烟草根际土壤中解钾细菌的分离与多样性分析

土壤中含有丰富的钾元素,但主要以缓效态形式存在于钾长石或云母等硅酸盐矿物中,不能被作物直接吸收利用。解钾微生物能溶解硅酸盐矿物中的钾,提高土壤中作物可利用钾的含量,有望缓解我国钾肥短缺的现状。本研究利用选择性培养基,从烟草根际筛选钾细菌,基于16S rDNA序列分析烟草根际土壤解钾细菌的多样性,通过测定解钾细菌的解钾效能及对烟草的促生作用,筛选有应用潜力的优良解钾细菌菌株。结果表明,从四川、湖北和山东烟区烟草根际土壤分离获得的27株解钾细菌,在解钾固体培养上溶钾圈直径为0.11~0.30 cm。16S rDNA序列分析表明,烟草根际土壤解钾细菌主要包括变形菌门γ亚群(Gammaproteobacteria,85.18%)、变形菌门α亚群(Alphaproteobacteria,3.70%)、变形菌门β亚群(Betaproteobacteria,3.70%)、放线菌门(Actinobacteria,3.70%)和拟杆菌门(Bacteroidetes,3.70%),其中克雷伯菌属(Klebsiella)为优势菌属(66.67%)。27个菌株均有一定的解钾能力,解钾活性为0.59~4.40 mg.L–1。参试菌株均对烟草有一定的促生作用,利用解钾细菌菌液处理烟株20 d后,与对照相比,株高增加0.97%~38.64%,最大叶长增加4.40%~31.02%。本研究筛选出的菌株XF11、GM2、JM19和GL7具有较高的解钾活性和促进植物生长的能力,展现了良好的应用潜力。     

适应玉米的溶磷细菌筛选及其对玉米生长的影响

从石灰性土壤中分离获得4株高效溶磷细菌X5、X6、Z4和Z8,研究其生物学特征,探索其单独及复合的溶磷促生潜能。研究发现菌株X5、X6、Z4和Z8均可以利用玉米根系分泌物作碳源生长。菌株X6和Z4均能产生吲哚乙酸(IAA)和铁载体,菌株Z8可产生IAA不产生铁载体,菌株X5可产生铁载体不产生IAA。盆栽试验结果表明,接种单一溶磷菌及4株菌复合处理均可促进玉米生长,但复合菌群的溶磷促生效果显著高于单一菌株。通过16S r RNA基因序列分析研究菌株的分类地位,初步鉴定X5、X6、Z4、Z8分别为荧光假单孢菌(Pseudomonas fluorescens)、草假单胞菌(Pseudomonas poae)、巨大芽孢杆菌(Bacillus megaterium)和枯草芽孢杆菌(Bacillus subtilis)。     

玉米和大豆根内生拮抗细菌筛选鉴定及生防能力评价

为获得对植物真菌病害有稳定作用效果的生防细菌,采用平板对峙培养法从大豆和玉米根部内生细菌中筛选获得12株菌株,这些细菌对大豆立枯丝核菌(Rhizoctonia solani)、大豆尖孢镰刀菌(Fusarium oxysporum f.sp.glycine)和番茄枯萎病菌(Fusarium oxgsporum f.sp.lycopersici)均具有明显拮抗作用。16S rRNA基因鉴定表明12株内生拮抗细菌分别属于芽孢杆菌属(Bacillus)、类芽孢杆菌属(Paenibacillus)和假单胞菌属(Pseudomonas)。选取5株细菌进行盆栽接种试验,结果表明,供试细菌均对大豆根腐病具有一定的防治效果,其中,菌株J1和J2防效分别达到20.4%和32.1%。     

