一株高效解磷细菌的紫外诱变选育及其在红壤稻田施用效果

红壤对磷有强大吸附固定力,磷肥易被土壤中活性铁、铝固定而使有效态磷转化为各种形态的非有效磷,从而大大降低磷肥的利用率。解磷菌能使土壤中难溶性或不溶性的磷转化为易于被植物吸收利用的磷。通过对江西鹰潭红壤分离筛选并经过紫外诱变获得一株性状稳定的高效解磷细菌Y8。经鉴定,菌株Y8为蜡状芽孢杆菌（Bacillus cereus）。通过与生产中应用的徐州华龙高效复合菌肥厂的解磷细菌X3相比,菌株Y8降解有机磷、溶解Ca3（PO4）2的能力均比较高,分别为155.3mg·L^-1和240.1mg·L^-1。研究各种理化因子对菌株Y8解磷能力的影响,确定了菌株Y8的最佳培养条件为葡萄糖20g·L^-1、NH4（SO4）21.5g·L^-1、pH7.0、温度为35℃,在该条件下菌株Y8溶解Ca3（PO4）2的量为295.6mg·L^-1。在江西鹰潭红壤稻田的施用试验表明,将菌株Y8制成微生物菌剂施用于水稻田可起到减施化肥的作用。     

钾细菌在剩余污泥资源化中的应用

通过对钾细菌在剩余污泥中的接种,利用钾细菌将土壤中难溶性钾转化为可溶性钾是增加作物钾素营养的一个途径,将含大量钾细菌的剩余污泥制成的微生物肥料应用于农田中,从而达到剩余污泥的资源化利用,为剩余污泥的科学处理提供依据。     

多菌灵降解菌NY97-1的鉴定及降解条件

分离筛选出一株能高效降解多菌灵的芽孢杆菌菌株NY97-1,经生理生化和序列同源性分析,将该菌株鉴定为短小芽孢杆菌Bacillus pumilus.该细菌降解多菌灵的最适pH值为6.0～10.0,最适温度为35.0～40.0℃.该菌在多菌灵浓度为10、30、50、100、300 mg·L-1的无机盐培养基中,30℃振荡培养24 h后,其对多菌灵的降解率分别为42.44％、48.97％、77.19％、78.66％和90.07％.添加少量有机氮源如酵母浸出粉、胰蛋白胨、酵母膏可促进菌株NY97-1对多菌灵的降解作用,添加少量无机氮源尿素会抑制菌株NY97-1对多菌灵的降解作用.     

Alga-lysing bioreactor and the dominant bacteria strain

Alga-lysing bacteria have been paid much attention to in recent years. In this study, the alga-lysing strain P05 which was isolated from an immobilizing biosystem was immobilized by coke and elastic filler, forming two biological reactors. The removal efficiencies of algae, NH3-N and organic matter using the two reactors were studied. The results showed that strain P05 was an ideal algal-lysing bacteria strain because it was easy to be immobilized by coke and elastic filler which are of cheap, low biodegradability and the simple immobilization procedure. After 7 d filming, the biological film could be formed and the reactors were used to treat the eutrophic water. These two reactors were of stability and high effect with low cost and easy operation. The optimal hydraulic retention time (HRT) of each reactor was 4 h. The algae removal rates were 80.38% and 82.1% (in term of Chl-a) of coke reactor and filler reactor, respectively. And that of NH3-N were 52.3% and 52.7%. The removal rates of CODMn were 39.03% and 39.64%. The strain P05 was identified as Bacillus sp. by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.     

利用玉米浸泡液制备苏云金杆菌生物杀虫剂

以玉米淀粉生产过程中的浸泡液作为原料培养基,在5L发酵罐中培养苏云金杆菌生物杀虫剂,并以常规黄豆粉培养基为对照,考察了苏云金杆菌在玉米浸泡液中的生长代谢状况(包括菌体形态、菌数增长与芽孢形成)以及48h发酵液的生物毒效.研究表明,无需添加其它成分及前处理,玉米浸泡液所含营养成分即可满足苏云金杆菌生长需求,其培养的总活菌数及芽孢数在27h可达最大值1.51 ×109CFU·mL-1和1.41×109CFU·mL-1,分别比常规培养基高出59%和85%,芽孢形成与晶体释放提前约9～12h;36h扫描电镜(SEM)观察表明,发酵36h浸泡液中大部分芽孢囊已经自溶,游离出卵圆形芽孢与菱形的晶体;而此时常规培养基正处于芽孢形成后期,只有极少数的游离晶体与芽孢;48h发酵终点的生物毒效结果显示,以浸泡液为培养基的苏云金杆菌发酵液毒效(891.51 IU·μL-1)比常规培养基高89%.本试验为浸泡液的再利用提供了一条崭新的途径,同时降低了杀虫剂的生产成本.     

Stability of green fluorescent protein using luminescence spectroscopy: is GFP applicable to field analysis of contaminants?

