四种不同生态系统的土壤解磷细菌数量及种群分布

通过分析农田、林地、草地和菜地土壤有机磷细菌和无机磷细菌的数量及种群结构,发现有机磷细菌数量比无机磷细菌多;有机磷细菌主要是芽孢杆菌属,其次是假单胞菌属;而无机磷细菌主要是假单胞菌属。菜地土壤解磷细菌的数理和种类最多,不同的细菌解磷能力差异很大,其解磷能力与培养介质的ｐＨ值有一定的关系;许多菌株在进一步的纯化过程中失去了解磷能力。     

一株耐盐解磷菌的解磷能力及对玉米敏感期生长的影响

从黄河三角洲盐碱化土壤中筛选了一株高效解磷真菌QL1501,经鉴定为草酸青霉菌,菌株QL1501对无机磷的解磷能力远大于对有机磷的解磷,解无机磷最大浓度达85.21 mg/L。菌株QL1501的最适生长pH值为8时菌体生长极好。当NaCl浓度为1%～5%时,菌株解磷能力变化不大,溶液中有效磷浓度为76.08～65.37 mg/L。当溶液中NaCl浓度高于7%时,菌体生长受到较大影响。接种解磷真菌QL1501处理的玉米株高、根干重和植株干重均显著高于未接种的对照处理,说明该解磷菌作为解磷生物肥料具有良好的效果。     

氮素形态对解磷细菌发酵液解磷效果及小白菜生长和品质的影响

采用盆栽试验,研究了施用硝态氮、铵态氮对解磷细菌发酵液不同组分解磷活性及其对小白菜产量、品质的影响。结果表明,施用相同氮肥时,浇灌解磷细菌原发酵液处理 T3的作用效果明显优于菌体悬浮液T2及菌液上清T1;以硝态氮为底肥时,施用发酵原液的土壤有效态磷含量较高,小白菜产量最高,品质明显优化,即硝态氮含量下降, Vc、可溶性糖、可溶性蛋白、纤维素以及植株磷含量显著增加,且增加变幅均高于以铵态氮为底肥的效果。由此可知,硝态氮与解磷细菌发酵原液配施时, W1菌株解磷效果更好,可能是解磷菌株W1最适的氮源。因此,种植小白菜时直接向其根际施用解磷细菌发酵液并配施硝态氮肥,更有利于提高小白菜的品质及产量。     

一株毒死蜱降解菌株Sphingomonas sp.Dsp-2的分离鉴定及降解特性

从长期受毒死蜱污染的污水处理池中分离到一株毒死蜱高效降解菌株,命名为Dsp-2。经生理生化和16S rDNA序列同源性分析,鉴定其为鞘胺醇单胞菌属(Sphingomonas sp.)细菌。该菌株能在24 h内完全降解100 mg L-1的毒死蜱,降解特性的研究表明:随着农药浓度的加大,绝对降解量也增大,但高浓度的毒死蜱会导致不能完全降解;起始接种量和降解毒死蜱的速率呈正相关;外加氮源营养能够明显促进降解;1 mmol L-1的Fe3 和Ni2 等对其降解性能有抑制作用。研究了Dsp-2在土壤中降解毒死蜱的效果。结果表明,Dsp-2在三种供试土壤中都能有效的降解毒死蜱,其中在潮土中降解的速率最快,且当毒死蜱的浓度范围在1~100 mg kg-1内Dsp-2都能有效的降解毒死蜱,7 d降解率达到85%~98%。     

