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。     

几种土壤生物因子对土壤养分的影响

试验研究了在果树—牧草间作模式下土壤中纤维分解菌、硅酸盐细菌以及蔗糖酶、脲酶、过氧化氢酶、纤维分解酶、多酚氧化酶对土壤中N、P、K全量养分、速效养分和有机质的影响,并通过通径分析发现:脲酶、硅酸盐细菌、纤维分解酶是促进有机质积累的主要生物因素,蔗糖酶是影响N、P、K速效养分的最主要因子,过氧化氢酶、多酚氧化酶、纤维分解菌只是选择性地对有机质的积存和N、P、K速效养分的形成起作用。     

紫色土硅酸盐细菌的遗传多样性研究

硅酸盐细菌是土壤中一类能分解硅酸盐类矿物，破坏其晶格结构，使矿物钾转化成有效钾释放出来供植物生长的细菌。它不仅能溶磷解钾，亦有固氮的作用。目前的种类主要有，环状芽孢杆菌（Bacillus circulans）、胶质芽孢杆菌（B．rmtcilaginosus）和土壤芽孢杆菌（B．edaphicus）三个种的菌株。由于硅酸盐细菌的独特作用，已广泛应用于采矿、冶金、微生物肥料、饲料工业上。我国在硅酸盐细菌的生物学功能方面已进行了大量的工作，尤其是在硅酸盐细菌作为生物肥料方面，但是对硅酸盐细菌的分类地位和遗传多样性研究较少。贺积强等研究表明，紫色土硅酸盐细菌的理化性质和解磷解钾特性存在差异，本研究应用RAPD、TP-PCR和16S rRNA PCR-RFLP进一步分析了这些硅酸盐细菌的遗传多样性及系统发育地位，以期弄清其遗传背景。     

硅酸盐细菌NBT菌株解钾效能及对钾的吸持作用

在摇瓶和土壤耗竭条件下研究了硅酸盐细菌NBT菌株的解钾作用以及对作物生长的促进作用。结果表明 ,在摇瓶条件下 ,培养 120h ,NBT菌株可以从钾长石中释放K 159.1mg/L ,比接灭活菌对照 (48.8mg/L)增加 226.02% ;耗竭条件下NBT菌株在未灭菌土壤中的解钾作用与在灭菌土壤中的解钾作用相当。在未灭菌土壤中 ,NBT菌株释放的矿物钾占植株吸钾量的 14.4 %～ 4 3.1% ;不接菌或接灭活菌处理土壤中矿物钾的释放量为零或极少。NBT菌株的解钾效能与土壤中速效性钾及有机质含量密切相关。土柱试验表明 ,供试土壤接种硅酸盐细菌后的土壤滤液中流失的钾比接种灭活硅酸盐细菌后的土壤滤液中流失钾减少 29.6 %～56.5% ;硅酸盐细菌NBT菌株荚膜多糖吸附钾的量占加入钾的量的 31.8%～ 69.4 %。NBT菌株的吸钾作用与NBT菌株本身及荚膜多糖的多少密切相关。     

