库布齐沙地生物土壤结皮中解磷菌的分离鉴定及解磷能力

解磷菌对生物土壤结皮(Biological soil crusts,BSCs)中不同形态磷之间的转化起重要的生物驱动作用.以无机磷和有机磷培养基分离筛选库布齐沙地BSCs中的解磷菌,基于16S r RNA基因进行分类鉴定;采用钼锑钪比色法测定菌株解无机磷能力;采用钒钼比色法测定其解植酸磷能力,以植酸酶活性表示.结果共分离得到33株可在无机磷(磷酸钙)培养基上生长迅速的细菌,以Bacillus属和Sphingomonas属为主;其中18株菌可以使PVK培养基变色,解无机磷能力较强,达0.422-3.531 mg/m L;这18株菌中有15株可在植酸钙培养基上生长,其植酸酶活力为2.45-20.84 IU/m L,主要为Sphingomonas属.本研究揭示了库布齐沙地BSCs中可培养解磷菌的类群组成及其解磷能力,结果可为认识和利用荒漠解磷菌提供理论依据和实践基础.     

磷细菌筛选及其对苗期玉米生长的影响

从潮土、水稻土、砂姜黑土、石灰土上植物根际土壤和根中分离了86株磷细菌,通过NBRIP液体摇瓶培养3 d,培养液水溶磷质量浓度为4.2～387.3 mg.L^-1,水溶磷质量浓度与培养液pH呈显著负相关（r^2=0.621 6）。筛选出3株磷细菌进行玉米盆栽试验,结果表明,1株磷细菌处理的玉米干物质量和吸磷量与对照（处理4）相比无显著差异,2株磷细菌处理的玉米干物质量和吸磷量与对照相比有明显增加,干物质量增加了19.6%～37.5%,吸磷量增加了22.7%～40.2%,其中编号为HCW115解磷菌株的效果相当于施用无机磷（P）10 mg.kg^-1处理。     

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。     

四种不同生态系统的土壤解磷细菌数量及种群分布

通过分析农田、林地、草地和菜地土壤有机磷细菌和无机磷细菌的数量及种群结构，发现有机磷细菌数量比无机磷细菌多；有机磷细菌主要是芽孢杆菌属，其次是假单胞菌属；而无机磷细菌主要是假单胞菌属。菜地土壤解磷细菌的数量和种类最多。不同的细菌解磷能力差异很大，其解磷能力与培养介质的ｐＨ值有一定的关系；许多菌株在进一步的纯化过程中失去了解磷能力。     

一株高效解磷细菌的筛选及其在红壤性水稻土中的施用效果

解磷菌能使土壤中被活性铁、铝等吸附固定的难溶性或不溶性的非有效态磷转化为易于被植物吸收利用的有效态磷,从而大大提高磷肥的利用率.通过对江西鹰潭红壤分离筛选,获得一株性状稳定的高效解磷细菌T4.经鉴定,菌株T4为伯克霍尔德菌属(Burkholderia sp.);T4溶解AlPO4、磷矿粉的能力均比较高,分别为334.2 mg L-1、193.1mg L-1;研究了各种理化因子对T4解磷能力的影响,确定了T4的最佳培养条件为乳糖15 g L-1,KNO3 1.0 g L-1,pH为7.0,温度为30℃,在该条件下T4溶解AlPO4的量为806.3 mg L-1.在江西鹰潭红壤性水稻土的施用试验表明,将菌株T4制成微生物菌剂施用于水稻田可起到减施化肥的作用.图6表2参28     

