Abstract:

Abstract:[Objective] Bacterial ncRNAs are a versatile class of non-coding RNA which plays an important role in the process of microbial life. In this study, we assess three ncRNA gene-prediction tools used frequently with the different bacterial genomes. [Methods] Prediction tools representing the position weight matrix method (sRNAscanner), comparative genomics method (sRNAPredict) and machine learning method (PORTRAIT) were tested by using 7 BSRD-archived bacterial genomes with low, middle and high G+C contents, each of which contains more than 30 experimentally verified ncRNA genes. A set of genomic G+C content-associated position weight matrixes of transcription initiation and termination regions of ncRNA genes was generated and employed to test sRNAscanner prediction. [Results] The sRNAPredict tool had higher specificity and positive prediction value, but lower sensitivity than PORTRAIT. The performance of both tools varied with the selected strains of different G+C contents. The obtained G+C content-associated matrix slightly improved the average accuracy of sRNAscanner. [Conclusion] The changing accuracy of the bacterial ncRNA gene detection tools under study was attributed to genomic G+C heterogeneity. Conserved sequence features of ncRNA gene promoters and terminators in genomes sharing similar G+C contents may be helpful to enhance bacterial ncRNA genes prediction.

Abstract:

Abstract:[Objective] Bacillus licheniformis is the predominant microbe producing Maotai-flavor in high-temperature Daqu which plays important roles in Maotai-flavor liquor making. Learing the machanisms of tolerance to this liquor making environment with hypertonic, acid and ethanol stress would provide an insight into characterisitcs of this liquor making. [Methods] The resistance to hypertonic, acid and ethanol of B. licheniformis CGMCC 3963 which produces Maotai-flavor was measured. the tolerance machanisms were analyzed on comparative transcriptomics level. [Results] The growth of B. licheniformis CGMCC 3963 in 15% KCl, 15% NaCl, pH 4.0 or 6% ethanol concentration was significantly better than that of B. licheniformis ATCC 14580. Comparative transcriptome analysis revealed a series of genes differently expressed in B. licheniformis CGMCC 3963. [Conclusion] Compared with B. licheniformis ATCC 14580, B. licheniformis CGMCC 3963 posessed a stronger tolerance ability and differently expressed a series of genes related to stress tolerance. Differently expressed genes encoding solute transport, potassium channel protein played important roles in imoproving the tolerance to osmotic stress; genes related to class II heat shock proteins, dehydrogenase, oxidative stress and pH homeostasis made significant contributions to the resistance to acid of B. licheniformis CGMCC 3963; class II and class III heat shock genes were important to ethanol tolerance ability of B. licheniformis CGMCC 3963.

Abstract:[Objective] Survey of cultivable bacterial diversity in the oil shale deposits is important to use untapped bacterial resources in the oil shale mines. [Methods] We isolated and purified bacteria in the three major oil shale mines (Fushun mine in Liaoning, Huadian mine in Jilin, and Maoning mine in Guangdong) in China on the nutrient agar medium by the dilution plate culture method. Then we determined 16S rRNA gene sequence and did phylogenetic analysis of the isolated bacteria. [Results] Ten genus were identified in the three mines, including nine OTUs (belonging to Pantoea, Brevibacillus, Paenibacillus, Microbacterium, Arthrobacter, Pseudomonas and Bacillus) in Fushun mine, five OTUs (belonging to Bacillus and Sinomona) in Maoming mine, and five OTUs (belonging to Staphylococcus, Acinetobacter, Bacillus and Arthrobacter) in Huadian mine. [Conclusion] The compositions of cultivable bacterial communities in the three mines were simple and similar at the phylum level, mainly including Firmicutes and Actinobacteria. Some Proteobacteria existed in Fushun mine and Huadian mine. However, their compositions in the three mines had quite differences at the genera level.

