Degradation of [8,9,-14C]endosulfan by soil microorganisms.

Twenty-eight soil fungi, 49 soil bacteria, and 10 actinomycetes were tested as to their ability to degrade the insecticide endosulfan. Using 14C-labeled material, the qualitative as well as the quantitative formation of metabolities, as well as of 14CO2, could be followed. Sixteen fungi, 15 bacteria, and 3 actinomycetes were found capable of metabolizing more than 30% of the applied endosulfan. The major metabolities detected were endosulfate, formed by oxidation of the sulfite group, and endodiol, formed by hydrolysis of the ester bond. The majority of highly active fungi formed endosulfate as the major metabolite, whereas the majority of active bacteria formed endodiol. In addition to endosulfate and endodiol, individual cultures contained small quantities of endohydroxyether and two unidentified products. The very small quantities of 14CO2 evolved from cultures indicated that an extensive mineralization of the carbon skeleton of endosulfan did not occur.     

桂林会仙喀斯特湿地芦苇群落土壤微生物数量动态分析

以广西桂林会仙喀斯特湿地典型芦苇植物群落为研究对象,于春、夏、秋、冬四个季节分别采集0~10cm,10~20 cm和20~30 cm不同层次的土壤样品,分析根际微生物与非根际微生物的数量特征及季节动态变化特点,探讨微生物数量对水热季节变化的响应规律。结果表明:不同季节的根际微生物与非根际微生物组成,均以细菌占绝对优势;微生物数量分布大小顺序为细菌放线菌真菌,细菌最高比例为96.62%,放线菌最高比例为35.38%,真菌的比例较低,最高仅为0.30%。细菌,真菌和放线菌的垂直变化明显,均随着土层的增加而呈现递减的趋势。不同土壤层次根际微生物与非根际微生物的季节变化一致,细菌数量表现为夏季秋季春季冬季,真菌数量表现为秋季夏季春季冬季,放线菌数量表现为秋季春季夏季冬季;细菌、放线菌、真菌的最大值分别为2.70×10~7、1.92×10~6、3.35×10~4cfu·g~-1,土壤微生物数量与土壤有机碳、全氮、全磷、全钾、速效氮、速效磷、速效钾等呈显著正相关。芦苇植物群落根际土壤微生物呈现出一定的根际效应,并与微生物数量、土壤深度、月平均降雨量和月平均气温变化等有关,而在冬季的根际效应则表现不显著。土壤养分含量是调节会仙喀斯特湿地土壤微生物数量变化的一个主要因素。     

Enrichment of an endosulfan-degrading mixed bacterial culture

An endosulfan-degrading mixed bacterial culture was enriched from soil with a history of endosulfan exposure. Enrichment was obtained by using the insecticide as the sole source of sulfur. Chemical hydrolysis was minimized by using strongly buffered culture medium (pH 6.6), and the detergent Tween 80 was included to emulsify the insecticide, thereby increasing the amount of endosulfan in contact with the bacteria. No growth occurred in control cultures in the absence of endosulfan. Degradation of the insecticide occurred concomitant with bacterial growth. The compound was both oxidized and hydrolyzed. The oxidation reaction favored the alpha isomer and produced endosulfate, a terminal pathway product. Hydrolysis involved a novel intermediate, tentatively identified as endosulfan monoaldehyde on the basis of gas chromatography-mass spectrometry and chemical derivatization results. The accumulation and decline of metabolites suggest that the parent compound was hydrolyzed to the putative monoaldehyde, thereby releasing the sulfite moiety required for growth. The monoaldehyde was then oxidized to endosulfan hydroxyether and further metabolized to (a) polar product(s). The cytochrome P450 inhibitor, piperonyl butoxide, did not prevent endosulfan oxidation or the formation of other metabolites. These results suggest that this mixed culture is worth investigating as a source of endosulfan-hydrolyzing enzymes for use in enzymatic bioremediation of endosulfan residues.     

