不同土壤中硅酸盐细菌生理生化特征及其解钾活性的研究

在以钾长石粉为唯一 K 源的硅酸盐细菌选择性培养基上,从我国部分省市土壤中筛选到 16 株硅酸盐细菌,以本实验室保藏 NBT 菌株为参照,对其生理生化特性、耐盐性、抗生素抗性、温度敏感性及释K 能力等生物学特性进行了测定。结果表明,17 株硅酸盐细菌菌体均为杆状,产生椭圆至圆形芽胞。其中 SB6、SB13 为短杆状,SB4、SB6、SB10、SB15 菌株是 G ,其余菌株是 G-。NH4 、NO3 为良好 N 源,且能在无 N -培养基上生长。菌株 SB13 和 NBT 解 K 能力较强,释放的 K 比接灭活菌对照分别增加 49.1 %和 45.3 %。菌株SB2、SB4、SB5、SB6 在 20 g/L NaCl 浓度的培养基上能生长,在温度为 10 ~ 40 范围内供试硅酸盐细菌能够良好生长。     

7株解有机磷细菌的分离和鉴定

从土壤中分离筛选出7株解有机磷的微生物。对这7株解磷细菌进行了形态、生理生化性状测定及16SrDNA序列分析(GenBankaccessionNo:S2,AY651922;S3,AY661923;X1,AY651925;Y1,AY651924;H1,AY663435;H2,AY663436andHe,AY663436)。其中S2、S3、X1和He属于假单胞菌属(Pseudomonas),Y1属于芽孢杆菌属(Bacillus),H1属于不动杆菌属(Acinetobacter),H2属于寡养单胞菌属(Stenotrophomonas)。进一步通过G C含量和DNA-DNA杂交研究,结果表明,S2、S3和X1为产碱假单胞菌(Pseudomonasalcaligenes),Y1为蜡状芽孢杆菌(Bacilluscereus)。     

ATP发光技术测定辐照前脱水蔬菜和调味品的含菌量

用ATP发光技术测定脱水蔬菜和调味品的初始含菌量,发现脱水蔬菜和调味品中细菌ATP的生物发光强度与其初始含菌量之间显著相关,从样品的制备到获得ATP发光强度结果的检测过程仅需1～2h。该技术可用于辐照灭菌前的微生物检测。     

活性光合细菌对改善水产养殖体系水质状况的作用初探

在鱼虾育苗、养殖，池塘淡水养鱼以及观赏鱼养殖中，使用以牛肉水提取液为主要培养基培养的活菌数大于50亿／mL的活性光合细菌，测定了使用前后水的pH值、COD值、NH3-N、亚硝酸盐、溶解氧等指标，结果表明：与市场出售的光合细菌(活菌数为25亿／mL)相比，活菌数大于50亿／mL活性光合细菌能明显改善水产养殖体系的水质状况。     

Survival of bacterial DNA and culturable bacteria in archived soils from the Rothamsted Broadbalk experiment

Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 10⁵ genomes g⁻¹ soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.     

高产水稻土细菌多样性的培养法与非培养法比较研究

利用细菌的通用引物扩增江西余江县高产水稻土红壤细菌总DNA和平板培养细菌混合总DNA的16S rDNA基因片段,在此基础上分别建立两种16S rDNA文库(文库a和文库b)。从两个文库中各随机挑选100个克隆,扩增出阳性克隆中的插入片段后选用HhaⅠ和RsaⅠ两种四碱基酶进行ARDRA(amplified rDNA restriction analysis)分析。统计比较分析发现,文库a的Shannon-Wienner指数、Simpson指数、丰富度、均一度分别为4.432、0.987、18.885和0.973,均高于文库b中相应的多样性参数(分别为2.271、0.758、5.736和0.501),即平板培养方法所展现的细菌群落结构多样性低于土壤中原始的多样性。结果表明,传统培养方法存在着很大的局限性,必须结合新的分子生物学技术手段才能更全面完善地认识土壤微生物群落结构多样性,以期充分利用其中丰富的微生物资源。     

几株光合细菌的分离鉴定及净水能力分析

从养猪场的排污池池水以及淤泥中分离纯化得到4株光合细菌,分别命名为P1、P2、P3、P4.根据菌株形态特征、对碳源利用能力以及活细胞光吸收特征的鉴定结果:P1为褐螺菌属莫氏褐螺菌(Phaeospirillum molischianum),P2为红螺菌属深红红螺菌(Rhodospirillum rubrum),P3为着色菌属小着色茵(Chromatium minus),P4为红菌属东方红茵(Rhodobium orientis).在实验室条件下,测定这些菌株对灭菌污水的化学需氧量(COD)去除率,以衡量它们的净水能力.结果显示,P1,P4对污水的COD降低作用最明显,COD降低率分别为55.2%和37.9%.     

湿地松幼苗根系微生物数量变化规律的研究

采用平板稀释法研究了湿地松幼苗1 a中微生物根系数量的变化规律,为合理的生物防治时期提供科学依据。结果表明:湿地松苗圃土壤中存在着大量的微生物。湿地松幼苗根系的生长和发展可以大大促进其周围微生物的生长,受湿地松根系生长和环境变化等因素的影响,在松苗移栽后的几个月内,根际细菌和真菌的种群数量迅速增加,细菌比真菌的种群增长速度要快。利用生物防治细菌进行苗木猝倒病防治的最佳时期应该是在芽苗移栽后1个月以内。     

水稻纹枯病菌胞壁降解酶的产生及致病作用

水稻纹枯病菌(Rhizoctonia solaniKühn)在改良的Marcus培养液中能产生多聚半乳糖醛酸酶(PG)、纤维素酶(Cx)、果胶甲基半乳糖醛酸酶(PMG)、多聚半乳糖醛酸反式消除酶(PGTE)和果胶甲基反式消除酶(PMTE)5种胞壁降解酶,其中PG、Cx和PMG活性较高,而PGTE和PMTE活性很低。病菌产生胞壁降解酶的最佳条件是:在pH5培养液中28℃下静置培养6 d。病菌接种水稻叶鞘后也能显著产生PG、Cx和PMG等胞壁降解酶,并且病斑外侧褪绿部酶活最高,平均为对照(健康部)的3.4倍;其次是褐色部,酶活总量平均为对照的2.9倍;病斑中央枯白部酶活较低,但明显高于对照。这一结果提示,在病菌侵染和扩展过程中胞壁降解酶起重要作用。果胶酶(PG、PMG)和纤维素酶(Cx)无论是单独处理还是混合处理,对水稻叶鞘都具有显著地损伤细胞膜和浸渍组织的作用。     

产低温脂肪酶假单胞菌的选育及产酶条件的优化

从取自华中农业大学的土样和长江流域及桂林桃花江水样中筛选到一株产低温脂肪酶的适冷菌,初步鉴定为假单胞菌,命名为Pseudomonas sp.YT34-8.对其产酶条件进行优化,得到其最佳发酵条件为:玉米浆2.00%,豆饼粉2.50%,蔗糖1.00%,橄榄油0.57%,MgSO4·7H2O0.05%,K2HPO4 0.10%,发酵温度8℃,初始pH值7.0,发酵时间48 h,250 mL摇瓶装液量21 mL;摇瓶转速182 r·min-1.在此条件下,其发酵酶活力可达到7.833 U·mL-1.对其所产低温脂肪酶进行初步研究,其最适作用pH值为8.0,最适作用温度为30℃,在pH值5.0～10.0范围内、40℃以下酶活力较稳定.