马铃薯内生固氮菌的分离及其促生特性研究

内生固氮菌是一类重要的植物促生菌,占据着植物组织内有利于营养供应和微环境适宜的生态位,能更好的发挥促生功能,促生机制包括固氮、解磷、产植物生长素、产ACC脱氨酶、产铁载体及拮抗病害等。本文从连作3年马铃薯根筛选获得8株内生固氮菌,其中短芽孢杆菌属(Brevibacterium)3株,芽孢杆菌属(Bacillus)3株和泛菌属(Pantoea)2株。促生特性研究发现:菌株Brevibacterium sp.GWR4具有较高固氮酶活性[16.206nmol C2H4/(mg蛋白质·h)],与圆褐固氮菌的固氮酶活性具有极显著性差异(P0.01);菌株Pantoea spp.GWR2、GWR3具有极高的产IAA能力,分别为186.07、162.21μg/m L,GWR2兼具产ACC脱氨酶活性[3.74μmol/(mg蛋白质·h)],GWR3兼具溶解无机磷的能力;菌株Brevibacterium sp.GWR5可以拮抗尖孢镰刀菌MLSOF(Fusarium xysporum)和茄病镰刀菌MLS-QB(Fusarium solani),兼具溶解有机磷和产IAA能力;Bacillus spp.GWR7、GWR8兼具固氮和产IAA能力。盆栽试验显示,分别接种Pantoea sp.GWR3,Brevibacterium spp.GWR4、GWR5和Bacillus spp.GWR7、GWR8后小白菜鲜重显著高于未接菌对照处理。可见,这5株菌兼具多种促生特性且对小白菜具有较好的促生效果,有望进一步研究开发成为微生物肥料生产菌种。     

马铃薯细菌多样性解析及促生高产 IAA菌株的筛选

采用 3种不同的培养基从马铃薯根际土壤和叶片中共分离出 78株细菌,优势菌属是 Pantoea,占菌株总数的 55.13%。马铃薯根际土采用 R2A培养基(27株优势菌,13个菌属)分离的细菌多样性相对较好,阿须贝培养基(31株优势菌,9个菌属)次之,土壤浸提液培养基(1株优势菌,1个菌属)最少;马铃薯叶片内生菌用阿须贝培养基(19株优势菌,3个菌属)分离的细菌多样性较差。采用 Salkowski比色液显色法定量测定菌株产 IAA能力,结果表明有 58株细菌具有分泌 IAA的能力,占测定菌株总数的 74.36%,从马铃薯根际土(42株优势菌,13个菌属)筛选的产 IAA菌数量及细菌多样性均高于马铃薯叶片内生菌(16株优势菌,2个菌属)。根据菌株产IAA能力强弱和分离部位及分离培养基的差异,选择 7株产 IAA菌进行促生特性和马铃薯盆栽幼苗促生能力等研究,结果显示:有 6株产 IAA菌具有 ACC脱氨酶能力,1株产 IAA菌具有溶解无机磷能力,2株产 IAA菌具有溶解有机磷能力;两轮温室促生实验结果显示,菌株 Pantoeasp.MLS-34-25对马铃薯幼苗具有明显的促生作用,是生产微生物肥料的潜在菌种。     

西北半干旱荒漠草原与耕地土壤可培养微生物多样性及分布特征比较

采用稀释涂布平板、分离培养和16S r DNA序列分析法对我国甘肃白银地区半干旱荒漠草原土壤可培养细菌、放线菌、真菌数量及群落分布特征进行了分析,比较了荒漠草原和耕地土壤微生物多样性。发现荒漠草原土壤可培养细菌、放线菌、真菌数量分别为1.23×106、0.19×106、0.18×106cfu·g-1,耕地三类微生物数量分别是3.03×106、0.53×106、0.05×106cfu·g-1。荒漠草原可培养细菌、放线菌数量明显低于耕地,而真菌数量高于耕地。从荒漠草原分离出14株细菌,分别属于γ-变形菌纲(γ-Proteobacteria)噬冷杆菌属(Psychrobacter),放线菌门(Actinobacteria)皮球菌属(Kytococcus),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus)、亮氨酸芽孢杆菌属(Lysinibacillus)、土壤芽孢杆菌属(Solibacillus)、气球菌属(Aerococcus),优势菌为芽孢杆菌属和噬冷杆菌属。耕地分离出可培养细菌19株,分别属于ɑ-变形菌纲(ɑ-Proteobacteri)根瘤菌属(Rhizobium)、中华根瘤菌属(Sinorhizobium),γ-变形菌纲(γ-Proteobacteria)假单胞菌属(Pseudomonas),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus),放线菌门(Actinobacteria)微杆菌属(Microbacterium)、节杆菌属(Arthrobacter)、微球菌属(Micrococcus)、考克氏菌属(Kocuria),以放线菌门细菌为主(占57.9%)。从荒漠草原分离放线菌共8株,分别属于链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)、间孢囊菌属(Intrasporangium),而耕地主要为链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)。荒漠草原真菌主要是交链孢霉属(Alternaria)、芽枝霉属(Cladosporium),耕地土壤真菌包括青霉属(Penicillium)、交链孢霉属(Alternaria)、曲霉属(Aspergillus)、毛霉属(Mucor)、链孢霉属(Coniothecium)。试验结果表明,荒漠草原与耕地土壤微生物都具有较丰富的多样性,但微生物群落结构存在一定差异,同一区域不同深度土壤中微生物数量和种类也存在差异,耕地土壤微生物多样性明显高于荒漠草原。     