Distribution of psychrophilic microorganisms in soils of Terra Nova Bay and Edmonson Point, Victoria Land and their biosynthetic capabilities

Microbiota of soil samples from Terra Nova Bay and Edmonson Point, Antarctica was observed by dilutions spread plate method. Variety of mesophilic and psychrophilic microorganisms was detected and isolated. Bacteria, actinomycetes, fungi, and microalgae occurred. Fungi genera Penicillium, Aspergillus, Paecilomyces, Cladosporium, Mortierella, Candida, Rhodotorula were found. By morphology and cell wall aminoacid composition the actinomycete genus Streptomyces was characterized. The bacteria and actinomycetes were screened for biologically active products. Some cultures formed enzymes, glycolipids and antibiotics. Psychrophilic strain Streptomyces sp. no. 8 was studied more detail and was established that it produced following antibiotics: azalomycin B, nigericin and non-polyenic macrolide antibiotic composed from two components that inhibited the growth of Gram-positive bacteria, yeasts, and phytopathogenic fungi.     

Enrichment of an Endosulfan-Degrading Mixed Bacterial Culture

An endosulfan-degrading mixed bacterial culture was enriched from soil with a history of endosulfan exposure. Enrichment was obtained by using the insecticide as the sole source of sulfur. Chemical hydrolysis was minimized by using strongly buffered culture medium (pH 6.6), and the detergent Tween 80 was included to emulsify the insecticide, thereby increasing the amount of endosulfan in contact with the bacteria. No growth occurred in control cultures in the absence of endosulfan. Degradation of the insecticide occurred concomitant with bacterial growth. The compound was both oxidized and hydrolyzed. The oxidation reaction favored the alpha isomer and produced endosulfate, a terminal pathway product. Hydrolysis involved a novel intermediate, tentatively identified as endosulfan monoaldehyde on the basis of gas chromatography-mass spectrometry and chemical derivatization results. The accumulation and decline of metabolites suggest that the parent compound was hydrolyzed to the putative monoaldehyde, thereby releasing the sulfite moiety required for growth. The monoaldehyde was then oxidized to endosulfan hydroxyether and further metabolized to (a) polar product(s). The cytochrome P450 inhibitor, piperonyl butoxide, did not prevent endosulfan oxidation or the formation of other metabolites. These results suggest that this mixed culture is worth investigating as a source of endosulfan-hydrolyzing enzymes for use in enzymatic bioremediation of endosulfan residues.     

低剂量混合稀土积累对黄褐土微生物主要类群的生态效应

采用田间小区试验和室内低剂量模拟叠加试验相结合的方法,研究低剂量混合稀土在黄褐土中积累对土壤微生物主要类群的生态效应．结果表明,低剂量稀土的持续积累对土壤细菌、放线菌产生刺激、抑制、再刺激的交替作用;对真菌也产生类似的作用,但抑制作用不显著,而刺激作用持续、明显．混合稀土对3类土壤微生物数量抑制程度顺序为：细菌>放线菌>霉菌．稀土积累至150mg·kg^-1时,土壤各类微生物的种群结构均发生显著的改变,耐稀土微生物数量大幅度增加,细菌中的G^-细菌、链霉菌的白孢类群、真菌中青霉分别成为优势种群．对低浓度稀土积累的田问土壤微生物学参数模拟计算结果表明,稀土对土壤细菌、放线菌和真菌的EC50(半抑制浓度)值分别为24．1、41．6～73．8和55．3～150．1mg·kg^-1,30mg·kg^-1值可以初步确定为稀土在黄褐土中积累的安全临界值．     

横断山北部高山区不同生态条件下土壤微生物数量及其与生态因子的相关性

采用全球定位系统(GPS)采集了横断山北部高山区35个样点的土样,用稀释平板法对该区细菌、真菌和放线菌进行分离计数,并分析了微生物数量与土壤养分、土壤水分、土地利用方式和pH等因子的相关性。结果表明,该区域3大类土壤微生物数量表现为细菌>放线菌>真菌。土壤微生物数量在不同区域表现为松林口>雀儿山>折多山;林地微生物比草地丰富,但不及草地微生物均衡。微生物数量与土壤理化因子间的相关分析表明,放线菌数量与pH呈显著正相关,真菌数量与土壤全钾含量呈极显著正相关。     

