陕甘宁地区根瘤菌数值分类与DNA同源性分析

在陕甘宁地区根瘤菌资源调查的基础上,选取其中与黄芪、岩黄蓍、木兰、鸡眼草及胡枝子共生的４０株快生根瘤菌和２５株参比菌株,对１３３项表型性状进行聚类分析,在８２％的相似性水平上构成５个不同于已知种的新亚群．对第１、２、３、４亚群ｘ（Ｇ＋Ｃ）／％测定及ＤＮＡ同源性分析结果表明,４个亚群ｘ（Ｇ＋Ｃ）／％含量均处于Ｒｈｉｚｏｂｉｕｍ属范围;各亚群内菌株ＤＮＡ同源性均在７５％以上,每个亚群中心菌株与１３个已知种模式菌株的ＤＮＡ同源性均小于６３％．从而证明,第１、２、３、４亚群为４个独立的快生根瘤菌新种群     

原生质体转化构建有机磷农药降解工程菌

降解有机磷农药甲胺磷、敌敌畏和对硫磷的菌株地衣芽孢杆菌 Ｐ１２（ Ｂａｃｉｌｌｕｓｌｉｃｈｅｎｉｆｏｒｍｉｓ） 经溶菌酶处理获得原生质体,加入降解乐果的供体菌 Ｇ１ Ｄ Ｎ Ａ,经ρ（ Ｐ Ｅ Ｇ６ ０００） ＝ ３００ ｇ Ｌ－ １ 诱导,在液体再生培养基中振荡培养ｔ＝ ２０ ｈ ,使其恢复细胞壁后,离心收集菌体,涂布于含乐果的基础培养基上,经筛选得一转化子 Ｊ Ｐ Ｚ．其菌落形态明显不同于出发菌株,乐果平板连续传代１０ 次,性状保持稳定．在θ＝ ３０ ℃,１００ ｒ／ｍｉｎ 的培养条件下,３ ｄ 内对ρ（ 甲胺磷） ＝ ０ ．５ ｇ Ｌ－ １ ,ρ（ 敌敌畏） ＝ ０ ．２ ｇ Ｌ－ １ ,ρ（ 对硫磷） ＝ ０ ．１ ｇ Ｌ－ １ ,ρ（ 乐果） ＝ ０ ．３ ｇ Ｌ－ １ 的降解率分别为 Ｒｄ ＝ ７９ ．１ ％ ,４６ ．７ ％ ,２９ ．４ ％ 和４６ ．４ ％ ．     

慢生型花生根瘤菌16S rRNA分析

以２１株分离自四川,云南不同花生产区的慢性型花生根瘤菌和６株参比菌株为材料,进行了１６ＳｒＤＮＡＰＣＲ－ＲＦＬＰ,结果除证实了慢性型花生根瘤菌同慢生型大豆根瘤菌（Ｂｒａｄｙｒｈｉｚｏｂｉｕｍｊａｐｏｎｉｃｕｍ）高度的遗传相似性外,还发现了一些菌株同Ｂｒａｄｙｒｈｉｚｏｉｂｉｕｍｌｉａｏｎｉｎｇｅｎｓｉｓ和Ｂｒａｄｙｒｈｉｚｏｂｉｕｍｅｌｋａｎｉｉ有较高的遗传相似性,菌株Ｓｐｒ１－２的１６ＳｒＲＮＡ     

培养条件对产酸克雷伯氏菌SG-11生物合成IAA影响的研究

研究了产酸克雷伯氏菌（Ｋｌｅｂｉｓｅｌｌａ ｏｘｙｔｏｃａ）ＳＧ－１１菌株的培养条件对其ＩＡＡ生物合成的影响。结果表明：该菌生长到稳定期时才开始大量产生ＩＡＡ；在基本培养基中，该菌对不同氮源的利用有别，在以（ＮＨ４）２ＳＯ４为氮源时，ＩＡＡ的产量最高，作为碳源的葡萄糖当加入浓度为６ｇ／Ｌ时，游离态ＩＡＡ的产量最高；在ＬＢ培养基中，当ｐ（Ｏ２）由正常值降低１／４时，ＩＡＡ的产生量陡然下降，灭菌前，把     