Green fluorescent protein (GFP) was first isolated in the early 1970s for experimental use from coelenterates or the Pacific jellyfish. Aequorea victoria (Morin and Hastings, 1971). GFP has since become a favored biomarker in the photophysical analysis of molecular and cell biology because of its strong intrinsic visible fluorescence and the feasibility of fusing it to other proteins without affecting their normal functions (Creemers et al., 2000). Here we report using Bacillus subtilis expressing GFP to evaluate the influence of different environmental pH conditions on GFP fluorescence. Emission acquisitions were configured to excite at 471 nm and detect at an emission from 490 to 650 nm at 1-nm increments. Fluorescence intensity was significantly better at pH 7 (4.2 x 105 cps; P-value < 0.01) than at acid or alkaline conditions. GFP is a good biomarker for environments near netural conditions: however, GFP may be unsuitable where soils or waters are below or above pH 7 because of loss in fluorescence intensity. Alternative fluorescent markers and delivery systems must be examined in different environments to optimize responses from bioreporter molecules.     

嗜热脂肪芽孢杆菌过氧化氢酶基因perA在大肠杆菌中的高效表达

利用PCR技术得到嗜热脂肪芽孢杆菌 (Bacillusstearothermophilus)过氧化氢酶基因 perA ,将该基因与表达载体 pKK2 2 3 3连接构建重组质粒pK perA ,转化大肠杆菌过氧化氢酶HPⅠ和HPⅡ双缺突变株UM 2 ,得到重组大肠杆菌UM 2 1.酶活测定结果表明 ,表达产物具有正常的生物学活性 .SDS PAGE电泳结果显示出明显的特异性表达条带 ,单体Mr =86× 10 3 ,与嗜热脂肪芽孢杆菌所产酶相同 .实验表明 ,重组质粒在宿主UM 2中有较好的稳定性 ,在无选择压力条件下传代 6 0次基本保持稳定 ,传代 10 0次重组质粒保留 80 %以上 .摇瓶实验确定重组菌的最佳表达条件为 :IPTG浓度 ,0 .75mmol/L ;诱导时间 3h ;培养基起始 pH 6 .5 ;诱导温度 37℃ ;装液量 5 0mL/ 2 5 0mL .在优化条件下 ,重组菌产生的过氧化氢酶占菌体总蛋白的 8% ,酶活力可达 35U/mL ,是原始菌株BacillusstearothermophilusIAM110 0 1的 11.7倍 .图 2表 1参 10     

Evaluation of microbially enhanced composting of sophora flavescens residues

The effects of inoculants on the composting of Sophora flavescens residues were evaluated based on several physical, chemical and biological parameters, as well as the infrared spectra. Compared to the control compost without inoculants, the treatment compost with inoculants (Bacillus subtilis strain G-13 and Chaetomium thermophilum strain GF-1) had a significantly longer thermophilic duration, higher cellulase activity and a higher degradation rate of cellulose, hemicellulose and lignin (P < 0.05). Thus, a higher maturity degree of compost with apparently lower C:N ratio (15.88 vs. 17.77) and NH₄-N:NO₃-N ratio (0.16 vs. 0.20) was obtained with the inoculation comparing with the control (P < 0.05). Besides, the inoculants could markedly accelerate the composting process and increase the maturity degree of compost as indicated by the germination index (GI) in which the treatment reached the highest GI of 133.2% at day 15 while the control achieved the highest GI of 125.7% at day 30 of the composting. Inoculation with B. subtilis and C. thermophilum is a useful method to enhance the S. flavescens residues composting according to this study.     

Identification and comprehensive evaluation of a novel biocontrol agent Bacillus atrophaeus JZB120050

Abstract

Bacillus spp. have long been used as biocontrol agents because of their efficient broad-spectrum antimicrobial activity. We identified a novel strain of Bacillus atrophaeus, named JZB120050, from soil. B. atrophaeus JZB120050 had a strong inhibitory effect against Botrytis cinerea and many other phytopathogens. Gas chromatography-mass spectrometry showed that B. atrophaeus JZB120050 produced many secondary metabolites, such as alkanes, alkenes and acids; some of which were related to pathogen inhibition. Enzyme activity analysis showed that B. atrophaeus JZB120050 secreted cell-wall-degrading enzymes, including chitinase, glucanase and protease, which degraded fungal cell walls. Both the novel glucanase gene bglu and chitinase gene chit1 were cloned and heterologously expressed in Escherichia coli and the products showed strong enzyme activity. In addition, B. atrophaeus JZB120050 secreted siderophores and formed a significant biofilm. Future studies should focus on these antimicrobial factors to facilitate widespread application in the field of agricultural biocontrol.     

Bacillus thuringiensis field applications: Effect of nozzle type,drop size,and application timing on efficacy against gypsy moth

Abstract

In a series of recent studies, we evaluated the influence of delivery systems, drop size and application timing on the efficacy of aerially applied Bacillus thuringiensis against gypsy moth infestations. Use of different nozzle systems including Micronair, Flat Fan or Twin Jet, did not appear to result in significant differences in Bt coverage efficiency, foliage protection or population reduction. Nor was there any significant difference in population reduction when Bt was applied at two different drop sizes with volume median diameters of 110 and 163 μm. The efficacy of different formulations on larval populations were similar when used against the younger 1st and 2nd instar but differed when treatment was delayed until the population matured to the 3rd and 4th instar stages.