Isolation and Characterization of ACC Deaminase Gene from Two Plant Growth-Promoting Rhizobacteria

Lowering of plant ethylene by deamination of its immediate precursor 1-aminocyclopropane-1-carboxylate (ACC) is a key trait found in many rhizobacteria. We isolated and screened bacteria from the rhizosphere of wheat for their ACC-degrading ability. The ACC deaminase gene (acdS) isolated from two bacterial isolates through PCR amplification was cloned and sequenced. Nucleotide sequence alignment of these genes with previously reported genes of Pseudomonas sp. strain ACP and Enterobacter cloacae strain UW4 showed variation in their sequences. In the phylogenetic analysis, distinctness of these two genes was observed as a separate cluster. 16S rDNA sequencing of two isolates identified them to be Achromobacter sp. and Pseudomonas stutzeri.     

Promotion of Plant Growth by Bacterial ACC Deaminase

To date, there has been only limited commercial use of plant growth-promoting bacteria in agriculture, horticulture, and silviculture. However, with recent progress toward understanding the mechanisms that these organisms utilize to facilitate plant growth, the use of plant growth-promoting bacteria is expected to continue to increase worldwide. One of the key mechanisms employed by plant growth-promoting bacteria to facilitate plant growth is the lowering of plant ethylene levels by the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase. This article reviews the published work on this enzyme, with an emphasis on its biochemistry, protein structure, genes, and regulation. In addition, this article provides some initial insights into the changes in both plants and ACC deaminase-containing plant growth-promoting bacteria as a consequence of plant-microbe interactions. Finally, a brief discussion of how bacterial ACC deaminase and indoleacetic acid (IAA) together modulate plant growth and development is included.     

含ACC脱氨酶的根际细菌提高植物抗盐性的研究进展

盐胁迫是抑制植物生长的主要非生物因素之一,高浓度的盐分不利于植物体的生长和发育,严重时会导致植物细胞及植物体死亡.已有大量实验结果显示含ACC脱氨酶的根际细菌可以缓解高盐对植物的危害.ACC脱氨酶可以降解乙烯的直接前体1-氨基环丙烷-1-羧酸(ACC),从而降低胁迫乙烯的合成量.胁迫乙烯是阻碍植物生长的主要原因.首先介...     

具有ACC脱氨酶活性的绞股蓝内生细菌的分离鉴定及其抑菌促生作用

采用富集筛选法从绞股蓝根中筛选得到6株具有ACC脱氨活性的细菌,其中菌株JDG-6、JDG-7、JDG-14、JDG-16、JDG-23均具有较强的分泌铁载体能力,但菌株JDG-32没有产铁载体能力。抑菌试验结果显示,菌株JDG-6、JDG-7、JDG-14和JDG16对一种或多种供试菌有抑菌作用,其中菌株JDG-14能抑制大肠埃希菌、藤黄八叠球菌和白色念球菌的生长。促生试验表明,菌株JDG-6、JDG-7和JDG-14均能促进水稻幼苗根的伸长,其中菌株JDG-14的促生作用最为明显,与对照组相比水稻幼苗的根长和根鲜重分别增长了26%和21%。通过形态特征观察、生理生化试验以及16S rDNA序列分析,菌株JDG-6、JDG-7、JDG-16和JDG-23属于假单胞杆菌属（Pseudomonas）,菌株JDG-14为木糖葡萄球菌（Staphylococcus xylosus）,而菌株JDG-32为枯草芽胞杆菌（Bacillus subtilis）。菌株JDG-6、JDG-7和JDG-14均具有ACC脱氨酶活性和抑菌活性的促生菌,具有农业应用潜力。     

