细菌铁载体拮抗植物病原真菌及促生作用研究进展

铁载体是微生物在缺铁条件下分泌的小分子有机化合物,以获取铁元素维持其生长。细菌分泌的铁载体在拮抗植物病原菌和促进植物生长方面具有重要作用。本文总结了细菌铁载体拮抗植物病原真菌的营养和生态位竞争、诱导植物诱导性系统抗性、扰乱病原菌铁稳态的机制,以及促进植物生长的作用,以解释细菌分泌的铁载体在多功能微生物菌剂研制中的重要作用。     

达乌里胡枝子产铁载体根瘤内生细菌的鉴定及其促生抗逆效应

【背景】根瘤是植物-微生物共生的特殊结构,蕴含丰富的微生物资源。产铁载体根瘤内生细菌对促进结瘤固氮具有重要作用,但目前关于其是否对植物具有促生作用尚不明确。【目的】筛选鉴定达乌里胡枝子(Lespedeza daurica)根瘤产铁载体内生细菌,明确其促生作用,为丰富植物促生菌资源提供新的途径。【方法】利用铬天青(chrome azurol S, CAS)培养基初筛产铁载体根瘤内生细菌,对目的菌株进行鉴定;通过液体发酵试验确定其产铁载体能力;利用促生功能培养基测定其他促生特性;利用不同盐浓度(0.01%-9.00%)、不同pH值(3.0-12.0)的酵母甘露醇(yeast mannitol,YMA)液体培养基和不同培养温度(4-60℃)确定其抗逆特性;通过盆栽试验初步验证其对植物的促生作用。【结果】48株根瘤内生细菌可产铁载体,其中9株为高产铁载体细菌,分别为TG4、TG9、TG13、TG15、TG28、TG32、TG38、TG76和TG78,铁载体相对表达量为26%-93%;经鉴定,除了TG28为巨大普里斯特氏菌(Priestiamegaterium),其余8株菌均为耐寒短杆菌(Bre...     

棉花黄萎病拮抗细菌产铁载体测定及其对抑菌活性的影响

【背景】棉花是新疆农业的支柱产业,棉花黄萎病严重影响棉花产量和纤维品质,有效的防治途径是目前解决的首要任务。【目的】探究BacillusvelezensisBHZ-29、Bacillusatrophaeus SHZ-24、Bacillus subtilis SHT-15、Bacillus vanillea SMT-24菌株产铁载体能力及其对抑菌活性的影响。【方法】MKB-CAS双层平板检测铁载体,Arnow法与高氯酸铁法鉴定铁载体类型,CAS染液定量铁载体,琼脂扩散法测定低铁条件下铁载体对抑菌物质的影响。【结果】MKB-CAS固体平板菌体周围出现橙紫色显色反应,Arnow实验4株拮抗菌反应液颜色为红色,高氯酸铁实验反应液颜色为黄色;BHZ-29、SHZ-24、SHT-15、SMT-24菌株铁载体定量活性单位最大值分别为8.27%±0.61%、31.80%±2.06%、16.06%±3.61%、15.53%±0.51%,吸光度值As/Ar比值最小分别为0.917±0.092、0.682±0.021、0.845±0.005、0.846±0.008。BHZ-29、SHZ-24、SMT-24菌株含铁离子浓度0、10、20μmol/L 3组处理发酵无菌滤液均未出现抑菌圈,SHT-15菌株3组处理出现抑菌圈,抑菌圈直径大小无差异性。【结论】4株拮抗菌株均产铁载体,铁载体类型为儿茶酚型,4株拮抗菌株整体产铁载体能力较低,最高达31.80%±2.06%,产铁能力As/Ar比值高于0.682时,BHZ-29、SHZ-24、SMT-24菌株含铁离子浓度0、10、20μmol/L 3组处理发酵无菌滤液对抑菌活性无影响,SHT-15菌株3组不同铁离子浓度处理对抑菌物质影响差异不显著。     