Abstract:[Objective] To study the diversity and antimicrobial activity of culturable actinomycetes isolated from Apis cerana cerana Fabricius, which will help to explore new microbial resources. [Methods] Seven media were used to isolate actinomycetes from Apis cerana cerana Fabricius samples. 16S rRNA PCR-RFLP analysis and 16S rRNA gene sequences analysis were utilized to study their genetic diversity. Pathogenic bacteria and fungus were used in antimicrobial test for exploring the antimicrobial activity of strains. [Results] Based on the morphology of the colonies and cells of the new isolates, 84 representative strains were selected from 180 isolated strains. They belonged to 3 orders, 4 families and 4 genera, and in which there were 6 strains identified as potential new novel species. The best surface disinfection method was by using 0.2% ClO2 for 60 s. The results of the antimicrobial test showed that 31.0%, 48.8%, 27.4% and 16.7% of the representative strains presented antimicrobial activity to Escherichia coli, Bacillus subtilis, Curvularia lunata and Fusarium oxysporum, and 71.4% of the representative strains presented antimicrobial activity to at least one pathogen. [Conclusion] The choice of the isolation method has a great impact on isolation result. There are great diverse actinomycetes inside Apis cerana cerana Fabricius and they have great antimicrobial activity, which show a great potential for researching new bioactive compounds.

Abstract:[Objective] The composition and diversity of microorganism during low rank coal authigenous microorganism methane conversion were explored in Xinjiang. [Methods] We evaluated the effect of low rank coal authigenous microorganism on methane conversion and organic acid content by anaerobic culture. The dynamic changes of microbial community were analyzed by terminal restriction fragment length polymorphism (T-RFLP) in Hami’s long flame coal. [Results] The results suggested that lang flame coal and brown coal had little influence on methane production of indigenous microorganism. As low rank coal biological methane conversion time went on, methane production showed an increasing tendency. The yield of long flame coal biological methane was 10.28 mL/g and the concentration of volatile fatty acid (VFA) were the lowest at 60 days. The diversity indexes of microbes did not change significantly. The major microbial groups were Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria in different low rank coal conversion period. The bacterial community structure was more complex, while the methanogens community structure was relatively simple. All samples harbored these phyla such as Methanosarcina, Methanohalobium, Methanolobus, Methanomethlovorans, which constituted a basic flora of community structure. [Conclusion] There was abundant microbial diversity in different low rank coal conversion period and the diversity was different in different period. The methanogens community structure was relatively simple, and the number of common groups was higher in methanogens community.

Abstract:[Objectives] To screen effective bacterial strains capable of decolorizing azo dyes containing wastewater from dewatered sludge. [Methods] nine bacterial strains (named T-1?T-9) capable of decolorizing mixed azo dyes were isolated from the dewatered sludge in municipal wastewater treatment system after proper acclimation. physiological and biochemical tests and 16S rRNA gene based bio-molecular identification were adopted to identify the isolates. Decolorization performances of pure cultures and mixed cultures of the isolates were determined and compared. Isolated strain T-8 was used to prepare dry powder and consequent determination of decolorization performance. [Results] Nine isolates belong to Bacillus sp., Exiguobacterium sp., Stenotrophomonas sp. and Paracoccus sp. according to preliminarily identification. The structure and concentration of azo dyes had a certain impact on the decolorization efficiency during domestication. T-8 had the best decolorization rate 85.9% for methyl orange and 86.2% for golden orange I in 40 h. The dry powder of T-8 was equally active to its origin isolate, which is confirmed by complete decolorization of golden orange I in 4 days without external carbon resource. Mixed culture of all isolates was obviously better in the decolorization (90.1%) of mixed zao dyes than pure culture did. [Conclusion] This study suggests that, as a new source for the functional flora of decolorizing azo dyes, the dewatered sludge and its isolates are of great value in zao dye containing wastewater.

Abstract:[Objective] Streptomyces ZM-16 has been proved to possess great anti-bacteria effect to many strains of bacteria. The main active component of the fermentation products is actinomycin D. This study aims at the investigation of the antifungal activities of the fermentation products of Streptomyces ZM-16 and the enhancement of the active substance yield of the strain by mutation breeding, in order to develop and improve its practical application in bio-control field of plant diseases. [Methods] Obtain the active fermentation products with liquid fermentation of the strain and conduct a cruel extraction; test the antimicrobial effect of the cruel extracts on 11 kinds of plant pathogenic fungi by inhibition zone method; improve the strain by microwave mutation breeding and antibiotics resistance screening. [Results] The cruel extracts solution showed antimicrobial effect on all the 11 kinds of plant pathogenic fungi. Among them, it possessed greatest inhibitive effect on Glomerella cingulated, Sclerotinia sclerotiorum and Fusarium graminearum. After induced mutation by microwave, a rifampin resistant mutant strain was screened out, whose yield of actinomycin D was enhanced 36.75%. The study on genetic stability of passage showed that the mutant strain was stable after 10 generations. [Conclusion] The fermentation products of Streptomyces ZM-16 have great inhibitive effect on most plant pathogen fungi; moreover, the yield of active products was enhanced after genetic improvement. These results demonstrated that the strain has a high value of practical application.

