小麥內生菌及其功能研究
發(fā)布時間：2020-10-25 17:43
內生菌對作物健康生長起著重要作用。本文分離、鑒定了大量小麥內生菌,研究了小麥內生菌的多樣性組成及其促生植物潛能和抗菌抗糖尿病潛能。1.從陜西、山西、河北、北京4省區(qū)采集到不同生長期的小麥植株樣品15個,從根、莖、葉中分離到細菌600株,放線菌2株、真菌16株。經過初步鑒定,新分離細菌系統發(fā)育地位屬于Achromobacter,Bacillus,Cedaceae,Citrobacter,Enterobacter,Flavobacterium,Flexibacter,Klebsiella,Kosakonia,Microbacterium,Ochrobactrum,Paenibacillus,Providencia,Pseudomonas,Rahnella,Rheinheimera,Rhizobium,Serratia,Sphingobacterium,Sphingobium,Stenotrophomonas,Xenophilus,Variovorax等23屬58種。其中12屬21種檢測到了nif H基因。2.高通量測序結果顯示小麥植株中內生細菌有500個屬,從高到低排列主要類群為Streptomyces,Sinorhizobium,Promicromonospora,Pantoea,Massilia,Cellulosimicrobium,Ohtaekwangia,Devosia,Kribbela,Micromonospora,Rhizobium,Nonomuraea,Kluyvera,Pseudomonas,Sphingomonas,Glutamicibacter,Niastella,Klebsiella。采用nif H基因高通量測序檢測到的小麥內生固氮菌100個屬。16S rRNA高通量測序檢測到大量放線菌,nif H基因高通量測序檢測到了大量稀有固氮菌群,而基于傳統培養(yǎng)技術的多樣性研究檢測到大量常見固氮菌群。3.對72株細菌,2株放線菌和15種真菌進行了促進植物生長和脅迫潛能測試分析。菌株固氮酶活(18.16±0.82~581.73±0.71nmol/mg蛋白·h),1-氨基環(huán)丙烷-1-羧酸脫氨酶(ACC deaminase;3.97±0.02~0.03±0.011μmolα丁酮酸/h·mg),IAA(1.00±0.1~49.37±0.005μgml~(-1))和PSI(2.17±0.02~5.16±0.36)活性。超過80%的分離株在鐵載體,NH_3和脲酶試驗中表現出陽性結果;只有13菌株產生了HCN。而在非生物脅迫測試中,Trichoderma菌株在高鹽(10%),重金屬(300μgml~(-1))和干旱(35%PEG)脅迫下表現出強耐受性,并對測試的抗生素(制霉菌素,酮康唑和伊曲康唑)具有抗性;菌株Talaromyces funiculosus耐高溫達到55~0C。盆栽試驗顯示,與對照相比,7株小麥內生固氮菌使小麥、玉米株高、干重和SPAD值增加。4.發(fā)現并命名了一株新的細菌Paenibacillusrhizophilus 7197~T。該菌株含有不同類型的脂肪酸(anteiso-C_(15:0),C_(14:0),C_(16:0)等)、極性脂(DPG,PG,PE等)、呼吸醌MK-7,Meso-DAP。含有4個nif H基因,顯示出高固氮酶活性(2526.41±34.9 nmol/mg蛋白·h)。ACC deaminase(1.53±0.01μmolα-丁酮酸/h·mg),IAA(26.12±0.004μg ml~(-1)),PSI(2.08±0.36),NH_3,鐵載體陽性,抗非生物脅迫[溫度(40~0C)、鹽度(2.5%NaCl)、干旱(20%PEG)、重金屬和抗生素]。盆栽試驗對小麥和玉米植物表現出生長促進作用(株高、干重和SPAD值高于對照處理)。
【學位單位】：中國農業(yè)科學院
【學位級別】：博士
【學位年份】：2019
【中圖分類】：S512.1;S182
【文章目錄】：
摘要
Abstract
CHAPTER Ⅰ INTRODUCTION
1.1 BACKGROUND
1.2 ENDOPHYTES
1.3 ENTRY OF ENDOPHYTES IN TO THE HOST PLANTS
1.4 BENEFICIAL FUNCTIONS OF ENDOPHYTES FOR PLANTS
1.4.1 ENHANCE PHYTOSTIMULATION
1.4.2 NITROGEN FIXATION
1.4.3 PHOSPHATE SOLUBILIZATION ABILITY OF ENDOPHYTES