盐胁迫对氢氧化细菌WMQ7浸种小麦萌发和生长的影响

从紫花苜蓿根际土壤分离得到的一株含有ACC脱氨酶以及铁载体的氢氧化细菌WMQ7,通过皿内试验,考查其在不同盐浓度下对小麦‘陕优225’的促生效应。结果表明,低盐胁迫(0.4%NaCl)下,用氢氧化细菌WMQ7菌悬液浸种的小麦发芽粒数、发芽率、发芽势、发芽指数、胚芽、胚根长有明显的增高;同时脯氨酸积累速度与对照组相比显著减缓,对照组脯氨酸含量于胁迫第11天时已达1 383μg.mL-1,高出接菌组3.8倍;丙二醛含量与对照组相比减少了69.28%,过氧化物酶活性显著升高,高出对照组44.44%。而高盐胁迫下(0.8%NaCl),与对照组相比,接菌组对小麦萌发指标有一定影响,丙二醛含量变化不显著,但脯氨酸含量及过氧化物酶活性变化显著。研究表明,氢氧化细菌WMQ7浸种小麦处理对低盐胁迫的伤害有一定的缓解作用,但在高盐胁迫下却无明显效果。     

适合高产铁载体细菌筛选、检测体系的改进与探析

采用pH6．8的磷酸缓冲液代替通用CAS固体检测培养基中的MM9缓冲体系,不加哌嗪二乙醇磺酸,得到颜色稳定、检测效果明显、配制方便,适合高产铁载体细菌筛选与检测的新型检测平板。根据CAS检测液连续吸收光谱比较分析结果,将CAS液体定量检测体系中的检测波长由630nm改为680nm,可以克服OD630读数偏高、易受干扰的问题,而且OD680与铁载体浓度线性关系不变,但采用OD680检测不同细菌产铁载体能力的差异更为明显,因此提高了CAS定量检测的灵敏度,更适合高产铁载体细菌筛选与检测。     

土壤产铁载体细菌的筛选及其对铁氧化物的活化与利用

【背景】土壤中铁主要以难溶态铁氧化物的形式存在,有效性较低,产铁载体细菌对铁氧化物的活化是提高铁利用效率的有效途径。【目的】从林木土壤筛选产铁载体细菌,并观察菌株对难溶性铁氧化物的利用效应,可为土壤微生物资源开发及其在养分调控中的作用提供理论依据。【方法】通过CAS检测法从林木根系附近表层土壤中分离产铁载体细菌,借助生物培养实验分析温度和pH对微生物生长和铁载体产生的影响,通过振荡平衡实验,探究细菌产铁载体对铁氧化物的活化效应。【结果】通过CAS检测法从林木根系附近表层土壤中分离得到12株产铁载体细菌,16S rRNA基因扩增子测序初步鉴定结果显示筛选细菌均为假单胞菌属。选取铁载体产生能力和生长活性较高的两株细菌ARSB02和CNRSB01作为重点研究对象,结果显示,不同条件下CNRSB01的生物量和铁载体产量均高于菌株ARSB02,22 h时菌株ARSB02和CNRSB01的铁载体活性单位分别达到67.07%和84.60%。pH 5.0-8.0的范围内两株细菌可以保持较好的铁载体产生能力,菌株ARSB02和CNRSB01在pH7.0时铁载体产生能力最强,铁载体活性单位分别达到38.9...     

CAS平板覆盖法检测氢氧化细菌铁载体

【目的】用CAS平板覆盖法检测氢氧化细菌铁载体,解决通用CAS琼脂平板法中十六烷基三甲基溴化铵对真菌和某些细菌的生长抑制问题。【方法】将改良的CAS检测培养基覆盖在长满菌落的无铁培养基上,生长抑制问题因微生物未与十六烷基三甲基溴化铵直接接触而解决。【结果】3株氢氧化细菌SDW-5、SDW-9和AaP-13均能产生单菌落,加入CAS检测培养基1 h后,菌落周围产生明显的铁载体晕圈。【结论】本方法成功解决了生长抑制问题,可以作为检测微生物铁载体的通用方法。     

小麦根圈细菌铁载体的检测

本文报道微生物铁载体定性与定量测定的方法,以及小麦根圈细菌铁载体的室内检测结果。２２株小麦根圈耐寒细菌和２株固氮菌在定性检测平板上产生铁载体,另有３株固氮菌不产。定量测定结果表明：３６株水溶性色素产生菌合成铁载体的量在０．７１５－０．１０６（Ａ／Ａｒ）之间,其中属于荧光假单胞菌群的２５个菌株铁载体产量的测定值（Ａ／Ａｒ）在０．１８４－０．１０６之间,达极高量水平。     

一株产铁载体内生细菌对尖孢镰刀菌的拮抗作用

通过改良蔗糖-天冬氨酸培养基筛选到一株产铁载体的内生细菌HS-4,测定了该菌在不同铁离子浓度下对棉花枯萎病致病菌尖孢镰刀菌(Fusarium oxysporum)的抑菌效果,并结合形态、生理生化、16S rDNA序列同源性和系统发育分析对菌株进行鉴定.结果表明:内生细菌HS-4在MSA培养基中产生荧光型铁载体,其铁载体相对含量为80%.该铁载体在低铁条件下对F.oxysporum具有抑制作用.内生细菌HS-4初步鉴定为萎缩芽孢杆菌(Ba-cillus atrophaeus).     

