Advantage of using PSIRB over PSRB and IRB to improve plant health of tomato

Twenty-one isolates of phosphate solubilizing-indole acetic acid producing rhizobacteria (PSIRB), 20 isolates of phosphate solubilizing rhizobacteria (PSRB) and 42 isolates of indole acetic acid producing rhizobacteria (IRB) were isolated from 49 rhizospheric soil samples of tomato (Lycopersicon esculentum Mill.) collected from tomato growing regions of Karnataka. A method combining Pikovskaya’s and Bric’s technique was developed to isolate PSRIB, PSRB and IRB’s. The selected isolates were further analyzed for their ability to solubilize calcium phytate. Based on the root colonization assays and the abilities of bacterial isolates to increase the seed germination and seedling vigor under laboratory conditions, five isolates were selected from each group for further studies. Under greenhouse conditions, all the selected rhizobacteria isolates significantly increased root length, shoot length, fresh weight, dry weight and total phosphorus content of 30-day-old-seedlings with respect to control. Isolate PSIRB1 and IRB36 significantly reduced the Fusarium wilt incidence over other isolates of same and other group, and the control. On the basis of results from laboratory and greenhouse studies, one bacterial isolate from each group was selected for plant growth and yield analysis studies. Isolate PSIRB2 showed increased plant height, fresh weight, number of fruits per plant and average weight of fruit over PSRB9, IRB36 and untreated controls. Studies on the nature of protection offered by these bacterial isolates following split-root technique revealed that the isolates PSIRB2 and PSRB9 had the ability to induce systemic resistance. One isolate, IRB36 appeared to protect the tomato seedlings through direct antagonism.     

Phosphate solubilizing rhizobacteria as alternative of chemical fertilizer for growth and yield of Triticum aestivum (Var. Galaxy 2013)

AimThe presence of Phosphorus as a macronutrient in soil is necessary for plant growth and its deficiency restricts crop yield. Therefore, the aim of current study is to isolate promising rhizospheric phosphate solubilizing bacteria presenting with plant growth promoting (PGP) traits and their utilization as biofertilizers to improve Triticum aestivum (Var. Galaxy 2013) growth and nutrition.MethodOut of 30 isolates obtained from rhizosphere of various plants of different regions, 10 best PSRB strains (WumS-3, WumS-4, WumS-5, WumS-11, WumS-12, WumS-21, WumS-24, WumS-25, WumS-26 and WumS-28) were selected based on their high P solubilization and good PGP (auxin, psiderphore, HCN, Nitrogen fixation) activities. Triticum aestivum (Var. Galaxy 2013) was used as an experimental crop under laboratory and field conditions.ResultsIn this study, P solubilization capacity of selected strains were found 4–7 solubilization index on agar plate and 30–246 µg/ml in liquid broth respectively. The optimum conditions for phosphate solubilization under in vitro condition were found 35 °C at pH 7, glucose as good carbon source and ammonium nitrate as a good nitrogen source. Furthermore, the selected strains had the ability to produces phytohormones (indole acetic acid), siderophore, ammonia and Hydrogen Cyanide. Finally, PSRB inoculum showed significant (p < 0.05) increase (50%–80%) in seed germination while 10–90% increase in root length and shoot length was found as compared to control in laboratory condition. Under natural conditions, 40–80% increase in seed germination while 5–34.8% increase in shoot length and 5–96% increase in seed weight was also observed.ConclusionIsolated strains are promising PSRB that enhance plant growth and this research is a base for recommending the use of these bacterial strains for biofertilizer, as an alternative of chemical fertilizer, for Triticum aestivum L. production.     

Rhizosphere mediated growth enhancement using phosphate solubilizing rhizobacteria and their tri-calcium phosphate solubilization activity under pot culture assays in Rice (Oryza sativa.)