耐盐植物根际促生菌筛选及促生效应研究

开展盐渍化土壤中耐盐植物根际促生菌研究,有助于利用根际促生菌改良盐碱土壤。以前期分离自宁夏银北盐碱区耐盐植物根际土壤的110株细菌为材料,测定了菌株解磷、产IAA、产ACC脱氨酶和铁载体等促生特性,通过高活性菌株的交互作用,筛选出11个互不拮抗的菌株进行了菌种鉴定和复合菌群的构建,并验证了高效菌群C3和C8对植物幼苗的促生效果。结果表明:不同菌株的促生能力差别较大,其中23株细菌能够溶解无机磷,解磷量在2.90—70.92 mg·L~(-1)之间;6株能够产生ACC脱氨酶,酶活性最高为1.56μmol·mg~(-1)·h~(-1);46株菌具有产IAA的能力,IAA产量在1.33—34.74 mg·L~(-1)之间;24株菌能够产生铁载体。筛选出的11个高活性菌株分别隶属于芽孢杆菌属、假单胞菌属和鞘氨醇杆菌属,每4个菌株为组合共构建出9组复合菌群,其中C8组合ACC脱氨酶活性最高,达到3.67μmol·mg~(-1)·h~(-1);其次是C3组合,为2.77μmol·mg~(-1)·h~(-1);产生的IAA和解磷量分别在4.62—13.30 mg·L~(-1)和3.52—56.96 mg·L~(-1)之间,均为C3组合最高;产铁载体能力表现为C1—C4组合较强。盆栽实验表明,接种复合菌群C3和C8能明显促进苜蓿和柳枝稷幼苗生物量的增长。C3的促生效果尤为显著,与对照相比,使苜蓿和柳枝稷幼苗的株高分别增长54.93%和50.96%,地上鲜质量/干质量分别增加113.8%/119.6%和124.6%/82.08%,具备开发为微生物菌剂的潜能。     

缓释肥料对烤烟旺长期解磷细菌的影响

研究了缓释肥料对烤烟旺长期解磷细菌的数量以及磷细菌的溶磷能力的影响．结果表明,施用缓释肥料提高土壤中解磷细菌的数量和磷细菌的溶磷能力．施用缓释肥料和普通肥料的土壤中磷细菌总量为分别为7．6×10^6个／g干土和2．3×10^5个／g干土,且磷细菌的总量都高于不施肥的处理．图2,表4,参11．     

四川盆地大豆根瘤内生细菌的分离鉴定及促生效果

以四川盆地大豆根瘤为材料,采用划线法分离内生细菌、16S rDNA PCR-RFLP分析其遗传多样性,并结合菌株促生特性和盆栽试验筛选优良促生菌.从分离获得的130株内生细菌中选取了40株细菌作为供试菌株,16S rDNA序列表明分属于芽孢杆菌属(Bacillus)、肠杆菌属(Enterobacter)、假单胞菌属(Pseudomonas)、中华根瘤菌属(Sinorhizobium)和慢生根瘤菌属(Bradyrhizobium).以大豆为供试作物筛选出了12株具有促生能力的细菌,所有菌株均能分泌吲哚-3-乙酸(IAA),浓度达到0.353-32.404μg/mL;7株能产铁载体,活性单位为7.35%-34.31%;有11株具有溶磷能力,溶磷量达到4.26-10.6μg/mL;6株具有固氮能力.接种12株供试菌株后,玉米的农艺性状、植株全氮和全磷含量均优于单施化肥处理,其中菌株DA16-5效果最好,表现出良好的促生潜力.综上,四川盆地大豆根瘤内生菌遗传多样性丰富并且普遍具有促生能力,是重要的生物资源.     

几种可利用多氯联苯和多溴联苯醚植物内生菌的分离及初步鉴定

采用PDA、NA以及高氏一号培养基,分别从毛白杨、绦柳、荻以及二穗短柄草植株内分离、纯化出37株内生菌,其中34株为细菌,3株为放线菌.以分离纯化得到菌株各自对应的营养培养基并添加不同浓度的污染物对获得的37株内生菌进行污染物的耐受筛选,发现24株分别对20 mg·L-1的aroclor1254和BDE209具有一定的耐受性.利用无机盐培养基并添加污染物作为微生物唯一碳源筛选,发现菌株CPY-4和菌株SGL-1可以在只提供4-BDE的无机盐培养基中存活,菌株CPY-4,SGL-1、菌株4、菌株13可以在提供BDE209和aroclor1254的无机盐培养基中存活.形态学观察和16Sr DNA序列同源性分析,初步确定菌株SGL-1为克雷伯氏菌属,菌株CPY-4为地衣芽胞杆菌,菌株4和菌株13为肠杆菌属.     