不同年龄段银杏树雌株根系土壤细菌群落演替特征分析

以幼年期(10年以下)、挂果期(20年)和盛年期(40年以上)的银杏树根系土壤为研究对象,探讨银杏雌株树龄与土壤中细菌菌群结构的关系。运用Illumina Mi Seq平台测序分析幼年期、挂果期以及盛年期银杏树根系土壤细菌的16S r RNA V3～V4区变异序列,对不同年龄阶段银杏树雌株根系土壤细菌的菌群多样性和结构进行研究。银杏树雌株根系土壤共获得612219条微生物的16S r RNA有效基因序列,在97%的相似水平下共得到OTU个数为23294。热图分析表明,各个年龄段银杏树雌株的土壤细菌群落差异较大,不同年龄段优势属不同。据9个土壤样品的多样性指数分析得出,在3组年龄段银杏雌株的土壤样本中,幼年期Shannon指数、Simpson指数、Chao1指数以及ACE指数都是最大,而挂果期皆最小,表明幼年期银杏雌株土壤的细菌菌群丰富度和多样性最高,挂果期银杏土壤的细菌菌群丰富度和多样性最低。变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)在土壤细菌群落中占绝对优势。挂果期和盛果期银杏雌株土壤中浮霉菌门(Planctomycetes)及变形菌门(Proteobacteria)和相对丰度普遍低于幼年期,挂果期银杏树的土壤与盛果期银杏树的土壤中的放线菌门(Actinobacteria)与绿弯菌门(Chloroflexi)的相对丰度明显高于幼年期银杏树的土壤。WS3在盛果期银杏土壤中的相对丰度最低。银杏树雌株树龄可对土壤细菌菌群的多样性及菌群结构产生影响。伴随着育种年限的增加,银杏雌株表层土壤中细菌多样性呈现先降低后增加的变化趋势,且银杏雌株树龄对其土壤细菌的多样性和结构有显著影响。     

不同施肥措施对棉田土壤磷细菌及磷转化强度的影响

研究了不同施肥措施对棉田土壤磷细菌的数量、分布、动态变化及磷转化强度的影响。结果表明，在有机肥合理配施N、P、K肥能有效的促进磷细菌的生长发育，磷细菌数量及活性增加，从而增强磷的有效性，改善土壤的供磷性能，有利于棉花生长。     

苹果树根际促生细菌种群分析

利用选择性培养基, 对多年生苹果树根际与连作幼树根际促生细菌进行了分离和测数, 并采用BOX-PCR技术进行聚类分析。结果表明: 多年生苹果树根际细菌总量及固氮细菌、解磷细菌、硅酸盐细菌、拮抗细菌4类根际促生细菌的数量均高于连作幼树根际。在多年生苹果树根际, 硅酸盐细菌的数量最大, 解磷细菌和固氮细菌的数量次之, 拮抗细菌的数量最小; 在连作幼树根际, 解磷细菌的数量最大, 硅酸盐细菌和固氮细菌的数量次之, 拮抗细菌的数量最小。从两种土壤中获得的促生细菌分离株的BOX-PCR图谱最大的相异百分数都在1.25以上, 说明这些细菌分离株的遗传进化距离比较接近。在细菌BOX-PCR图谱相异百分数为0.25的水平上, 多年生苹果树根际促生细菌分为79个聚类群, 其中固氮细菌18个聚类群, 解磷细菌29个聚类群, 硅酸盐细菌19个聚类群, 拮抗细菌18个聚类群; 连作幼树根际促生细菌分为46个聚类群, 其中固氮细菌15个聚类群, 解磷细菌19个聚类群, 硅酸盐细菌8个聚类群, 拮抗细菌9个聚类群。多年生苹果树4类根际促生细菌的多样性、丰富度和均匀度指数均高于连作幼树根际, 而优势度指数低于连作幼树根际。与连作幼树相比, 多年生苹果树根际促生细菌具有丰富的种属多样性。     

饲用玉米连作土壤可培养细菌多样性与肥效菌株监测

为深入了解饲用玉米连作对土壤微生物多样性的影响,对甘肃区采集的连续7年种植玉米土壤中可培养细菌的多样性和促生细菌资源进行全面地分析和挖掘。分别利用富营养(NA)和选择性(Ashby)培养基进行可培养细菌的分离,对获得菌株进行16S rRNA基因测序和比对分析,评价其产植物生长素(IAA)、溶磷、固氮等能力。自不同年份土壤中共分离得到各类细菌224株。菌株功能评价结果表明,34株具有产IAA的能力,25株具溶解有机磷的能力,19株具溶解无机磷的能力,27株具有较高的固氮酶活性,有5株可同时具有产IAA和溶磷的功能。可培养细菌分离信息分析结果表明,种植玉米土壤的细菌及肥效菌株的丰富度及多样性随着连作年限的增加呈逐渐上升趋势,优势菌属均为Pseudomonas,广泛分布于各年份的土壤中,且表现出优秀的产IAA、溶磷及固氮的能力。研究表明,玉米7年连作可明显影响土壤菌群结构,土壤中蕴藏着丰富的有益功能细菌资源。分离的功能菌株为研制适用于甘肃玉米绿色生产的微生物肥料提供信息与优良菌种支撑。     