Rhizobacteria of Populus euphratica Promoting Plant Growth Against Heavy Metals

The heavy metal-resistant bacteria from rhizospheric soils of wild Populus euphratica forest growing in arid and saline area of northwestern China were investigated by cultivation-dependent methods. After screening on medium sparked with zinc, copper, nickel and lead, 146 bacteria strains with different morphology were isolated and most of them were found to be resistant to at least two kinds of heavy metals. Significant increase in fresh weight and leaf surface area of Arabidopsis thaliana seedlings under metal stress were noticed after inoculated with strains especially those having multiple-resistance to heavy metals such as Phyllobacterium sp. strain C65. Investigation on relationship between auxin production and exogenous zinc concentration revealed that Phyllobacterium sp. strain C65 produced auxin, and production decreased as the concentration of zinc in medium increased. For wheat seedlings treated with zinc of 2 mM, zinc contents in roots of inoculated plants decreased by 27% (P < 0.05) compared to the uninoculated control. Meanwhile, zinc accumulation in the above-ground tissues increased by 22% (P < 0.05). The translocation of zinc from root to above-ground tissues induced by Phyllobacterium sp. strain C65 helped host plants extract zinc from contaminated environments more efficiently thus alleviated the growth inhibition caused by heavy metals.     

Isolation and Characterization of Salt Tolerant Plant Growth Promoting Rhizobacteria from Plants Grown in Tsunami Affected Regions of Andaman and Nicobar Islands

In order to explore salt tolerant plant growth promoting rhizobacteria (PGPR), 121 isolates were recovered from Tsunami affected area of Andaman and Nicobar Islands, India, out of which 23 were grown at 10% NaCl. Out of total isolates grown at 10% NaCl concentration, 14 solubilized phosphate, 13 showed siderophore production, five showed Indole acetic acid (IAA) production and 16 produced one or more extra cellular enzymes. Isolates SJ10 and CR5 showed the highest enzyme production and PGP properties and five isolates showed antagonistic activity against Sclerotium rolfsii. Screening of potential isolates for nitrogen fixing gene (nifH) showed only three isolates (MM3, MM6 and MRC11) possessed nifH gene. Salt tolerant Bacillus PGPR could be used as a bio-inoculant to enhance the crop growth under Tsunami affected soils.     

Isolation and Characterization of Three Plant Growth-Promoting Rhizobacteria for Growth Enhancement of Rice Seedling

Journal of Plant Growth Regulation - Biofertilizers are currently considered the only alternative of chemical fertilizers. In addition, improving the effect of biofertilizers can make chemical...     

Genetic Diversity of Plant Growth Promoting Rhizobacteria Isolated from Rhizospheric Soil of Wheat Under Saline Condition

In this study, a total of 130 rhizobacteria was isolated from a saline infested zone of wheat rhizosphere, and screened for plant growth promoting (PGP) traits at higher salt (NaCl) concentrations (2, 4, 6, and 8%). The results revealed that 24 rhizobacterial isolates were tolerant at 8% NaCl. Although all the 24 salt tolerable isolates produced indole-3-acetic acid (IAA), while 10 isolates solubilized phosphorus, eight produced siderophore, and six produced gibberellin. However, only three isolates showed the production of 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Diversity was analyzed through 16S rDNA-RFLP, and of these isolates with three tetra cutter restriction enzymes (HaeIII, AluI, and MspI), the representative cluster groups were identified by 16S rDNA sequencing. Bacillus and Bacillus-derived genera were dominant which showed PGP attributes at 8% NaCl concentration. Out of 24 isolates, nitrogen fixing ability (nif H gene) was detected in the two isolates, SU18 (Arthrobacter sp.) and SU48.     