沼液与化肥配施对冬小麦根际土壤微生物数量和酶活性的影响

研究了等氮量条件下沼液与化肥配施对冬小麦根际土壤微生物(细菌、真菌和放线菌)数量及土壤酶(脲酶、蛋白酶和过氧化氢酶)活性的影响.结果表明:随着生育期的推进,不同施肥处理小麦根际土壤微生物数量和土壤脲酶、过氧化氢酶活性均呈先降后升的变化趋势,而蛋白酶活性则呈"S"形曲线变化.沼液与化肥合理配施能显著增加根际土壤微生物数量,提高土壤脲酶和蛋白酶活性,以基施50%沼液氮基础上追施50%化学氮处理和基施25%沼液氮基础上追施75%化学氮处理较好,传统尿素处理和单一沼液处理效果均较差.土壤过氧化氢酶活性以基施25%沼液氮基础上追施75%化学氮处理及单施沼液处理在所有测定时期均表现最高,其他处理在各生育时期间差异很大.沼液与尿素配合施用可以提高小麦根际土壤微生物数量和酶活性.     

秸秆和生草覆盖对桃园土壤养分含量、微生物数量及土壤酶活性的影响

以秸秆(覆盖重量分别为小麦(Triticum aestivum)秸3.25 kg·m^-2、玉米(Zea mays)秸1.97 kg·m^-2、禾本科杂草3.67 kg·m^-2)和生草(白三叶草(Trifolium repens)、高羊茅(Festuca arundincea)和紫花苜蓿(Medicago sativa), 播种量均为50 kg·hm^-2)为覆盖材料, 以不覆盖为对照, 研究了不同覆盖材料对桃园土壤微生物数量和酶活性的影响, 及其与土壤养分的关系。结果表明, 与对照相比, 除覆盖生草根际和非根际土壤全磷和速效磷含量差异均不显著外, 其他处理根际和非根际土壤碱解氮、速效钾、全氮、全钾和有机质含量差异均达到显著水平; 所有处理根际和非根际土壤氨化细菌、真菌和放线菌数量、土壤含水率和pH值、土壤脲酶和磷酸酶活性差异均达到显著水平。白三叶草处理的根际和非根际土壤碱解氮、速效钾、全氮、全钾、有机质含量, 土壤氨化细菌和真菌数量, 土壤脲酶和磷酸酶活性的平均升幅均最高, 分别为99%、270%、267%、117%、272%、158%、141%、156%和64%。氨化细菌、真菌、放线菌、脲酶和磷酸酶分别与土壤碱解氮、速效钾(放线菌和磷酸酶除外)、全氮、全钾和有机质呈显著或极显著的正相关。通径分析表明, 在3种土壤微生物和2种酶对养分含量的影响中, 脲酶是影响土壤碱解氮、速效钾、全氮、全钾和有机质的主要因子。     

Selective medium with nalidixic acid for isolating antibiotic-producing Actinomyces

The majority of actinomycetes belonging to various genera proved to be resistant to nalidixic acid concentrations having an inhibitory effect on bacteria with trailing growth i.e. B. subtilis and B. mycoides. The bacteria prevented isolation of actinomycetes as pure cultures. The use of a selective medium with nalidixic acid for isolation of soil actinomycetes resulted in 20 per cent increase in the number of the actinomycetes isolated as pure cultures. Preliminary treatment of the soil samples with calcium carbonate under moist conditions followed by the inoculation to the medium with nalidixic acid made it possible to increase isolation of actinomycetes at most 100-fold. With this complex method 495 actinomycete cultures were isolated, their antibiotic properties were studied and their taxonomic position at the genus level was determined. The complex method including the preliminary treatment of soil samples with calcium carbonate followed by inoculation to the selective medium with nalidixic acid is efficient and may be recommended for screening organisms producing new antibiotics.     

祁连山东段高原鼢鼠洞道土壤微生物和土壤酶

高原鼢鼠繁杂的洞道系统对高寒草甸土壤的理化性质有着重要影响。本研究采集高原鼢鼠活动洞道土壤样本,并以无鼢鼠扰动的同深度土层土壤作为对照。对比分析了2个处理土壤微生物（细菌、真菌、放线菌）、土壤酶（脲酶、蔗糖酶、碱性磷酸酶）活性以及土壤全氮、有机质、速效磷含量的差异。结果表明：高原鼢鼠洞道土壤速效磷和全氮的含量均高于对照,其中春季和秋季的速效磷达到显著水平（P<0.05）;秋季无鼢鼠扰动的土壤有机质和真菌的含量显著高于洞道土壤（P<0.05）;3种酶活性各处理间无显著差异（P>0.05）;对照区春季土壤的细菌和秋季真菌的含量显著高于洞道土壤（P<0.05）;洞道和对照区土壤的细菌、放线菌、速效磷、全氮和有机质的含量均在夏季达到最大值,各种指标其它月份差异不显著（P>0.05）。基于以上结果,本研究发现高原鼢鼠活动洞道土壤微生物和土壤酶与对照区土壤无显著差异（P>0.05）。     

pH dependence and thermostability of lipases from cultures from the ARS culture collection

