发根农杆菌A4菌株诱导三分三毛状根的研究

利用发根农杆菌A4菌株侵染三分三组培苗叶片诱导毛状根,并对其毛状根进行继代增殖培养;采用PCR技术检测毛状根。结果表明:发根农杆菌A4菌株Ri质粒的rolB基因已整合到三分三叶肉细胞的基因组中,进而成功地从叶片诱导出毛状根,诱导率可达77%以上;毛状根除菌后,能在无激素的MS固体培养基上快速生长。三分三遗传转化体系的建立为实现基于毛状根的三分三产托品烷类生物碱的代谢工程奠定基础。

康氏木霉的原生质体诱变

康氏木霉作为出发菌株,用溶壁酶对菌丝体进行破壁,提取原生质体再用紫外线做诱变处理,经多次筛选,从中选出一株酶活较高菌株,其FP酶活为36.2g/minmL,是原菌酶活的2.41倍。

Study on the Fermentation Conditions of a Mucopolysaccharide-producing Bacterium LV-1

bacterium LV-1 which isolated from soil sample were studied bacterium was isolated by serial dilution method, the effects of carbon source, nitrogen source, the initial pH and temperature on producing polysaccharide by it were discussed to confirm the optimum fermentation conditions. [ Result] The physicochemical properties showed that the polysaccharide was water-soluble, but insoluble in organic solvents including ethanol, butanol, and chloroform. It was neutral polysaccharide with negative charge and without reducing terminal. The pH of its solution was pH =7.5. There were no protein, fructose, uronic acid, sulphate and starch-like structure included in po/ysacchadde molecules. The optimum fermentation conditions for po/ysaccharide produc- tion were 3% mannitol as carbon source, 0.25% yeast extract as nitrogen source, culture temperature 28 ~（3 and pH =7.5. [Coadu^en] The re- search could provide basis for development and utilization of LV-1 and industrialized production of mucopolysaccharide.

Isolation and Phylogenetic Analysis of Plant Pathogen-Inhibiting Strains from Southern Ocean

[Objective] The aim was to isolate the strains resistant to plant pathogenic fungi from Southern Ocean and study their phylogenetic relationship and antimicrobial spectrum. [Method] Agar diffusion method was adopted to screen antimicrobial strains and determine the antimicrobial spectrum. Phylogenetic relationship of the strains was analyzed by neighbor-joining method of the Mega 4.0 software. [Result] Twenty antimicrobial strains were screened from seawater of Southern Ocean collected during the 27^th Chinese Antarctic Scientific Expedition. Molecular identification and phyloge- netic analysis indicated that two antimicrobial strains were members of Pseu- domonas, two strains were members of Psychrobacter, and the other 16 trains were members of Pseudoalteromonas. The antimicrobial spectrum of four strains which had higher antimicrobial activity indicated that the strains 312, 83-1 and 195 greatly inhibited the growth of Fusarium oxysporum, Rhizoctonia solani K（Jhn, Phytophthora capsici Leonian, Verticillium dahliae, Alternaria solani, Thanatephoru scucumeris and Phomopsis asparagi （Sacc）; strain 312-1 had obvious antimicrobial effect on the six of the plant pathogens except R. solani. [Conclusion] Four strains which had higher antimicrobial effect were obtained and should be further studied for development and application.

Study on Identification and Fermentation Condition of Strain ZY-19-2 with Inhibitory Effect against Tobacco Black Shank (Phytophtora parasitica var. nicotianae Tucker)

[Objective] The paper was to identify stain ZY-19-2 with inhibitory effect against tobacco black shank （Phytophtora parasitica var.nicotianae Tucker）, and study the fermentation condition of the strain. [Method]A strain ZY-19-2 with strong inhibitory effect against P. parasitica were isolated and screened from tobacco rhizosphere soil samples, and identified according to its morphological characteristics. The chitinase production activity of the strain under different culture conditions was also studied. [Result] For stain ZY-19-2 Paecilomyces lilacinus, the optimal fermentation conditions were as follows： 1.2% colloidal chitin as carbon source, 1% peptone as nitrogen source, 0.1% Tween 80 as surfactant, initial pH of fermentation broth at 6.0, the fermentation time of 60 h, inoculum amount at 1%, shaker speed at 120 r/min. The highest enzyme activity reached 0.216 U/ml. [Conclusion]The optimization of fermentation condition of strain ZY-19-2 lay foundation for large-scale production of cheap and efficient chitinase and chitin oligosaccharides, as well as application of the strain for control of tobacco black shank.