一株根际解磷菌的筛选鉴定及促生作用研究

磷在植物生长发育过程中起着重要的作用。为获得优质的解磷促生菌,以北京平谷桃园根际土壤为原料,从中筛选具有较高解磷促生能力的菌株。通过平板试验初筛可产解磷圈的菌株,再进行液态培养,采用钼锑抗比色法测定其最大的解磷能力,并探索菌株解磷能力与pH变化的关系,同时借助盆栽试验研究其促生特性。从桃树根际土共分离筛选出3株解磷菌,编号分别为JP01、JP03和PG62,初步鉴定分别为黑曲霉、杰氏假单胞菌和苍白杆菌。其中,黑曲霉JP01解磷能力优异,其溶解磷酸三钙的能力为112.52 mg/L,对卵磷脂的转化量为145.50 mg/L,最大解磷矿粉能力为95.74 mg/L,在培养过程中,该解磷菌对应的培养液pH显著下降。盆栽试验表明,黑曲霉能增加玉米幼苗株高、茎粗和地下部干重,还可使植株全磷含量增加。本研究结果可为开发解磷微生物菌剂提供优良的菌株资源。     

两株土壤分离菌的解磷能力及对玉米的促生作用

从植物根际的土壤样品中分离筛选出两株高效解磷细菌P_9和P_(28),分别被鉴定为粘质沙雷氏菌和产气肠杆菌.研究了不同环境条件对解磷能力的影响,结果表明,P_(28)耐 3%NaCl(w/v,下同)、28℃是两菌的最适温度、在初始pH 5.5～10.0条件下培养时能够高效溶解Ca_3(PO_4)_2.盆栽试验结果表明,单接种、双接种解磷菌处理的玉米株高、植株干重和磷含量显著高于不接菌对照;单接种P_9处理的玉米明显强于单接种P_(28)处理;将有机肥(M)和磷矿石(RP)作为载体和解磷菌一同混合施入土壤的处理,玉米苗干重,磷含量较单施解磷菌显著增加.     

Cd2+ 对无机磷细菌生长及溶磷作用的影响

无机磷细菌(phosphorus solubilization bacteria,PSB)是重要的土壤有益微生物,它们将植物不能吸收的难溶性无机磷转化为有效磷,对土壤磷素供应十分重要。镉是污染土壤的常见重金属,研究Cd~(2+)胁迫下PSB的生长和溶磷能力,可揭示镉污染对土壤无机磷转化供应的影响,为改善植物磷素供应提供有益信息。试验以重庆市缙云山国家森林公园毛竹林地分离的4株假单胞菌为供试菌株,比较它们的溶磷能力,研究了不同浓度Cd~(2+)对PSB生长和溶磷作用的影响。结果表明:供试菌株溶磷能力PSB05PSB09PSB07PSB01,溶磷指数和溶磷率最高值是最低值的2.08倍和4.14倍。摇瓶培养120 h后,培养液pH表现为PSB01PSB07PSB09PSB05,并与培养液中的无机磷含量(y)呈显著负相关(y=–83.148 pH+581.96,R2=0.9052,n=25)。在固体培养时,PSB05的溶磷圈和菌落直径随培养基中Cd~(2+)浓度提高而降低。在Cd~(2+)浓度0.5~2.0 mg/L液体培养基中,PSB05数量较无Cd~(2+)对照减少22.09%~68.18%,草酸、乙酸、柠檬酸和氢离子分泌量显著下降,溶磷量降低5.27%~38.45%。供试菌株的溶磷能力差异显著;高浓度Cd~(2+)抑制PSB05生长、有机酸和氢离子分泌及溶磷作用,不利于土壤供给磷素。     