1.4.4 IRON HOMEOSTASIS
1.4.5 ROLE IN INDUCED SYSTEMIC TOLERANCE
1.5 GENES RELATED TO NITROGEN FIXATION
1.6 MIRACULOUS DIAZOTROPHIC BACTERIA
1.6.1 BIOLOGICAL NITROGEN FIXATION OF DIAZOTROPHS
1.6.2 SYMBIOTIC NITROGEN FIXERS PIONEERS FOR BNF
1.6.2.1 IMPORTANCE OF SYMBIOTIC NITROGEN FIXATION IN LEGUMES
1.6.2.2 ROLE OF SYMBIOTIC NITROGEN FIXATION IN NON-LEGUMES
1.6.3 EFFECT OF ASSOCIATIVE DIAZOTROPHIC NITROGEN FIXERS
1.6.4 ENDOPHYTIC DIAZOTROPHIC NITROGEN FIXERS
1.6.5 FREE LIVING NITROGEN FIXERS
1.7 NATURAL PRODUCTS FROM ENDOPHYTES
1.7.1 ANTIBIOTICS
1.7.2 ANTIVIRAL COMPOUNDS
1.7.3 ANTICANCER COMPOUNDS
1.7.4 ANTIOXIDANT PROPERTY
1.7.5 ANTIDIABETIC ACTIVITY
1.7.6 IMMUNOSUPPRESIVE COMPOUNDS FROM ENDOPHYTES
1.8 MICROBIAL DIVERSITY RESEARCH METHODS AND PROGRESS
1.9 JUSTIFICATION OF THE STUDY
1.10 RESEARCH PLAN
1.11 SCIENTIFIC QUESTIONS
1.12 HYPOTHESIS
1.13 OBJECTIVES
CHAPTER Ⅱ MATERIALS AND METHODS
2.1 PLANT SAMPLING AND ISOLATION OF ENDOPHYTES
2.2 MOLECULAR IDENTIFICATION OF THE ISOLATED ENDOPHYTES
2.3 ANALYSIS OF THE DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA
2.3.1 ENDOPHYTIC BACTERIA16S r DNA V5-V7 REGION AMPLIFICATION
2.4 ANALYSIS OF THE DIVERSITY OF nif HCONTAINING DIAZOTROPHS
2.5 DATA PROCESSING METHODS
2.6 GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF WHEAT ENDOPHYTES
2.6.1 EVALUATION OF SOME IMPORTANT PGP PROPERTIES
2.6.1.1 QUALITATIVE AND QUANTITATIVE ESTIMATION OF ACC DEMINASEACTIVITY
2.6.1.2 QUALITATIVE AND QUANTITATIVE SCREENING OF NITROGEN FIXING PROPERTY OF THE ISOLATED ENDOPHYTIC BACTERIA
2.6.1.3 QUANTIFICATION OF IAA
2.6.1.4 SIDEROPHORE PRODUCTION
2.6.1.5 QUALITATIVE SCREENING OF PHOSPHATE SOLUBILIZATION POTENCY·
2.6.1.6 AMMONIA（NH3）PRODUCTION
2.6.1.7 CYANOGENESIS-HYDROGEN CYANIDE（HCN）PRODUCTION
2.6.1.8 UREASE TEST
2.6.1.9 CATALASE TEST
2.6.2 ABIOTIC STRESS TOLERANCE PROPERTIES
2.6.2.1 SALT TOLERANCE TEST
2.6.2.2 HEAVY METAL RESISTANCE TEST
2.6.2.3 DROUGHT TOLERANCE TEST
2.6.2.4 TEMPERATURE TOLERANCE TEST
2.6.2.5 ANTIBIOTIC SENSITIVITY TEST