玉米各品种育种对比与策略分析

玉米是非常重要的粮食作物,还可以用作能源和饲料。在禾谷类农作物中占有重要地位并且伴随技术的不断发展玉米的增产增收空间巨大,玉米的育种技术更是其中的关键技术。本文针对当前国内外的多种玉米育种技术进行对比与分析,将玉米的品种和不同的育种策略相互对照,总结出优势与不足。同时也介绍了如何通过观察形态来判断育种成果的方法。为玉米种植因地制宜,合理选用育种技术提供了参考建议。

Fermentation optimization to improve production of antagonistic metabolites by Bacillus subtilis strain BS501a

To improve antagonistic metabolites production of Bacillus subtilis strain BS501a, physical parameters of fermentation and metal inorganic salts in medium, namely initial pH value, culture temperature, fermentation time, concentrations of CaC12, FeSO4, ZnSO4, MnSO4 and MgSO4, were optimized using one-factor-at-a-time and orthogonal tests. The results show that the optimal physical parameters of fermentation are an initial pH of 7.0, a culture temperature of 30 ~C, and a fermentation time of 48 h. The optimal concentrations of metal inorganic salts in basal medium are 10.2 mmol/L CaCl2, 0.4 mmol/L FeSO4, 3.5 mmol/L ZnSO4, 0.6 mmol/L MnSO4 and 2.0 mmol/L MgSO4. Among the metal inorganic salts, MgSO4 and MnSO4 play important roles in the improvement of the antagonistic metabolites production of B. subtilis strain BS501a; especially, MgSO4 contributes a highly significant effect. The average diameter of inhibition zone of the BS501a filtered fermentation supernatant （FFS） cultured in the optimal fermentation conditions against Magnaporthe grisea DWBJ329 reaches 71.4 mm, and there is 2.4-fold increase in antifungal activity as compared with 21.2 mm under the pre-optimized conditions.

Partial Characteristics of Hydrogen Production by Fermentative Hydrogen-producing Bacterial Strain B49

To investigate the characteristics of hydrogen production by a novel fermentative hydrogen-producing bacterial strain B49 (AF481148 in EMBL), batch experiments are conducted under different conditions. Hydrogen production has a correlation with cell growth and the consumption of glucose and soluble protein. The optimum pH for cell growth is 4.5±0.15. At acidic pH 4.0±0.15, the bacteria has the maximum accumulated hydrogen volume of 2382 ml/L culture and the maximum hydrogen evolution rate of 339.9 ml/L culture·h with 1% glucose. The optimum temperature for cell growth and hydrogen production is 35℃. In addition, fermentative hydrogen-producing bacterial strain B49 can generate hydrogen from the decomposition of other organic substrates such as wheat, soybean, corn, and potato. Moreover, it can also produce hydrogen from molasses wastewater and brewage wastewater, and hydrogen yields are 137.9 ml H 2/g COD and 49.9 ml H 2/g COD, respectively.