Summary Previously we used a simple, sensitive agar plate method to screen lipase activity from 1229 selected cultures including 508 bacteria, 479 yeasts, 230 actinomycetes and 12 fungi covering many genera and species. About 25% of the cultures tested were lipase-positive. These lipase-positive strains were further classified as good, moderate or weak enzyme producers. We have expanded our screening method to focus specifically on the pH dependence and thermostability of these lipase activities. The lipases exhibited various pH sensitivities and were divided into three groups: (i) lipases which are active at pH 5.5 but not at pH 7.5—produced by 36 bacteria, 23 yeasts and four actinomycetes; (ii) lipases which are active at pH 7.5 but not at pH 5.5—produced by 17 bacteria, four yeasts, two actinomycetes and one fungus; and (iii) lipases which are active at both pH 5.5 and pH 7.5—produced by 112 bacteria, 90 yeasts, 15 actinomycetes and five fungi. By screening at 60°C and pH 9.0, we further identified 50 bacteria and 26 yeasts that produce thermostable alkali-tolerant lipases. Product analyses confirmed our screening results. Lipases with specific pH dependency and thermostability have potential to be developed into industrial enzymes.     

Studies on Antiviral and Antitumor Antibiotics

A screening for antiviral antibiotics was carried out using paper-disc agar-diffusion method. The microorganisms tested were unidentified soil fungi and the type cultures of our laboratory including actinomycetes, fungi, yeasts and bacteria. Mycelia or cells were extracted with acetone and the antiviral activity of the acetone extracts was determined. The extracts of actinomycetes mycelia showed the highest frequency of the appearance of antiviral activity against Newcastle disease virus. The frequencies of the appearance of antiviral activity in fungal and bacterial type cultures were the same degree and that of yeasts was low. Antiviral activity of the principles thus obtained was studied by microscopic observation in tube cultures using HeLa cells as a host.     

免耕高留茬抛秧对稻田土壤肥力和微生物群落的影响

通过大田试验,研究了不同秸秆还田和耕作方式（免耕+秸秆还田、免耕、常耕+秸秆还田、常耕）对稻田不同层次土壤肥力和主要微生物类群数量的影响.结果表明：上层土壤中,免耕+秸秆还田处理的有机质含量分别比免耕、常耕+秸秆还田和常耕处理高5.33、2.79和5.37 g·kg^-1;全氮、全磷、全钾、碱解氮、速效磷和速效钾含量也均以免耕+秸秆还田处理最高,免耕和常耕+秸秆还田处理次之,常耕处理最低.下层土壤中,各肥力指标以常耕+秸秆还田处理较高.秸秆还田各处理微生物类群数量较高,上层土壤以免耕+秸秆还田处理的细菌、真菌和放线菌数量最高,成熟期其纤维分解强度分别比常耕+秸秆还田、免耕和常耕处理高26.44%、79.01%和98.15%;下层土壤以常耕+秸秆还田处理的细菌、真菌和放线菌数量最高.免耕+秸秆还田处理的土壤养分和微生物呈表层富集特征.细菌、放线菌和纤维分解强度与土壤肥力各指标呈显著或极显著正相关关系.     

Antagonistic activity of soil acidophilic actinomycetes

It has been shown that soil acidophilic actinomycetes (mycelial prokaryotes with a growth optinum between pH 3 and 7) markedly differ from neutrophilic actinomycetes in antimicrobial activity: the former are more active against fungi and yeasts, whereas the latter effectively suppress Gram-positive bacteria. Acidophilic streptomycetes actively inhibit the growth of phytopathogenic fungi, especially on acidic media.     

