Study of mechanisms for plant growth promotion elicited by rhizobacteria in Arabidopsis thaliana

Plant growth-promoting rhizobacteria (PGPR) colonize plant roots and exert beneficial effects on plant health and development. We are investigating the mechanisms by which PGPR elicit plant growth promotion from the viewpoint of signal transduction pathways within plants. We report here our first study to determine if well-characterized PGPR strains, which previously demonstrated growth promotion of various other plants, also enhance plant growth in Arabidopsis thaliana. Eight different PGPR strains, including Bacillus subtilis GB03, B. amyloliquefaciens IN937a, B. pumilus SE-34, B. pumilus T4, B. pasteurii C9, Paenibacillus polymyxa E681, Pseudomonas fluorescens 89B-61, and Serratia marcescens 90-166, were evaluated for elicitation of growth promotion of wild-type and mutant Arabidopsis in vitro and in vivo. In vitro testing on MS medium indicated that all eight PGPR strains increased foliar fresh weight of Arabidopsis at distances of 2, 4, and 6 cm from the site of bacterial inoculation. Among the eight strains, IN937a and GB03 inhibited growth of Arabidopsis plants when the bacteria were inoculated 2 cm from the plants, while they significantly increased plant growth when inoculated 6 cm from the plants, suggesting that a bacterial metabolite that diffused into the agar accounted for growth promotion with this strain. In vivo, eight PGPR strains promoted foliar fresh weight under greenhouse conditions 4 weeks after sowing. To define signal transduction pathways associated with growth promotion elicited by PGPR, various plant-hormone mutants of Arabidopsis were evaluated in vitro and in vivo. Elicitation of growth promotion by PGPR strains in vitro involved signaling of brassinosteroid, IAA, salicylic acid, and gibberellins. In vivo testing indicated that ethylene signaling was involved in growth promotion. Results suggest that elicitation of growth promotion by PGPR in Arabidopsis is associated with several different signal transduction pathways and that such signaling may be different for plants grown in vitro vs. in vivo.     

Bidirectional plant‐mediated interactions between rhizobacteria and shoot‐feeding herbivorous insects: a community ecology perspective

1. Plants interact with various organisms, aboveground as well as belowground. Such interactions result in changes in plant traits with consequences for members of the plant‐associated community at different trophic levels. Research thus far focussed on interactions of plants with individual species. However, studying such interactions in a community context is needed to gain a better understanding.
2. Members of the aboveground insect community induce defences that systemically influence plant interactions with herbivorous as well as carnivorous insects. Plant roots are associated with a community of plant‐growth promoting rhizobacteria (PGPR). This PGPR community modulates insect‐induced defences of plants. Thus, PGPR and insects interact indirectly via plant‐mediated interactions.
3. Such plant‐mediated interactions between belowground PGPR and aboveground insects have usually been addressed unidirectionally from belowground to aboveground. Here, we take a bidirectional approach to these cross‐compartment plant‐mediated interactions.
4. Recent studies show that upon aboveground attack by insect herbivores, plants may recruit rhizobacteria that enhance plant defence against the attackers. This rearranging of the PGPR community in the rhizosphere has consequences for members of the aboveground insect community. This review focusses on the bidirectional nature of plant‐mediated interactions between the PGPR and insect communities associated with plants, including (a) effects of beneficial rhizobacteria via modification of plant defence traits on insects and (b) effects of plant defence against insects on the PGPR community in the rhizosphere. We discuss how such knowledge can be used in the development of sustainable crop‐protection strategies.