Characterization and identification of enzyme-producing microflora isolated from the gut of sea cucumberApostichopus japonicus

Gut microorganisms play an important role in the digestion of their host animals. The purpose of this research was to isolate and assess the enzyme-producing microbes from the Apostichopusjaponicus gut. Thirty-nine strains that can produce at least one of the three digestive enzymes （protease, amylase, and cellulase） were qualitatively screened based on their extracellular enzyme-producing abilities. The enzyme-producing strains clustered into eight groups at the genetic similarity level of 100% by analyzing the restriction patterns of 16S rDNA amplified with Mbo L Phylogenetic analysis revealed that 37 strains belonged to the genus Bacillus and two were members of the genus Virgibacillus. Enzyme-producing capability results indicate that the main enzyme-producing microflora in the A.japonicus gut was Bacillus, which can produce protease, amylase, and cellulase. Virgibacillus, however, can only produce protease. The high enzyme-producing capability of the isolates suggests that the gut microbiota play an important role in the sea cucumber digestive process.

Optimization of Culture Conditions for Production of Cellulase by a Thermophilic Bacillus Strain

The production of cellulase in Bacillus amyloliquefaciens UNPDV-22 was optimized using response surface methodology （RSM）. Central composite design （CCD） was used to study the interactive effect of culture conditions （temperature, pH, and inoculum） on cellulase activity. Results suggested that temperature and pH all have significant impact on cellulase production. The use of RSM resulted in a 96% increase in the cellulase activity over the control of non-optimized basal medium. Optimum cellulase production of 13 U/mL was obtained at a temperature of 42.24 ℃, pH of 5.25, and inoculum size of 4.95% （v/v） in a fermentation medium containing wheat bran, soybean meal and malt dextrin as major nutritional factors.

Establishment of Kinetics Models for Batch Fermentation Process of β-mannase with Bacillus licheniformis HDYM-04

In order to improve the yield of β-mannase and to investigate the rules of fermentation production, a high-yield β-mannase producing strain, Bacillus licheniformis HDYM-04, was used to investigate the kinetics models based on the optimal fermentation conditions： HDYM-04 strain was fermented at 37℃ for 30 h with agitation speed at 300 r/min and aeration rate at 3 L/min in a 5 L fermenter, the initial addition amount of konjac flour was 2%（w/v）, the initial pH of medium was 8.0, and the inoculum concentration was 6.7%（v/v）. Three batch fermentation kinetic models were established （cell growth kinetic model, substrate consumption kinetic model, product formation kinetic model） bases on Logistic and Luedeking-Piret equations. To be specific, cell growth kinetic model was dX/dt =0.431X （1- X/ 15.522 ）, substrate consumption kinetic model was -ds/dt =1.11 dX/dt ＋0.000 2 dP/dt ＋0.000 8X, and product formation kinetic model was dP/dt=133.1 dX ＋222.87X. The correlation coefficients R^2 of the three equations were 0.990 21, 0.989 08 and 0.988 12, respectively, which indicated a good correlation between experimental values and models. Therefore, the three equations could be used to describe the processes of cell growth, enzyme synthesis and substrate consumption during batch fermentation using B. licheniformis strain HDYM-04. The establishment of batch fermentation kinetic models （cell growth kinetic model, substrate depletion kinetic model, product formation kinetic model） could lay the theoretical foundation and provide practical reference for the applica- tion of HDYM-04 in fermentation industry.

Study on the Monascus Pigment of Monascus Strains JF-02 Fermentation

[ Objective] The aim was to study the fermentation technology of monascus pigment of monascus strains JF-02. [ Method] Single factor experiment was carried out to study the influence of temperature, initial pH, culture time, different agricultural byproduct, and nitrogen source on monascus pigment in fermentation solution. Meanwhile, orthogonal experiment was conducted to get the optimal culture medium and cultivation condition. [ Resultl The optimal gene in the pigment of monascus pigment was 200 g/L of rice, 30 g/L of sweet potato powder, 10 g/L of glucose, 15 g/L of monosodium glutamate, 0.1% of zinc sulfate, and 0.1% of magnesium sulfate. The optimal culture condition was 30 ℃ and initial pH was 6.0. Fermentation time was 72 h, but when 24-L fermentation pot was used, culture time can last to 84 h. [ Condusion] The study provided theoretical basis for the development and application of monascus strains.