2.6.3 EFFECT OF ENDOPHYTIC INOCULATION ON WHEAT AND MAIZE GROWTH
2.7 ANTIDIABETIC AND ANTIMICROBIAL POTENTIALS OF WHEAT ENDOPHYTES·
2.7.1 SECONDARY METABOLITE EXTRACTION
2.7.2 PHYTOCHEMICAL ANALYSIS
2.7.2.1 TEST FOR PHENOLS
2.7.2.2 TEST FOR TERPENOIDS
2.7.2.3 TEST FOR FLAVONOIDS
2.7.2.4 TEST FOR SAPONINS
2.7.2.5 TEST FOR TANNINS
2.7.2.6 TEST FOR ALKALOIDS
2.7.2.7 TEST FOR CARBOHYDRATES
2.7.2.8 TEST FOR STEROIDS
2.7.3 ANTIDIABETIC SCREENING OF THE ENDOPHYTIC METABOLITES
2.7.3.1 α-AMYLASE INHIBITION ASSAY
2.7.3.2 α-GLUCOSIDASE INHIBITION ASSAY
2.7.4 ANTIMICROBIAL TEST OF CRUDE METABOLITES OF ENDOPHYTES
2.7.4.1 TEST STRAINS AND CULTURE MEDIA
2.7.4.2 ANTIFUNGAL ASSAY
2.7.4.3 ANTIBACTERIAL ASSAY
T（Paenibacillus rhizophilus）'> 2.8 CHARACTERIZATION OF THE ISOLATED NOVEL ENDOPHYTIC BACTERIAUM7197T（Paenibacillus rhizophilus）
2.8.1 GENOME SEQUENCING
2.8.2 16S rRNA GENE SEQUENCING AND ANALYSIS
2.8.3 MORPHOLOGICAL CHARACTERISTICS
2.8.4 PHENOTYPICAL AND BIOCHEMICAL CHARACTERISTICS
2.8.5 NITROGENASE ACTIVITY OF7197T
2.8.6 CYTOCHEMICAL COMPONENTS
T'> 2.8.7 Growth Promoting and Abiotic Stress Tolerance Properties of 7197T
2.8.8 INOCULATION EFFECT OF7197T ON WHEAT AND MAIZE GROWTH
2.9 STATISTICAL ANALYSIS
CHAPTER Ⅲ ANALYSIS OF THE DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA
3.1 INTRODUCTION
3.2 RESULTS AND ANALYSIS
3.2.1 DIVESRSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA WITH CULTURE DEPENDENT METHOD
3.2.2 ANALYSIS OF DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA BY CULTURE INDEPENDENT METHOD
3.2.2.1 DATA QUALITY CONTROL AND OTU BASIS ANALYSIS
3.2.2.2 GENUS ABUNDANCE HEAT-MAP
3.2.2.3 ANALYSIS OFΑLPHA（α）DIVERSITY OF N-FIXING ENDOPHYTIC BACTERIA IN WHEAT
3.2.2.4 ANALYSIS OF BETA（β）DIVERSITY OF N-FIXING ENDOPHYTIC BACTERIA IN WHEAT
3.2.2.5 ANALYSIS OF N-FIXING ENDOPHYTIC BACTERIA CORE FLORA
3.2.2.6 ANALYSIS OF N FIXING ENDOPHYTIC BACTERIA ON TAXONOMIC LEVELS·
3.3 Comparison of Biodiversity of Endophytic Bacteria with Culture Dependent and Culture Independent Methods