甘蔗黑穗病菌拮抗细菌的筛选及鉴定

从甘蔗根际土壤及甘蔗不同组织内分离到的928个细菌菌株中，对甘蔗黑穗病菌有拮抗作用的细菌菌株有301个．占32．4％．其中拮抗能力强(拮抗带大于10mm)的菌株有18个，占1．9％。经在KBA培养基上培养．发现具有拮抗作用的细菌主要是荧光菌和非荧光菌中的白色菌群。在18个拮抗性强的菌株中，13个菌株来自甘蔗的茎、芽(生长点)，占72％；5个菌株来自根际土壤，占28％；12个菌株为革兰氏阳性细菌中的芽孢杆菌属；6个菌株为革兰氏阴性细菌，其中4个菌株分别为假单胞杆菌属、不动杆菌属、伯克氏菌属及沙雷氏菌属，其余2个菌株有待进一步鉴定。     

山西矿区复垦土壤中解磷细菌的筛选及鉴定

【目的】矿区复垦土壤贫瘠、 有效磷含量低。解磷细菌能够将有机磷和难溶性无机磷转化为可溶性磷,促进植物对磷素的利用。因此筛选和鉴定具有解磷能力的菌株,可为解决矿区生态恢复使用的微生物肥料提供菌种资源。【方法】采用平板分离法初筛菌株,得到D/d1.5的菌株,然后以磷酸钙为磷源,通过液体发酵试验复筛菌株,挑选出解磷率高于巨大芽孢杆菌(Bacillus megaterium)As1.223的菌株。以磷矿粉和卵磷脂为磷源,液体发酵试验测定菌株的解磷能力及磷酸酶活性。进行菌株的生长试验以测定菌株温度适宜性、 耐盐性及耐酸碱性。通过形态学、 基因序列分析及脂肪酸组成分析综合进行菌株鉴定。 菌落形态观察用营养琼脂平板培养基培养;菌体形态即细胞形态及其大小采用扫描电镜观察;基因序列分析采用16S rDNA序列测定,基因在线比对采用EzTaxon数据库;使用美国MIDI公司的Sherolock全自动细菌鉴定系统对菌株进行脂肪酸组成分析。【结果】利用无机磷和有机磷平板培养基,从山西省矿区复垦区土壤样品中筛选出19株解磷微生物,其中D/d1.5的有7株。在以磷酸钙为磷源的液体培养试验中,4株菌的解磷率高于巨大芽孢杆菌As1.223,解磷率为7.89%～12.61%,最高的为菌株Y14。4株菌对磷矿粉的解磷率为0.81%～1.21%,最高的为菌株Y14。在以卵磷脂为磷源的液体培养试验中,4株菌的解磷率与酸性磷酸酶活性分别为1.79%～3.07%和24.3～28.4U/L,均高于巨大芽孢杆菌As1.223; 碱性磷酸酶活性为11.9～50.2U/L;菌株Y14的解磷率与磷酸酶活性均最高。4株菌均有较强的环境适应能力,以Y14的适应性最强。H22、 Y11和Y34与假单胞菌属(Pseudomonas sp.)同源性在99%以上,Y14与泛菌属(Pantoea sp.)有99.79%的同源性; H22、 Y11和Y34的细胞脂肪酸组成特征峰与假单胞菌属(Pseudomonas sp.)相一致,Y14与泛菌属(Pantoea sp.)相一致;H22、 Y11和Y34被鉴定为假单胞菌(Pseudomonas sp.),Y14为泛菌属(Pantoea sp.)。【结论】分离、 筛选到4株高效解磷菌,对于磷酸钙和卵磷脂的解磷率均高于巨大芽孢杆菌As1.223。4株菌分别隶属于假单胞菌属(Pseudomonas sp.)和泛菌属(Pantoea sp.)。菌株Y14无机磷与有机磷平板的D/d值分别为3.28与1.59,降解磷酸钙、 磷矿粉、 卵磷脂的解磷率分别为12.61%、 1.21%、 3.07%,酸性与碱性磷酸酶活性分别为28.4 U/L和50.2 U/L,均为4株菌里最高的,且环境适应能力最强,生长温度为20～60℃,能耐受pH 4～11的酸碱梯度和2%～7%的盐分梯度,Y14被鉴定为泛菌属(Pantoea sp.)。4株菌均具有良好的解磷能力及较强的环境适应能力,可望进一步研发成为微生物肥料生产菌种。综合D/d值、 解磷率、 磷酸酶活性和生长试验,本试验最终确定适合山西矿区复垦农田推广的高效解磷菌菌株为Y14。     