     

Plant Growth-Promoting Rhizobacteria Associated with Ancient Clones of Creosote Bush (Larrea tridentata)

Plant growth-promoting rhizobacteria (PGPR) are common components of the rhizosphere, but their role in adaptation of plants to extreme environments is not yet understood. Here, we examined rhizobacteria associated with ancient clones of Larrea tridentata in the Mohave desert, including the 11,700-year-old King Clone, which is oldest known specimen of this species. Analysis of unculturable and culturable bacterial community by PCR-DGGE revealed taxa that have previously been described on agricultural plants. These taxa included species of Proteobacteria, Bacteroidetes, and Firmicutes that commonly carry traits associated with plant growth promotion, including genes encoding aminocyclopropane carboxylate deaminase and β–propeller phytase. The PGPR activities of three representative isolates from L. tridentata were further confirmed using cucumber plants to screen for plant growth promotion. This study provides an intriguing first view of the mutualistic bacteria that are associated with some of the world’s oldest living plants and suggests that PGPR likely contribute to the adaptation of L. tridentata and other plant species to harsh environmental conditions in desert habitats.     

Plant growth promoting bacteria from Crocus sativus rhizosphere

Present study deals with the isolation of rhizobacteria and selection of plant growth promoting bacteria from Crocus sativus (Saffron) rhizosphere during its flowering period (October–November). Bacterial load was compared between rhizosphere and bulk soil by counting CFU/gm of roots and soil respectively, and was found to be ~40 times more in rhizosphere. In total 100 bacterial isolates were selected randomly from rhizosphere and bulk soil (50 each) and screened for in-vitro and in vivo plant growth promoting properties. The randomly isolated bacteria were identified by microscopy, biochemical tests and sequence homology of V1–V3 region of 16S rRNA gene. Polyphasic identification categorized Saffron rhizobacteria and bulk soil bacteria into sixteen different bacterial species with Bacillus aryabhattai (WRF5-rhizosphere; WBF3, WBF4A and WBF4B-bulk soil) common to both rhizosphere as well as bulk soil. Pseudomonas sp. in rhizosphere and Bacillus and Brevibacterium sp. in the bulk soil were the predominant genera respectively. The isolated rhizobacteria were screened for plant growth promotion activity like phosphate solubilization, siderophore and indole acetic acid production. 50 % produced siderophore and 33 % were able to solubilize phosphate whereas all the rhizobacterial isolates produced indole acetic acid. The six potential PGPR showing in vitro activities were used in pot trial to check their efficacy in vivo. These bacteria consortia demonstrated in vivo PGP activity and can be used as PGPR in Saffron as biofertilizers.This is the first report on the isolation of rhizobacteria from the Saffron rhizosphere, screening for plant growth promoting bacteria and their effect on the growth of Saffron plant.     

Enhancing the growth of Vicia faba plants by microbial inoculation to improve their phytoremediation potential for oily desert areas

Two experiments were conducted to investigate the effect of inoculating Vicia faba plants (broad beens) raised in clean and oily sand with nodule-forming rhizobia and plant-growth-promoting rhizobacteria (PGPR) on growth of these plants in sand and to test whether this can improve the phytoremediation potential of this crop for oily desert areas. It was found that crude oil in sand at concentrations < 1.0% (w/w) enhanced the plant heights, their fresh and dry weights, the total nodule weights per plant, and the nitrogen contents of shoots and fruits. Similar enhancing effects were recorded when roots of the young plants were inoculated with nodule bacteria alone, PGPR alone, or a mixture of one strain of nodule bacteria and one of the PGPR. Such plant growth effects were associated with a better phytoremediation potential of V. faba plants for oily sand. The total numbers of oil-utilizing bacteria increased in the rhizosphere and more hydrocarbons were eliminated in sand close to the roots. The nodule bacteria tested were two strains of Rhizobium leguminosarum and the PGPR were Pseudomonas aeruginosa and Serratia liquefaciens. The four strains were found to use crude oil, n-octadecane, and phenanthrene as sole sources of carbon and energy. It was concluded that coinoculation of V. faba plant roots in oily sand with nodule bacteria and PGPR enhances the phytoremediation potential of this plant for oily desert sand through improving plant growth and nitrogen fixation.     