微生物降解蔬菜残留农药研究

从富集培养物中筛选到以甲胺磷为唯一碳源生长的菌株ＮＭＪ５和以乐果为唯一碳源生长的菌株ＮＭＬ３,经鉴定分别为芽孢杆菌属（Ｂａｃｉｌｕｓｓｐ．）和假单胞菌属（Ｐｓｅｕｄｏｍｏｎａｓｓｐ．）．它们利用甲胺磷、乐果生长的最适条件及在无机盐培养基中农药最大耐受浓度分别为ｐＨ７．５,ｔ＝３５℃,ρ＝１０００ｍｇ／Ｌ和ｐＨ７．５,ｔ＝３０℃,ρ＝２０００ｍｇ／Ｌ．在无碳基础培养基内单菌株培养８ｄ,５２４ｍｇ／Ｌ的甲胺磷好气降解４２．５％,厌气降解３５．９％,２５０ｍｇ／Ｌ的乐果好气降解５０．２％,厌气降解１６．４％．小区试验表明,ＮＭＪ５、ＮＭＬ３的菌液制剂对普通白菜的变种南农矮脚黄（Ｂｒａｓｉｃａｃａｍｐｅｓｔｒｉｓｓｐ．ｃｈｉｎｅｎｓｉｓＬ．ｖａｒ．ｃｏｍｍｕｎｉｓＴｓｅｎｅｔＬｅｅ）中残留的甲胺磷、乐果有明显的去除作用．     

一株拮抗烟草青枯病菌的真菌的筛选与鉴定

为寻找烟草青枯病的生防微生物,研究采用平板涂布法从重庆烟田健康土壤中分离根际微生物,纯化后得到了59株真菌。用滤纸片法从中筛选出1株对青枯雷尔氏菌（Ralstonia solanacearum）有稳定拮抗效果的真菌,其抑菌圈为10.0 mm。其抑制活性优于或接近于近期发现的其他生防微生物。测定该株活性真菌的全细胞脂肪酸组成,进行菌落和分生孢子形态学观察,以及rDNA ITS区测序,结果表明：该菌株全细胞脂肪酸属性未被收录于TSBA6脂肪酸数据库,而根据形态学观察结果,以及对所获rDNA ITS区序列的分析,鉴定该株真菌为烟曲霉（Aspergillus fumigatus）。     

孔雀绿染料的微生物脱色研究

从印染废水处理池的活性污泥中分离到五株对孔雀绿染料有脱色能力的细菌,其中编号为３＃的菌株人苍白杆菌（Ｏｃｈｒｏｂａｃｔｒｕｍ ａｎｔｈｒｏｐｉ）的脱色能力较强。对３＃菌株的脱色反应条件进行初步研究表明,３＃细菌的脱色反应的ｐＨ为５－１１,最佳范围ｐＨ６－７;温度１０－３０℃,最佳温度３０℃;有氧气民政部下的脱色效果较缺氧情况下好;在最佳条件下该菌对孔雀绿染料的脱色作用较强,处理４、１２、２４ｈ后,     

有机磷农药降解菌的紫外诱变育种

有机磷农药降解菌地衣芽孢杆菌（ Ｂａｃｉｌｌｕｓｌｉｃｈｅｎｉｆｏｒｍｉｓ） 经紫外线诱变后,筛选出突变菌株Ｐ１２ ．在θ＝３０℃,溶解氧ρ（Ｏ２） ＝２ ．５ ｍｇ Ｌ－１ 的培养条件下,３ ｄ 内对甲胺磷的降解率为８０ ．１％ ,比出发菌株提高了将近１０％的降解率．农药斜面连续传代１０ 次,降解活力保持稳定．     

二氯苯在盐水溶液中活度系数的研究

本文采用紫外分光光度法测定了２５．００±０．００１℃下间位和对位二氯苯在ＨＣＯＯＮａ,ＣＨ３ｃＯＯＮａ,（ＮＨ４）２ＳＯ４,ＭｇＳＯ４和Ｎａ２ＳＯ４水溶液中活度系数（ｆ）,它们的ｌｇｆ－Ｃ关系符合Ｓｅｔｓｃｈｎｏｗ盐效应公式。     