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。     

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)。     

Effects of Plant Growth Promoting Rhizobacteria on Yield and Some Fruit Properties of Strawberry

Effects of plant growth promoting rhizobacteria (PGPR) [(Pseudomonas BA-8 (biological control agent), Bacillus OSU-142 (N₂-fixing), and Bacillus M-3 (N₂-fixing and phosphate solubilizing)] on yield and some fruit properties of strawberry cultivar ‘Selva’ in the province of Erzurum, Turkey in 2002–2003. Foliar + root application of PGPR strains significantly increased yield per plant as compared with the control. Root application of PGPR strains significantly increased total soluble solids, total sugar and reduced sugar, but decreased titratable acidity. It was also determined that bacteria applications have no important effect on the average fruit weight and pH. The results of this study suggested that Pseudomonas BA-8, Bacillus OSU-142 and Bacillus M-3 have potential for increasing yield in strawberry plant.     

Response of alfalfa (Medicago sativa L.) to single and mixed inoculation with phosphate-solubilizing bacteria and Sinorhizobium meliloti

The objectives of this work were to phenotypically and genetically characterize alfalfa rhizosphere bacteria and to evaluate the effect of single or mixed inoculation upon nodulation and biological nitrogen fixation. Thirty-two strains showed tricalcium phosphate solubilization ability, and two of them caused bigger or equal solubilization halos than the control strain P. putida SP22. The comparison of the 16S ribosomal DNA sequences indicated that these strains are phylogenetically related to Bacillus spp. and Pseudomonas spp. A beneficial effect of both isolates on alfalfa growth was observed in coinoculation assays. Pseudomonas sp. FM7d caused a significant increase in root and shoot dry weight, length, and surface area of roots, number, and symbiotic properties of alfalfa plants. The plants coinoculated with Sinorhizobium meliloti B399 and the Bacillus sp. M7c showed significant increases in the measured parameters. Our results indicating that strains Pseudomonas sp. FM7d and Bacillus sp. M7c can be considered for the formulation of new inoculants.     

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.     

Evaluation of the effects of spores and their heat-treated residues from different Bacillus strains on the initial growth of rice plants

This study evaluated the effects of the spores and their heat-treated residues from different strains of Bacillus species (B. pumilus, B. altitudinis, and B. megaterium) on the early growth of paddy rice cultivars, including Hitomebore (short-grain japonica rice), Takanari (high-yielding indica rice), and two new lines, TULK-143-6 and LTAT-29. The spores of seven Bacillus strains positively affected Hitomebore root growth, while, the root volume of TULK-143-6 with inoculation of B. pumilus TUAT1 and JM52, and root length and root surface area of LTAT-29 with inoculation of B. megaterium MAFF301694 were increased significantly. In contrast with Hitomebore, Takanari root growth was significantly inhibited by the spores of six Bacillus strains. Surprisingly, inoculations with the spore residues from all tested Bacillus strains increased the root dry weight of Hitomebore, with the effects of four bacterial strains being significant. Furthermore, there were more Bacillus spores than vegetative cells at different time points during the initial rice growth stage, and most plant samples mainly consisted of Bacillus spores. Thus, the spores of Bacillus species likely promote rice root development.     