Plant Growth Promoting Characterization of Indigenous Azotobacteria Isolated from Soils in Iran

It has been well known that the bacteria of the genus Azotobacter, in addition to the beneficial N₂-fixing activity, are able to improve plant growth by a number of direct and indirect mechanisms. To identify this potential in indigenous azotobacteria, the efficiency of 17 isolates of Azotobacter from the rhizosphere of wheat and barley plants cultivated in salt- and/or drought-affected soils in Iran were evaluated for their ability to dissolve inorganic and organic phosphates, siderophore secretion, indole acetic acid (IAA) production; and protease, chitinase, and ACC deaminase (ACCD) activities. First, they were biochemically characterized and one isolate (strain) was identified by 16S rDNA sequencing. Eight isolates were designated as Azotobacter vinelandii and the remaining isolates were identified as A. chroococcum. All isolates hydrolyzed the organic and inorganic phosphate compounds and effectively produced IAA. Fifteen isolates produced siderophore, but only one isolate showed protease activity which is being reported for the first time in relation to Azotobacter. None of the 17 isolates was capable of producing ACCD or chitinase. However, polymerase chain reaction amplification of the ACCD coding genes, by the use of the gene-specific primers, indicated that not all contain the ACCD gene. The standard screening methods with slight modifications, especially in the case of ACCD assay, were applied. The results showed that the use of specific screening methods, modified according to bacterial nutritional requirements, are the efficient methods for precise evaluation of the plant growth promoting rhizobacteria activity.     

Plant Growth Promoting Rhizobacteria Protect Wheat Plants Against Temperature Stress Through Antioxidant Signalling and Reducing Chloroplast and Membrane Injury

Journal of Plant Growth Regulation - In India and other tropics, wheat plants suffer from exposure to high temperature with heat increase above 35&nbsp;°C causing stress-like conditions....     

Growth Enhancement of Chickpea in Saline Soils Using Plant Growth-Promoting Rhizobacteria

Bacterial isolates with the ability to tolerate salinity and plant growth-promoting features were isolated from the saline areas of Gujarat, India, that is, Bhavnagar and Khambat. A total of 176 strains of rhizobacteria were isolated out of which 62 bacterial strains were able to tolerate 1 M NaCl. These were then further studied for their potential plant growth-promoting rhizobacteria characteristics like phosphate solubilization, siderophore production, and IAA production. Twenty-eight isolates of the 62 strains showed good tricalcium phosphate solubilization in solid medium in the range of 9–22 mm and 15 isolates showed good phosphate solubilization in liquid medium in the range of 9–45 μg/ml. Siderophore production was checked in all 15 isolates, and 13 were screened out that produced the hydroxamate type of siderophore in the range of 11–50 mM. Among the 13 isolates, 10 were able to produce indole acetic acid in the range of 10–26 μg/ml after 72 h of incubation. Pot trials were carried out on chickpea under 300 mM NaCl stress using the best five isolates. Plants inoculated with MSC1 or MSC4 isolates showed an increase in the parameters that evaluate plant growth when compared to uninoculated controls. Strains MSC1 and MSC4 were identified as Pseudomonas putida and Pseudomonas pseudoalcaligens, respectively, according to sequence analysis of the 16S rRNA gene.     

Cloning and Characterization of a Plasmid Encoded ACC Deaminase from an Indigenous Pseudomonas fluorescens FY32

In addition to the characterized mechanisms responsible for many direct effects of plant growth promoting bacteria (PGPB) on plants, it has been suggested that a number of PGPB contain the enzyme ACC deaminase that catalyzes degradation of 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, into α-ketobutyrate and ammonia. As part of an effort to obtain an ACC deaminase encoding gene from a collection of soil samples, only one bacterial isolate, Pseudomonas fluorescens FY32 was capable of growing on ACC as a sole source of nitrogen. The ACC deaminase gene was amplified from the above isolate by polymerase chain reaction (PCR) giving an expected DNA fragment, 1017 bp. Sequence analysis of the fragment showed that it was highly homologous (94% and 98% identities at nucleotide and amino acid levels, respectively) to the previously characterized acdS gene from Pseudomonas sp. 6G5. Furthermore, fusion of the ACC deaminase ORF with lacZ gene resulted in the expression of active enzyme in Escherichia coli. In addition, further analyses revealed that the acdS gene was plasmid-encoded so that a large plasmid (pFY32) with almost 50 kb in size was identified from this bacterium. Furthermore, transfer of pFY32 into E. coli DH5α proved its ACC deaminase activity. This result was in accordance with previous reports suggesting horizontal transfer of the acdS gene. However, it needs more investigation to identify whether this pFY32 plasmid has undergone lateral gene transfer during the evolutionary process.     