PGPR regulate caspase-like activity,programmed cell death,and antioxidant enzyme activity in paddy under salinity

The response of two root associated bacteria Pseudomonas pseudoalcaligenes and Bacillus pumilus were studied in the (salt-sensitive) rice GJ17 cultivar to salinity under controlled environmental growth conditions for protection of plant from adverse effect of salinity. Salinity affects the growth of salt-sensitive cultivar, but inoculation of plant growth promoting rhizobacteria (PGPR) reduces the harmful effect of salinity. The present study states that PGPR helps to reduce lipid peroxidation and superoxide dismutase activity in salt-sensitive GJ17 cultivar under salinity and play an important role in the growth regulation for positive adaptation of plants to salt stress. This study shows that inoculation of paddy (Oryza sativa) with such bacteria could provide salt-tolerant ability by reducing the toxicity of reactive oxygen species by reducing plant cell membrane index, cell caspase-like protease activity, and programmed cell death and hence resulted in increase cell viability. As these isolates remain associated with the roots, the effects of tolerance against salinity are observed here. Results also indicate that isolated PGPR strain help in alleviating up to 1.5 % salinity stress as well as improve tolerance.     

Plant growth promoting potential of bacteria isolated on N free media from rhizosphere of Cassia occidentalis

Plant growth promoting rhizobacteria (PGPR) are an attractive eco-friendly alternative to chemicals in agriculture. While the rhizospheres of crop plants have been well studied with the objective of screening PGPR, weeds, which play an important role in maintaining ecological balance, have largely been ignored. The rhizosphere of a luxuriantly growing, medicinal weed, Cassia occidentalis was analysed by enumerating PGPR on N free media from the most diverse stage of plant (determined by profiles obtained on denaturing gradient gel electrophoresis). Each isolate was tested for other plant growth promotion assays including production of cellulase, indole acetic acid (IAA), ammonia, HCN, siderophore and chitinase to select for ones possessing multi-trait plant growth promoting (PGP) properties. Selected isolates were used for bacterization of Vigna radiata and Vigna mungo to evaluate their efficacy in promoting plant's growth in seedling germination and axenic pot conditions. Thirty five isolates were analysed further for the array of PGP properties they exhibit. A total of 6 isolates were shortlisted on the basis of maximum traits positive, amount of phosphate solubilized and IAA produced. V. radiata responded well to seed bacterization during seedling germination. A maximum increase of approximately 36 and 60?% was observed for shoot and root length, respectively in V. radiata in axenic pot culture over control plants. Extensive branching of roots was also observed with isolate NL, which produced the maximum amount of IAA. Present study investigated the plant growth promoting isolates obtained on N free media in the rhizosphere of C. occidentalis, which have the potential to be used as inoculants for other crops. This provides a new dimension to the significance of weeds in agricultural ecosystems. The study opens up possibilities for utilization of this property of weeds in plant growth promotion, and subsequent enhancement of yield for agricultural crops.     

Potential plant growth-promoting activity of <Emphasis Type="Italic">Serratia nematodiphila</Emphasis> NII-0928 on black pepper (<Emphasis Type="Italic">Piper nigrum</Emphasis> L.)

A potential bacterial strain designated as NII-0928 isolated from Western ghat forest soil with multiple plant growth promoting attributes, and it has been identified and characterized. Plant growth promoting traits were analyzed by determining the P-solubilization efficiency, Indole acetic acid production, HCN, siderophore production and growth in nitrogen free medium. It was able to solubilize phosphate (76.6 μg ml⁻¹), and produce indole acetic acid (58.9 μg ml⁻¹) at 28 ± 2°C. Qualitative detection of siderophore production and HCN were also observed. At 5°C it was found to express all the plant growth promotion attributes except HCN production. The ability to colonize roots is a sine qua non condition for a rhizobacteria to be considered a true plant growth-promoting rhizobacteria (PGPR). 16S rRNA gene sequencing reveals the identity of the isolate as Serratia nematodiphila with which it shares highest sequence similarity (99.4%). Seed bacterization with black pepper cuttings in greenhouse trials using Sand: Soil: FYM with three individual experimental sets with their respective control showed clearly the growth promoting activity. Hence, Serratia nematodiphila NII-0928 is a promising plant growth promoting isolate showing multiple PGPR attributes that can significantly influence black pepper cuttings. The result of this study provides a strong basis for further development of this strain as a bioinoculants to attain the desired plant growth promoting activity in black pepper growing fields.     