Phosphate solubilizing rhizobacteria are considered as an important alternative to increase the availability of accumulated phosphates through solubilization. These increase the growth of plant by enhancing the efficiency of fixing biological nitrogen. This was studied through a pot experiment involving two Phosphate Solubilizing Rhizobacteria (PSRB) isolates, Pseudomonas aeruginosa and Bacillus subtilis along with Tri-calcium phosphate (TCP) on availibity of nutrients, biological composition of soil and yield attributes of rice crop at its growth stages. Experiment was laid in factorial completely randomized design (CRD) comprising of eight treatments replicated thrice with two factors viz. factor 1 with or without TCP (1 g^?1soil) and factor 2 with single or combined inoculation of PSRB isolates. Considerable enhancement in available content of potassium (K), phosphorous (P), nitrogen (N) in soil was found with TCP 1 g^?1soil (P₁) and consortium of Pseudomonas aeruginosa and Bacillus subtilis broth culture at crop growth stages. Highest increase in available N (17.13% and 19.1%), available P (232% and 265%), available K (19.6% and 29.2%) over control were recorded in B₃ (consortium of Pseudomonas aeruginosa and Bacillus subtilis broth culture). Similarly, maximum nutrient uptake N (6.4%), P (15.8%) and K (8.9%) were recorded with same treatment. A considerable growth in soil microbial biomass carbon and dehydrogenase activity at crop growth stages was recorded on application of TCP 1 g^?1soil (P₁) and consortium of PSRB isolates' Pseudomonas aeruginosa and Bacillus subtilis (B₃). Highest increase in microbial biomass carbon (16.4% and 16.5%) and dehydrogenase activity 34.7% and 43.8% over control were recorded in B₃ (consortium of PSRB isolates Pseudomonas aeruginosa and Bacillus subtilis) and was found best among all treatments in terms of yield (63.2%) and yield attributes; number of panicles^?1plant (54.8%), number of grains^?1panicle (156%) and average panicle length (63.9%).     

Identification,characterization and optimization of phosphate solubilizing rhizobacteria (PSRB) from rice rhizosphere

Two billion people worldwide take rice (Oryza sativa L.) as a staple food. Phosphorus (P) and Nitrogen (N) are the major requirements of rice; although these are available in limited concentrations within rice growing regions. Among different types of Plant growth-promoting rhizobacteria (PGPR), Phosphate solubilizing rhizobacteria (PSRB) constitute an important class. These are known for plant growth promotion by enhancing P and N uptake. PSRB are nowadays used as biofertilizers to restore the soil health. Under the present investigation identification, characterization and optimization of phosphate solubilizing activity of these microbes at different pH, temperature and salt concentrations was carried out. Thirty-seven isolates were recovered from different regions of rice rhizosphere on Pikovskaya (PVK) agar among which 15 isolates were recovered from R.S. Pura, 12 isolates from Bishnah and 10 isolates were recovered from Akhnoor sector of Jammu, India. A prominent halo zone of clearance was developed around the colonies of 12 different isolates, indicating phosphate solubilization activity. Four distinct isolates were amplified, cloned and sequenced for taxonomic identification using 16S primers. The results indicated that PS 1, PS 2, PS 3, PS 4 were related to Pseudomonas aeruginosa, Bacillus subtilis strain 1, B. subtilis strain 2, B. subtilis strain 3, respectively. These strains when grown at a wide range of ecological factors showed maximum growth at pH between 6.8 and 8.8, temperature between 28 °C and 37 °C and salinity between 1% and 2%. Screening for phosphate solubilization activity revealed that the halo zone diameter formed by these isolates extended from 2.1 to 3.2 mm. The phosphate solubilizing efficiency (SE) ranged from 35.4 to 50.9 with highest value of 50.9 by PS4 and maximum P solubilization of 10.22 µg/ml was recorded by PS4 at 7th day. Phosphate solubilization activity of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus. MIC value for zinc sulphate heptahydrate in 12 isolates varied from 1 mg/ml to 6 mg/ml. Phosphate solubilization activity and MIC of these identified PSRB strains can be utilized and explored in the rice growing belts of Jammu region which are deficient in phosphorus.     