Abstract:[Objective] To isolating the Bacillus strains from the rhizosphere of Lactuca sativa, which have a variety of biological characteristics, the growth-promoting ability and effects of biocontrol. [Methods] In the study, high-temperature method was used to separate Bacillus spp.. The strains were identified according to morphological characteristics, physiological and biochemical responses, 16S rRNA and gyrB gene analysis. The phosphate solubilization, IAA syntheses, siderophore syntheses, the inhibition against the plant pathogenic fungis of two strains were studied. Planted the lettuce seeds soaked with the suspension liquid of two strains, and evaluated the effects of the growth-promoting after 30 days. Using the strain WXD 3-2 treated with wheat, evaluated the effects of inoculum concentrations of WXD 3-2. [Results] The strain WXD 3-1 was identified as Bacillus megaterium and WXD 3-2 was confirmed as Bacillus subtilis. Two strains had phosphate solubilization, IAA syntheses, siderophore syntheses, and the growth-promoting ability. The strain WXD 3-2 could inhibit varieties of plant pathogenic fungis. The height of the lettuce, the width of leaves, the fresh weights, and the dry weights were increased 21.51% and 8.88%, 31.93% and 14.51%, 41.30% and 13.58%, 42.76% and 26.35% compared with control. The results showed that the strain WXD 3-2 could reduce wheat root rot disease and wheat root lesion. [Conclusion] Two strains had phosphate solubilization, IAA syntheses, siderophore syntheses, and growth promotion ability. The strain WXD 3-2 had effects of biocontrol.

Abstract:[Objective] The research is to study the effects of endophytic fungi re-inoculation on folia endogenous flora structure of grape. [Methods] Eight strains of endophytic fungi which isolated from wine grape leaves were re-inoculated to the healthy leaves of grapevine (variety: Rose-honey), and the effects on the folia endophytic fungi structure of grape were analyzed. Meanwhile, effects differences on the endophytic fungi structure to both groups of grapevine with and without pesticide after endophytic fungi re-inoculation, were also studied. We describe the folia endophytic fungi structure by using the isolation rate, dominance index and diversity index three months after re-inoculation. [Results] The results indicated that among all the treatments, five strains of the inoculated endophytic fungi (Xylaria sp., Nigrospora sp., Alternaria sp. 1, Alternaria sp. 2 and Colletotrichum sp.) can be relatively higher frequently isolated. Significant differences of folia endophytic fungi structures were found between one fungus re-inoculation and another. [Conclusion] Compared with the controls, fungi re-inoculation can increase the isolation rate, but decrease the diversity index significantly of folia endophytes. And the using of pesticide could decrease the isolation rate, but shares a higher diversity of folia endophytic fungi, compared with that of the pesticide free group.

Abstract:[Objective] To analyze yeast community structure and its influence on brewing during the fermentation process of Chinese Maotai-flavour liquor. [Methods] The yeast community in the fifth round (higher yield and quality of the liquor) and the seventh round of fermentation process (lower yield and quality of the liquor) were analyzed using denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR (RT-qPCR). The metabolic components in the fermented grains were investigated through headspace-solid phase microextraction gas chromatograph-mass apectrometer (HS-SPME-GC-MS) and high performance liquid chromatography (HPLC). Finally, the effects of yeast communities on the yield and quality of Maotai-flavor liquor were analyzed. [Results] At least nine yeasts were detected in the fermented grains, including Issatchenkia orientalis, Torulaspora delbrueckii, Pichia galeiformis, Schizosaccharomyces pombe, Galactomyces geotrichum, Trichosporon asahii, Zygosaccharomyces bailii, Saccharomyces cerevisiae and Pichia fabianii. Among them, G. geotrichum and S. cerevisiae was dominant yeasts of the fifth round, while the other two yeast, I. orientalis and Z. bailii, were also advantage in seventh round and Sc. pombe was not detected. In addition, five kinds of yeasts declined significantly during latter period of the seventh round fermentation process, but the yeast community structure during the fifth round was relatively more stable. The total yeasts during the early stage of the fifth round fermentation is 2?5 times more than the seventh round’s, while ethanol content of fifth round fermented grains was 2.45 times as higher as the seventh’s at the end of fermentation. Besides, volatile metabolic component of fermented grains was rich in species. The content of esters of the fifth round was remarkably higher than the seventh round, while organic acids and fusel oils were lower than the seventh round. [Conclusion] During Maotai-flvour liquor brewing process, there is rich diversity in yeast species, and the dynamics of yeast community structure affect liquor’s yield and quality obviously, which provides the basis for the study of Maotai-flavour liquor brewing mechanism.