轮作与休闲对日光温室黄瓜连作土壤微生物和酶活性的影响

采用盆栽试验,研究了不同作物轮作和休闲方式对日光温室黄瓜连作土壤微生物数量、酶活性及后茬黄瓜生长和产量的影响.结果表明： 与连作相比,轮作有利于改善土壤微生物结构,增加细菌和放线菌数量,减少真菌数量;轮作与休闲有利于提高土壤转化酶、脲酶、过氧化氢酶和多酚氧化酶活性.在不同作物轮作和休闲方式下,后茬黄瓜结果期的细菌、放线菌数量和土壤转化酶活性均呈先升后降趋势,在盛瓜期达到最大值;真菌数量随生育期延长而增加;脲酶、过氧化氢酶和多酚氧化酶活性则随生育期延长而降低,在初瓜期最高.不同栽培模式下,以大葱-黄瓜轮作和糯玉米-黄瓜轮作的效果更佳,明显改善了后茬黄瓜的生长,提高了产量.     

巨桉连栽对土壤微生物生物量和数量的影响

为探讨巨桉连栽对土壤微生物生物量和数量的影响,采用时空互换法,研究了马尾松(Pinus massoniana)林和不同连栽代次的巨桉(Eucalyptus grandis)人工林的微生物生物量、数量与土壤理化性质的关系。结果表明,巨桉一代林的土壤微生物生物量碳、生物量氮和土壤真菌、细菌数量与马尾松林的差异不显著,放线菌数量则显著增加。随巨桉连栽代次增加,土壤微生物生物量碳、生物量氮和土壤细菌、放线菌、真菌数量均递减。回归分析表明,土壤全磷能独立解释微生物生物量氮、细菌和真菌数量71.7%、86.1%和63.0%的变异,与总孔隙度共同解释微生物生物量碳87.9%的变异,与全氮共同解释放线菌数量89.6%的变异。可见,土壤全磷较大程度解释了微生物的变化。     

福建和溪亚热带雨林地表微生物的数量动态

分枯枝落叶层、腐解层和表土层（0—20cm深）三个层次研究福建和溪亚热带雨林的细菌、真菌和放线菌三大微生物类群的数量和月变化。结果表明：三个层次的微生物数量均以细菌最多,次为真菌,后为放线菌。比较三个层次,各微生物类群在各月份都是枯枝落叶层和腐解层数量较多。同一微生物类群在三个层次的数量均有明显而相似的季节变化;不同类群变化规律不尽相同,但基本只有一个高峰。细菌的高峰值在4月份,真菌约滞后一个月在5月份,放线菌则在8月份才出现高峰。各层次的细菌数和真菌数与月凋落物量、月降雨量和月平均气温各有显著的线性正相关（p＜0．01或0．05）,而放线菌则仅与月平均气温显著相关。     

有机物料厌氧发酵液中拮抗苹果再植障碍病原真菌的细菌筛选及其防治效果

有机物料厌氧发酵液(AFOF)能显著改善苹果再植障碍.本研究对AFOF中能拮抗苹果再植障碍主要病原菌(腐皮镰刀菌、层出镰刀菌、尖孢镰刀菌、串珠镰刀菌)的细菌进行了分离筛选,并对其作用效果进行了盆栽验证.结果表明: AFOF能显著抑制病原真菌的生长繁殖;对峙试验共得到4株具有较强拮抗作用的细菌(L11、L12、L13、L14),最高抑菌率达到57.3%,鉴定发现这4株细菌均属于芽孢杆菌属,相互之间没有明显的拮抗作用;在盆栽条件下,与连作土相比,溴甲烷熏蒸处理和拮抗菌菌液处理对平邑甜茶幼苗的生物量均有不同程度的促进作用;在幼苗的长势上,溴甲烷熏蒸处理效果要好于拮抗菌菌液处理;在根系活力上,拮抗菌菌液处理效果要好于溴甲烷熏蒸处理,根系长度、根尖数分别增加了25.1%、70.9%.与连作土处理相比,拮抗菌菌液和溴甲烷熏蒸均能显著降低土壤中的真菌数量,分别降低了71.2%和64.2%,拮抗菌菌液处理能显著增加土壤中的细菌和放线菌数量,分别增加了48.0%和140.2%,使土壤微生物结构向“细菌型”转化;而溴甲烷熏蒸处理显著降低了土壤中的细菌和放线菌数量,说明拮抗菌的确能够抑制土壤中病原真菌的生长.