Screening for fluorescent pseudomonades,isolated from the rhizosphere of tomato,for antagonistic activity toward <Emphasis Type="Italic">Clavibacter michiganensis</Emphasis> subsp<Emphasis Type="Italic">. michiganensis</Emphasis>

Bacterial canker of tomato, caused by Clavibacter michiganensis subsp. michiganensis, continues to be a problem for tomato growers in the Souss-Massa Draa valley, South of Morocco. Assuming that biological control is an alternative for the management of this disease, a total of 303 fluorescent pseudomonads strains isolated from roots and rhizospheric soil of tomato plants were in vitro tested against C. michiganensis subsp. michiganensis. Fluorescent pseudomonads strains which showed the highest antagonistic properties were thereafter investigated for their ability to colonize tomato roots. Our results showed that fluorescent pseudomonads are more represented in rhizospheric soils. However, the most efficient fluorescent pseudomonads isolates were found in the rhizoplane soil and the endorhizosphere. Among 42 spontaneous antibiotic resistant mutants obtained by treatment of the wild-type isolates with five antibiotics (rifampicine, nalidixic acid, ampicilline and chloramphenicol), 28 completely colonized the roots of all tomatoes seedlings used in this investigation. The 42 wild type isolates were then used for in vivo screening with the cotyledon test. Using this test, eight isolates from 42 tested induced a significant decrease of disease incidence and disease symptoms. The eight efficient isolates were then tested for their effectiveness in the protection of tomato plants in pots under greenhouse conditions. Results obtained showed that all tested isolates applied as seed and root treatments reduced significantly (P ≤ 0.001) the incidence of bacterial canker.     

Diversity of rhizospheric and endophytic bacteria isolated from dried fruit of Ficus carica

There is currently an increasing demand for the characterization of endophytic bacteria isolated from different parts of plants (rhizosphere, roots, fruit, leaf) in order to improve the organic agriculture practices. The current research was performed to identify both rhizospheric bacteria isolated from the rhizosphere of Ficus carica in three different sites in the north of Tunisia and endophytic bacteria isolated from dried figs. We then characterized them for a diversity of plant growth-promoting (PGP) activities. A collection of 120 isolates from rhizospheric soil and 9 isolates from dried figs was obtained and purified. 16SrDNA gene amplification of rhizospheric bacteria revealed significant diversity and allowed for the assigning of the isolates to 6 phyla: Gammaproteobacteria, Alphaproteobacteria, Betaproteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. Representative strains of the collection (90 strains) were tested for numerous PGP activities and resistance to abiotic stresses. The most common PGP trait for all bacteria from the three regions was siderophore production (62%), followed by cellulase (38%), then protease activity (37%), then by lipases activity (17%) and lastly by solubilization of phosphates (9%). Twenty -three strains that showed most PGP traits were selected, 8 strains presented 12 or more, and 15 strains displayed between 7 and 11 of 17 PGP activities. The majority of the isolates manifested a possible adaptation to abiotic stress and unfavorable environments. PCR-DGGE analysis of soil rhizosphere of the three sites allowed also for the acquisition of a Cluster analysis of rhizospheric bacterial communities. Our current study identified and characterized for the first time in Tunisia rhizospheric and endophytic bacteria from dried fruit of Ficus carica.     

Antagonistic bacteria of composted agro-industrial residues exhibit antibiosis against soil-borne fungal plant pathogens and protection of tomato plants from Fusarium oxysporum f.sp. radicis-lycopersici

