东北地区四种农作物根际磷细菌的分布

从小麦、水稻、玉米,大豆根际土壤中磷细菌数量及其群落结构、多样性和相似性等几个方面,对东北地区代表性农作物根际磷细菌生态分布规律进行了系统研究.结果表明,4种作物根际土壤中分布大量解磷菌和溶磷菌,其中解磷菌达到2.15～6.68×105cfu·g-1、溶磷菌达到0.7～3.9×104cfu·g-1,解磷菌数量多于溶磷菌;作物根际土壤有效磷含量与磷细菌数量问无直接相关性;东北地区农作物根际磷细菌种群结构复杂,小麦、玉米、水稻、大豆根际土壤解磷菌分别涉及9个、11个、13个、12个菌属,溶磷菌分别涉及9个、8个、9个、7个菌属:4种主要农作物根际土壤中分布的磷细菌具有较高的多样性指数,且一般来说根际土壤中解磷菌多样性更高;作物根际磷细菌组成相似性系数在0.50～1.00之间,总体处于较高水平,且溶磷菌的相似性系数高于解磷菌.     

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。     

接种自生固氮菌对玉米根际土壤酶活性和细菌群落功能多样性的影响

为了探索玉米根际土壤微生态特征对接种自生固氮菌的响应机理,在盆栽实验条件下,研究了3株自生固氮菌褐球固氮菌（Azotobacter chroococcum YCYS）、芸苔叶杆菌（Phyllobacterium brasssicacearum QL54）和类芽孢杆菌（Paenibacillus sabinae MX31）接种盆栽玉米（Zea mays L.）之后,玉米根际土壤酶活性和细菌群落功能多样性的变化。结果表明,接种自生固氮菌对玉米根际土壤酶活性和细菌群落功能多样性产生了一定的影响,而且不同自生固氮菌之间的接种效果有一定的差异。接种褐球固氮菌（A. chroococcum YCYS）和类芽孢杆菌（P. sabinae MX31）玉米根际土壤脲酶活性分别比对照高20.55%和9.58%。然而接种处理对玉米根际土壤碱性磷酸酶活性的影响差异不显著（P0.05）。BIOLOG结果显示,接种自生固氮菌可以提高细菌总代谢活性,其中接种褐球固氮菌（A. chroococcum YCYS）处理的AWCD是对照的1.8倍,并且细菌群落丰富度指数（R）显著高于对照（P0.01）。不同接种处理土壤根际细菌生理碳代谢优势群落结构不同。主成分分析（PCA）表明接种自生固氮菌可以调控根际土壤细菌群落功能多样性。     

黄河三角洲刺槐根际与非根际细菌结构及多样性

为精确分析黄河三角洲刺槐根际与非根际土壤细菌群落定殖情况,本研究采用高通量测序方法对刺槐根际与非根际土壤细菌结构及多样性进行了研究。研究表明,根际土壤细菌共有36门214属,非根际土壤细菌共有33门153属。变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)细菌丰度超过15%,是根际与非根际土壤中的优势菌落。根际与非根际土壤中酸杆菌门、硝化螺旋菌门(Nitrospirae)丰度差异显著。根际土壤中红游动菌属(Rhodoplanes)、溶杆菌属(Lysobacter)、热单胞菌属(Thermomonas)、链霉菌属(Streptomyces)及非根际土壤中红游动菌属、溶杆菌属、链霉菌属、Kaistobacter细菌丰度超过4%。根际土壤中固氮菌丰度显著高于非根际土壤,解磷、解钾细菌丰度差异不显著。根际与非根际土壤细菌Chao丰富度分别为2 054、2 376,差异显著。根际与非根际土壤细菌之间的权重(Weighted Unifrac)距离在0.12~0.25之间。综上所述,黄河三角洲刺槐根际与非根际土壤细菌结构具有一定差异,多样性差异显著。     

土壤叶杆菌和红球菌菌株的分离鉴定及其自生固氮作用

非光合自生固氮细菌是一类自由生活在土壤有机质中不具有光合作用的固氮菌,也是促进植物生长根际细菌(PGPR)的重要组成分。自生固氮菌的固氮效率受土壤环境中有效碳源和种群数量限制。植物根分泌的许多简单有机化合物,可能被用作自生固氮菌的有效碳源,因而在植物根际存     