3.4 SUMMARY OF THE CHAPTER
CHAPTER Ⅳ DIVERSITY OF nif H CONTAINING WHEAT DIAZOTROPHS
4.1 INTRODUCTION
4.2 RESULTS AND ANALYSIS
4.2.1 DIVERSITY OF nif H CONTAINING DIAZOTROPHS WITH CULTURE DEPENDENT METHOD
4.2.2 ANALYSIS OF DIVERSITY OF WHEAT ENDOPHYTIC NITROGENFIXINGDIAZOTROPHSIN WHEAT BY CULTURE INDEPENDENT METHOD
4.2.2.1 DATA QUALITY CONTROL AND OTU BASIS ANALYSIS
4.2.2.2 GENUS ABUNDANCE HEAT-MAP
4.2.2.3 ANALYSIS OFΑLPHA（α）DIVERSITY OF NITROGEN FIXING DIAZOTROPHS IN WHEAT
4.2.2.4 ANALYSIS OF BETA（β）DIVERSITY OF NITROGEN FIXING DIAZOTROPHS IN WHEAT
4.2.2.5 ANALYSIS OF CORE FLORA OF ENDOPHYTIC NITROGEN FIXING DIAZOTROPHS
4.2.2.6 ANALYSIS OF ENDOPHYTIC DIAZOTROPHS ON TAXONOMIC LEVELS
4.3 COMPARISON OF DIAZOTROPHBIODIVERSITYWITH CULTURE DEPENDENTAND CULTURE INDEPENDENT METHODS
4.4 SUMMARY OF THE CHAPTER
CHAPTER Ⅴ GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF WHEAT ENDOPHYTES
5.1 INTRODUCTION
5.2 RESULTS
5.2.1 PGP TRAITS OF THE ISOLATED ENDOPHYTIC MICROBES
5.2.1.1 QUALITATIVE AND QUANTITATIVE SCREENING FOR NITROGENFIXATION ABILITY OF THE ISOLATED ENDOPHYTES
5.2.1.2 QUALITATIVE AND QUANTITATIVE ESTIMATION OF ACC DEAMINASE ACTIVITY
5.2.1.3 IAA PRODUCTION
5.2.1.4 SIDEROPHORE PRODUCTION
5.2.1.5 PHOSPHATE SOLUBILIZATION POTENCY
5.2.1.6 AMMONIA PRODUCTION
5.2.1.7 CYANOGENESIS
5.2.1.8 UREASE TEST
5.2.1.9 CATALASE TEST
5.2.2 ABIOTIC STRESS TOLERANCE
5.2.2.1 SALT TOLERANCE TEST
5.2.2.2 HEAVY METAL RESISTANCE TEST
5.2.2.3 DROUGHT RESISTANCE TEST
5.2.2.4 TEMPERATURE TOLERANCE TEST
5.2.2.5 ANTIBIOTIC SENSITIVITY TEST
5.3 INOCULATION EFFECT OF ENDOPHYTES ON WHEAT AND MAIZE
5.4 SUMMARY OF THE CHAPTER
CHAPTER Ⅵ ANTIDIABETIC AND ANTIMICROBIAL POTENTIALS OF WHEATENDOPHYTES
6.1 INTRODUCTION
6.2 RESULTS
6.2.1 PHYTOCHEMICAL ANALYSIS
6.2.2 ANTIDIABETIC ACTIVITY OF THE MICROBIAL EXTRACTS
6.2.3 ANTIMICROBIAL ACTIVITY OF THE MICROBIAL EXTRACTS
6.3 SUMMARY OF THE CHAPTER
CHAPTER Ⅶ CHARACTERIZATION OF THE ISOLATED NOVEL ENDOPHYTIC BACTERIUM7197T（Paenibacillus rhizophilus）