耐盐碱细菌DQSA1的分离鉴定及盐碱胁迫下对绿豆的促生作用

[背景]近年来,大庆地区土壤盐碱化程度逐渐加剧,而微生物改良盐碱土是当今的研究热点.[目的]从大庆盐碱土中筛选出耐盐碱菌株,验证其促生作用,为改良大庆盐碱土壤提供微生物资源.[方法]采用筛选培养基从大庆盐碱土中获得耐盐碱促生菌,对其进行形态学观察、生理生化和16S rRNA基因鉴定,并测试菌株在盐碱胁迫下对绿豆植株和土...     

Isolation and characterization of plant growth promoting endophytic diazotrophic bacteria from Korean rice cultivars

We have isolated 576 endophytic bacteria from the leaves, stems, and roots of 10 rice cultivars and identified 12 of them as diazotrophic bacteria using a specific primer set of nif gene. Through 16S rDNA sequence analysis, nifH genes were confirmed in the two species of Penibacillus, three species of Microbacterium, three Bacillus species, and four species of Klebsiella. Rice seeds treated with these plant growth-promoting bacteria (PGPB) showed improved plant growth, increased height and dry weight and antagonistic effects against fungal pathogens. In addition, auxin and siderophore producing ability, and phosphate solubilizing activity were studied for the possible mechanisms of plant growth promotion. Among 12 isolates tested, 10 strains have shown higher auxin producing activity, 6 isolates were confirmed as strains with high siderophore producing activity while 4 isolates turned out to have high phosphate-solubilizing activity. These results strongly suggest that the endophytic diazotrophic bacteria characterized in this study could be successfully used to promote plant growth and inducing fungal resistance in plants.     

一株海洋放线菌的鉴定及其促生作用机理

【背景】海洋放线菌BM-2是本实验室从连云港海域分离得到的一株具有抗菌和促生作用的优良菌株,具有良好的开发应用前景。【目的】明确海洋放线菌BM-2的分类地位,揭示该菌株的促生作用机理,为菌株的开发应用提供理论依据。【方法】通过形态观察、生理生化特性和16S rRNA基因序列分析,对海洋放线菌BM-2菌株进行种属鉴定;采用透明圈法、平板划线法测定BM-2菌株解磷、解钾作用、固氮作用和产植酸酶、1-氨基环丙烷-1-羧基(1-aminocyclopropane-1-carboxylate,ACC)脱氨酶的能力;运用沙尔科夫斯基反应(Salkowski法)和铬天青(chromeazurol S,CAS)法分别测定菌株产吲哚乙酸(indole acetic acid,IAA)和产铁载体的能力。【结果】培养特征、菌落形态观察及生理生化试验结果表明,BM-2菌株符合链霉菌属(Streptomyces)的特征,16SrRNA基因序列与GenBank中栗褐链霉菌(Streptomycesbadius)的序列相似性为99.72%;BM-2菌株具有固氮和解有机磷活性,能够产生ACC脱氨酶、铁载体和IAA。【...     

有机磷降解菌的筛选及其促生特性

【目的】从环境中筛选高效有机磷降解菌及研究其促生机制。【方法】利用蒙金娜有机磷培养基筛选有机磷降解菌,用生化实验对其促生特性进行研究,且通过盆栽实验筛选对黄瓜苗有促生作用的高效有机磷降解菌。【结果】从草坪根周筛选到35株有机磷降解菌,选择5株代表性菌进行黄瓜盆栽实验。结果表明G3-6有机磷降解能力最强,溶磷圈直径(HD)与菌落直径(CD)比值为3.28,且对黄瓜苗的促生效果优于其他菌株。与CK相比,G3-6可提高黄瓜苗鲜重71.53%、干重69.78%和株高33.55%;与阳性对照枯草芽孢杆菌F-H-1相比,G3-6可提高黄瓜苗鲜重2.52%、干重21.14%和株高8.27%。相关分析结果表明降解有机磷能力在促进植物生长过程中可能发挥着比其他功能更重要的作用。16S r RNA序列分析初步鉴定G3-6为假单胞菌属。【结论】假单胞菌G3-6除具有较强的有机磷降解、分泌IAA和铁载体能力,对黄瓜苗也有较好的促生作用,是1株潜在的具有广阔市场应用价值的高效促生菌。     