Abstract:[Objective] This study aimed to clone the ferritin gene Abferritin from Acinetobacter baumannii and identify its anti-oxidative activity. [Methods] Relative expression of Abferritin under oxidative stress was analyzed by real-time PCR. The gene encoding sequence of Abferritin was inserted into the pET28a vector to generate the pET28a-Abferritin recombinant plasmid. This plasmid was transformed into the E. coli BL21(DE3) to create transformed strain of BL/pET28a-Abferritin. The Abferritin protein was expressed by IPTG induction and was purified by Ni2+-affinity chromatography. Kinetics of Fe2+ oxidation which catalyzed by Abferritin protein was determined by spectrophotometric analysis. Anti-oxidative activity of Abferritin was examined by the radical scavenging assay. Survival rations of the recombinant and control E. coli under the oxidative stress of H2O2 were also measured. [Results] Expression level of Abferritin was up-regulated in A. baumannii under oxidative stress. Abferritin protein was expressed in E. coli BL21(DE3) and was purified successfully. Ferroxidase activity assay demonstrated that the Abferritin protein could convert Fe2+ to Fe3+. The hydroxyl radicals were scavenged by the Abferritin protein in vitro and the ectopic expression of Abferritin could increase the survival ratios of E. coli cells under the oxidative stress. [Conclusion] Abferritin was strongly up regulated under oxidative stress and the Abferritin protein exhibited ferroxidase and anti-oxidative activity.

Abstract:[Objective] Control of the spread of tobacco black shank caused by Phytophthora nicotianae in tobacco production. [Methods] With Phytophthora nicotianae used as the assay fungus, bacterial strains antagonistic to P. nicotianae were isolated from tobacco rhizosphere soil obtained from Bijie District, Guizhou Province using dilution-plate methods and identified based on morphology, Biolog identification and 16S rRNA gene sequence analysis. The antimicrobial spectrum of these strains was further tested against Ralstonia solanacearum, Botrytis cinerea, Alternaria alternate and Colletotrichum destructivum in vitro. Optimization of growth conditions was also investigated through single factor variable analysis. [Results] Of 44 antagonistic strains obtained, strain 21b showed an inhibition rate of 78.33% to P. nicotianae and was identified as Bacillus subtilis. The size of inhibition zone of this strain against Ralstonia solanacearum, Botrytis cinerea, Alternaria alternate, and Colletotrichum destructivum were 19.5, 18.2, 14.6 and 13.4 mm, respectively. The optimal fermentation conditions for 21b was a temperature of 30 °C, pH 7.0?8.0, media/flask volume 12%, and 0.5% salt. [Conclusion] One bacterial strain with antagonistic activity against P. nicotianae isolated in this study could provide microbiological resources for the development of biocontrol of tobacco black shank in the future.

Abstract:[Objective] The current study aimed at studying the diversity of gut microbiota on young people living in rural Harbin and urban Harbin. [Methods] A combination of polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) was used to examine the diversity of gut microbiota of the subjects. Based on the DGGE profile, the similarity was performed by analysis of clustering and PCA; the analysis of diversity was evaluated by the Shannon-Weine index, richness and eveness; and the composition of gut microbiota was characterized by sequencing of the common and special bands of the lanes in DGGE profile. Based on the PCR, the species diversity of Lactobacillus and Bifidobacterium in the gut of rural subjects and urban subjects were analyzed. [Results] There was a separated tendency in structure of gut microbiota between rural and urban residents of Harbin, and the similarity of intestinal flora between rural and urban residents was lower than the similarity among rural people or urban people. The differences of diversity of gut microbiota between urban people and rural persons were not significant. The results of sequencing showed that the compositions of gut microbiota of urban subjects and rural subjects were same in phylum level, but different in the level of species and genus. The results of PCR showed that Lactobacillus and Bifidobacterium were prevalence bacteria in the gut of rural and urban young subjects of Harbin, and there was no significant difference in detection rate on the species of Lactobacillus and Bifidobacterium between rural subjects and urban subjects. The detection rate of L. plantarum, L. casei and L. salivarius was nearly 100%, and B. longum and B. breve was about 90%. [Conclusion] By the present research, the conclusion can be made that the diversity of gut microbiota was not significantly difference between the young subjects from rural Harbin and urban Harbin.