一株人参根区解磷细菌的筛选、鉴定及对人参生长的影响

人参是我国传统名贵中药材,也是吉林省对外交往中的代名词。吉林参畦土壤有效磷含量低,但大量施用磷肥又会导致人参品质的下降,因此,本研究从人参根区土壤中筛选高效解磷细菌,接种到人参根部,研究其对人参生长、品质、土壤速效养分、土壤酶活性的影响,以期为人参解磷微生物肥料提供优良菌株,从而促进人参产业绿色发展。采用PKO平板初筛及摇瓶复筛从野山参根区土壤筛选高效解磷细菌。盆栽试验以3年生人参种苗为材料,每盆种植6棵单株重相同的人参种苗,设对照(CK)和处理组,待完全展叶后间苗,留取4棵长势一致的人参苗,采用滴灌的方式将菌液接种在人参苗根部附近,生长至红果期(90 d)取样,测定植物生长量和叶绿素含量、土壤速效养分、参根和参叶中NPK的含量、土壤酶活性、参根中微量元素和8种皂苷的含量。结果表明,1)平板初筛和摇瓶复筛得到P1、P7菌株,摇瓶培养中,P1可溶性磷为472.85μg/m L,P7可溶性磷为437.33μg/m L。2)P1处理的人参参根鲜重、干重显著高于P7、BM、CK处理,株高显著高于对照,叶绿素含量显著高于P7、BM、CK(P0.05)。3)P1处理促进人参对土壤钾元素的吸收,提高磷素在参根中的积累量。4)接种菌剂可以不同程度的提高土壤酶活性,以P1处理的效果最显著,P1处理人参根际土壤脲酶、酸性磷酸酶、过氧化物酶活性均为最高。5)P1处理显著提高了人参根中Fe、Mn、Cu的含量,分别较对照增加了52.74%、127.03%、28.96%,P1处理Zn与对照相比增加19.96%;人参根中8种单体皂苷P1处理均显著高于P7、BM、CK。6)经16S rRNA分析结合P1形态初步鉴定为假单胞菌。     

磷细菌P17对不同来源磷矿粉的溶磷作用及机制

比较了一株磷细菌P17对来自多种产地磷矿粉的溶解能力。通过摇瓶试验、扫描电镜观察提出磷细菌P17对不同来源磷矿粉生物风化的证据 ,选出了P17最适合作用的磷矿粉类型。从摇瓶试验看出 ,磷细菌P17对来源于黄麦岭、黄金卡黄的变质岩型磷矿粉有较好的溶解能力。经过P17长达 70d的溶解 ,磷矿粉的难溶磷逐渐被P17菌株溶解下来。连续 10次接种培养后 ,P17菌株能溶解黄麦岭磷矿粉全磷的81 0 2 % ;而对于黄金卡黄磷矿粉 ,P17溶解了全磷总量的 78 97%。试验结果表明磷细菌P17能够提高黄麦岭磷矿粉和黄金卡黄磷矿粉的持续利用率。另外 ,培养 7d后 ,磷细菌P17能够在以黄麦岭磷矿粉、黄金卡黄磷矿粉为唯一磷源的发酵液中产生 3 10mmolL-1的挥发性有机酸 ,同时能分别产生 4 0 5mmolL-1、5 1 1mmolL-1的难挥发性有机酸。经过气相色谱检测 ,磷细菌P17能够产生柠檬酸、琥珀酸、乳酸以及乙酸等有机酸 ,可能螯合磷矿粉中的金属离子 ,使磷游离出来。初步探讨了磷细菌P17的溶磷机制     

两株抗铜细菌的筛选、鉴定及对碳酸铜的溶解作用

本研究从长期受铜污染的铜矿废弃地土壤中分离得到两株对重金属铜具有较强抗性的菌株F16a和Fw17a。对菌株形态、生理生化特性及其16S rDNA系统进化进行分析,将F16a鉴定为肠杆菌属(Enterobacter),Fw17a鉴定为假单胞菌属(Pseudomona)。F16a和Fw17a对氨苄青霉素、头孢他啶均有抗性;此外,Fw17a还对氯霉素、四环素及低浓度的卡那霉素具有抗性。在有氮培养基(含500 mg/L CuCO3)液体培养48 h后,F16a使培养基上清液中铜浓度增加了300%左右;相反,Fw17a使培养基上清液中铜浓度降低了60%左右。     