披碱草根际促生菌筛选及其接种剂的促生作用

【目的】体外促生能力是衡量微生物菌株作用的一个重要指标,测定获取的植物根际促生菌并明确其对披碱草的促生效果,可为其在生产中的应用提供依据。【方法】2014年9月从西藏阿里地区采集披碱草根系及根际土壤,以常规方法分离出其中的溶磷菌、 固氮菌和分泌3-吲哚乙酸(IAA)细菌的10株菌株。测定其溶磷量、 固氮酶活性及分泌生长素能力,并将其制成植物根际接种剂,测定接种剂对披碱草生长的影响及其在根际的定殖能力。【结果】菌株PWXZ10溶磷能力较好,达40.89 mg/L; 菌株003PWXZ6固氮酶活性较强,达421.21 nmol/(mL·h); 菌株NXP17分泌生长素能力较强,达31.33 μg/mL。与对照菌株(Pseudomonas sp. Jm92)相比,菌株003PWXZ6和NXP17制备的接种剂可显著增加披碱草株高、 地上生物量和地下生物量(P0.05),但两者之间差异不显著(P0.05); 接种剂003PWXZ6对披碱草根总长、 根表面积、 根体积、 根直径、 含磷量、 含氮量和粗蛋白含量增加显著(P0.05),分别较对照菌株(Pseudomonas sp. Jm92)增加了330%、 199%、 118%、 187%、 70%、 15%和19%,并且该菌株在根际定殖能力很强。 【结论】植物根际促生菌003PWXZ6和NXP17对披碱草具有良好促生效果,可为开发经济环保的生物肥料提供了菌种资源。     

氧化尾矿与白茅根际尾矿中可培养溶磷菌比较研究

溶磷菌是根际促生菌类群中重要组成部分,它转化的磷成为植物生长所需的重要磷源。研究分别从氧化尾矿和白茅根际尾矿中筛选溶磷菌,采用DNA分子技术确定它们的分类地位,并比较它们的溶磷能力和培养液pH的降低值。结果表明,白茅根际尾矿中筛选出的溶磷菌在数量和种类上均多于氧化尾矿,白茅根际尾矿中的筛选的溶磷菌具有更高的溶磷能力,放线菌门溶磷菌的溶磷能力显著低于其他类群的溶磷细菌。白茅根际尾矿中筛选出的隶属于伯克氏菌属的菌株N5-4溶磷能力最强,溶磷量达到536.40 mg L~(-1)。培养过程中,所有溶磷菌培养液的pH均有所降低,除放线菌门的微生物外,培养液pH降低值与溶磷菌的溶磷量存在着显著的正相关性(P0.01,R=0.76)。     

水稻根际促生菌的筛选鉴定及促生能力分析

植物根际促生菌(Plant growth-promoting rhizobacteria,PGPR)可分泌植物生长激素,促进土壤养分循环,是生物肥料重要的种质资源。本文从水稻根际土壤分离纯化根际促生菌,进行菌株鉴定,测定其促生能力。经过16S r DNA测序比对,筛选得到解磷菌4株(Bacillus pumilus LZP02,Bacillus aryabhattai LZP08,Staphylococcus epidermidis LZP10,Bacillus ginsengisoli LZP05),溶磷菌3株(Bacillus megaterium LZP03,Bacillus oryzaecorticis LZP04,Bacillus ginsengisoli LZP07),解钾菌3株(Bacillus aryabhattai LZP01,Bacillus subtilis LZP06,Bacillus licheniformis LZP09)。养分转化能力测试结果表明,Bacillus aryabhattai LZP01和Bacillus subtilis LZP06解钾能力较好;Bacillus pumilus LZP02和Bacillus huizhouensis LZP05解磷能力较强;Bacillus megaterium LZP03和Bacillus ginsengisoli LZP07溶磷能力较好。对养分转化能力较强的菌株进行激素分泌能力测定,结果表明6种菌株均能产生生长素、赤霉素,均具有合成铁载体的能力。综合分析菌株养分转化与激素分泌能力发现,Bacillus megaterium LZP03、Bacillus huizhouensis LZP05和Bacillus subtilis LZP06促生能力较强,具有较强的开发利用潜力。研究成果为水稻微生物肥料的开发与生产提供了理论和技术支持。     

解磷菌株YR-P31的分离、鉴定、生物学特性及其促生作用

从水稻等作物根际土壤中分离到1株解磷菌。该菌株能够以Ca3(PO4)2为唯一的磷源良好地生长。经过对其形态特征、生理生化分析,初步鉴定为巨大芽孢杆菌属细菌。该菌株利用无机磷培养基生长的最适温度和pH值分别为35℃和7.0,其解磷作用是通过溶磷圈及在液体培养基内可溶性磷的增加来证实的。此外该菌株在固体培养基上还能促进大肠杆菌的生长。该菌株具有解磷能力和促生作用,在微生物肥料的进一步开发中具有很大的潜力。     