Tales from the underground: molecular

Colonization of the rhizosphere by micro‐organisms results in modifications in plant growth and development. This review examines the mechanisms involved in growth promotion by plant growth‐promoting rhizobacteria which are divided into indirect and direct effects. Direct effects include enhanced provision of nutrients and the production of phytohormones. Indirect effects involve aspects of biological control: the production of antibiotics and iron‐chelating siderophores and the induction of plant resistance mechanisms. The study of the molecular basis of growth promotion demonstrated the important role of bacterial traits (motility, adhesion and growth rate) for colonization. New research areas emerge from the discovery that molecular signalling occurs through plant perception of eubacterial flagellins. Recent perspectives in the molecular genetics of cross‐talking mechanisms governing plant–rhizobacteria interactions are also discussed.     

Applications of free living plant growth-promoting rhizobacteria

Free-living plant growth-promoting rhizobacteria (PGPR) can be used in a variety of ways when plant growth enhancements are required. The most intensively researched use of PGPR has been in agriculture and horticulture. Several PGPR formulations are currently available as commercial products for agricultural production. Recently developing areas of PGPR usage include forest regeneration and phytoremediation of contaminated soils. As the mechanisms of plant growth promotion by these bacteria are unravelled, the possibility of more efficient plant-bacteria pairings for novel and practical uses will follow. The progress to date in using PGPR in a variety of applications with different plants is summarized and discussed here.     

Potential of Rhizobium and Bradyrhizobium species as plant growth promoting rhizobacteria on non-legumes: Effect on radishes (Raphanus sativus L.)

Bradyrhizobia and rhizobia are symbiotic bacterial partners forming nitrogen fixing nodules on legumes. These bacteria share characteristics with plant growth promoting rhizobacteria (PGPR). Nodule inducing bacteria, like other PGPR, are capable of colonizing the roots of non-legumes and produce phytohormones, siderophores and HCN. They also exhibit antagonistic effects towards many plant pathogenic fungi. The potential of nodule inducing bacteria to function as PGPR, was examined by using radish as a model plant. Three percent of the 266 strains tested were found to be cyanogens, while a majority (83%) produced siderophores. Fifty eight percent of the strains produced indole 3-acetic acid (IAA) and 54% solubilized phosphorus. Some of the bacterial species examined were found to have a deleterious effect while others were neutral or displayed a stimulatory effect on radishes. Bradyrizobium japonicum strain Soy 213 was found to have the highest stimulatory effect (60%), and an arctic strain (N44) was the most deleterious, causing a 44% reduction in radish dry matter yield. A second plant inoculation test, performed in growth cabinets, revealed that only strain Tal 629 of B. japonicum significantly increased (15%) the dry matter yield of radish. This indicates that specific bradyrhizobia have the potential to be used as PGPR on non-legumes.     

Effect of inoculation with putative plant growth-promoting rhizobacteria isolated from Pinus spp. on Pinus pinea growth, mycorrhization and rhizosphere microbial communities