Rhizospheric and root-associated/endophytic (RAE) bacteria were isolated from tomato plants grown in three suppressive compost-based plant growth media derived from the olive mill, winery and Agaricus bisporus production agro-industries. Forty-four (35 rhizospheric and 9 RAE) out of 329 bacterial strains showed in vitro antagonistic activity against at least one of the soil-borne fungal pathogens, Fusarium oxysporum f.sp. radicis-lycopersici (FORL), F. oxysporum f.sp. raphani, Phytophthora cinnamomi, P. nicotianae and Rhizoctonia solani. The high percentage of total isolates showing antagonistic properties (13%) and their common chitinase and β-glucanase activities indicate that the cell wall constituents of yeasts and macrofungi that proliferate in these compost media may have become a substrate that favours the establishment of antagonistic bacteria to soil-borne fungal pathogens. The selected bacterial strains were further evaluated for their suppressiveness to tomato crown and root rot disease caused by FORL. A total of six rhizospheric isolates, related to known members of the genera Bacillus, Lysinibacillus, Enterobacter and Serratia and one RAE associated with Alcaligenes faecalis subsp. were selected, showing statistically significant decrease of plant disease incidence. Inhibitory effects of extracellular products of the most effective rhizospheric biocontrol agent, Enterobacter sp. AR1.22, but not of the RAE Alcaligenes sp. AE1.16 were observed on the growth pattern of FORL. Furthermore, application of cell-free culture extracts, produced by Enterobacter sp. AR1.22, to tomato roots led to plant protection against FORL, indicating a mode of biological control action through antibiosis.     

Comparison among bacterial communities present in arenized and adjacent areas subjected to different soil management regimes

Aims

The aims of this work were to characterize the soil bacterial communities in an arenized area in southern Brazil subjected to different management regimes through cultivation-dependent and cultivation-independent methods and to evaluate the potential of selected plant growth-promoting (PGP) bacteria to improve the growth of native Lupinus albescens plants.

Methods

Bulk soil samples from an arenized site and rhizospheric soil and roots of L. albescens grown in this arenized site as well as samples from soils of the same region outside of the arenized area and rhizospheric soil and roots of L. albescens grown in non-arenized sites were evaluated. Phosphate solubilization, indolic compound and siderophore production abilities of the isolates were screened and compared. Some isolates were selected for in vivo plant growth promotion in greenhouse experiment.

Results

The samples from the arenized area presented less microbial biomass and less diverse bacterial communities compared with those from non-arenized areas. The PGP characteristics produced by the bacterial isolates showed differences among arenized and non arenized areas. A growth chamber experiment with L. albescens showed that phosphate-insoluble conditions coupled with bacterial inoculation resulted in the best PGP effect.

Conclusions

Culture-dependent and culture-independent methods showed converging results regarding diversity indices and the rhizospheric environments increased bacterial diversity and biomass when compared to bulk soils. The PGP traits analyzed in this work were affected by environmental conditions.     

盐碱地柠条根围土中黑曲霉的分离鉴定及解磷能力测定

在盐碱滩地的改良过程中,柠条具有提升土壤供氮、供磷、供钾的潜力.以盐碱滩地上建植的柠条灌木林为研究对象,以柠条根围土壤为培养基质,采用无机磷培养基筛选,用平板溶菌圈法分离获得1株具有溶磷能力的真菌.将测得的ITS基因序列在NCBI上进行同源性检索,结果表明,所测序列与黑曲霉(Aspergillus niger)同源性为100％.综合形态特征和ITS基因序列同源性两方面分析,该菌株鉴定为黑曲霉(Aspergillus niger).168h连续监测无机磷培养液pH值、速效磷含量、菌丝重量和菌体吸磷量,研究该菌株的解磷能力.研究结果表明:随着培养时间的延长,培养液pH值从7.0下降到2.0左右,溶液中速效磷含量逐渐增加到4.7 mg,菌体自身吸磷量由5.4 mg下降到0.5mg,在36-48h后各项指标达到稳定状态.可见,黑曲霉菌体可以有效利用难溶性磷源,并将其转化成可被植物吸收利用的有效磷.     