玉米根际无机磷溶解菌的筛选与促生特性

从玉米根际土中筛选对无机磷具有高效溶解作用和促生功能的菌株,以向微生物肥料的开发提供菌株资源.采用溶磷圈法从玉米根际土中筛选溶磷菌株,采用钼锑抗比色法进一步测定菌株的溶磷能力.通过对菌株产吲哚-3-乙酸(IAA)、产铁载体、固氮、产几丁质酶能力及抗菌活性进行检测,综合评价溶磷菌的促生能力,并通过盆栽实验探索单一菌株及复合菌系对玉米促生的潜能.共筛选获得34株溶磷菌,其溶磷量为33.74-273.44 mg/L;34株菌中有10株菌(29.41%)同时具有3种及以上的促生功能,菌株PSM16、PSM27、PSM30同时具有4种促生功能;18株菌(52.94%)对不同供试病原菌表现出不同程度的抗性,菌株PSM12、PSM22、PSM25、PSM27、PSM34对5种目标病原菌同时具有抑菌作用;盆栽结果显示在土壤有效磷缺乏的条件下,经菌株PSM01、PSM12、PSM16及复合菌系处理后的玉米植株的株高、物质量、根系发达程度均显著提高(P 0.05),促生效果显著,且能提高土壤有效磷含量,复合菌系的效果更优. 16S rRNA基因测序结果显示,PSM01、PSM12、PSM16分别属于Bacillus aryabhattai、Paenibacillus silvae、Pseudomonas moorei.本研究为溶磷菌剂的开发提供了菌种资源,所构建的复合菌系具有较好的应用前景.(图4表4参43)     

驯化菌株降解油制气废水能力的比较

从3种污泥中驯化筛选出10种菌株,研究了各菌株对油制气废水不同污染指标的处理能力差别,以及各菌株对废水中芳烃化合物的降解能力.结果表明,各菌株可在初始阶段提高废水的BOD值,在高低废水浓度条件的降解能力基本一致;酚的去除率可达93.9%,但对废水中氨氮的去除率小于27.3%;实验采用的3种菌株对废水中的芳烃化合物都能降解,但其对芳环数≤3的芳烃化合物的降解能力强于对芳环数为4~6的芳烃化合物的降解能力.图4表2参10     

Quantification of polyphenols during retting and characterization of bacteria from the Kadinamkulam Backwaters, Kerala

The retting environment which provides a competitive niche for specialized microbes is speculated to harbour a variety of microbes with high biodegradation potential. In this context, an effort has been made to isolate and identify bacterial species having high tolerance to phenol In vitro. Maximum polyphenol (1.897 mg l(-1)) as observed during the initial period of retting, which decreased as retting proceeded. Based on biochemical characterization, the isolated bacterial strains were identified as Micrococcus sp., Moraxella sp. strain MP1, Moraxella sp. strain MP2 and Moraxella sp. strain MP3, Pseudomonas sp. strain PP1 and Pseudomonas sp. strain PP2, Amphibacillus sp., Brucella sp. strain BP1 and Brucella sp. strain BP2, Aquaspirillum sp., Escherichia coli strain EP1 and Escherichia coli strain EP2, Campylobacter sp., Aeromonas sp., Neisseria sp., Vibrio sp., Erwinia sp. and Mesophilobacter sp. These strains were found to tolerate maximum concentration of phenol viz. 200 to 1000 mg l(-1). Plasmid analysis of phenol resistant bacterial isolates showed that almost all the cultures had at least one plasmid of size > 1Kb. Studies on the protein profile of isolated bacterial cultures showed the presence of proteins with molecular sizes ranging from 10 to 85 KDa with exception of Mesophilobacter and Neisseria having still high molecular weight protein (95 KDa). Bacterial strains isolated from coir-ret-liquor showed tolerance to high phenol concentration.     