Variability in Chemical Composition and Antimicrobial Activity of Mentha pulegium L. Essential Oil,Cultivated under Different Plant Growth Promoting Rhizobacteria

The aim of present work was to study the essential oil chemical composition and antimicrobial activity of cultivated Mentha pulegium L. under different plant growth promoting rhizobacteria (Pseudomonas fluorescens, Bradyrhizobium sp. and Sinorhizobium meliloti) individually and in consortium. Yield, in plants inoculated with Bradyrhizobium sp. and S. meliloti in consortium, increase significantly relative to control plants. GC and GC/MS analyses pointed to a qualitative and quantitative variability of components. The investigated essential oils were clustered into three chemotypes: piperitenone/1,8-cineol (40.9/29.4 %) chemotype in plants inoculated with Bradyrhizobium sp. individually, S. meliloti individually, and Bradyrhizobium sp. and S. meliloti in consortium, piperitone/menthone (41.8/33.8 %) chemotype in plants inoculated with P. fluorescens individually, P. fluorescens and Bradyrhizobium sp. in consortium, and P. fluorescens and S. meliloti in consortium and pulegone/menthol (47.9/31.5 %) chemotype in control plants. The antimicrobial activity, carried out by the disc diffusion method and the determination of the Minimum Inhibitory Concentration (MIC) against ten microorganisms, varied significantly according to the tested microorganism and the rhizobacterial species used individually or in consortium (inhibition zone: 8.5–33.5 mm; MIC: 0.25–2.5 μL/mL). Our findings provided useful indications to select interesting chemotype within M. pulegium, especially in perspective of its cultivation.     

Differential Effects of Plant Growth-Promoting Rhizobacteria on Maize Growth and Cadmium Uptake

Maize is a plant known for food, feed, and energy value, but being a greater biomass, it may also be utilized to extract pollutants from soil. Plant growth-promoting rhizobacteria (PGPR) may act as biofertilizer to improve plant health and indirectly may enhance metal extraction. This study focuses on five bacterial strains isolated from the vegetable (Bitter gourd) rhizosphere irrigated with industrial effluent and characterized for various plant growth-promoting activities. Based on 16S rRNA gene sequencing, bacterial strains belonging to the genera, Bacillus (CIK-517, CIK-519), Klebsiella (CIK-518), Leifsonia (CIK-521), and Enterobacter (CIK-521R), were tested for their ability to promote maize growth in axenic conditions. Results showed negative and positive regulation of maize growth by the exogenous application of Cd and PGPR, respectively. Seed germination assays revealed significant reduction in relative seedling growth of maize cultivars upon Cd exposure (0–80 mg Cd L^?1). The tested strains showed tolerance to Cd (1.78–4.45 mmol L^?1) and were positive for catalase, oxidase, phosphate solubilization, exopolysaccharide (EPS), and auxin production, whereas CIK-518, CIK-519, and CIK-521R were negative for EPS, phosphate solubilization, and oxidase activities, respectively. Bacterial strains significantly increased shoot/root growth and their dry biomass in normal and Cd-contaminated soil as compared to their respective controls. None of the strains showed significant effects on relative water content or membrane permeability; however, Cd uptake significantly increased in plant tissues upon bacterial inoculation. Bacterial strains CIK-518 and CIK-521R are effective colonizers and thus can be potential inoculants to promote maize growth and Cd extraction/stabilization in Cd-contaminated soil.     

Growth and Yield Response of Upland Rice to Application of Plant Growth-Promoting Rhizobacteria

Journal of Plant Growth Regulation - This study evaluated the effects of plant growth-promoting rhizobacteria (PGPR) isolates in enhancing upland rice growth and yield. Bacteria were isolated,...     