Genetic variability of rhizobacteria from wild populations of four Lupinus species based on PCR-RAPDs§

The metabolic capacities of rhizosphere bacteria can depend on intraspecific genetic variability at strain level. We sampled bacteria from the rhizosphere of three populations of four different Lupinus species at two growth stages (flowering: GS1 and fruiting: GS2). Isolates were identified to the genus level by classical biochemical tests. The most abundant genera found were Bacillus, Aureobacterium, Cellulomonas, Pseudomonas, and Arthrobacter. Genetic divergence of rhizobacteria was tested by PCR-RAPDs. The genetic distances were low, with mean values of 37 % for Bacillus, 25 % for Aureobacterium, 46 % for Cellulomonas, 16 % for Pseudomonas, and 23 % for Arthrobacter. Aureobacterium, the most abundant genus, predominated in the rhizosphere of all populations and at both growth stages (GS1 and GS2) of L. angustifolius. The Aureobacterium strains consisted of 11 groups with 90 % similarity indexes. The cluster analysis of these groups shows that strains isolated from different lupin species and sampling times have extraordinary low diversity indexes, or are even identical. This fact, together with the low genetic distance detected in the rhizosphere, reveals a clear specificity in the plant-bacteria interaction. This specificity could be related with several aspects of plant physiology.     

砒砂岩中植物促生芽孢杆菌的筛选及其对土壤的改良作用

[目的]筛选砒砂岩土壤中植物促生芽孢杆菌,为微生物强化植物改良砒砂岩土壤的科学设想提供资源和技术基础。[方法]以产生植物激素IAA(indole acetic acid)、铁载体和生物膜为筛选指标,从内蒙古砒砂岩区土壤和植物样品中筛选植物促生芽孢杆菌,采用盆栽试验探究植物促生芽孢杆菌改良砒砂岩土壤特性和促进苜蓿和黑麦草生长的作用。[结果]筛选到的12株芽孢杆菌产生IAA、铁载体和生物膜的能力不同,分别属于Bacillus halotolerans,B.atrophaeus,B.siamensis和B.zhangzhouensis种群。与对照相比,B.halotolerans P75能够显著增加砒砂岩土壤的有机质含量(24.7%)、速效磷含量(11.9%)和速效钾含量(21.0%)等养分指标,每1 g土壤可培养细菌达到7.4 lg CFU,土壤蔗糖酶活性显著增强(58.8%)。接种B.halotolerans P75后,在砒砂岩土壤中生长的苜蓿和黑麦草干重增加22.3%～81.5%,[结论]从生长于砒砂岩土壤的苜蓿根内筛选到B.halotolerans P75,可提高砒砂岩土壤肥力,促进苜蓿和黑麦草生长,具有强化植物改良砒砂岩土壤的潜力。     

Effects of Culture Filtrates of Rhizobacteria Isolated from Wild Lupine on Germination,Growth, and Biological Nitrogen Fixation of Lupine Seedlings

Abstract

In order to select potential Plant Growth Promoting Rhizobacteria (PGPRs), a selection of strains from the predominant genera in the rhizosphere of four lupine species, based on genetic divergence criteria, carried out in a previous study, yielded 11 Aureobacterium (Aur), four Cellulomonas (Cell), two Arthrobacter (Arth), two Pseudomonas (Ps), and six Bacillus (Bc) strains. Cell‐free culture filtrates of each bacterium were assayed for effects on germination, growth, and biological nitrogen fixation (BNF) of Lupinus albus L. cv. Multolupa seeds or seedlings. Four (Aur 6, Aur 9, Aur 11, and Cell 1) of the twenty‐five strains assayed promoted germination. Aureobacterium 6 and Aur 9 also increased root surface, total nitrogen content, and BNF. As a result of the screening, and considering all the variables studied, Aur 6 can be considered a plant growth promoting rhizobacterium and is suitable for further field trials in other plants and in different production systems.     

