial community in the rhizosphere of grape at Kefeng vineyard in the southeast of China was significantly lower than that of Zixuan vineyard in the northwest of China),the differences in fungal community abundances were not obvious,but there was a significant difference in the composition and diversity of fungal communities between the two regions(there was a significant difference between the genus and species of fungal communities),infered that fungi were not the main factors affecting the difference in the rhizosphere microorganisms between the two places.According to the analysis of community structure,the number of actinomycetes in root soil at Zixuan vineyard was higher than that of Kefeng vineyard.Acidobacteria,Proteobacterium and Verrucomicrobia were more abundant in the rhizosphere soil of grape at Kefeng vineyard,including species of Bradyrhizobium,Phenylobacterium,Devosia,Cellvibrio,Mycoplana and Prosthecobacter.According to the index of diversity of the rhizosphere microbial community,the richness was low,but the diversity was uncertain.According to the community structure and the difference analysis,the structure and the difference of soil microbial community in the rhizosphere soil of the two vineyards were obviously greater,which may be related to the region and the soil environment,and inferred that the Gemmatimonadetes,the Acidobacteria and the Pezizomycetes were one of the factors that needed for the growth of the wine grape.Based on the prediction of function of gene,bacterial communities in rhizosphere soil of grape in the function(Cell Motility,Signal Transduction Function,Translation,Transcription and Degradation,Polysaccharide Synthesis and Metabolism,Enzyme Metabolism Function and so on)showed the strongest correlation,indicated that the rhizosphere soil bacterial community of grapes at Kefeng vineyard was abundance,had a higher potential of resource utilization.3.The main physical and chemical properties of different grapevine varieties were analyzed at Zixuan vineyard.The results showed that:(1)The varieties of grapevine were one of the main factors affecting the contents of fruit conponents,and the fruit qualities of the wine grape with longer planting year were lower than that of the shorter planting year.The test found that qualities of varieties of red grape(N08 Merlot and E05 Cabernet Sauvignon)and white grape varieties(F06 Vidal and F02 Riesling)were of good quality in accordance with the selection criteria of brewing fruit,such as higher sugar acid ratio,and higher content total phenolics and tannin,suitable for cultivating at Zixuan vineyard.(2)Comprehensive test results showed that the number of actinomycetes and fungi in soil may be related to the soil nutrient elements competing with grapes,the contents of TDS in soil also influence on the quality of grape fruit,and the amount of Zn,Mn and Fe in the soil could be increased in the color of the soil,which may be beneficial to the improvement of fruit quality.4.Isolation,purification and identification of endophytes from the root of vitis vinifera of the Zixuan vineyard,the strains with IAA activity and the characteristics of phosphorus degradation were obtained.The results showed that:(1)A total of 27 endophytes isolates(19 of bacteria and 8 of fungi)were isolated to further analyses.Based on the morphological and molecular identification methods,the isolates of microorganisms were grouped into 9 genera and 5 genera,respectively.The genera of bacteria are Amycolatopsis,Microbacterium,Streptomyces,Agrobacterium,Rhizobium,Pseudomonas,Enterobacter,Serratia,Staphylococcus and Bacillus,respectively.The fungi genera are Penicillium,Aspergillus,Achaetomium,Fusarium and Mucor,respectively.(2)All the purified strains were screened on the basis of IAA secretion and phosphorus dissolving activity,4 strains of endophytic bacteria were isolated from 27 strains of endophytic bacteria,7 strains of inorganic phosphate dissolving bacteria and 10 strains of IAA secreting bacteria,and the strain XZ11 belonging to the genus Agrobacterium showed the potential to promote the growth of grapevine.