溶磷真菌的筛选及耐盐特性分析

【背景】土壤盐渍化已成为影响土壤质量和作物产量的重要因素之一,利用微生物改良盐渍化土壤是既经济又环保的方法。【目的】从不同土壤样品和生物肥料中筛选溶磷能力较强的真菌并讨论其耐盐能力,为盐渍化土壤改良提供菌种资源。【方法】采用平板培养法筛选有一定溶磷能力的真菌,经ITS序列分析初步确定菌株的分类地位。以溶磷能力为考察指标,以NaCl梯度和磷酸钙梯度为考察因素,分析不同菌株利用无机磷源的能力,以及溶磷能力与pH的关系。【结果】共筛选得到16株具有较强溶磷能力的真菌,其中4株真菌对水稻发芽有显著的促进作用,它们是1株长枝木霉(MF-1)和3株踝节菌属菌株(SD-2、XJ-7和HLJ-3)。菌株SD-2和XJ-7生长的耐盐能力显著好于菌株MF-1和HLJ-3。当NaCl浓度为5%时,菌株XJ-7的溶磷能力最好,溶磷率可达9.50%;当NaCl浓度为1%时,菌株HLJ-3的溶磷能力较好,溶磷率为6.93%;当NaCl浓度为0时,菌株SD-2和MF-1的溶磷能力较强,溶磷率分别为9.07%和3.73%。进一步研究发现踝节菌属真菌的溶磷能力与菌液pH呈显著负相关关系。【结论】筛选获得的4株真菌其溶磷能力在不同盐环境中有显著差异,为今后土壤改良和生物肥料中菌种的选择提供理论依据和试验基础。     

Potential of endophytic Streptomyces spp. for biocontrol of Fusarium root rot disease and growth promotion of tomato seedlings

Sixteen endophytic actinobacteria isolated from roots of native plants were evaluated for their antagonistic potential against soil-borne phytopathogenic fungi. Among them, three strong antagonistic isolates were selected and characterised for in vitro plant-growth-promoting and biocontrol traits, including production of hydrogen cyanide, indole-3-acetic acid and siderophores, chitinase and β-1,3-glucanase activities, and inorganic phosphate solubilisation. In all trials, the strain Streptomyces sp. SNL2 revealed promising features. The selected actinobacteria were investigated for the biocontrol of Fusarium oxysporum f. sp. radicis lycopersici and for growth promotion of tomato (Solanum lycopersicum L. cv. Aïcha) seedlings in autoclaved and non-autoclaved soils. All seed-bacterisation treatments significantly reduced the root rot incidence compared to a positive control (with infested soil), and the isolate SNL2 exhibiting the highest protective activity. It reduced the disease incidence from 88.5% to 13.2%, whereas chemical seed treatment with Thiram® provided 14.6% disease incidence. Furthermore, isolate SNL2 resulted in significant increases in the dry weight, shoot and root length of seedlings. 16S rDNA sequence analysis showed that isolate SNL2 was related to Streptomyces asterosporus NRRL B-24328^T (99.52% of similarity). Its interesting biocontrol potential and growth enhancement of tomato seedlings open up attractive uses of the strain SNL2 in crop improvement.     

Biodiversity of phosphate solubilizing bacteria in rhizosphere of chickpea,mustard and wheat grown in different regions of Haryana

The native population of phosphate solubilizing bacteria (PSB) was studied in the rhizosphere of chickpea, mustard and wheat grown in different regions of Haryana. A total of 193 PSB were isolated from 245 rhizospheric samples collected from south-west and north-east zones. The PSB count showed large variations (3−67 × 10⁵cfu/g) and biodiversity within the crop and place of sampling. Using biochemical analysis, the isolates were tentatively identified as belonging to four genera, Pseudomonas, Aeromonas, Klebsiella and Enterobacter. Phosphate solubilization of these isolates varied from 5.9 to 123.8% and 2.2 to 227.2 μg/ml in solid and liquid Pikovskaya’s medium, respectively. Based on their morphological traits, all the isolates were placed into 20 groups, majority of them falling in the group having white, round and gummy colonies, irrespective of the crop or the region. The intrinsic antibiotic resistance pattern showed large variations among the isolates and most of the isolates were resistant to streptomycin, ampicillin and penicillin. The highest PSB number and greatest variability were found in the rhizosphere of chickpea, followed by wheat and then mustard.     