红壤溶磷菌的筛选及溶磷机制

采用以磷酸铝为磷源的蒙金娜(PVK)液体培养基研究了从红壤土中筛选出的4种溶磷菌的溶磷效果,选出其中的优势菌株B1,并对其溶磷机理做出初步探讨。结果表明,所筛选出的4株溶磷菌在液体培养条件下均有显著的溶磷效果,其中菌株B1在培养4 d后有效溶磷量最大,达到292.8 mg L-1。各处理培养液pH在培养期间均有显著下降,pH从7.0下降至3.2~4.7。高效液相色谱测定发现,各菌株培养液中有机酸的种类与含量随培养时间变化而不同,其中菌株B1主要分泌草酸和苹果酸,培养1 d后有机酸总量可达到5 mmol L-1;通过添加有机酸对磷酸铝活化的试验表明,分泌有机酸溶磷仅是菌株B1溶磷机制之一,可能还存在其他溶磷机制。菌株B1生长的适宜pH范围为5~9,最适培养温度为30℃,100 ml三角瓶的最适装液量为30~40 ml。经鉴定,菌株B1与苏云金芽孢杆菌有99.9%的相似性。     

Isolation and characterization of mineral phosphate-solubilizing bacteria naturally colonizing a limonitic crust in the south-eastern Venezuelan region

With the aim to explore the possible role of mineral phosphate-solubilizing bacteria (PSB) in phosphorus (P) cycling in iron-rich, acidic soils, we conducted a survey of PSB naturally colonizing a limonitic crust in the south-east region of Venezuela (Bolívar State). A total of 130 heterotrophic bacterial isolates showing different degrees of mineral tri-calcium phosphate (Ca₃(PO₄)₂)-solubilizing activities were isolated from NBRIP plates. In contrast, no isolates showing iron phosphate (FePO₄)- or aluminum phosphate (AlPO₄)-solubilizing activities were detected by this experimental approach. The 10 best Ca₃(PO₄)₂-solubilizers were selected for further characterization. These isolates were shown to belong to the genera Burkholderia, Serratia, Ralstonia and Pantoea by partial sequencing analysis of their respective 16S rRNA genes. All the PSB isolates were able to mediate almost complete solubilization of Ca₃(PO₄)₂ in liquid cultures; in contrast, the PSB isolates were less effective when solubilizing FePO₄. Two groups of PSB isolates were clearly differentiated on the basis of their Ca₃(PO₄)₂ solubilization kinetics. Acidification of culture supernatants seemed to be the main mechanism for P solubilization. Indeed, gluconic acid was shown to be present in the supernatant of five isolates. Furthermore, detection of genes involved in the production of this organic acid was possible in three isolates by means of a PCR protocol.     

Soil organic carbon and phosphorus availability regulate abundance of culturable phosphate-solubilizing bacteria in paddy fields

Low availability of phosphorus(P) is a major constraint for optimal crop production, as P is mostly present in its insoluble form in soil. Therefore,phosphate-solubilizing bacteria(PSB) from paddy field soils of the Indo-Gangetic Plain, India were isolated, and their abundance was attempted to be correlated with the physicochemical characteristics of the soils. Ninety-four PSB were isolated on Pikovskaya's agar medium, and quantitative phosphate solubilization was evaluated using NBRIP medium. The isolates solubilized P up to a concentration of 1 006 μg mL~(-1) from tricalcium phosphate with the secretion of organic acids. These isolates were identified by 16 S rRNA gene sequence comparison, and they belonged to Gammaproteobacteria(56 isolates),Firmicutes(28 isolates), Actinobacteria(8 isolates), and Alphaproteobacteria(2 isolates). Phylogenetic analysis confirmed the identification by clustering the isolates in the clade of the respective reference organisms. The correlation analysis between PSB abundance and physicochemical characteristics revealed that the PSB population increased with increasing levels of soil organic carbon, insoluble P, K~+, and Mg~(2+). The promising PSB explored in this study can be further evaluated for their biofertilizer potential in the field and for their use as potent bio-inoculants.     