Mixed planting with a leguminous plant outperforms bacteria in promoting growth of a metal remediating plant through histidine synthesis

The effectiveness of plant growth promoting bacteria (PGPB) in improving metal phytoremediation is still limited by stunted plant growth under high soil metal concentrations. Meanwhile, mixed planting with leguminous plants is known to improve yield in nutrient deficient soils but the use of a metal tolerant legume to enhance metal tolerance of a phytoremediator has not been explored. We compared the use of Pseudomonas brassicacearum, Rhizobium leguminosarum, and the metal tolerant leguminous plant Vicia sativa to promote the growth of Brassica juncea in soil contaminated with 400 mg Zn kg^–1, and used synchrotron based microfocus X-ray absorption spectroscopy to probe Zn speciation in plant roots. B. juncea grew better when planted with V. sativa than when inoculated with PGPB. By combining PGPB with mixed planting, B. juncea recovered full growth while also achieving soil remediation efficiency of >75%, the maximum ever demonstrated for B. juncea. μXANES analysis of V. sativa suggested possible root exudation of the Zn chelates histidine and cysteine were responsible for reducing Zn toxicity. We propose the exploration of a legume-assisted-phytoremediation system as a more effective alternative to PGPB for Zn bioremediation.     

Role of halotolerant phosphate‐solubilising bacteria on growth promotion of peanut (Arachis hypogaea) under saline soil

Soil salinity is a major abiotic stress that limits plant growth, and inoculating plant growth‐promoting rhizobacteria is a well‐known strategy to reduce stressors under adverse soil conditions. This study was conducted to assess the effect of halotolerant phosphate‐solubilising bacteria (PSB) on protecting peanut against salt stress. Four candidate strains: Bacillus megaterium (YM13), Enterobacter sp. (YM14), Providencia rettgeri (TPM23) and Ensifer adhaeren (TPMX5) showed strong tolerance to NaCl and high phosphate‐solubilising ability even at a NaCl concentration of 1.4 M. In addition, all four strains demonstrated variable levels of phosphate solubilisation activity in the presence of various carbon and nitrogen sources, indicating high phosphate‐solubilising efficacy. Germination and radicle length of peanut seedlings increased with inoculation of PSB under both control and saline conditions. Statistically significant increase in the root length (range: 25.71–49%), stem length (19–48%), number of leaves (12.5–37%) and root/shoot biomass were observed. This could be attributed to plant hormones (i.e., indole acetic acid [IAA], abscisic acid [ABA] and gibberellic acid [GA3]) and successful root colonisation by bacterial inoculants. Root colonisation was positively correlated to plant growth and shown to be influenced by soil conditions. In addition, the PSB also improved the levels of available P in soil. The most pronounced beneficial effect on the growth of peanut plants and soil available P content was observed in the inoculation of the PSB isolates with Ca₃(PO₄)₂ addition. This is the first report that describes Providencia rettgeri as a plant growth‐promoting bacterium that may be utilised to alleviate the negative effects of salt stress on peanut plants. This bacterial species may thus be potentially used as a biofertilizer for sustaining the growth of peanut in salt‐stressed soil and in mitigating soil stress conditions.     

Experimental and mathematical simulation of plant growth promoting rhizobacteria and plant interaction under cadmium stress