Antifungal and Plant Growth Promoting Activities of Indigenous Rhizobacteria Isolated from Maize (Zea mays L.) Rhizosphere

Plant growth promoting rhizobacteria (PGPR) enhance the plant growth directly by assisting in nutrient acquisition and modulating plant hormone levels, or indirectly by decreasing the inhibitory effects of various pathogens. The aim of this study was to select effective PGPR from a series of indigenous bacterial isolates by plant growth promotion and antifungal activity assays. This study confirmed that most of the isolates from maize rhizosphere were positive for PGPR properties by in vitro tests. Azotobacter and Bacillus isolates were better phosphate solubilizers and producers of lytic enzymes, hydrocyanic acid (HCN), and siderophores than Pseudomonas. Production of indole-3-acetic acid (IAA) and antifungal activity were the highest in Azotobacter, followed by Bacillus and Pseudomonas. The most effective Azotobacter isolates (Azt₃, Azt₆, Azt₁₂) and Bacillus isolates (Bac_10, Bac₁₆) could be used as PGPR agents for improving maize productivity. Further selection of isolates will be necessary to determine their efficiency in different soils.     

Seed inoculation with <Emphasis Type="Italic">Bacillus</Emphasis> spp. improves seedling vigour in oil-seed plant <Emphasis Type="Italic">Jatropha curcas</Emphasis> L.

Jatropha (Jatropha curcas L.) is a non-edible oil-seed plant with adaptability to marginal semi-arid lands and wastelands. The Indian Government is promoting jatropha to reduce dependence on the crude oil and to achieve energy independence by the year 2012, under the National Biodiesel Mission. Selected strains of Bacillus spp., either supplemented with or without chitin, were tested for their ability to promote growth of jatropha seedlings in pot culture studies. The strains supported growth of jatropha seedlings up to 42 days after sowing. Among all strains, Bacillus pumilus (IM-3) supplemented with chitin showed over all plant growth promotion effect resulting in enhanced shoot length (113%), dry shoot mass (360%), dry root mass (467%), dry total plant mass (346%), leaf area (256%), and chlorophyll content (74%) over control. Treating seeds with strain IM-3 without chitin resulted in enhanced dry shoot mass (473%), dry total plant mass (407%), and chlorophyll content (82%). However, Bacillus polymyxa (KRU-22) with chitin supported maximum root length (143%). Either strain IM-3 alone or in combination with other promising strains could be promoted further for enhanced initial seedling growth of jatropha.     

Isolation and characterization of phosphate-solubilizing bacteria from walnut and their effect on growth and phosphorus mobilization

The objectives of this work were to isolate and characterize walnut phosphate-solubilizing bacteria (PSB) and to evaluate the effect of inoculation with the selected PSB stains to walnut seedlings fertilized with or without insoluble phosphate. Thirty-four PSB strains were isolated and identified under the genera Pseudomonas, Stenotrophomonas, Bacillus, Cupriavidus, Agrobacterium, Acinetobacter, Arthrobacter, Pantoea, and Rhodococcus through a comparison of the 16S ribosomal DNA sequences. All isolated PSB strains could solubilize tricalcium phosphate (TCP) in solid and liquid media. Phosphate-solubilizing activity of these strains was associated with a drop in the pH of medium. A significantly negative linear correlation was found between culture pH and phosphorus (P) solubilized from inorganic phosphate. Three isolates Pseudomonas chlororaphis (W24), Bacillus cereus (W9), and Pseudomonas fluorescens (W12) were selected for shade house assays because of their higher phosphate-solubilizing abilities. Under shade house conditions, application of W24 or W12 remarkably improved plant height, shoot and root dry weight, and P and nitrogen (N) uptake of walnut seedlings. These increases were higher on combined inoculation of PSB with TCP addition. The most pronounced beneficial effect on growth of walnut plants was observed in the co-inoculation of the three PSB strains with TCP addition. In comparison, the isolate of W9 failed to increase available soil P, nutrient levels in plants, or to promote plant growth, suggesting that more insoluble phosphate compounds than tricalcium phosphate should be used as substrates to assess the phosphate-solubilizing ability of PSB under greenhouse conditions. The present results indicated that strains P. chlororaphis or P. fluorescens could be considered for the formulation of new inoculants of walnut, even of more woody plants.