Aims: In this study, 10 putative plant growth-promoting rhizobacteria (PGPR) were assayed for their ability to improve Pinus pinea growth and mycorrhization. Methods and Results: After an inoculation assay, except for two, all strains stimulated plant growth. All bacteria altered rhizosphere microbial communities as revealed by phospholipid fatty acid analysis; associating plant growth promotion with a decrease in biological diversity. Three strains were tested for their ability to enhance pine mycorrhization with wild fungi species. Only strain BB1 increased the total number of mycorrhizal root tips. Mycorrhizas present in the roots of each treatment were identified by ribosomal RNA sequencing and denaturing gradient gel electrophoresis analysis, detecting specificity between mycorrhizal species colonizing the roots and the inoculated PGPR. Conclusions: In conclusion, BB1 appears to be a good candidate to be developed into a biofertilizer directed to enhance pine growth and mycorrhization, which should result in a better establishment rate for plants used in reforestation. Significance and Impact of the Study: This study shows the potential of PGPR to improve fitness of forest tree specie. Moreover, the specificity between the bacteria inoculated and the mycorrhiza that the plant selects involve a potential biotechnological use in production of value-added fungi.     

The role of mycorrhizae and plant growth promoting rhizobacteria (PGPR) in improving crop productivity under stressful environments

Both biotic and abiotic stresses are major constrains to agricultural production. Under stress conditions, plant growth is affected by a number of factors such as hormonal and nutritional imbalance, ion toxicity, physiological disorders, susceptibility to diseases, etc. Plant growth under stress conditions may be enhanced by the application of microbial inoculation including plant growth promoting rhizobacteria (PGPR) and mycorrhizal fungi. These microbes can promote plant growth by regulating nutritional and hormonal balance, producing plant growth regulators, solubilizing nutrients and inducing resistance against plant pathogens. In addition to their interactions with plants, these microbes also show synergistic as well as antagonistic interactions with other microbes in the soil environment. These interactions may be vital for sustainable agriculture because they mainly depend on biological processes rather than on agrochemicals to maintain plant growth and development as well as proper soil health under stress conditions. A number of research articles can be deciphered from the literature, which shows the role of rhizobacteria and mycorrhizae alone and/or in combination in enhancing plant growth under stress conditions. However, in contrast, a few review papers are available which discuss the synergistic interactions between rhizobacteria and mycorrhizae for enhancing plant growth under normal (non-stress) or stressful environments. Biological interactions between PGPR and mycorrhizal fungi are believed to cause a cumulative effect on all rhizosphere components, and these interactions are also affected by environmental factors such as soil type, nutrition, moisture and temperature. The present review comprehensively discusses recent developments on the effectiveness of PGPR and mycorrhizal fungi for enhancing plant growth under stressful environments. The key mechanisms involved in plant stress tolerance and the effectiveness of microbial inoculation for enhancing plant growth under stress conditions have been discussed at length in this review. Growth promotion by single and dual inoculation of PGPR and mycorrhizal fungi under stress conditions have also been discussed and reviewed comprehensively.     

Root Colonization by Inoculated Plant Growth-Promoting Rhizobacteria

Certain rhizobacteria referred to as 'plant growth-promoting rhizobacteria' (PGPR) can contribute to the biological control of plant pathogens and improve plant growth. They enhance root development either directly by producing phytohormones, or indirectly by inhibiting pathogens through the synthesis of different compounds. PGPR are likely to be of great interest in sustainable crop protection and have drawn much attention in recent years. However, the use of these bacteria to protect crops sometimes fails because rhizobacteria are unable to recolonize the rhizosphere of inoculated plants. The colonization of roots by inoculated bacteria is an important step in the interaction between beneficial bacteria and the host plant. However, it is a complex phenomenon influenced by many biotic and abiotic parameters, some of which are now apparent. This paper summarises knowledge on rhizosphere colonization by PGPR.     

Bacillus subtilis strain L1 promotes nitrate reductase activity in Arabidopsis and elicits enhanced growth performance in Arabidopsis,lettuce, and wheat

Journal of Plant Research - Plant growth promoting rhizobacteria (PGPR) are a group of bacteria that promote plants growth in the rhizosphere. PGPRs are involved in various mechanisms that...     