Abstract:High-throughput bacterial species identification is an important research topic in the area of microbiology, which plays a pivotal role in the disease diagnosis and environmental monitoring. Compared to the traditional phenotypic identification methods, molecular methods have the features of higher stability, shorter detection period and lower cost, which are becoming the main trend in the bacterial species identification. Especially, next generation sequencing technique, the detection chips based on nucleic acid identification, the protein profile analyses based on mass spectrometry provide the valid means for high-throughput, fast, accurate and quantitative bacterial species identification.

Abstract:Animal probiotics agent is a sort of viable bacteria formulation regulated under animal micro-ecological theory. This paper generally reviewed the current situation, research progress and the unsolved puzzles of probiotics agents in the domain of pet application by combination of technical data from overseas.

Abstract:A variety of physical, chemical and biological methods have been used to synthesize nanoparticles in which biological methods are green chemistry approachs, interconnecting organism with nanotechnology. Microorganisms are an important group of organism which can be used in nanoparticles synthesis, because they are growing fast, easy to culture, inexpensive and widely available. The diversity of microorganisms and nanomaterials determines the diversification of its synthetic mechanism. According to researches of home and abroad, this paper focus on summarizing mechanism of synthesis nanoparticles using microorganisms. The prospects of biosynthesis nanoparticles using microorganism have also been addressed.

Abstract:It is a practice that we foster students’ innovation ability and explore innovation experiment in microbiology experimental teaching. Students were assigned to isolation of bacteria, actinomycetes and molds from campus’s soil and Shennongjia’s soil, which they had collected during summer filed practice. The experiment is aimed at helping students to learn the method for determination of microorganism numbers in soil and master the basic techniques about plate count. The subsequent experiments involve techniques, such as aseptic technique, microscopical technique, pure culture technique, separation and purification methods of microorganisms, and some methods of molecular biology. The experiment has three innovation points. Firstly, the resources of soil were various, which could be collected from campus, Shennongjia, etc. Secondly, the isolates of microorganisms in soil could be used to apply National University or Wuhan University Student Innovation Program, e.g., isolating and identifying of a new bacterial species. The last, it combined common education with personalized teaching.

Abstract:The reform practice of the course “Food Microbiology Analysis” based on employment position and working task-driven was discussed from the aspects of course objectives, teaching contents, teaching implementation and teaching assessment. As a result, the course reform has a large impact on improving students study motivation and many abilities. These reform measures are helpful to improve the teaching quality in higher vocational education.

Abstract:[Objective] A Bifidobacteria genus-specific T-RFLP technique was developed to differentiate Bifidobacterium strains in microbial communities. [Methods] A Bifidobacteria genus-specific primer (g-bifid-F), based on the bacterial 16S rRNA gene sequence, was 5′-labeled with hexachlorofluorescein (HEX), followed by PCR with a 16S universal primer (1510r). The PCR amplicon was then digested by restriction enzyme Hae III and Alu I before terminal sequencing to achieve T-RFLP profiles. Moreover, combined with a Lactobacillus genus-specific T-RFLP technique in our lab’s previous study, this technique was used to detect bacterial composition and the compositional dynamic of a commercial probiotic product—Biostime. [Results] This technique was more specific than the traditional T-RFLP using universal primers, which could be used to detect or semi-quantitate different species of Bifidobacteria. [Conclusion] A Bifidobacteria genus-specific T-RFLP technique was well-established, and a combined system of these two T-RFLP techniques was successfully applied to commercial probiotic product detection.