黄土高原不同植被坡地土壤微生物区系季节性变化

应用稀释平板法对夏季和秋季黄土高原不同植被坡地土壤0~20 cm土层的细菌、真菌和放线菌的分布特征进行了研究。结果表明:(1)该区域不同植被下土壤中微生物总体比较丰富,数量差异较大,排序为刺槐林地>荒坡地>柠条林地>苜蓿地>耕地;同一植被下各类菌群数量排序为细菌>放线菌>真菌。(2)夏季微生物在土壤中的总量明显比秋季多。(3)秋季土壤中的真菌数量比夏季有明显的增长趋势;夏季土壤中细菌数量比秋季土壤中细菌数量多。     

土壤矿物钾活化途径

针对我国钾肥资源短缺问题,分析了土壤钾资源的开发潜力,概述了土壤矿物钾的存在状态及其释放特点,综述了硅酸盐细菌、富钾植物和有机酸活化土壤矿物钾的能力与机理,并展望了活化土壤矿物钾的三种途径在生产中的应用前景。     

中国太湖流域水稻土中的反硝化细菌

This study attempted to determine the characteristics of the communities, the ecological factors, and the denitrifying enzyme activity for denitrifying bacteria found in the paddy soils of the Taihu Lake Basin, China. Samples of the six main soil types of the basin were taken from paddy fields with different fertilities. The total numbers of bacteria and denitrifying bacteria in the high fertility soils were much more than those in low fertility soils, and the number of denitrifying bacteria accounted for 49% to 80% of the total number of bacteria. The O2 content was an important ecological factor that affected denitrification. Of test the strains isolated from the paddy soils in the Taihu Lake Basin, some (e.g., Pseudomonas spp.) grew well under low oxygen partial pressure, while others (e.g., Bacillus spp.) had no strict predilection with O2 content. Another critical ecological factor was the nitrogen concentration. Three selected denitrifying bacteria grew better in aculture medium with 135 instead of 276mg L^-1 nitrogen. At the same time 67% of the test strains were able to reduce NO3^- to NO2^- and 56% had N2O reductase.     

测定土壤pH的新混合指示剂及其使用

土壤酸碱性是影响植物生长的重要因素之一。在林业生产中,往往根据土壤的pH值决定树种适生的土壤类型。在测定土壤pH的方法中,电位法的精度较高。混合指示剂比色法的精度较差,一般认为其误差在0.5个pH单位左右。关于我国常用的几种混合指示剂在使用上的问题和优缺点,袁可能和朱祖祥(1956)已作了研究。     

解钾细菌C6X对不同富钾矿物含量土壤钾素迁移的影响

为了改善黄土高原地区煤炭开采引起土壤质量急剧退化的现状,该文以玉米为供试植物,通过日光温室短期盆栽的方式,系统研究解钾细菌C6X和玉米生长对土壤钾素迁移的影响.结果表明:1)玉米生长条件下,解钾细菌在富钾矿物质量分数45％上层土壤(0～20 cm)中对速效钾增量的促进作用最佳.2)解钾细菌和玉米生长协同提高上层土壤钾素固定能力,缓效钾增量在土壤富钾矿物质量分数68％为最大值.3)解钾细菌和玉米生长协同促进土壤钾素上移能力,在富钾矿物质量分数45％水平,土壤上移速效钾呈最大值;同时,解钾细菌促进土壤上移速效钾和玉米钾素积累量二者趋于线性稳定,利于土壤钾肥长期管理.因此,解钾细菌和玉米生长协同促进土壤钾素的释放和固定,并促进土壤钾素上移.     