延边烟区不同土壤的根际土壤微生物生态效应研究

为揭示烤烟根际微生物数量及酶活性的变化规律,阐明烤烟-土壤-微生物之间的协同作用机理,在延边烟区采集不同土壤类型的烤烟根际土壤,研究了烤烟根际土壤微生物生态效应.结果表明:不同土壤条件下烤烟各时期根际微生物区系和微生物生理菌群数量存在着较大的差异,其中暗棕壤黏土和暗棕壤黑砂土比白浆土更适合细菌和解钾菌的生长,白浆土真菌数的变化幅度较大可能是引起烟叶质量变差的原因之一,暗棕壤黏土和暗棕壤黑砂土在烤烟生育期内放线菌数量的变化较大,明显大于白浆土;自生固氮菌在移栽后 60 天前数量均较低,60 天后急剧升高,100 天时暗棕壤黏土和暗棕壤黑砂土自生固氮菌数量显著高于白浆土;氨化菌生长的最适宜期出现的早晚顺序为:暗棕壤黏土＞暗棕壤黑砂土＞白浆土;生育期内根际土壤解磷菌数变化较大,暗棕壤黏土烟株在旺盛生长时期的解磷菌数显著高于其他 2 种类型土壤.过氧化氢酶活性的变化规律很大程度上受土壤类型的影响,而转化酶和多酚氧化酶变化受土壤类型影响较小,但两者的大小受土壤类型的影响.     

Phosphate-solubilizing rhizobacteria enhance the growth and yield but not phosphorus uptake of canola (Brassica napus L.)

The ability of phosphate-solubilizing rhizobacteria to enhance the growth and phosphorus uptake of canola (Brassica napus L., cv. Legend) was studied in potted soil experiments in the growth chamber. One hundred and eleven bacteria isolated from the rhizosphere of field-grown plants, and a collection of nine bacteria known to be effective plant growth-promoting rhizobacteria (PGPR), were screened for P-solubilization in vitro. All rhizobacteria were identified using whole-cell fatty acids methyl ester (FAME) profiles. The best P-solubilizing isolates were two Bacillus brevis strains, B. megaterium, B. polymyxa, B. sphaericus, B. thuringiensis, and Xanthomonas maltophilia (PGPR strain R85). The P-solubilizers were tested for their effects on growth and P-uptake of canola plants in a P-deficient soil amended with rock phosphate. Although some of the P-solubilizing rhizobacteria significantly increased plant height or pod yield, none increased P-uptake. The most effective inoculant was a B. thuringiensis isolate which significantly increased the number and weight of pods and seed yield without rock phosphate. Xanthomonas maltophilia increased plant height, whereas the other bacilli increased the number on weight of pods. These results demonstrate the potential use of these P-solubilizing rhizobacteria as inoculants for canola, but indicate that P-solubilization was not the main mechanism responsible for positive growth response. Received: 8 February 1996     

施氮量对玉米/大豆套作系统土壤微生物数量及土壤酶活性的影响

为揭示玉米/大豆套作体系下土壤氮素转换的调控机理和根际微生态效应,以种植模式为主因素[设玉米单作(MM)、大豆单作(SS)和玉米/大豆套作(IMS)3种处理],以玉米、大豆施氮总量(玉米、大豆施氮比例为3∶1)为副因素[设不施氮(NN,0 kg?hm~(-2))、减量施氮(RN,180 kg?hm~(-2))和常量施氮(CN,240 kg?hm~(-2))3个处理],研究了玉米/大豆套作系统下不同施氮量对作物根际土壤微生物数量及土壤酶活性的影响。结果表明:与相应单作相比,套作下玉米根际土壤真菌、放线菌数量分别提高25.37%和8.79%;套作大豆根际土壤真菌、放线菌、固氮菌数量高于单作大豆;套作玉米根际土壤蛋白酶、脲酶活性和套作大豆根际土壤蛋白酶活性均显著升高。各施氮水平间,减量施氮下玉米、大豆根际土壤真菌数量较常量施氮和不施氮均有所提高;施氮提高了玉米、大豆根际土壤放线菌数量;大豆根际土壤固氮菌数量以减量施氮最高,比不施氮和常量施氮高17.78%和5.67%;玉米根际土壤蛋白酶活性、脲酶活性和大豆根际土壤脲酶活性均以减量施氮为最高。适宜的施氮量不仅能增加玉米/大豆套作土壤中真菌、放线菌、固氮菌的数量,还能提高土壤蛋白酶、脲酶活性,调节土壤氮素的转化,促进玉米/大豆对土壤中氮素的吸收,实现节能增效。     