Abstract:[Objective] In order to better understand the role of filamentous fungi in Chinese liquor fermentation process, a rapid and accurate method to monitor the change of fungal biomass is necessary. This study established a real-time qPCR method to detect and quantify Aspergillus tubingensis, which is widely used in Chinese liquor fermentation. [Methods] The methods of extracting genome from fermented grains have been optimized, the specific primers for A. tubingensis has also been designed and validated. [Results] The total DNA extracting from fermented grains can reach 1.060×105 ng/g by in situ mechanical crushing method. The applicability of real-time qPCR method to detect A. tubingensis in solid-state matrix has been validated in liquor fermentation process. [Conclusion] Real-time qPCR method can rapidly and accurately detect the fungal biomass in solid-state matrix, which provides a powerful tool for related researches.

Abstract:[Objective] To establish a kind of technology which is able to measure the distribution of the intracellular and extracellular Pb2+ of Fusarium sp.. [Methods] Soaked the mycelium pellet in EDTA solution to elute and measure the extracellular Pb2+, then digested the soaked mycelium pellet to measure the intracellular Pb2+. [Results] EDTA can elute the extracellular Pb2+, but the cells will not be damaged in 99 minutes; and that it is feasible to measure the concentrations of Pb2+ ions within cells, on the surface of cells and in the medium, with EDTA as reaction medium and titrant and XO as the indicator. On the basis of this experimental method, the growth curve of the strain in medium with 500 mg/L Pb2+ was determined, and the Pb2+ concentrations of medium, and intracellular and extracellular lead contents were measured. [Conclusion] Pb2+ will be adsorbed on the surface of cells at first, and then transported into cells. Results show that the Pb2+ adsorbance on the surface of fusaria cells is 1.37 mg/g, and the Pb2+ adsorption site on the surface of fusaria cells are about 3.97×1018/g.

Abstract:[Objective] Considering of the extremely special biochemical characteristics of Acidithiobacillus sp., the efficient double-layers screening, fed-batch high-density fermentation strategy and preservation method was respectively established and optimized for enhancing the utilization and reservation efficiency of these microbial resources. [Methods] The heterotrophic microorganisms such as Sacchromyces ellipsoideu and Rhodotorula sp. was employed as bottomed culture when the transmission electronic technology (TEM) was employed for observing the morphological differences. Based on the designed sulfide medium and elemental sulfur fed-batch strategy, the logarithmic phase of Acidithiobacillus thiooxidans was extended when the specific growth rate was improved. The effects of different preserved methods on cell survival rates were also investigated. [Results] Using heterotrophic microorganisms——Rhodotorula sp. as the underlying culture in the double-layers culture, the screening cycle was reduced by 1/3, while the plating efficiencies of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans were improved by about 3 times. More regular cell morphology was shown by TEM in the double-layers culture. The average specific growth rate of Acidithiobacillus thiooxidans was improved via fed-batch fermentation strategy in the Starkey-sulfide medium. The biomass conversion rate of sulfur and productivity were improved by 31.1% and 187.9% respectively, compared to the batch fermentation. The simple and efficient 4 °C cryopreservation perseveration was more suitable for preserving Acidithiobacillus sp. with the validity period about 1-3 months. [Conclusion] The screening efficiency of Acidithiobacillus sp. could be effectively improved by the double-layers culture with the auxiliary culture-Rhodotorula sp.. High-density fermentation of Acidithiobacillus thiooxidans was achieved via fed-batch fermentation strategy in the Starkey-sulfide medium. Simple and efficient 4 °C cryopreservation preservation was more suitable for preserving Acidithiobacillus sp. in the short-term.

Abstract:[Objective] To estimate the reference range of internal standard amount of unknown samples and optimize the method of data processing. [Methods] The different amount use of Methyl Nonadecanoate as internal standard was studied and discussed. The data analysis model of fatty acids was established to optimize the data processing of PLFA. [Results] The number and response values of detected PLFA were advanced with the increased of amount of internal standard, but both reduced when the amount over 16 nmol/g. Twenty-five sets of PLFA data showed that the PLFA contents calculated by response value were higher than by percentage value. A data analysis model of PLFA was established using Dot Net C# language. [Conclusion] The reference range of internal standard amount of unknown samples was preliminary estimated and optimized. Calculation of PLFA content by response value can avoid the error caused by the lack of percentage value. The use of calibration coefficient can reduce the system error. Data processing was automated by the data analysis mode. It improves the efficiency and accuracy of data analysis.