甲基对硫磷对红壤地区土壤微生物数量的影响

有机P农药是我国最重要一种农药类型,在农业生产上使用广泛。采用室内模拟方法,研究了甲基对硫磷对红壤地区不同类群土壤微生物数量的影响。研究结果表明,甲基对硫磷对土壤微生物数量的影响随甲基对硫磷添加的浓度、微生物类群和培养时间的不同而变化。添加100 mg/L和500 mg/L浓度甲基对硫磷能明显增加土壤细菌的数量,细菌数量的最大值出现在第10天左右;低浓度甲基对硫磷对土壤微生物数量影响不大。平板混合菌体培养实验证明,甲基对硫磷通过抑制或者杀灭某些种类土壤细菌,从而大大促进土壤生态系统中部分种类细菌数量的增殖。     

Effects of lead on the soil bacterial microflora

The sensitivity of soil bacteria towards Pb was investigated. Soil suspensions from fourteen different soil types with a high or low Pb content were plated out on soil extract agar containing various concentrations of PbCl₂.In agar with a high Pb content, higher bacterial counts were found with suspensions from Pb-containing soils than with those of soils with a low Pb content. In the Pb-containing soils, proportionally more gram-negative rods were present while coryneform bacteria decreased. In an additional experiment, in which Pb was added to a sandy soil, more Pb-tolerant bacterial strains were found 3 years later than in the same soil without Pb.When pure cultures of the bacteria isolated from the soils were tested in liquid media for Pb tolerance, a higher proportion of tolerant strains was found in Pb-containing soils. Among strains of gram-negative bacteria isolated from these soils a higher proportion of tolerant strains was found than in corneform bacteria.It was concluded that as a consequence of Pb pollution of soil a selection of Pb-tolerant bacteria may take place.     

东北地区保护地土壤拮抗放线菌的筛选

利用稀释分离法从东北蔬菜保护地37份土壤样品中共分离到各种放线菌菌株543株,采用活体对峙培养法共获得对植物病原菌具有拮抗作用的放线菌菌株19株,将其中抑菌圈直径超过15mm的菌株进行液体发酵试验,以叶霉病菌作为指示菌,采用杯碟法测定各菌株发酵液的抑菌活性,其中发酵液抑菌圈直径超过15mm的菌株有5株。对编号为MY02、MY04的菌株,进行了进一步的验证试验,测得其对叶霉病菌的抑菌圈直径分别为24 mm、23 mm。同时进行了这两个菌株的抑菌谱试验,结果表明无论其活体还是其发酵液对番茄叶霉病菌、番茄灰霉病菌等多种植物病原真菌均表现强烈的抑菌效果。菌株MY02除茄子黄萎病菌外,对试验的其他10种病原真菌抑制率均接近或超过90%。同时菌株MY02、MY04对植物病原细菌也具有很好的抑菌活性,试验结果表明,对番茄溃疡病菌、水稻白叶枯病菌、大白菜软腐病菌、果树根癌病菌、烟草野火病菌的抑菌圈直径分别可达20mm、16mm以上。     

Soil microorganisms antagonistic towards Rhizobium japonicum

Success in introducing Rhizobium japonicum strains into soil is related to their interaction with native microorganisms including some that are antagonistic. Actinomycetes, bacteria, fungi and rhizobiophages antagonistic towards strains of R. japonicum were counted directly using soil samples from field plots under different crop and soil management systems. The antagonistic actinomycete population varied from 1.3 × 10³ to 4.5 × 10⁵ g^?1 dry soil and ranged up to 90% of total actinomycetes. Soybean rhizosphere soil samples included antagonistic actinomycetes ranging up to 70% of total actinomycetes. The antagonistic bacterial population was less than 10% of total bacteria and the proportion did not vary significantly with crop or soil management practices. Antagonistic fungi were observed for many of the soils examined but they could not be counted. There were few rhizobiophages and they were found most frequently in soybean rhizospheres. Occasional bacterial and actinomycete colonies that stimulated growth of R. japonicum were randomly observed among the soil samples tested.     

