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Survival and colonization of rhizobacteria in a tomato transplant system.

Plant-growth-promoting rhizobacteria (PGPR) are used on crops most often as seed treatments; however, an alternative application method for transplanted vegetables is mixing PGPR into the soilless medium in which the transplants are grown. Studies were undertaken to compare root colonization and persistence of rifampicin-resistant mutants of PGPR strains Bacillus pumilus SE34 and Pseudomonas fluorescens 89B61, SE34r and 89B61r, on tomato as a function of application method. When the bacteria were incorporated into Promix soilless medium at log 6, 7, and 8 colony- forming units/g, populations of strain SE34r per gram of medium maintained the initial inoculum densities, while populations of 89B61r decreased approximately one to two orders of magnitude by 4 weeks after planting. The populations of each PGPR strain colonizing roots after application into the soilless medium showed a similar pattern at 6 weeks as that at 4 weeks after planting, with higher populations on the whole roots and lateral roots than on the taproots. Strain SE34r but not 89B61r moved upwards and colonized the phyllosphere when incorporated into the soilless medium. Following application as seed treatment, populations of SE34r were significantly higher on upper roots and on the taproot than were populations following application through the soilless medium. Conversely, populations were higher on lower roots and lateral roots following application through the soilless medium than were populations following application as seed treatment. While strain SE34 enhanced plant growth with application both to the medium and as seed treatment, the level of growth promotion was significantly greater with application in the soilless medium. The results indicate that PGPR can be successfully incorporated into soilless media in vegetable transplant production systems.

Bacillus↗

[Isolation and phenotypic characteristics of growth-stimulating rhizobacteria (PGPR), with high root-colonizing and phytopathogenic fungi inhibiting abilities].

Some bacterial strains isolated from the plant rhizosphere showed high root-colonizing ability and antiphytopathogenic activity against 6 fungal species. The antifungal activity was species-specific, which could be accounted for by the fact that the isolates differed in the ability to produce lytic enzymes (chitinases, proteases, and lipases) and to secrete cyanide. The possibility of using there rhizobacteria to control phytopathogens is discussed.

Fungi↗

Single-Cell Force Spectroscopy Uncovers Root Zone- and Bacteria-Specific Interactions.

Understanding root-bacteria interactions with plant growth-promoting rhizobacteria (PGPR) is key to developing effective biofertilizers for sustainable agriculture. We performed single-cell force spectroscopy using the atomic force microscope (AFM) to study the primary attachment of two PGPR, Bacillus velezensis and Pseudomonas defensor, to different regions of Arabidopsis thaliana roots. Force measurements with individual cells uncovered distinct attachment strategies by each strain, involving binding via micrometer-long polymers from both bacteria and root surfaces. Flagella differentially affected the binding interactions of each PGPR; their removal altered binding characteristics differently for each strain, highlighting the importance of flagella in early root colonization. Using silica beads to mimic the negatively charged bacteria, we demonstrated the influence of electrostatic forces on root-bacteria interactions. We also examined interactions with abiotic surfaces of varying surface energies, revealing the roles of hydrophilic and hydrophobic forces in initial binding. Our measurements show that differences in the physicochemical properties of bacteria and roots are responsible for variations in primary attachment strategies between PGPR strains and root regions. Parallel fluorescence measurements corroborated our AFM single-cell analysis. Overall, our results provide a nanoscale view of bacterial attachment to roots, offering key insights into how beneficial bacteria colonize roots, crucial for enhancing biofertilizer effectiveness.

Plant Roots↗

Root colonization of maize and lettuce by bioluminescent Rhizobium leguminosarum biovar phaseoli.