Rhizosphere: its structure,bacterial diversity and significance

Sustainable agricultural practices are the answer to multifaceted problems that have resulted due to prolonged and indiscriminate use of chemical based agronomic tools to improve crop productions for the last many decades. The hunt for suitable ecofriendly options to replace the chemical fertilizers and pesticides has thus been aggravated. Owing to their versatile and unmatchable capacities microbial agents offer an attractive and feasible option to develop the biological tools to replace/supplement the chemicals. Exploring the microorganisms that reside in close proximity to the plant is thus a justified move in the direction to achieve this target. One of the most lucrative options is to look into the rhizosphere. Rhizosphere may be defined as the narrow zone of soil that surrounds and get influenced by the roots of the plants. It is rich in nutrients compared to the bulk soil and hence exhibit intense biological and chemical activities. A wide range of macro and microorganisms including bacteria, fungi, virus, protozoa, algae, nematodes and microarthropods co-exist in rhizosphere and show a variety of interactions between themselves as well as with the plant. Plant friendly bacteria residing in rhizosphere which exert beneficial affect on it are called as plant growth promoting rhizobacteria (PGPR). Here we review the structure and bacterial diversity of the rhizosphere. The major points discussed here are: (1) structure and composition of the rhizosphere (2) range of bacteria found in rhizosphere and their interactions with the plant with a particular emphasis on PGPR (3) mechanisms of plant growth promotion by the PGPR (4) rhizosphere competence.     

Effects of inoculation with plant growth promoting rhizobacteria (PGPRs) andSinorhizobium fredii on biological nitrogen fixation, nodulation and growth ofGlycine max cv. Osumi

We investigated the effects of three plant growth promoting rhizobacteria (PGPR), on Biological Nitrogen Fixation (BNF), nodulation and growth promotion by soybean (Glycine max) var. Osumi plants. The strains, Aur 6, Aur 9 and Cell 4, belong toPsedomonas fluorescens, Chryseobacterium balustinum andSerratia fonticola, respectively. Inoculation modes for the PGPRs andSinorhizobium fredii (carried out through irrigation), were examined. In the first mode, PGPRs andS. fredii were co-inoculated. In the second mode, we first inoculatedS. fredii and after the PGPRs, which were added 5 or 10 days later (each inoculation being an independent treatment). In the third mode, the PGPRs were inoculated first, and theS. fredii was inoculated 5 days later. We also included treatments inoculated with only the PGPRs (one PGPR per treatment) and only withS. fredii. Plants were maintained in a greenhouse under controlled environmental conditions, and were sampled 3 months after sowing. The results obtained showed the effects of the inoculation sequence. The most significant effects on growth parameters (stem plus leaf weight and fresh root weight) were found when inoculations with PGPR andS. fredii were at different times or when we inoculated only with PGPR and the plants were watered with nitrogen. Co-inoculation had no positive effects on any parameter, probably due to competition between the PGPR andS. fredii. Our results indicate that the inoculation modes with PGPR and rhizobia play a very important role in the effects produced. Thus, although plant growth promoting rhizobacteria may interact synergistically with root-nodulating rhizobia, plant growth promoting rhizobacteria selected for one crop should be assessed for potentially hazardous effects on other crops before being used as inoculants.     

Plant growth promoting rhizobacteria as biofertilizers

Numerous species of soil bacteria which flourish in the rhizosphere of plants, but which may grow in, on, or around plant tissues, stimulate plant growth by a plethora of mechanisms. These bacteria are collectively known as PGPR (plant growth promoting rhizobacteria). The search for PGPR and investigation of their modes of action are increasing at a rapid pace as efforts are made to exploit them commercially as biofertilizers. After an initial clarification of the term biofertilizers and the nature of associations between PGPR and plants (i.e., endophytic versus rhizospheric), this review focuses on the known, the putative, and the speculative modes-of-action of PGPR. These modes of action include fixing N₂, increasing the availability of nutrients in the rhizosphere, positively influencing root growth and morphology, and promoting other beneficial plant–microbe symbioses. The combination of these modes of actions in PGPR is also addressed, as well as the challenges facing the more widespread utilization of PGPR as biofertilizers.