7.1 INTRODUCTION
7.2 RESULTS AND ANALYSIS
7.2.1 PHYLOGENETIC ANALYSIS OF16S r RNA GENE SEQUENCES
7.2.2 DETERMINATION OF DNA G+C MOL%AND DNA HOMOLOGY
7.2.3 MORPHOLOGICAL CHARACTERISTICS
7.2.4 PHENOTYPICAL AND BIOCHEMICAL CHARACTERISTICS
7.2.5 NITROGENASE ACTIVITY OF7197T
7.2.6 CYTOCHEMICAL COMPONENTS
7.2.6.1ANALYSIS OF FATTY ACID COMPONENTS
7.2.6.2 ANALYSIS OF POLAR VALUE COMPONENTS
7.2.6.3 ANALYSIS OF BISMATH COMPONENT
7.2.6.4 ANALYSIS OF AMINO ACID COMPOSITION IN CELL WALL
7.3 GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF7197T
7.4 INOCULATION EFFECT OF7197T ON WHEAT AND MAIZE PLANTS
7.5 SUMMARY OF THE CHAPTER
CHAPTER Ⅷ DISCUSSION
8.1 ANALYSIS OF DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA.
8.2 ANALYSIS OF DIVERSITY OF nif H CONTAINING WHEAT DIAZOTROPHS
8.3 GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF WHEAT ENDOPHYTES
8.4 ANTIDIABETIC AND ANTIMICROBIAL POTENTIALS OF WHEAT ENDOPHYTES
T （Paenibacillus rhizophilus）'> 8.5 Characterization of the Isolated Novel Endophytic Bacterium 7197T （Paenibacillus rhizophilus）
CONCLUSION
REFERENCES
APPENDIX
ACKNOWLEDGEMENTS
RESUME
本文編號：2855745
【學位單位】：中國農業(yè)科學院
【學位級別】：博士
【學位年份】：2019
【中圖分類】：S512.1;S182
【文章目錄】：
摘要
Abstract
CHAPTER Ⅰ INTRODUCTION
1.1 BACKGROUND
1.2 ENDOPHYTES
1.3 ENTRY OF ENDOPHYTES IN TO THE HOST PLANTS
1.4 BENEFICIAL FUNCTIONS OF ENDOPHYTES FOR PLANTS
1.4.1 ENHANCE PHYTOSTIMULATION
1.4.2 NITROGEN FIXATION
1.4.3 PHOSPHATE SOLUBILIZATION ABILITY OF ENDOPHYTES
1.4.4 IRON HOMEOSTASIS
1.4.5 ROLE IN INDUCED SYSTEMIC TOLERANCE
1.5 GENES RELATED TO NITROGEN FIXATION
1.6 MIRACULOUS DIAZOTROPHIC BACTERIA
1.6.1 BIOLOGICAL NITROGEN FIXATION OF DIAZOTROPHS
1.6.2 SYMBIOTIC NITROGEN FIXERS PIONEERS FOR BNF
1.6.2.1 IMPORTANCE OF SYMBIOTIC NITROGEN FIXATION IN LEGUMES
1.6.2.2 ROLE OF SYMBIOTIC NITROGEN FIXATION IN NON-LEGUMES
1.6.3 EFFECT OF ASSOCIATIVE DIAZOTROPHIC NITROGEN FIXERS
1.6.4 ENDOPHYTIC DIAZOTROPHIC NITROGEN FIXERS
1.6.5 FREE LIVING NITROGEN FIXERS
1.7 NATURAL PRODUCTS FROM ENDOPHYTES
1.7.1 ANTIBIOTICS
1.7.2 ANTIVIRAL COMPOUNDS
1.7.3 ANTICANCER COMPOUNDS
1.7.4 ANTIOXIDANT PROPERTY
1.7.5 ANTIDIABETIC ACTIVITY
1.7.6 IMMUNOSUPPRESIVE COMPOUNDS FROM ENDOPHYTES
1.8 MICROBIAL DIVERSITY RESEARCH METHODS AND PROGRESS
1.9 JUSTIFICATION OF THE STUDY
1.10 RESEARCH PLAN
1.11 SCIENTIFIC QUESTIONS
1.12 HYPOTHESIS
1.13 OBJECTIVES
CHAPTER Ⅱ MATERIALS AND METHODS
2.1 PLANT SAMPLING AND ISOLATION OF ENDOPHYTES
2.2 MOLECULAR IDENTIFICATION OF THE ISOLATED ENDOPHYTES
2.3 ANALYSIS OF THE DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA
2.3.1 ENDOPHYTIC BACTERIA16S r DNA V5-V7 REGION AMPLIFICATION
2.4 ANALYSIS OF THE DIVERSITY OF nif HCONTAINING DIAZOTROPHS
2.5 DATA PROCESSING METHODS
2.6 GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF WHEAT ENDOPHYTES
2.6.1 EVALUATION OF SOME IMPORTANT PGP PROPERTIES