Phosphate solubilization by a wild type strain and UV-induced mutants of Aspergillus tubingensis

Phenotypic mutants of Aspergillus tubingensis were obtained by UV irradiation and phosphate solubilization ability of these mutants were studied and compared with the wild type strain. Low phosphate solubilizing mutant was also selected in this study. Among the different mutants, AtM-5 and AtM-2 showed highest P solubilization when tri-calcium phosphate and rock phosphate were used as P source compared to the wild type strain and other mutants. These mutants also showed maximum acid phosphatase and phytase activity. These results suggest that P solubilization by these isolates is due to lowering of pH of the culture filtrate and also the activity of acid phosphatase and phytase.     

Isolation and characterization of phosphate solubilizing bacteria from the rhizosphere of crop plants of Korea

Whole-cell fatty acids methyl ester (FAME) profile and 16S rDNA sequence analysis were employed to isolate and identify the bacterial groups that actively solubilized phosphates in vitro from rhizosphere soil of various crops of Korea. Out of several hundred colonies that grew on Pikovskaya's medium 13 best isolates were selected based on the solubilization of insoluble phosphates in liquid culture and further characterized and identified. They were clustered under the genera Enterobacter, Pantoea and Klebsiella and the sequences of three representative strains were deposited in the GenBank nucleotide sequence data library under the accession numbers AY335552, AY335553, AY335554.     

Single and double inoculation with Azospirillum/Trichoderma: the effects on dry bean and wheat

A plant growth-promoting rhizobacterium (Azospirillum brasilense Sp7) and a bio-control fungus, which can solubilize insoluble phosphorus (Trichoderma harzianum Rifai 1295-22), were evaluated for their single and combined effects on dry bean (Phaseolus vulgaris) and wheat (Triticum aestivum L.) grown in soil. A pot experiment with bean and a field experiment with both bean and wheat were established. In contrast to single inoculation of Trichoderma, the single inoculation of Azospirillum and the double inoculation did not significantly (P >0.05) increase nodule numbers and nodule mass at 45 days after planting in pot grown beans. However, the Azospirillum inoculation with supplementary phosphorus significantly (P <0.05) increased nodule mass. There were no significant (P >0.05) differences among the inoculation treatments for plant dry weight, total plant nitrogen, and total plant phosphorus at 45 days after planting in both pot and field experiments with bean. However, the combined inoculation and rock phosphate application at 1 Mg ha^–1 significantly (P <0.05) increased bean seed yield, total seed nitrogen and phosphorus in the bean field trial. This treatment more than doubled the mentioned properties compared to the control. The microbial inoculations, with the exception of the combined inoculation, significantly (P <0.05) increased total seed nitrogen, but never affected seed yield in the wheat field trial (P >0.05). The combined inoculation improves many plant and yield parameters and, therefore, has some advantages over single inoculation provided that rock phosphate was supplied at an amount not exceeding 1 Mg ha^–1. Higher rock phosphate application rates decreased many plant and yield parameters in our study.This work was carried out at Gaziosmanpaa University, Tokat, Turkey.     

一株分离自葛藤根际高效溶磷细菌特性研究及菌株鉴定

对分离自葛藤(Pueraria lobata L.)根际的一株高效溶磷细菌GTR15进行促生特性、主要生理生化指标测定和16S rDNA序列分析。结果表明,菌株GTR15的HD/CD值(溶磷圈直径HD,菌落直径CD)为2.22,28℃液体振荡培养7 d后对磷酸钙的溶解量为138.72 mg.L-1,分泌IAA(3-吲哚乙酸)及有机酸量分别为14.44 mg.L-1、46.00 mmol.L-1。菌株革兰氏染色为阴性,细胞短杆状;淀粉水解、吲哚、V-P(二乙酰试验)、苯丙氨酸脱氨酶、明胶液化及M-R(甲基红)试验呈阴性;柠檬酸盐、过氧化氢酶、硫化氢及硝酸盐还原等试验呈阳性,结合菌株16S rDNA序列分析结果,初步鉴定为肠杆菌(Enterobacter sp.)。该菌株在研制高效微生物磷肥接种剂方面可能具有较大潜力。     

Identification of uncultured bacteria tightly associated with the intestine of the earthworm Lumbricus rubellus (Lumbricidae; Oligochaeta)