Selection of Yeast Strains Containing β-Glucosidase for Improving Wine Aroma

Aroma is an important component for characteristic of wine. β-glucosidase is an enzyme for enhancing wine aroma during fermentation and aging. The aim of this study was to evaluate β-glucosidase activities from 17 yeast strains. It was found that strain 71 B-1122 exhibited highest activity of β-glucosidase. The purified enzyme of this strain was characterized. The purified enzyme was added in Muscat must before wine making process. The must and wine were determined for volatile compounds by headspace-SPME/GC-MS and found that the hexyl acetate, linalool, phenethyl, nerol and geraniol compounds were significantly increased after treated with the enzyme. Sensory evaluation of the combined alcoholic fermentation and flavour enrichment of Muscat juice by adding of purified β-glucosidase enzyme showed well oenological properties especially aroma. Therefore, in producing wine, yeast strain selection and addition of β-glucosidase enzyme should be considered because they were important factors affecting wine aroma.

Experimental Design as a Tool for the Development of Citric Acid Production by Yarrowia lipolytica Using Glycerol as a Feedstock

Due to the scarcity of fossil fuels in the world, there is increasing interest in the commercial production of biodiesel, which leads to obtaining large amounts of glycerol as a byproduct. If not disposed of properly, glycerol can generate environmental impact. One of the promises, the application of the crude glycerol is the production of citric acid by microbial fermentation. Citric acid is industrially produced by a submerged fermentation process with Aspergillus niger, using sucrose as carbon source, but due to increased demand for citric acid, alternative processes using renewable sources or waste materials as substrates and the cultivation of yeast strains are being studied. The aim of the study was to determine the best culture condition for maximum citric acid synthesis and lower isocitric acid production from crude glycerol through experimental design tool. For this purpose, the yeast strain Yarrowia lipolytica IMUFRJ-50682 was cultivated in nitrogen-limited glycerol-based media. Therefore, glycerol and yeast extract concentrations and agitation speed were evaluated as independent variables. With pure glycerol, the highest citric acid production achieved was 16.5 g/L with an isocitric acid production of 7.7% （in relation to citric acid）. With crude glycerol, citric acid production reduced to 6.7 g/L because of higher biomass yield. Therefore, an increase in the initial carbon to nitrogen molar ratio from 714 to 1,561 was necessary to increase citric acid production to 9.2 g/L, reducing isocitric acid production and to achieve a yield of 0.41 g of citric acid per glycerol consumed. In this condition, less nitrogen source was used, reducing production costs.

Chryseobacterium chengduensis sp. nov. isolated from the air of captive giant panda enclosures in Chengdu, China

A Gram-negative, aerobic, non-motile, rod-shaped bacterial strain, designated 25-1T, was isolated from the air inside giant panda enclosures at the Chengdu Research Base of Giant Panda Breeding, China. Strain 25-1T grew optimally at pH 7.0-8.0, at 28-30 ℃ and in the presence of NaCI concentrations from 0.0% to 0.5%. 16S rRNA gene sequence analysis indicated that strain 25-1m belongs to the genus Chryseobacterium within the family Flavo- bacteriaceae and is related most closely to C. camis G81m （96.4% similarity）, C. lathyri RBA2-6T （95.8% similarity）, and C. zeae JM1085T （95.8% similarity）. Its genomic DNA G＋C molar composition was 36.2%. The major cellular fatty acids were iso-C15：0 （44.0%）, iso-C17：0 3OH （19.8%） and C16：1 ω7c/16：1 ω6c （12.7%）. The only isoprenoid quinone was menaquinone 6 （MK-6）. The major polar lipids were phosphatidylethanolamine, two unidentified amino lipids and two unidentified lipids. The DNA-DNA relatedness between strain 25-1m and C. lathyri RBA2-6T was 38%. Phenotypic, genotypic, and phylogenetic characteristics indicated that strain 25-1T is a novel member of the genus Chryseobacterium, for which the name C. chengduensis sp. nov. is proposed. The type strain is 25-1T （ccmoc AB2015133T=DSM 100396T）.