2.6.1.1 QUALITATIVE AND QUANTITATIVE ESTIMATION OF ACC DEMINASEACTIVITY
2.6.1.2 QUALITATIVE AND QUANTITATIVE SCREENING OF NITROGEN FIXING PROPERTY OF THE ISOLATED ENDOPHYTIC BACTERIA
2.6.1.3 QUANTIFICATION OF IAA
2.6.1.4 SIDEROPHORE PRODUCTION
2.6.1.5 QUALITATIVE SCREENING OF PHOSPHATE SOLUBILIZATION POTENCY·
2.6.1.6 AMMONIA（NH3）PRODUCTION
2.6.1.7 CYANOGENESIS-HYDROGEN CYANIDE（HCN）PRODUCTION
2.6.1.8 UREASE TEST
2.6.1.9 CATALASE TEST
2.6.2 ABIOTIC STRESS TOLERANCE PROPERTIES
2.6.2.1 SALT TOLERANCE TEST
2.6.2.2 HEAVY METAL RESISTANCE TEST
2.6.2.3 DROUGHT TOLERANCE TEST
2.6.2.4 TEMPERATURE TOLERANCE TEST
2.6.2.5 ANTIBIOTIC SENSITIVITY TEST
2.6.3 EFFECT OF ENDOPHYTIC INOCULATION ON WHEAT AND MAIZE GROWTH
2.7 ANTIDIABETIC AND ANTIMICROBIAL POTENTIALS OF WHEAT ENDOPHYTES·
2.7.1 SECONDARY METABOLITE EXTRACTION
2.7.2 PHYTOCHEMICAL ANALYSIS
2.7.2.1 TEST FOR PHENOLS
2.7.2.2 TEST FOR TERPENOIDS
2.7.2.3 TEST FOR FLAVONOIDS
2.7.2.4 TEST FOR SAPONINS
2.7.2.5 TEST FOR TANNINS
2.7.2.6 TEST FOR ALKALOIDS
2.7.2.7 TEST FOR CARBOHYDRATES
2.7.2.8 TEST FOR STEROIDS
2.7.3 ANTIDIABETIC SCREENING OF THE ENDOPHYTIC METABOLITES
2.7.3.1 α-AMYLASE INHIBITION ASSAY
2.7.3.2 α-GLUCOSIDASE INHIBITION ASSAY
2.7.4 ANTIMICROBIAL TEST OF CRUDE METABOLITES OF ENDOPHYTES
2.7.4.1 TEST STRAINS AND CULTURE MEDIA
2.7.4.2 ANTIFUNGAL ASSAY
2.7.4.3 ANTIBACTERIAL ASSAY
T（Paenibacillus rhizophilus）'> 2.8 CHARACTERIZATION OF THE ISOLATED NOVEL ENDOPHYTIC BACTERIAUM7197T（Paenibacillus rhizophilus）
2.8.1 GENOME SEQUENCING
2.8.2 16S rRNA GENE SEQUENCING AND ANALYSIS
2.8.3 MORPHOLOGICAL CHARACTERISTICS
2.8.4 PHENOTYPICAL AND BIOCHEMICAL CHARACTERISTICS
2.8.5 NITROGENASE ACTIVITY OF7197T
2.8.6 CYTOCHEMICAL COMPONENTS
T'> 2.8.7 Growth Promoting and Abiotic Stress Tolerance Properties of 7197T
2.9 STATISTICAL ANALYSIS
CHAPTER Ⅲ ANALYSIS OF THE DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA
3.1 INTRODUCTION
3.2 RESULTS AND ANALYSIS
3.2.1 DIVESRSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA WITH CULTURE DEPENDENT METHOD
3.2.2 ANALYSIS OF DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA BY CULTURE INDEPENDENT METHOD
3.2.2.1 DATA QUALITY CONTROL AND OTU BASIS ANALYSIS
3.2.2.2 GENUS ABUNDANCE HEAT-MAP
3.2.2.3 ANALYSIS OFΑLPHA（α）DIVERSITY OF N-FIXING ENDOPHYTIC BACTERIA IN WHEAT
3.2.2.4 ANALYSIS OF BETA（β）DIVERSITY OF N-FIXING ENDOPHYTIC BACTERIA IN WHEAT
3.2.2.5 ANALYSIS OF N-FIXING ENDOPHYTIC BACTERIA CORE FLORA
3.2.2.6 ANALYSIS OF N FIXING ENDOPHYTIC BACTERIA ON TAXONOMIC LEVELS·
3.3 Comparison of Biodiversity of Endophytic Bacteria with Culture Dependent and Culture Independent Methods
3.4 SUMMARY OF THE CHAPTER
CHAPTER Ⅳ DIVERSITY OF nif H CONTAINING WHEAT DIAZOTROPHS
4.1 INTRODUCTION
4.2 RESULTS AND ANALYSIS
4.2.1 DIVERSITY OF nif H CONTAINING DIAZOTROPHS WITH CULTURE DEPENDENT METHOD