盐碱土耐盐碱细菌筛选及其植物促生能力研究

  目的  为寻找耐盐碱促生细菌资源。  方法  采用传统定向筛选培养基培养、摇瓶培养和16S rDNA鉴定的方法,自盐碱土筛选细菌并对其促生特性进行测定。  结果  筛选得到具有促生特性的细菌14株;其中,不动杆菌属6株,肠杆菌属3株,假单胞菌属5株。在筛选出的14株细菌,均具有解钾能力,培养液速效钾含量与对照12.86 ± 0.30 mg L?相比增加16.24 ~ 36.21 mg L?1;12株细菌具有溶磷能力,培养液磷含量对照组为0.8 ± 0.1 mg L?1,接种细菌菌株的为59.1 ~ 233.2 mg L?1,培养基pH值由6.86 ± 0.34下降至4.06 ~ 5.69,线性相关分析表明溶磷培养液磷含量与培养液pH值呈负相关;7株细菌具有产植物生长激素吲哚乙酸（IAA）能力,在底物色氨酸（L-Trp）浓度为500 mg L?1时产IAA能力为5.57 ~ 21.11 mg L?1·OD₆₀₀;9株细菌具有产铁载体能力,能力最强的Su值达到95%。14株细菌均能在pH值为8、NaCl浓度为0.5 mol L?1的培养液中生长;有4株能在NaCl浓度为1.5 mol L?1培养基中生长,且B1421生长良好;有5株细菌可在pH为10的培养基中生长,为B2222生长最为良好。  结论  松嫩平原盐碱土中存在具有不同促生能力的细菌,能够耐受高pH值和较高NaCl含量的胁迫,是可供以耐盐碱植物利用改良盐碱土的菌株资源。     

Effects of regenerating vegetation on soil enzyme activity and microbial structure in reclaimed soils on a surface coal mine site

The aim of this study was to evaluate the influence of reclaimed scenarios on soil enzyme activities and microbial community in a reclaimed surface coal mine on the Northwest Loess Plateau of China. Soil samples were collected from a bare land (CK), and a plantation (PL) and four mixed forests (MF1-4). Soil physicochemical characteristics, four enzyme activities and microbial abundance and T-RFLP (terminal restriction fragment length polymorphism) profiles were measured. Effects of reclaimed scenarios on soil nutrients content, enzyme activities and microbial community were pronounced. Soil organic carbon could be well used to predict the major differences in enzyme activities, and microbial abundance and composition. Soil enzyme activities were more significantly correlated with fungal abundance than bacterial and archaeal ones. The higher soil nutrient content, enzyme activities, and microbial abundance and diversity were from mixed forests and the lowest ones were from CK, which suggested mixed forests would be feasible scenarios in semi-arid Loess Plateau. Soil bacteria, archaea and fungi evolved with reclaimed process, but the influences of reclaimed scenarios on each domain of microbial abundance, diversity and composition were different. These findings suggested that soil bacteria, archaea and fungi play different ecological roles during restoration process.     

Gram-negative bacterial flora on the root surface of wheat (Triticum aestivum) grown under different soil conditions

We identified 161 Gram-negative bacterial strains isolated from the root surface of wheat grown under different soil conditions. The strains were divided into seven groups based on major morphological and physiological properties. Taxonomic allocation of the groups was verified by guanine+cytosine contents of DNA. Except for one group, which may be assumed to include bacteria belonging to the genera Flavobacterium and Cytophaga, the various groups were taxonomically united. The distribution of the groups changed with soil improvement. Pseudomonads predominated in unimproved soil, but Flavobacterium and Cytophaga spp. were predominant in the most improved soil. As all the strains were non-fermentative by Hugh and Leifson's test, API 20NE identification was applied. However, many strains were misidentified by this system, especially in the Flavobacterium and Cytophaga spp. group. For ecological studies, the strains were classified to species level by the API 20 NE system and by the results of a combination of guanine+cytosine (mol%) and isoprenoid quinone data. The pattern of distribution of the bacteria on the root surface of wheat varied at species level within one genus depending on soil conditions.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday