Ion-exchange aspects of toxic metal uptake by Indian mustard

Uptake of lead (Pb), copper (Cu), zinc (Zn), and cadmium (Cd) as +2 ions by excised roots of Indian mustard was demonstrated to be an ion-exchange process with existing Ca or protons released to the solution. This initial reaction at the root-aqueous interface is important in the uptake of these toxic metals from contaminated soil. Ethylene diamine tetraacetic acid (EDTA)-amended soil for phytoremediation has Pb in anionic form as [Pb-EDTA]2-, which was not taken up by excised roots. In nonliving B. juncea, Pb2+ was translocated from a solution through a cut stem to petiole and leaves much more quickly than anionic [Pb-EDTA]2-. However, in living plants [Pb-EDTA]2- was more quickly translocated from a solution through roots and petiole to leaves than Pb2+. The final amount of uptake on roots of the living plants was the same for both forms of Pb. The present results are important toward understanding the mechanism of phytoremediation of toxic metal-contaminated soil for two reasons: 1) the initial process, uptake of metal ions by roots, was shown to occur by cation exchange and 2) since [Pb-EDTA]2- was not sorbed by excised roots, other factors such as transpiration and active transport are important in applications using EDTA-amended soils contaminated by Pb.     

沙棘根瘤内生细菌中抑菌促生菌株的筛选和鉴定

【背景】植物内生细菌可产生具有抑菌和促生活性的物质,既能抑制植物病原菌对寄主植物的侵染,也能促进植物的生长。沙棘根瘤内生细菌是根瘤内除共生固氮的弗兰克氏菌外,与沙棘共生的一大类微生物。研究具有抑菌和促生活性的植物内生菌,可为微生物菌肥的研究提供理论基础。【目的】筛选具有优良抑菌和促生活性的沙棘根瘤内生细菌,初步研究其抑菌和促生活性,并对菌株进行鉴定。【方法】采用双层琼脂法、琼脂扩散法、双层平板对峙法、牛津杯法进行沙棘根瘤拮抗性内生细菌的筛选。选取抑菌活性较高的内生细菌,分别采用Salkowski比色法、ChromeAzurolS(CAS)平板检测法和钼锑抗比色法进行产吲哚乙酸、铁载体及溶磷能力的测定。采用发酵液灌根法测定沙棘根瘤内生细菌SR308对黄瓜促生作用的盆栽效果。通过形态和培养特征、生理生化试验及16S rRNA基因序列分析法对菌株TT201和SR308进行鉴定。【结果】从131株沙棘根瘤内生细菌中筛选出9株具有较强抑菌活性的内生细菌,其中菌株TT201抑菌性最佳、抑菌谱广;菌株SR308的促生活性最好,其发酵液对黄瓜的生长具有较强的促进作用。对具有较强抑菌和促生活性的菌株TT201和SR308进行鉴定的结果表明,菌株TT201为侧孢短芽孢杆菌(Brevibacilluslaterosporus),菌株SR308为蕈状芽孢杆菌(Bacillusmycoides)。【结论】获得2株具有优良抑菌和促生活性的沙棘根瘤内生细菌,为进一步开发微生物农药及菌肥提供了资源。     

Characterization of plant-growth promoting diazotrophic bacteria isolated from field grown Chinese cabbage under different fertilization conditions

Diazotrophic bacteria isolated from the rhizosphere of Chinese cabbage were assessed for other plant growth promoting characteristics viz., production of IAA, ethylene, ACC deaminase, phosphate solubilization, and gnotobiotic root elongation. Their effect on inoculation to Chinese cabbage was also observed under growth chamber conditions. A total of 19 strains that showed higher nitrogenase activity identified by 16S rRNA gene sequence analysis were found to be the members of the genera Pseudomonas and Agrobacterium belonging to α- and γ-Proteobacteria groups. These strains were also efficient in producing IAA and ACC deaminase though they produced low levels of ethylene and no phosphate solubilization. In addition, inoculation of selected diazotrophic bacterial strains significantly increased seedling length, dry weight, and total nitrogen when compared to uninoculated control. The colonization of crop plants by diazotrophic bacteria can be affected by many biotic and abiotic factors, and further studies are oriented towards investigating the factors that could influence the establishment of a selected bacterial community.     

西双版纳保护区植物根际细菌的筛选及其促生能力研究

【背景】西双版纳保护区具有丰富的生物多样性,而该区域植物根际细菌特别是放线菌及其促生能力相关报道较少。【目的】从西双版纳保护区根际土壤中筛选出植物根际促生菌,并检测其促生能力。【方法】采用5种不同培养基筛选出植物根际促生菌并通过16S rDNA序列分析进行分类学鉴定,运用Salkowski法测定菌株产IAA的能力,CAS法测定菌株产铁载体能力,钼锑抗显色法测定菌株的解磷能力,CMC-Na法测定菌株产纤维素酶能力和改良的Young法测定产淀粉酶能力,综合评价所得菌株的促生能力。【结果】从土样中分离纯化得到14株典型促生菌,经鉴定分别归属于链霉菌属(Streptomyces)、诺卡菌属(Nocardi)、杆菌属(Bacillus)、中华根瘤菌属(Ensifer)、中慢生根瘤菌属(Mesorhizobium)、固氮螺菌属(Azospirillum)和狭单胞菌属(Stenotrophomonas)。其中菌株B433产吲哚乙酸的能力在培养12 d时达到最大值9.23 mg/L;菌株B351、B453、B546这3株菌株产铁载体的能力较强,其Su80%,最高可达86.67%,强度为+++++;菌株B541的解磷能力最强,磷酸根的浓度达到9.79 mg/L;菌株B442综合产纤维素酶能力最强为31.86 U/mL;菌株B412淀粉酶活力为16.07 U/mL。【结论】西双版纳保护区植物根际土壤促生细菌种类丰富,且具有较强的广谱促生能力,有潜在的开发价值,本研究可为此地的微生物资源开发提供可靠的菌株资源依据。     

Isolation of phytase-producing bacteria from Himalayan soils and their effect on growth and phosphorus uptake of Indian mustard (Brassica juncea)

Phytase-producing bacteria (PPB) is being investigated as plant growth promoting rhizobacteria (PGPR) to improve the phosphorus (P) nutrition and growth of plants grown in soil with high phytate content. Phytate is dominant organic P forms in many soils and must be hydrolyzed to be available for plants. Indian mustard (Brassica juncea) is a plant with economic importance in agriculture and phytoremediation, therefore biotechnological tools to improve growth and environmental stress tolerance are needed. In this study, we isolated and characterized PPB from Himalayan soils and evaluated their effect on growth and P uptake by B. juncea under greenhouse conditions. Sixty five PPB were isolated and based on phytate hydrolysis, three efficient PPB were chosen and identified as Acromobacter sp. PB-01, Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13. Selected PPB showed ability to grow at wide range of pH, temperature and salt concentrations as well as to harbour diverse PGPR activities, such as: solubilization of insoluble Ca-phosphate (193–642 μg ml^?1), production of phytohormone indole acetic acid (5–39 μg ml^?1) and siderophore. Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 showed 50 and 70 % inhibition of phytopathogen Rhizoctonia solani, respectively. Greenhouse potting assay also showed that the bacterization of B. juncea seeds with Tetrathiobacter sp. PB-03 and Bacillus sp. PB-13 significantly increased the biomass and P content in 30 days old seedlings. This study reveals the potential of PPB as PGPR to improve the growth of B. juncea.     

两株具促生作用的苜蓿内生菌的分离纯化与鉴定

【目的】分离具有植物促生功能的苜蓿内生菌。【方法】从苜蓿的新鲜组织中分离纯化菌株,进行16S r RNA基因序列分析,生理生化鉴定并检测内生菌对苜蓿的促生特性。【结果】获得两株苜蓿内生菌,成团泛菌ASR16和短小芽孢杆菌ALR33。两株菌株均能产生植物生长激素IAA和嗜铁素,能溶解磷,对苜蓿的促生作用明显。【结论】获得两株具有植物促生特性的苜蓿内生菌ASR16和ALR33,可作为微生物菌肥的研究基础。     

Non-contiguous finished genome sequence of plant-growth promoting Serratia proteamaculans S4

Serratia proteamaculans S4 (previously Serratia sp. S4), isolated from the rhizosphere of wild Equisetum sp., has the ability to stimulate plant growth and to suppress the growth of several soil-borne fungal pathogens of economically important crops. Here we present the non-contiguous, finished genome sequence of S. proteamaculans S4, which consists of a 5,324,944 bp circular chromosome and a 129,797 bp circular plasmid. The chromosome contains 5,008 predicted genes while the plasmid comprises 134 predicted genes. In total, 4,993 genes are assigned as protein-coding genes. The genome consists of 22 rRNA genes, 82 tRNA genes and 58 pseudogenes. This genome is a part of the project “Genomics of four rapeseed plant growth-promoting bacteria with antagonistic effect on plant pathogens” awarded through the 2010 DOE-JGI’s Community Sequencing Program.     

Nodulation and plant-growth promotion by methylotrophic bacteria isolated from tropical legumes

The nitrogen fixing methylotrophic bacteria were isolated from the nodules of tropical legumes. Two isolates CMCJ317 and CMSA322 isolated from Crotalaria juncea and Sesbania aculeata possessing high nitrogenase activities under pure culture conditions and able to form nodules under inoculated conditions were further characterized. The biochemical characteristics revealed their close relationship with Methylobacterium nodulans type strain ORS2060. The PCR amplification of nodA and mxaF genes showed the expected 584 and 555 bp products, respectively, similar to M. nodulans ORS2060 and digestion with restriction enzymes revealed that the two isolates differed. The strains showed significantly higher nitrogenase activity and also improved nodulation and shoot nitrogen of the plants when inoculated to Macroptilum atropurpureum. CMCJ317 and CMSA322 formed nodules on C. juncea and M. atropurpureum under green house conditions and also significantly increased the nitrogen concentration in shoots. These findings show that the ability to establish symbiosis with legumes is more widespread in Methylobacterium.     

Characterization of plant growth-promoting Bacillus edaphicus NBT and its effect on lead uptake by Indian mustard in a lead-amended soil

The plant growth promotion characteristics of a heavy-metal-resistant strain of Bacillus edaphicus NBT was characterized. The strain was also evaluated for promoting plant growth and lead (Pb) uptake of Brassica juncea L. Czern (Indian mustard) in soil artificially contaminated with 0, 400, and 800 mg Pb.kg-1 soil. Atomic absorption spectrometer analysis demonstrated that strain NBT could release water-soluble Pb from lead carbonate in the solution. Strain NBT had the capacity to produce indole acetic acid, siderophores, and 1-aminocyclopropane-1-carboxylate deaminase. Low and high Pb treatments significantly decreased the growth of Indian mustard. Inoculation with strain NBT was found to increase root dry mass (ranging from 16% to 22%) and above-ground tissue dry mass (ranging from 24% to 30%) of Indian mustard in the Pb-amended soil. Strain NBT was able to mobilize Pb efficiently in plants in Pb-amended soil. In the soil treated with 400 and 800 mg Pb.kg-1 soil, the increase in Pb uptake varied from 18% to 46% in live bacterium-inoculated Indian mustard plants compared with dead bacterium-inoculated control. The strain was also able to colonize and develop in the rhizosphere soil of Indian mustard after root inoculation.     

Effects of three environmental variables on sulfate uptake by aerobic bacteria.

The effects of various concentrations of sulfate, organic sulfur, and organic carbon on sulfate uptake by aerobic bacteria were studied using pure cultures growing in a defined medium. Cultures of Pseudomonas fluorescens and Corynebacterium striatum took up sulfate faster when young, but sulfate uptake by Serratia marcescens was faster in older cultures. Organic sulfur was found to decrease sulfate uptake, but at concentrations somewhat higher than occurs in most natural freshwater ecosystems. Low levels of sulfate can theoretically directly limit bacterial biomass production but such limitation probably does not occur in natural systems. Evidence is presented which indirectly links the uptake of sulfate and organic carbon, adding credibility to the proposal that sulfate uptake can be used as an indicator of microbial biomass production in freshwater ecosystems.     

Invasion ecology applied to inoculation of plant growth promoting bacteria through a novel SIMPER-PCA approach

Plant and Soil - Plant growth promoting bacteria (PGPB) have been used on crops for years, but inoculants that are efficient in some locations may not be efficient in others. Here, we applied...     

Selenium assimilation and volatilization from dimethylselenoniopropionate by Indian mustard

Earlier work from our laboratory on Indian mustard (Brassica juncea L.) identified the following rate-limiting steps for the assimilation and volatilization of selenate to dimethyl selenide (DMSe): (a) uptake of selenate, (b) activation of selenate by ATP sulfurylase, and (b) conversion of selenomethionine (SeMet) to DMSe. The present study showed that shoots of selenate-treated plants accumulated very low concentrations of dimethylselenoniopropionate (DMSeP). Selenonium compounds such as DMSeP are the most likely precursors of DMSe. DMSeP-supplied plants volatilized Se at a rate 113 times higher than that measured from plants supplied with selenate, 38 times higher than from selenite, and six times higher than from SeMet. The conversion of SeMet to selenonium compounds such as DMSeP is likely to be rate-limiting for DMSe production, but not the formation of DMSe from DMSeP because DMSeP was the rate of Se volatilization from faster than from SeMet and SeMet (but no DMSeP) accumulated in selenite- or SeMet-supplied wild-type plants and in selenate-supplied ATP-sulfurylase transgenic plants. DMSeP-supplied plants absorbed the most Se from the external medium compared with plants supplied with SeMet, selenate, or selenite; they also accumulated more Se in shoots than in roots as an unknown organic compound resembling a mixture of DMSeP and selenocysteine.     

Effects of Ni(II) on respirometric oxygen uptake

The effects of Ni(II), substrate and initial biomass concentrations on biochemical oxygen demand (BOD) were studied by using an electrolytic respirometer. The effects of Ni(II) (2.5, 5.0, 10.0, 25.0 mg/l) and substrate (325, 650, 1300 mg/l as chemical oxygen demand) in a synthetic wastewater with differing initial biomass concentrations (1, 10, 100 mg/l) were investigated. The biomass-to-metal ratio was found to be the most important parameter affecting the measured BOD values. The maximum specific growth rates were calculated and the results of batch respirometric experiments were analysed both by graphical and statistical methods. In statistical analyses, a factorial experimental design approach was followed and results were treated by multiple regression techniques. A mathematical model was developed to express the maximum oxygen uptake in terms of nickel, substrate and initial biomass concentrations and their magnitudes of their effects were compared. The biomass-to-metal ratio was found to be very significant so that another model that expresses oxygen uptake in relation to the biomass-to-metal ratio and also to substrate concentration was developed. Finally, the effect of Ni(II) was demonstrated to depend on both substrate and initial biomass concentrations. This effect was stimulatory at low concentrations of Ni(II), and complete inhibition was never observed even at the highest concentration of Ni(II) studied, which was 25.0 mg/l. Received: 4 January 1997 / Received revision: 10 June 1997 / Accepted: 29 June 1997     

Total nitrogen uptake by maize with Azotobacter inoculation

Summary The economic feasibility of using cultures of nitrogen fixing microorganisms in programmes to increase crop production, as a selfgenerating source of nitrogen, has been proved beyond doubtviz. Legume-Rhizobium symbiosis and blue green algal ‘fertilizer’ for rice. The extent to which the free living, N-fixing, aerobic, heterotropicAzotobacter chroococcum could replace the application of nitrogenous fertilizer to maize was investigatedin vivo. Total nitrogen uptake (kg ha⁻¹) by maize after inoculation with Azotobacter combined with moderate applications of nitrogen fertilizer and farmyard manure was influenced significantly and resulted in a higher nitrogen concentration in grain and stover along with a higher yield. Part of Ph.D. Thesis     

Identification of rhizobacteria from maize and determination of their plant-growth promoting potential

During the growing season of 1986, the rhizobacteria (including organisms from the ectorhizosphere, the rhizoplane and endorhizosphere) of 20 different maize hybrids sampled from different locations in the Province of Quebec were inventoried by use of seven different selective media. Isolates were characterized by morphological and biochemical tests and identified using the API20E and API20B diagnostic strips.Pseudomonas spp. were the prominent bacteria found in the rhizoplane and in the ectorhizosphere.Bacillus spp. andSerratia spp. were also detected, but in smaller numbers. In the endorhizosphere,Bacillus spp. andPseudomonas spp. were detected in order of importance. Screening for plant growth-promoting rhizobacteria was carried out in three soils with different physical and chemical characteristics. The results depended on the soil used, but two isolates (Serratia liquefaciens andPseudomonas sp.) consistently caused a promotion of plant growth.Contribution no. 350 of the Research Station, Agriculture Canada, Sainte-Foy, Quebec.     

Effect of inoculation on element uptake by plants grown on saline soils

A relationship between inoculation and elemental uptake ofMedicago sativa inoculated withRhizobia meliloti (isolated from a saline area) was found. The plant uptake of the elements with atomic number between 19 and 42 was significantly higher in plants grown on inoculated soils, with the exception of molybdenum. Preliminary evidence shows that the concentration of some elements was affected by inoculation.     

Effects of non-ionic surfactants on the uptake and hydrolysis of fluoresceindiacetate by alkane-oxidizing bacteria

Biological effects of non-ionic surfactants on alkane-oxidizing bacteria were studied by assessing their influence on the uptake of prefluorochrome fluoresceindiacetate (FDA) and its intracellular hydrolysis to fluorescein. Both decreasing and increasing rates of hydrolysis as a consequence of the presence of surfactants were observed. The surfactants influenced the uptake of FDA, but not its intracellular hydrolysis. The effects of the surfactants on the uptake rate depended strongly on the structure and physico-chemical properties of the surfactants. There was no qualitative or significant quantitative difference in surfactant susceptibility between induced (alkane grown) and non-induced bacteria (acetate grown), even though the induced cells possess greater cell surface hydrophobicity.     

Distinct uptake of tellurate from selenate in a selenium accumulator, Indian mustard (Brassica juncea)

Tellurium (Te) is widely used in industry because of its unique chemical and physical properties, and has recently become a part of everyday life as a component of phase-change optical magnetic disks. However, the recovery of Te from the environment has not been discussed yet. In this regard, we evaluated the potential use of Indian mustard (Brassica juncea), a selenium (Se) accumulator, for the phytoremediation of Te. The Indian mustard plant was exposed to selenate and tellurate and the concentrations of Se and Te and the chemical species in the plant were determined. The Indian mustard plant accumulated less Te than Se, and the amount of Te accumulated in the plant was approximately 1/69 of that of Se. Although the incorporation of selenate was reduced by increasing sulfate concentration in the medium, the incorporation of Te was not affected by it, suggesting that this plant was able to discriminate tellurate from selenate in the roots. Three Te species were detected in the plant. The major species was tellurate. The other two species were not identical to available Te standards and thus could not be identified. Consequently, the Indian mustard plant is inappropriate for the phytoremediation of Te because it can strictly distinguish tellurate from selenate.     

接种丛枝菌根真菌对枳吸收利用磷酸铝的影响

在温室沙培条件下,以枳实生苗为试材,研究了丛枝菌根真菌摩西球囊霉和地表球囊霉对枳吸收利用难溶性磷酸盐(Al-P)的影响.结果表明：接种菌根真菌显著增加了枳的干物质量、含磷量及磷吸收量,且随Al-P施用量的提高,菌根贡献率、全株磷吸收量、真菌磷吸收量及真菌磷吸收贡献率显著增加;接种处理显著增加了枳根系与菌丝磷酸酶的分泌量,特别是酸性磷酸酶和中性磷酸酶的分泌量,但二者随施磷量的增加而降低.真菌磷吸收贡献率与酸性磷酸酶、中性磷酸酶、碱性磷酸酶和总磷酸酶含量呈极显著正相关.