南方红豆杉内生及根际放线菌多样性及其生物活性

【目的】研究药用植物南方红豆杉内生及根际土壤放线菌的多样性及其抑菌、抗肿瘤等重要生物活性并获得一些具有强抑制植物病原真菌以及抗肿瘤等重要生物活性的菌株。【方法】选择7种培养基从南方红豆杉及其根际土壤中分离放线菌,对链霉菌进行形态学分类,去重复后对其进行抑制植物病原真菌以及抗肿瘤活性的筛选并对高活性菌株进行初步鉴定。对部分菌株进行16S rRNA基因测序分析研究其多样性。【结果】研究共分离得到277株放线菌,经去重复后剩余111株放线菌,可归类到6个亚目、7个科、8个属。其中链霉菌可分为10个类群。生物活性研究结果显示:30.9%的菌株具有抑制植物病原真菌活性,其中6株放线菌对多种植物病原真菌显示了强的抑菌活性。分别有44.1%和33.3%的菌株对胃癌肿瘤细胞株SGC-7901和肺癌肿瘤细胞株NCI-H460的抑制率在40%以上。【结论】药用植物南方红豆杉及其根际土壤蕴含种类丰富的放线菌资源,具有良好的生物学活性。菌株KLBMP 2170具有显著的抑菌以及抗肿瘤活性,值得我们去进一步研究。     

Genotyping Assessment of Phosphate Solubilizing Bacteria Isolated from Rhizospheric Soil from Central Regions of Lucknow

Abstract

Phosphate solubilizing bacteria (PSB) can convert insoluble form of phosphorous (P) to an available form which is a major concern in Indian agriculture. In this study, 21 isolates having phosphate solubilizing capability were isolated from different regions of Lucknow, India. Among all, six efficient PSB were confirmed by using in vitro P estimation and 16S rRNA universal primers. The similarity detection was done using random amplified polymorphic DNA (RAPD) finger printing for genotyping the PSB isolates and to determine genetic relatedness between them. Twenty different OPA primers were tested among which four primers produced prominent, highly reproducible, and polymorphic bands. An average of 10.5 polymorphic bands per primer with the amplified DNA fragments ranging from 200 to 2000?bp in size. A dendrogram constructed from these data indicated 25–76% homology. Highest similarity was found in between Bacillus anthracis and Bacillus cereus with 33.8% similarity while least dissimilarity was found in B. anthracis and Pseudomonas fragi with 12% of similarity. These findings provide that there is a great genetic diversity between bacterial isolates from different geographical regions and RAPD can be used as a specific, time consuming and also proves as a reliable molecular tool which helps in strain level discrimination.     

青海地区解磷微生物的筛选及对小油菜生长的影响

为了筛选出能适应青海省冷凉气候环境的解磷菌株,以磷酸钙、卵磷脂、植酸为单一磷源,对胶冻样芽孢杆菌、巨大芽孢杆菌、蜡状芽孢杆菌、紫变异链霉菌、肉桂褐链霉菌、黄团孢链霉菌进行固体平板培养基初筛和液体培养基复筛,通过综合比较固体平板培养基中解磷圈的大小和液体培养基可溶性磷含量,初步筛选出解磷效果较好的3株解磷菌,分别为紫变异链霉菌、肉桂褐链霉菌和胶冻样芽孢杆菌。将这3株解磷菌制成液体菌剂,在9月冷凉气候下采用两种不同肥力土壤进行小油菜盆栽试验。结果表明:与对照相比,施加紫变异链霉菌剂后,高肥力耕地土的小油菜收获时株高、鲜重、根长、根重分别增加了35.5%、191.0%、26.2%、282.7%,植株磷吸收量、根际土壤全磷、速效磷分别增加了968.9%、5.1%、2.1%;低肥力自然土的小油菜收获时株高和鲜重分别增加了45.8%和61.3%,根长和根重分别减少了2.6%和4.4%,植株磷吸收量、根际土壤全磷、速效磷分别增加了91.5%、29.1%和213.7%。其他两种解磷菌剂效果不如紫变异链霉菌明显,表明紫变异链霉菌为适合青海地区冷凉气候环境的解磷菌株。     