Plant growth-promoting rhizobacteria as transplant amendments and their effects on indigenous rhizosphere microorganisms

Field trials were conducted in Florida on bell pepper (Capsicum annuum) to monitor the population dynamics of two plant growth-promoting rhizobacteria (PGPR) strains (Bacillus subtilis strain GBO3 and Bacillus amyloliquefaciens strain IN937a) applied in the potting media at seeding and at various times after transplanting to the field during the growing season. In-field drenches of an aqueous bacterial formulation were used for the mid-season applications. The effects of the applied PGPR and application methods on bacterial survival, rhizosphere colonization, plant growth and yield, and selected indigenous rhizosphere microorganisms were assessed. The Gram-positive PGPR applied to the potting media established stable populations in the rhizosphere that persisted throughout the growing season. Additional aqueous applications of PGPR during the growing season did not increase the population size of applied strains compared to treatments only receiving bacteria in the potting media; however, they did increase plant growth compared to the untreated control to varying degrees in both trials. Most treatments also reduced disease incidence in a detached leaf assay, indicating that systemic resistance was induced by the PGPR treatments. However, treatments did not result in increased yield, which was highly variable. Application of the PGPR strains did not adversely affect populations of beneficial indigenous rhizosphere bacteria including fluorescent pseudomonads and siderophore-producing bacterial strains. Treatment with PGPR increased populations of fungi in the rhizosphere but did not result in increased root disease incidence. This fungal response to the PGPR product was likely due to an increase in nonpathogenic chitinolytic fungal strains resulting from the application of chitosan, which is a component of the PGPR formulation applied to the potting media.     

覆盖模式及小麦根系对土壤微生物区系的影响

采用平皿分离培养法研究了5种栽培模式和小麦根系对土壤细菌、真菌及放线菌数量的影响。连续2年的定位测定结果表明:覆膜有利于土壤微生物数量增加。5种栽培模式中,小麦根区、根外土壤细菌数量均以覆膜模式下最高,分别为116.8×106cfu·g-1和86.7×106cfu·g-1;土壤真菌和放线菌数量均以垄沟覆膜(垄上覆膜、垄沟播种)模式下最高,分别为3.0×103cfu·g-1、1.4×103cfu·g-1和18.9×105cfu·g-1、19.7×105cfu·g-1。不同模式下小麦根系对土壤细菌和真菌数量影响较大,表现为根区高于根外;而根系对放线菌影响较小,只有补灌和覆膜2种模式为根区高于根外。多重比较结果显示,覆膜与其他模式之间细菌数量差异极显著,根区土壤细菌和真菌数量与根外存在显著差异。覆盖和根系能大幅度增加根区细菌、真菌和放线菌的数量,强化小麦根区根外细菌和真菌的数量差异。     

Molecular characterization and PCR detection of a nitrogen-fixing Pseudomonas strain promoting rice growth

Nitrogen-fixing plant growth-promoting rhizobacteria (PGPR) from the genus Pseudomonas have received little attention so far. In the present study, a nitrogen-fixing phytohormone-producing bacterial isolate from kallar grass (strain K1) was identified as Pseudomonas sp. by rrs (16S ribosomal RNA gene) sequence analysis. rrs identity level was high with an uncharacterized marine bacterium (99%), Pseudomonas sp. PCP2 (98%), uncultured bacteria (98%), and Pseudomonas alcaligenes (97%). Partial nifH gene amplified from strain K1 showed 93% and 91% sequence similarities to those of Azotobacter chroococcum and Pseudomonas stutzeri, respectively. The effect of Pseudomonas strain K1 on rice varieties Super Basmati and Basmati 385 was compared with those of three non-Pseudomonas nitrogen-fixing PGPR (Azospirillum brasilense strain Wb3, Azospirillum lipoferum strain N4 and Zoogloea strain Ky1) used as single-strain inoculants. Pseudomonas sp. K1 was detected in the rhizosphere of inoculated plants by enrichment culture in nitrogen-free growth medium, which was followed by observation under the microscope as well as by PCR using a rrs-specific primer. For both rice varieties, an increase in shoot biomass and/or grain yield over that of noninoculated control plants was recorded in each inoculated treatment. The effect of Pseudomonas strain K1 on grain yield was comparable to those of A. brasilense Wb3 and Zoogloea sp. Ky1 for both rice varieties. These results show that nitrogen-fixing pseudomonads deserve attention as potential PGPR inoculants for rice.     