The bacteria associated with the intestine and casts of the earthworm Lumbricus rubellus Hoffmeister, 1843, were examined by direct counts, culturability studies, 16S rRNA gene clone libraries, and fluorescent in situ hybridization (FISH). A significant fraction, 24-47%, of the total numbers of prokaryotes remaining in the intestine after casting were tightly associated with the intestinal wall. Bacterial 16S rRNA gene clone libraries constructed from washed earthworm intestinal tissue suggested that the bacterial community was dominated by a few phylotypes that were either absent from, or in low abundance, in the casts. The specific phylotypes present depended on the date of sampling and included representatives of the Acidobacteria, Firmicutes, β-Proteobacteria, and one phylogenetically deep, unclassified group. Juvenile earthworms subsequently collected contained three of the four phylotypes observed in the intestine clone libraries. The Firmicutes phylotype was examined by FISH and was found to be a short rod that represented only a small fraction of the total population of the juvenile samples. These results suggested that the microbial community tightly associated with the intestine was dominated by a small number of phylotypes and that this association was opportunistic rather than obligate.     

Effect of plant growth-promoting bacteria and soil compaction on barley seedling growth, nutrient uptake, soil properties and rhizosphere microflora

Inoculants are of great importance in sustainable and/or organic agriculture. In the present study, plant growth of barley (Hordeum vulgare) has been studied in sterile soil inoculated with four plant growth-promoting bacteria and mineral fertilizers at three different soil bulk densities and in three harvests of plants. Three bacterial species were isolated from the rhizosphere of barley and wheat. These bacteria fixed N₂, dissolved P and significantly increased growth of barley seedlings. Available phosphate in soil was significantly increased by seed inoculation of Bacillus M-13 and Bacillus RC01. Total culturable bacteria, fungi and P-solubilizing bacteria count increased with time. Data suggest that seed inoculation of barley with Bacillus RC01, Bacillus RC02, Bacillus RC03 and Bacillus M-13 increased root weight by 16.7, 12.5, 8.9 and 12.5% as compared to the control (without bacteria inoculation and mineral fertilizers) and shoot weight by 34.7, 34.7, 28.6 and 32.7%, respectively. Bacterial inoculation gave increases of 20.3–25.7% over the control as compared with 18.9 and 35.1% total biomass weight increases by P and NP application. The concentration of N and P in soil was decreased by increasing soil compaction. In contrast to macronutrients, the concentration of Fe, Cu and Mn was lower in plants grown in the loosest soil. Soil compaction induced a limitation in root and shoot growth that was reflected by a decrease in the microbial population and activity. Our results show that bacterial population was stimulated by the decrease in soil bulk density. The results suggest that the N₂-fixing and P-solubilizing bacterial strains tested have a potential on plant growth activity of barley.     

Genetic diversity and phylogeny of pink-pigmented facultative methylotrophic bacteria isolated from the phyllosphere of tropical crop plants

Genetic diversity of pink-pigmented facultative methylotrophic bacteria belonging to the genus, Methylobacterium, was assessed using 16S ribosomal RNA (rRNA) gene sequencing, amplified ribosomal DNA restriction analysis (ARDRA), and differential carbon-substrate utilization profile in the phyllosphere of cotton, maize, sunflower, soybean, and mentha plants. Methylobacterium populi, Methylobacterium thiocyanatum, Methylobacterium suomiense, M. aminovorans, and Methylobacterium fujisawaense were identified to colonize the phyllosphere of these crop plants. Among these, M. populi found to be the dominating species followed by M. aminovorans. The diversity indices like Shannon index of diversity, Pielou index of evenness, and Margalef index of richness calculated based on combined data of ARDRA, 16S rRNA gene sequence, and differential carbon-substrate utilization revealed that sunflower leaves showed richest methylobacterial diversity, followed by soybean, while cotton and mentha leaves recorded the lowest diversity. The outcome of this study indicates that diversity of Methylobacterium in phyllosphere depends upon the host plant species colonized.