紫云英根瘤分离菌株质粒多样性的研究

从广东、广西、湖南、湖北、江西、浙江、江苏７省紫云英（ＡｓｔｒａｇａｌｕｓｓｉｎｉｃｕｓＬ．）主要分布区４６个采集点的紫云英根瘤中分离到１０１个菌株．检测发现有１６种不同质粒类型,所含质粒１～４个不等,含两个质粒的菌株占绝大部分,质粒Ｍｒ为３６×１０６～３６４×１０６．用克隆Ｍ．ｈｕａｋｕｉ７６５３Ｒ的ｎｏｄＤＢＣ４．２ｋｂ片段为探针（α３２ｐｄＣＴＰ）进行的分子杂交结果表明,所有这些菌株均含有共生质粒,通常为第一大质粒,但Ｃ、Ｌ、Ｍ、Ｎ和Ｏ型为第二大质粒,共生质粒Ｍｒ在１３１×１０６～３６４×１０６之间．比较质粒类型与１６Ｓ２３ＳｒＤＮＡＩＧＳ（ｐＨｒ,ｐ２３ＳＲ０１）ＰＣＲＲＦＬＰ分析结果表明,ＩＧＳ类型不同,即使在同一植株分离所得的根瘤其质粒类型常不同,但在同一植株上分离菌株的质粒类型相同,则其ＩＧＳ常表现相同．表明质粒的类型是细菌染色体背景和细菌外环境（如共生条件等）共同作用的结果     

Antibacterial effects of Anodonta cygnea fluids on Escherichia coli and enterococci multi-drug-resistant strains: environmental implications

The persistence of bacteria in Anodonta cygnea largely depends on the hemolymph bactericidal activity against aquatic micro-organisms.

The aim of this study was to assess the in vitro bactericidal activity of whole hemolymph against multi-drug-resistant Escherichia coli and enterococci strains. Four mussels were injected with multi-resistant strains of E. coli in order to determine their in vivo bactericidal reactivity. In vitro experiments showed that the hemocytes viability decreased almost 70% in 4 h, the same happened in the control hemolymph. Enterococci tested in in vitro experiments were more susceptible than E. coli to whole hemolymph and hemocytes fractions. None of the enterococci bacteria was detected in the hemolymph fractions from 24 to 72 h, while E. coli was still detectable 72 h after inoculation both in the control and respective plasma fractions. The microbial inhibition on both hemolymph and cell pellets suggest that hemocytes may be the main responsibles for that process.

In vivo experiments showed that hemolymph inhibited the E. coli load injected and it was correlated with a high tendency for the increase in hemocytes counts, after 72 h. High adherence of E. coli and an intense pseudopods manifestation, mainly to the granulocytes, suggest a clear phagocytosis process.     

Sulphur-oxidising bacteria isolated from deep caves improve the removal of arsenic from contaminated harbour sediments

Acidophilic S-oxidising bacteria isolated from sulphur-rich deep caves (Frasassi, Italy), characterised by relatively low temperature, were tested for their ability to mobilise (semi-)metals from contaminated sediments. Sediment samples from two commercial Italian seaports were used to set up bioleaching experiments. The effect of different growth substrates was also investigated. Our experiments revealed that S-oxidising bacteria isolated from Frasassi caves have a high potential to remove As from contaminated marine sediments, as never reported before. Although As solubilisation efficiency was quite low (i.e. about 30%), only a small amount of As was associated with non-residual fractions of the sediment. On the contrary, the solubilisation efficiencies of Zn and Ni (20% and 10%, respectively) were lower than those previously obtained by the use of other acidophilic bacteria and mainly influenced by the experimental conditions rather than by the presence of the S-oxidising bacteria. Results presented here open new perspectives in bioleaching applications for the remediation of contaminated sediments. Indeed, microbial strains adapted to relatively low-temperature environments could improve sediment bioleaching at temperature regimes where mesophilic and thermophilic strains are not favoured. Such strains could be exploited for developing selective bioremediation procedures for sediments contaminated with As, to be applied in multistep biotreatment processes.     