Bacterial inoculants of the commercially available plant growth promoting rhizobacteria (PGPR) Arthrobacter mysorens 7, Flavobacterium sp. L30, and Klebsiella mobilis CIAM 880 were selected to obtain ecologically safe barley crop production on cadmium (Cd) polluted soils. All the PGPR immobilized 24–68% soluble cadmium from soil suspension. A. mysorens 7 and K. mobilis CIAM 880 were highly resistant to Cd and grew in up to 1 and 3 mmol CdCl₂ on DAS medium respectively. All PGPR were able to fix nitrogen (276–1014 nmol mg^–1 bacterial DW) and to produce indole acetic acid (IAA) (126–330 nmol mg^–1 bacterial DW) or ethylene (4.6–13.5 nmol bacterial DW). All the PGPR actively colonized barley root system and rhizosphere and significantly stimulated root elongation of barley seedlings (up to 25%), growing on soil containing 5 or 15 mg Cd kg^–1 of soil. Created in the simulation mathematical model confirms our hypothesis that PGPR beneficial effect on barley growing under Cd-stress is a complex process. One of mechanisms underlying this effect might be increase of bacterial migration from rhizoplane to rhizosphere, where PGPR bind soluble free Cd ions in biologically unavailable complex forms. Among the studied PGPR K. mobilis CIAM 880 was the most effective inoculant. Inoculation with K. mobilis CIAM 880 of barley plants growing on Cd contaminated soil (5 mg Cd kg^–1 of soil) under field conditions increased by 120% grain yield and 2-fold decreased Cd content in barley grain. The results suggest that the using K. mobilis CIAM 880 is an effective way to increase the plant yield on poor and polluted areas.     

Synthesis and biological activity of saccharide based lipophilic siderophore mimetics as potential growth promoters for mycobacteria

Siderophores based on sugar backbones substituted at the 2,3,4- or 2,3,6 positions with hydroxamic or retro-hydroxamic acid chelating units were synthesized and characterized. The alkyl terminus of the iron-coordinating side chain units facilitate lipophilic interactions. Iron coordination properties and complex stability were investigated by ESI-MS and the CAS-Test. The results were correlated to structure activity relationships determined by microbial growth promotion studies under iron limited conditions using wild type strains and iron transport mutants of Mycobacterium smegmatis.     

盐胁迫下解钾菌对枸杞幼苗的促生效应

以'宁杞1号'枸杞为研究对象,采用单因素随机区组试验设计,研究不添加NaCl和菌剂(CK)、100 mmol·L-1 NaCl胁迫(NaCl)、100 mmol·L-1 NaCl胁迫下添加解钾菌KSB-GY01单菌液(NaCl-M1)、KSBGY02单菌液(NaCl-M2)及其混合菌液(NaCl-M3)对枸杞幼苗叶片叶...     

Gene expression profiling through microarray analysis in Arabidopsis thaliana colonized by Pseudomonas putida MTCC5279, a plant growth promoting rhizobacterium

Plant growth promotion is a multigenic process under the influence of many factors; therefore an understanding of these processes and the functions regulated may have profound implications. Present study reports microarray analysis of Arabidopsis thaliana plants inoculated with Pseudomonas putida MTCC5279 (MTCC5279) which resulted in significant increase in growth traits as compared with non-inoculated control. The gene expression changes, represented by oligonucleotide array (24652 genes) have been studied to gain insight into MTCC5279 assisted plant growth promotion in Arabidopsis thaliana. MTCC5279 induced upregulated Arabidopsis thaliana genes were found to be involved in maintenance of genome integrity (At5g20850), growth hormone (At3g23890 and At4g36110), amino acid synthesis (At5g63890), abcissic acid (ABA) signaling and ethylene suppression (At2g29090, At5g17850), Ca⁺² dependent signaling (At3g57530) and induction of induced systemic resistance (At2g46370, At2g44840). The genes At3g32920 and At2g15890 which are suggested to act early in petal, stamen and embryonic development are among the downregulated genes. We report for the first time MTCC5279 assisted repression of At3g32920, a putative DNA repair protein involved in recombination and DNA strand transfer in a process of rapid meiotic and mitotic division.     

Antagonistic control of microbial pathogens under iron limitations by siderophore producing bacteria in a chemostat setup

Certain bacteria develop iron chelation mechanisms that allow them to scavenge dissolved iron from the environment and to make it unavailable to competitors. This is achieved by producing siderophores that bind the iron which is later liberated internally in the cell. Under conditions of iron limitation, siderophore producing bacteria have therefore an antagonistic growth advantage over other species. This has been observed in particular in agricultural and aquacultural systems, as well as in food microbiology. We investigate here the possibility of a probiotic biocontrol strategy to eradicate a well established, often pathogenic, non-chelating population by supplementing the system with generally regarded as safe siderophore producing bacteria. Set in a chemostat setup, our modeling and simulation studies suggest that this is indeed possible in a finite time treatment.