4.2.2 ANALYSIS OF DIVERSITY OF WHEAT ENDOPHYTIC NITROGENFIXINGDIAZOTROPHSIN WHEAT BY CULTURE INDEPENDENT METHOD
4.2.2.1 DATA QUALITY CONTROL AND OTU BASIS ANALYSIS
4.2.2.2 GENUS ABUNDANCE HEAT-MAP
4.2.2.3 ANALYSIS OFΑLPHA（α）DIVERSITY OF NITROGEN FIXING DIAZOTROPHS IN WHEAT
4.2.2.4 ANALYSIS OF BETA（β）DIVERSITY OF NITROGEN FIXING DIAZOTROPHS IN WHEAT
4.2.2.5 ANALYSIS OF CORE FLORA OF ENDOPHYTIC NITROGEN FIXING DIAZOTROPHS
4.2.2.6 ANALYSIS OF ENDOPHYTIC DIAZOTROPHS ON TAXONOMIC LEVELS
4.3 COMPARISON OF DIAZOTROPHBIODIVERSITYWITH CULTURE DEPENDENTAND CULTURE INDEPENDENT METHODS
4.4 SUMMARY OF THE CHAPTER
CHAPTER Ⅴ GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF WHEAT ENDOPHYTES
5.1 INTRODUCTION
5.2 RESULTS
5.2.1 PGP TRAITS OF THE ISOLATED ENDOPHYTIC MICROBES
5.2.1.1 QUALITATIVE AND QUANTITATIVE SCREENING FOR NITROGENFIXATION ABILITY OF THE ISOLATED ENDOPHYTES
5.2.1.2 QUALITATIVE AND QUANTITATIVE ESTIMATION OF ACC DEAMINASE ACTIVITY
5.2.1.3 IAA PRODUCTION
5.2.1.4 SIDEROPHORE PRODUCTION
5.2.1.5 PHOSPHATE SOLUBILIZATION POTENCY
5.2.1.6 AMMONIA PRODUCTION
5.2.1.7 CYANOGENESIS
5.2.1.8 UREASE TEST
5.2.1.9 CATALASE TEST
5.2.2 ABIOTIC STRESS TOLERANCE
5.2.2.1 SALT TOLERANCE TEST
5.2.2.2 HEAVY METAL RESISTANCE TEST
5.2.2.3 DROUGHT RESISTANCE TEST
5.2.2.4 TEMPERATURE TOLERANCE TEST
5.2.2.5 ANTIBIOTIC SENSITIVITY TEST
5.3 INOCULATION EFFECT OF ENDOPHYTES ON WHEAT AND MAIZE
5.4 SUMMARY OF THE CHAPTER
CHAPTER Ⅵ ANTIDIABETIC AND ANTIMICROBIAL POTENTIALS OF WHEATENDOPHYTES
6.1 INTRODUCTION
6.2 RESULTS
6.2.1 PHYTOCHEMICAL ANALYSIS
6.2.2 ANTIDIABETIC ACTIVITY OF THE MICROBIAL EXTRACTS
6.2.3 ANTIMICROBIAL ACTIVITY OF THE MICROBIAL EXTRACTS
6.3 SUMMARY OF THE CHAPTER
CHAPTER Ⅶ CHARACTERIZATION OF THE ISOLATED NOVEL ENDOPHYTIC BACTERIUM7197T（Paenibacillus rhizophilus）
7.1 INTRODUCTION
7.2 RESULTS AND ANALYSIS
7.2.1 PHYLOGENETIC ANALYSIS OF16S r RNA GENE SEQUENCES
7.2.2 DETERMINATION OF DNA G+C MOL%AND DNA HOMOLOGY
7.2.3 MORPHOLOGICAL CHARACTERISTICS
7.2.4 PHENOTYPICAL AND BIOCHEMICAL CHARACTERISTICS
7.2.5 NITROGENASE ACTIVITY OF7197T
7.2.6 CYTOCHEMICAL COMPONENTS
7.2.6.1ANALYSIS OF FATTY ACID COMPONENTS
7.2.6.2 ANALYSIS OF POLAR VALUE COMPONENTS
7.2.6.3 ANALYSIS OF BISMATH COMPONENT
7.2.6.4 ANALYSIS OF AMINO ACID COMPOSITION IN CELL WALL
7.3 GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF7197T
7.4 INOCULATION EFFECT OF7197T ON WHEAT AND MAIZE PLANTS
7.5 SUMMARY OF THE CHAPTER
CHAPTER Ⅷ DISCUSSION
8.1 ANALYSIS OF DIVERSITY OF WHEAT ENDOPHYTIC NITROGEN FIXING BACTERIA.
8.2 ANALYSIS OF DIVERSITY OF nif H CONTAINING WHEAT DIAZOTROPHS
8.3 GROWTH PROMOTING AND STRESS TOLERANCE PROPERTIES OF WHEAT ENDOPHYTES
8.4 ANTIDIABETIC AND ANTIMICROBIAL POTENTIALS OF WHEAT ENDOPHYTES
T
CONCLUSION
REFERENCES
APPENDIX
ACKNOWLEDGEMENTS
RESUME
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