慢生根瘤菌交叉结瘤及其系统发育关系的研究

选用包括6株花往根瘤菌[Bradyrhizobium sp.(Arachis)]在内的13株慢生根瘤菌，进行了6种豆科植物结瘤试验及共系统发育分析。研究结果表明，慢生根瘤菌与试验的豆科宿主植物间普遍存在交叉结瘤，还发现有些菌株在原宿主外的豆科植物根部形成类根瘤的现象。16S rDNA和23S rDNA的PCR－RFLP分析揭示出这些菌株rDNA基因的差异性，不同宿主来源的慢生根瘤菌株可以同属一个rDNA类型，而从同一宿主中分离的根瘤菌菌株可分属多种不同的rDNA类型，研究结果表明，采用rDNA PCR-RFLP等遗传背景分析的手段，可能筛选出广谱的根瘤菌力株，表4参8。     

温度和溶菌酶敏感菌株的L-谷氨酸合成

用温度敏感型菌株发酵生产L-谷氨酸不存在生物素亚适量问题,因此该方法在国际上被广泛使用.通常采用对出发菌株进行传统诱变的方法获得温度敏感型菌株.以谷氨酸棒杆菌CICC 10226为出发菌株,先克隆其ItsA基因,然后通过基因敲除的方法构建了突变菌株Corynebacterium glutamicum WT ΔL,该菌株同时具有温度敏感性和溶菌酶敏感性.经透射式电子显微镜观察发现,于38℃培养的突变株细胞与在30℃培养的同一种细胞相比,细胞明显增大,而出发菌株无该现象.发酵试验表明,在生物素过量的情况下,在发酵进入细胞产酸期后通过将发酵温度从原来的30℃提高到38℃,温度敏感突变株的产酸量增加近5倍.如果在发酵培养基巾添加适量的琥珀酸和乙酸,该菌株与不添加的在30℃培养的对照相比,产酸量增加近6.5倍.对于野生型出发菌株而言,在生物素过量的情况下,无论是否采用变温发酵方法都几乎不产酸.说明温度敏感突变株即使在生物素过量的情况下也能通过变温发酵诱导其合成并分泌产生L-谷氨酸.图10表1参13     

杂合抗菌肽LPCB在毕赤酵母中的表达条件优化

为获取能表达具有天然活性的杂合抗菌肽的重组酵母菌,确定其最佳表达条件,将构建好的重组酵母表达载体pPICZα-A-CecropinB(1～10)-Magainin2(1～12)(简称pPICZα-A-CBMA)和pPICZα-A-Lfcin-Pro-CecropinB(简称pPICZα-A-LPCB)经SacⅠ线性化处理,分别电转入巴斯德毕赤酵母Pichia pastoris X-33,利用抗生素Zeocin筛选阳性克隆,PCR扩增验证,甲醇诱导表达.采用抑菌圈法初步测定产物活性,优化杂合肽LPCB诱导时间、诱导剂浓度、诱导温度、培养基pH等表达条件,传代检测其质粒稳定性.结果显示,基因工程菌P.pastoris X-33/pPICZα-A-CBMA和P.pastoris X-33/pPICZα-A-LPCB成功构建.表达产物前者抑菌活性微弱,后者抑菌活性良好.杂合肽LPCB的最佳表达条件为:25℃,pH中性培养,每24 h添加0.5%(V/V,φ)甲醇,诱导表达72 h.重组酵母菌pPICZα-A-LPCB在培养超过100代后,未发生质粒丢失.     