In vitro evaluation of some bacterial isolates as biofertilizers and biocontrol agents against the second stage juveniles of <Emphasis Type="Italic">Meloidogyne incognita</Emphasis>

Representative soil samples were collected from soil apart and the rhizosphere of different plant varieties grown in five Egyptian Governorates. The presence of plant parasitic nematodes (PPN) and free living nematodes (FLN) in the collected samples were estimated. In addition, one hundred and 65 bacterial cultures which well known as biofertilizers (some are able to fix nitrogen and the others solubilize either phosphate or potassium) were isolated. The isolated bacteria were screened depending on their rate of growth. Thirty-five cultures of fast growing nitrogen fixing bacteria (NFB), phosphate solubilizing bacteria (PSB) and potassium solubilizing bacteria (KSB) and their cultural filtrates were tested in vitro as biocontrol agents against the second stage juvenile (J_2s) of the Meloidogyne incognita. In general, higher mortality percentages of nematodes were recorded by bacterial cultures than their comparative cultural filtrates. The highest mortality percentages were recorded by cultures of NFB7, PSB2 and KSB2 (100% at dilution 1/10), while it was 99.3, 99 and 97.8% at dilution 1/100, respectively. NFB7 exhibited a high nitrogen fixation rate (4.2 μmole N₂/mL/h), while PSB2 and KSB2 effectively solubilized phosphate and potassium comparing with the control treatments (1.94 fold of available phosphate and 2.0 fold of available potassium, respectively). NFB7, PSB2 and KSB2 isolates showed the highest protease, gelatinase and chitinase activities which were thought responsible for their nematicidal effect. The three bacterial isolates were identified as Paenibacillus polymyxa, Bacillus megaterium and Bacillus circulans, respectively.     

防风根际真菌的分离鉴定及其生防潜力评价

作为世界性分布的镰刀属常见病原真菌,尖孢镰刀菌（Fusarium oxysporum）和木贼镰刀菌（F. equiseti）对包括经济作物及药用植物等的生长均有较大危害。利用源自于植物根际土壤的有益微生物防控尖孢镰刀菌和木贼镰刀菌引起的真菌性病害,是目前较为理想的植物病害管理策略。为获得可有效抑制尖孢镰刀菌和木贼镰刀菌的生防菌源,于防风根际土壤中分离纯化真菌104株,基于平板对峙法筛选获得1株对尖孢镰刀菌和木贼镰刀菌具有显著抑制效果的菌株MR-43,结合形态特征、ITS序列分析,将其确定为Sirastachys castanedae（GenBank登录号：OK287148.1）,隶属于Sirastachys组进化分支,发现其可宿生于植物根际土壤。基于盆栽试验法,探究MR-43在防风栽培土壤中的定殖能力,评价MR-43对由尖孢镰刀菌引起的防风枯萎病和由木贼镰刀菌引起的防风根腐病的防病能力,及其对防风植株的促生能力。结果显示,Sirastachys castanedae MR-43对尖孢镰刀菌、木贼镰刀菌的抑菌率为57%以上,且经多次验证抑菌效果稳定;菌株MR-43可稳定定殖于防风栽培土壤并实现有效扩繁;菌株MR-43的孢子悬液可有效控制防风枯萎病和防风根腐病,平均防效达68.52%,与多菌灵、代森锰锌的防病效果无显著性差异（P>0.05）;此外,MR-43对防风植株生长具有明显促进作用。因此,Sirastachys castanedae MR-43在防风枯萎病、根腐病等真菌性病害管理方面具有较好的开发价值及应用潜力。     