Rhizobacteria Strain from a Hypersaline Environment Promotes Plant Growth of Kengyilia thoroldiana

Microbiology - Kengyilia thoroldiana is a nutritionally rich grass species of the Qinghai-Tibet Plateau. Here, to improve its quality and biomass via biological fertilization, we sought out...     

Effects of Plant Growth Promoting Rhizobacteria (PGPRs) on the Biological Nitrogen Fixation,Nodulation, and Growth of Lupinus albus l. cv. Multolupa

The effects of five plant growth promoting rhizobacteria on the biological nitrogen fixation (BNF), nodulation, and growth promotion of plants of Lupinus albus cv. Multolupa were investigated. The plants were selected for their capacity to use 1‐aminocyclopropane‐1‐carboxylic acid (ACC) as the sole source of nitrogen. Four strains belonged to the genus Pseudomonas (Luc 1, Luc 2, Luc 3, and Luc 4) and one (Luc 5) belonged to the genus Bacillus. Three patterns of inoculation were examined. In the first pattern, PGPRs were inoculated seven days before being inoculated with B. japonicum. In the second pattern, PGPRs and B. japonicum were co‐inoculated, and in the last pattern, PGPRs were inoculated seven days after being inoculated with B. japonicum. The plants were sampled 30 and 45 days after being inoculated with B. japonicum (T1 and T2). In the first pattern of inoculation, Luc 5 significantly increased the biological nitrogen fixation compared to the control at the first sampling time, as did Luc 1 and Luc 4 at the second sampling time. In the second pattern of inoculation, Luc 5 negatively effected the biological nitrogen fixation at both sampling times. In the third pattern of inoculation, all PGPRs caused a decrease in the nitrogen content of the plants compared to the control. The results obtained according to the patterns of inoculation showed that the mechanisms of action of the effects and routes used by Gram‐negative and Gram‐positive strains were clearly different. Competition between PGPRs and B. japonicum, competition for the niches in the rhizoplane, production of auxins, and induction of systemic resistance (ISR) by the production of siderophores or by lipopolysaccharides present in the outer membrane (LPS) are discussed as probable reasons for the effects observed.     

Plant Growth Promoting Rhizobacteria and Soybean [ Glycine max (L.) Merr.] Nodulation and Nitrogen Fixation at Suboptimal Root Zone Temperatures

Co-inoculation of plant growth promoting rhizobacteria (PGPR)withBradyrhizobium has been shown to increase legume nodulationand nitrogen fixation at optimal soil temperatures. Nine rhizobacteriaco-inoculated withBradyrhizobium japonicum532C were tested fortheir ability to reduce the negative effects of low root zonetemperature (RZT) on soybean [Glycine max(L.) Merr.] nodulationand nitrogen fixation. Three RZTs were tested: 25 (optimal),17.5 (somewhat inhibitory), and 15°C (very inhibitory).At each temperature some PGPR strains increased the number ofnodules formed and the amount of fixed nitrogen when co-inoculatedwithB. japonicum,but the stimulatory strains varied with temperatures.The strains that were most stimulatory varied among temperaturesand were as follows: 15°C,Serratia proteamaculans 1-102;17.5°C,S. proteamaculans 1-102andAeromonas hydrophilaP73;25°C,Serratia liquefaciens2-68. Bradyrhizobium japonicum ; Glycine max; plant growth promoting rhizobacteria; suboptimal root zone temperatures     

Plant Growth Promoting Metabolites of Sclerotinia sclerotiorum

In the research on the plant growth regulators produced by phytopathogenic fungi, two active metabolites, sclerotinin A and B, in addition to sclerin have been isolated from the culture filtrate of S. sclerotiorum. Sclerotinin A and B have been shown to be 3,6,8-tri-hydroxy-3,4,5,7-tetramethyl-3,4-dihydroisocoumarin and 3,6,8-trihydroxy-3,5,7-trimethyl-3,4- dihydroisocoumarin, respectively.