Isolation of culturable phosphobacteria with both phytate-mineralization and phosphate-solubilization activity from the rhizosphere of plants grown in a volcanic soil

Chilean volcanic soils contain large amounts of total and organic phosphorus, but P availability is low. Phosphobacteria [phytate-mineralizing bacteria (PMB) and phosphate-solubilizing bacteria (PSB)] were isolated from the rhizosphere of perennial ryegrass (Lolium perenne), white clover (Trifolium repens), wheat (Triticum aestivum), oat (Avena sativa), and yellow lupin (Lupinus luteus) growing in volcanic soil. Six phosphobacteria were selected, based on their capacity to utilize both Na-phytate and Ca-phosphate on agar media (denoted as PMPSB), and characterized. The capacity of selected PMPSB to release inorganic P (Pi) from Na-phytate in broth was also assayed. The results showed that from 300 colonies randomly chosen on Luria–Bertani agar, phosphobacteria represented from 44% to 54% in perennial ryegrass, white clover, oat, and wheat rhizospheres. In contrast, phosphobacteria represented only 17% of colonies chosen from yellow lupin rhizosphere. This study also revealed that pasture plants (perennial ryegrass and white clover) have predominantly PMB in their rhizosphere, whereas PSB dominated in the rhizosphere of crops (oat and wheat). Selected PMPSB were genetically characterized as Pseudomonas, Enterobacter, and Pantoea; all showed the production of phosphoric hydrolases (alkaline phosphatase, acid phosphatase, and naphthol phosphohydrolase). Assays with PMPSB resulted in a higher Pi liberation compared with uninoculated controls and revealed also that the addition of glucose influenced the Pi-liberation capacity of some of the PMPSB assayed.     

Phosphate-solubilizing microorganisms associated with the rhizosphere of mangroves in a semiarid coastal lagoon

 The phosphate-solubilizing potential of the rhizosphere microbial community in mangroves was demonstrated when culture media supplemented with insoluble, tribasic calcium phosphate, and incubated with roots of black (Avicennia germinans L.) and white [Laguncularia racemosa (L.) Gaertn.] mangrove became transparent after a few days of incubation. Thirteen phosphate-solubilizing bacterial strains were isolated from the rhizosphere of both species of mangroves: Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus atrophaeus, Paenibacillus macerans, Vibrio proteolyticus, Xanthobacter agilis, Enterobacter aerogenes, Enterobacter taylorae, Enterobacter asburiae, Kluyvera cryocrescens, Pseudomonas stutzeri, and Chryseomonas luteola. One bacterial isolate could not be identified. The rhizosphere of black mangroves also yielded the fungus Aspergillus niger. The phosphate-solubilizing activity of the isolates was first qualitatively evaluated by the formation of halos (clear zones) around the colonies growing on solid medium containing tribasic calcium phosphate as a sole phosphorus source. Spectrophotometric quantification of phosphate solubilization showed that all bacterial species and A. niger solubilized insoluble phosphate well in a liquid medium, and that V. proteolyticus was the most active solubilizing species among the bacteria. Gas chromatographic analyses of cell-free spent culture medium from the various bacteria demonstrated the presence of 11 identified, and several unidentified, volatile and nonvolatile organic acids. Those most commonly produced by different species were lactic, succinic, isovaleric, isobutyric, and acetic acids. Most of the bacterial species produced more than one organic acid whereas A. niger produced only succinic acid. We propose the production of organic acids by these mangrove rhizosphere microorganisms as a possible mechanism involved in the solubilization of insoluble calcium phosphate. Received: 21 April 1999     

Inoculation of Earthworms and Plant Growth–Promoting Rhizobacteria (PGPR) for the Improvement of Vegetable Growth via Enhanced N and P Availability in Soils

A pot trial was conducted to investigate the single, dual, and triple inoculation of earthworms or plant growth–promoting rhizobacteria (PGPR), including nitrogen-fixing bacteria (NFB) (Azotobacter chroococcum HKN-5) and phosphate-solubilizing bacteria (PSB) (Bacillus megaterium HKP-1), on the growth of Brassica parachinenesis and nitrogen (N) and phosphorus (P) availability in soils. All of the five inoculation treatments significantly (P < 0.05) increased the shoot growth of B. parachinenesis. The greatest shoot and root biomass were recorded in the triple inoculation of earthworm, NFB, and PSB. All of the five inoculation treatments significantly (P < 0.05) increased the concentrations of ammonium (NH₄ ⁺)-N, NO_x-N, and sodium bicarbonate (NaHCO₃)–extractable P in soils. Based on plant growth and availability of N and P in soils, the present study suggested that the triple inoculation may be a promising approach for reducing the need for chemical fertilizers in growing vegetables.