Sol-gel immobilisation of methyl parathion degrading bacteria isolated from agricultural areas of Pakistan

The following areas are discussed in this paper: immobilisation of bacterial consortium in sol-gel; methyl parathion degradation and bioremediation applications; evaluation of indigenous bacterial isolates of contaminated soils. Bacterial strains were isolated from agricultural areas of Pakistan which were contaminated with methyl parathion. A bacterial consortium of seven (out of 64) Enterobacteriaceae isolates including Citrobacter, Enterobacter and Proteus vulgaris capable of degrading methyl parathion (enzyme activity ranging 410–675 mU mL^?1 for individual isolates and 982 mU/mL for consortium) was selected and subsequently immobilised in tetraethyl orthosilicate (TEOS) and sodium-silicate-based sol-gel matrices. Cell viability of suspended and immobilised bacterial consortium was monitored using a minimal salt medium supplemented with methyl parathion. The results indicate that sol-gel immobilisation can be helpful to increase the shelf life of methyl parathion degrading bacterial strains along with preservation of biological activity for bioremediation applications in field.     

Phylogenetic diversity of epibiotic bacteria in the accessory nidamental glands of squids (Cephalopoda: Loliginidae and Idiosepiidae)

Bacterial communities were identified from the accessory nidamental glands (ANGs) of European and Western Pacific squids of the families Loliginidae and Idiosepiidae, as also in the egg capsules, embryo and yolk of two loliginid squid species, and in the entire egg of one idiosepiid squid species. The results of phylogenetic analyses of 16S RNA gene (rDNA) confirmed that several phylotypes of α-proteobacteria, γ-proteobacteria and Cytophaga–Flavobacteria–Bacteroides phylum were present as potential symbiotic associations within the ANGs. Several identified clones were related to reference strains, while others had no known close relatives. Gram positive strains were rare in loliginid squids. Several bacterial groups may play important roles in the function of the ANGs, such as production of the toxic compounds involved in egg protection and carotenoid pigments. Within the eggs, no bacteria were associated with embryo or yolk of Loligo vulgaris and Sepioteuthis lessoniana, but α- and γ-proteobacteria were present in the egg capsules. Most bacterial strains detected in the egg capsules were the same as those found in the ANGs. The cephalopods of temperate regions (European cuttlefishes and the squid L. vulgaris) appear to be associated with one Agrobacterium strain (Agro2) while tropical-subtropical strains (Asian and Australian loliginids) are associated with Silicibacter-related strains, suggesting a biogeographic clustering for the Agrobacterium-like strains.     

Bioactive potential of seagrass bacteria against human bacterial pathogens

Study of marine organisms for their bioactive potential, being an important part of marine ecosystem, has picked up the rhythm in recent years with the growing recognition of their importance in human life. Investigation was carried out to isolate 32 strains of endo and epiphytic bacteria in 2 seagrass species viz., Syringodium isoetifolium and Cymodocea serrulata. Morphologically different bacterial strains were tested against 5 antibiotic resistant human bacterial pathogens, of which 10 associated bacteria shown inhibitory activity against one or more bacterial pathogens. Minimum inhibitory concentration (MIC) and Minimum bacterial concentration (MBC) determination with extracellular bioactive compounds from the associated bacteria reveals that, the strain ENC 5 showed inhibitory activity against all the bacterial pathogens with the maximum sensitivity against Pseudomonas aeruginosa at the MIC value of 500 microg ml(-1).     

Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.     

Effect of hydrostatic pressure and temperature on the activity and synthesis of chitinases of Antarctic Ocean bacteria

The formation of chitinases by psychrophilic and psychrotrophic marine Antarctic bacteria and the activity of these extracellular enzymes were investigated under simulated deep-sea conditions. The formation of the chitinases was affected by hydrostatic pressure of 400 bars. However, the extent of pressure inhibition differed with the bacterial strains tested and was considerably less with the extreme psychrophilic bacteria isolated from sediments of greater depth. Growth of these psychrophilic strains had a moderately barophilic character at 400 bars, whereas growth of the psychrotrophic strains was clearly restricted under simulated deep-sea conditions. With regard to the activity of the extracellular chitinases of various bacterial strains, a relatively uniform response was found. All chitinases were highly barotolerant at near neutral pH and were active up to 1000 bars. Low temperatures reduced their activity but not their barotolerance. A low pH of 5.1 diminished the barotolerance of some chitinases. The results suggest that the indigenous deep-sea bacteria are capable of decomposing chitin settled to or produced in the depth of the Antarctic Ocean.     

采用生物发光检测法进行Ames试验的研究

将自行构建的含有青海弧菌荧光酶基因的质粒pACYCL184引入Ames试验的4个菌株TA97、TA98、TA100、TA102中,构建成能够生物发光的工程菌,分别命名为TAL97、TAL98、TAL100、TAL102.实验表明,该4株工程菌保持原出发菌株所具有的与Ames试验相关的性状,对各类致突变剂有良好的反应,且可用平皿计数法进行致突变试验.其发光值与致突变剂浓度存在良好的量效关系,故可用发光检测取代原Ames试验的平皿计数,使检测更简单、方便、快速.图3表3参11     

Larval settlement in the serpulid polychaete Hydroides elegans in response to bacterial films: an investigation of the nature of putative larval settlement cue

Larval settlement in the marine polychaete Hydroides elegans (Haswell) is induced by certain bacteria in marine biofilms. The exact nature of the settlement cue that larvae of H. elegans receive from these bacteria remains unknown. In this study, we revealed some properties of the bacterially derived larval settlement cue by investigating the larval settlement inductive activity of two bacterial strains after various treatments. These two bacterial strains, Roseobacter sp. and an α-subclass Proteobacteria, are highly inductive to larval settlement of H. elegans. The larvae responded similarly to Roseobacter and Proteobacteria in all the larval settlement bioassays, suggesting that the larval settlement-inducing substances produced by these bacteria may share common characteristics. First of all, the larvae did not settle in the seawater conditioned by the bacteria attached as a film or by the bacteria that were freely suspended in seawater. The results suggest that the putative larval settlement cue is not released into seawater and, therefore, should be associated with the surface of the bacteria. Secondly, formaldehyde treatment entirely eliminated the larval settlement induction activity of the bacterial films, and streptomycin treatment reduced the percentage of larval settlement on the bacterial films in a concentration-dependent manner. Since both treatments can kill bacteria with little damage to the surface chemistry of bacterial cells, the decline in larval settlement is suggested be due to a reduction of the viable bacterial population in the bacterial films. In fact, the reduction of larval settlement in the streptomycin treatments coincided with the decrease in viable bacterial populations in broth cultures containing respective concentrations of streptomycin. These results suggest that the viability of Roseobacter and Proteobacteria is important to their settlement induction effect. Since the larval settlement induction activity of the bacterial strains appears to correlate with their viability, we suggest that the putative larval settlement cue is derived from a metabolic pathway in the bacteria and that the cue is exported to and concentrated at the extracellular polymer matrix of the bacterial cell, at which the larvae establish contact with the bacteria. The larval settlement cue may be highly susceptible to degradation so that a metabolically active bacterial film is needed to maintain the putative cue at a concentration that surpasses the threshold for induction of larval settlement. Received: 14 October 1998 / Accepted: 5 September 2000     

Degradation of chitin and antagonism in Phomopsis asparagi by Bacillus amyloliquefaciens MY001北大核心CSCD

About 10 billion tons of chitin, a natural polymer, is generated annually. However, the utilization of chitin was hindered because of the lack of a clean and effective degradation process. In this study, bacterial strains with chitosanase-producing ability were isolated from soil and identified using 16S rDNA, gyrA, and gyrB gene sequence analysis. In addition, the degradation of insoluble chitin and antifungal capability of the bacterial strains were investigated. One selected isolate was identified as Bacillus amyloliquefaciens strain MY001. It showed antagonistic effect on Phomopsis asparagi (Sacc.) Bubak and could degrade insoluble chitin. The combination of chitin/chitosan and MY001 strain could enhance the inhibition on the growth of P. asparagi. In conclusion, strain B. amyloliquefaciens MY001 can directly degrade chitin. The combination of MY001 strain with chitin shows more effective inhibition on P. asparagi growth. These results suggest the direct utilization of chitin against fungal diseases of crops. © 2018 Science Press. All rights reserved.