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。     

ERIC-PCR:一种快速鉴别环境细菌菌株的方法

以 Ｅ Ｒ Ｉ Ｃ（ 肠杆菌基因间共有重多序列） 序列设计的特异引物对３ 株大肠杆菌、３ 株软腐欧氏杆菌、２ 株草生欧氏杆菌、１ 株梨火疫欧氏杆菌、１ 株催娩克氏菌、１ 株阴沟肠杆菌和９ 株具有降酚能力的细菌等２０ 种细菌的基因组 Ｄ Ｎ Ａ 进行 Ｐ Ｃ Ｒ 分析．每种菌株均存在数目不等的各自独特的带型,能够区分同一种类中极为相近的菌株．经多次重复,各特异性扩增的主要带型能重复稳定出现．聚类分析的结果表明,供试菌株多数按照分类地位归到相应的组中．９ 种具有降酚能力的细菌中,２ 种分离自处理焦化废水池活性污泥,７ 种来自土壤样品． Ｅ Ｒ Ｉ Ｃ Ｐ Ｃ Ｒ 指纹图显示,前２ 种属于一类,都有一条大小约１ ．１ ｋｂ 的主带,其它７ 种土壤中分离出来的细菌除其中１ 种有１ 条约１ ．２ ｋｂ 大小的主带外,其余６ 种都有１ 条大小约１ ．４ ｋｂ 的带．聚类分析将从土壤中分离出来的细菌归为３ 种不同的类型． Ｅ Ｒ Ｉ Ｃ Ｐ Ｃ Ｒ 可以从分子水平对细菌基因组进行快速指纹图分析,在对环境细菌分离物进行快速鉴定和归类等方面具有实用价值     

Role of phosphate solubilizing bacteria on rock phosphate solubility and growth of aerobic rice

Use of phosphate-solubilizing bacteria (PSB) as inoculants has concurrently increased phosphorous uptake in plants and improved yields in several crop species. The ability of PSB to improve growth of aerobic rice (Oryza sativa L.) through enhanced phosphorus (P) uptake from Christmas island rock phosphate (RP) was studied in glasshouse experiments. Two isolated PSB strains; Bacillus spp. PSB9 and PSB16, were evaluated with RP treatments at 0, 30 and 60 kg ha(-1). Surface sterilized seeds of aerobic rice were planted in plastic pots containing 3 kg soil and the effect of treatments incorporated at planting were observed over 60 days of growth. The isolated PSB strains (PSB9 and PSB16) solubilized significantly high amounts of P (20.05-24.08 mg kg(-1)) compared to non-inoculated (19-23.10 mg kg(-1)) treatments. Significantly higher P solubilization (24.08 mg kg(-1)) and plant P uptake (5.31 mg plant(-1)) was observed with the PSB16 strain at the highest P level of 60 kg ha(-1). The higher amounts of soluble P in the soil solution increased P uptake in plants and resulted in higher plant biomass (21.48 g plant(-1)). PSB strains also increased plant height (80 cm) and improved root morphology in aerobic rice. The results showed that inoculation of aerobic rice with PSB improved phosphate solubilizing activity of incorporated RP.     

挂膜生长的白腐真菌处理草浆造纸黑液废水

比较了几株白腐真菌在造纸黑液废水中的挂膜生长状况及其对黑液废水的处理效果．结果表明，在pH6．0的废水中添加葡萄糖1．0g／L，酒石酸铵0．2g／L及适量无机盐时，黄孢原毛平革菌(Phanerochaete chrysosporium)和侧耳菌(Pleurotus ostieatus)以及本实验室自选的白腐真菌S22的挂膜状况和对黑液废水的处理效果最好．废水中添加的葡萄糖和酒石酸铵的浓度分别为1．0g／L和0．2g／L时，侧耳菌的挂膜和对黑液废水的处理效果最佳．S22菌在pH10．0时其木质素降解率和COD去除率最高，分别可达84％和69％．黄孢原毛平革菌、侧耳菌和S22菌能够在碱性较强的废水中生长挂膜并显著降解木质素，表现出对废水很强的适应能力．生物膜对黑液废水的半连续化处理结果表明了生物膜法的优越性．图8表1参14     

Protoplast fusion between Saccharomyces cerevisiae and Rhodobacter sphaeroides for the treatment of soybean wastewater

The kinetic parameters and the purification rates of the hybrid cell Foaz in soybean processing wastewater (SPW) were measured through a shaking reaction and in two automatic regulated control stable fermentation systems (ARCSFS). The maximum specific growth rate of Foaz was 0.576 h^‐1, higher than that of one of its parental strains Rhodobacter sphaeroides P9479 and lower than that of another parental strain Saccharomyces cerevisiae Y9407. The BOD₅ removal rate of Foaz in the No. 1 system was 61.3%, higher than those of both its parental strains when the influent BOD₅ concentration was 4600 mg/L. The results of this study suggest that the hybrid Foaz has a better capacity of the degradation of organic pollutants in SPW than its parental strains and it may be applicable to the treatment of high concentration organic wastewater.