Impact of long-term application of industrial wastewater on the emergence of resistance traits in Azotobacter chroococcum isolated from rhizospheric soil

A total of 57 (36 and 21) Azotobacter chroococcum were isolated from wheat (Triticum aestivum) rhizospheric soil irrigated with industrial wastewater (about a decade) and ground water (uncontaminated) and characterized on the basis of morphological, cultural and biochemical characteristics. Rhizospheric soils were analyzed for metal concentrations by atomic absorption spectrophotometery and the test soil samples were contaminated with Fe, Zn, Cu, Cr, Ni and Pb. All the isolates of A. chroococcum were tested for their resistance against Hg2+, Cd2+, Cu2+, Cr3+, Cr6+, Zn2+, Ni2+ and Pb2+. Among 36 isolates of Azotobacter from soil irrigated with industrial wastewater, 94.4% were resistant to Pb2+ and Hg2+ and 86.1%, 77.5% and 63.8% were resistant to Zn2+, Cr6+ and Cr3+ respectively. The highest minimum inhibitory concentration of 200 microg/ml for Hg2+ and 1600 microg/ml for other metals were observed against these bacteria from soil. The incidences of metal resistance and MICs of metals for A. chroococcum from wastewater irrigated soil were significantly different to those of uncontaminated soil. All A. chroococcum isolates were tested for their resistance against 11 commonly used antibiotics/drugs. 91.6% were found to be resistant against nitrofurantoin while 86.4% and 80.5% were found to be resistant against polymyxin-B and co-trimoxazole respectively. Agarose gel electrophoresis using the miniprep method for plasmid isolation revealed that these isolates harboured plasmids of molecular weights 58.8 and 64.5 kb using EcoRI and HindIII digests of X DNA and undigested X DNA as standard markers.     

Isolation and evaluation of bacteria and fungi as biological control agents against Rhizoctonia solani

Rhizoctonia solani is one of the most important limiting factors for potato production and storage in Belgium and worldwide. Its management is still strongly dependent on chemical treatments. The aim of this work was to evaluate the possibility of exploiting bacteria and fungi in order to control this pathogen. Among a collection of 220 bacterial strains isolated from different organs of healthy potato plants and rhizospheric soils, 25 isolates were selected using screening methods based on in vitro dual culture assays. The mycelial growth inhibition rate of the pathogen was ranged from 59.4 to 95.0%. Also seven fungal strains isolated from the rhizospheric soil and potato roots showed a highly mycelial growth inhibition of R. solani. The mycelial growth inhibition rate obtained with these fungi was included between 60.0 and 99.4%. From this preliminary study, the further investigations will be planned to determine the bacterial isolates systematic, species of fungal strains by using molecular tools and to assess their efficacy against R. solani in greenhouse trials.     

Ability of <Emphasis Type="Italic">Emericella rugulosa</Emphasis> to mobilize unavailable P compounds during Pearl millet [<Emphasis Type="Italic">Pennisetum glaucum</Emphasis> (L.) R. Br.] crop under arid condition

Phosphate solubilizing microorganisms are ubiquitous in soils and could play an important role in supplying P to plants where plant unavailable P content in soil was more. A phosphatase and phytase producing fungus Emericella rugulosa was isolated and tested under field condition (Pearl millet as a test crop) in a loamy sand soil. In the experimental soil 68% organic phosphorous was present as phytin; less than 1% of phosphorous was present in a plant available form. The maximum effect of inoculation on different enzyme activities (acid phosphatase, alkaline phosphatase, phytase, and dehydrogenase) was observed between 5 and 8 weeks of plant age. The depletion of organic P was much higher than mineral and phytin P. The microbial contribution was significantly higher than the plant contribution to the hydrolysis of the different P fractions. A significant improvement in plant biomass, root length, seed and straw yield and P concentration of root and shoot resulted from inoculation. The results suggest that Emericella rugulosa produces phosphatases and phytase, which mobilize P and enhance the production of pearl millet.