Two strains of Rhizobium leguminosarum bv. phaseoli and three other plant growth-promoting rhizobacteria (PGPR) were examined for the potential of maize and lettuce root colonization. All of these strains were selected in vitro for their phosphate-solubilizing abilities. Maize and lettuce seeds were treated with derivatives of all strains marked with lux genes for bioluminescence and resistance to kanamycin and rifampin prior to planting in nonsterile Promix and natural soil. The introduced bacterial strains were quantified on roots by dilution plating on antibiotic media together with observation of bioluminescence. Rhizobia were superior colonizers compared with other tested bacteria; rhizobial root populations averaged log 4.1 CFU/g (fresh weight) on maize roots 4 weeks after seeding and log 3.7 CFU/g (fresh weight) on lettuce roots 5 weeks after seeding. The average populations of the recovered PGPR strains were log 3.5 and log 3.0 CFU/g (fresh weight) on maize and lettuce roots, respectively. One of the three PGPR was not recovered later than the first week after seeding in Promix. Bioluminescence also permitted visualization of in situ root colonization in rhizoboxes and demonstrated the efficiency of rhizobial strains to colonize and survive on maize and lettuce roots.

Lactuca↗

[Factors affecting colonization of introduced microorganisms on plant roots].

Microorganisms such as biological control agents (BCA), plant growth-promoting rhizobacteria (PGPR) and yield increasing bacteria (YIB) were introduced along growing roots. The colonization process of introduced bacteria was proved that they attached root tipfirst, then distributed along roots, multiplicated there, and survived as certain population size. The colonization location was closely related with root exudates, which was usually at the junction between cortex cells or at the base of lateral roots or root hairs. The variation of colonization by introduced microorganisms in the rhizosphere was caused by biotic and abiotic factors. Biotic factors included the physiological characters of introduced microorganisms and interactions between introduced microorganisms and native microbes. The more important factors were plant genotypes which associated with introduced beneficial microbes and regulated the population and community of those microbes affecting the colonization of introduced microorganisms. Abiotic factors here referred to soil environmental conditions, e.g., soil texture, water content, soil temperature and pH value.

Ecosystem↗

Microarray analysis of the gene expression profile induced by the endophytic plant growth-promoting rhizobacteria, Pseudomonas fluorescens FPT9601-T5 in Arabidopsis.

Pseudomonas fluorescens FPT9601-T5 was originally identified as an endophytic plant growth-promoting rhizobacteria (PGPR) on tomato. To perform a molecular dissecttion of physiological and biochemical changes occurring in the host triggered by P. fluorescens FPT9601-T5 colonization, the model plant Arabidopsis was used in this study. Root colonization of Arabidopsis with P. fluorescens FPT9601-T5 promoted plant growth later than three weeks after inoculation and partially suppressed disease symptoms caused by Pseudomonas syringae pv. tomato DC3000, indicating that P. fluorescens FPT9601-T5 acted as a PGPR on Arabidopsis. To obtain a global view on transcript modification during the Arabidopsis-FPT9601-T5 interaction, we performed microarray analysis using Affymetrix Genechip probe arrays representing approximately 22,800 genes. The results showed that 95 and 105 genes were up- or down-regulated, respectively, more than twofold in FPT9601-T5-treated Arabidopsis plants as compared with control plants. Those up-regulated included genes involved in metabolism, signal transduction, and stress response. Noteworthy, upon FPT9601-T5 colonization, putative auxin-regulated genes and nodulin-like genes were up-regulated, and some ethylene-responsive genes were down-regulated. Our results suggest that P. fluorescens FPT9601-T5 triggered plant responses in a manner similar to known PGPR and, at least in some aspects, to rhizobia.

Arabidopsis↗

Colonization of Bacillus spp. on seeds and in plant rhizoplane.

Seed coating, dipping and Scanning Electron Microscopy (SEM) were employed to study bacterial and fungal colonization of the seeds and rhizoplane of maize (Zea mays L.) during the early stages of growth. Isolation of Bacillus spp. entailed screening soil bacteria with potential growth stimulation and plant pathogen suppressive abilities isolated from the rhizospheres and rhizoplanes of vegetable crops. The bacterial colonization of the spermosphere was 90%. When the coated seeds were fully germinated, bacteria moved to the emerging radicle. Virtually no bacteria occurred on the root tip both for the treated and untreated. However, colonization was 20% in the basal portion of the roots close to the seed-root junction. SEM observations showed that the bacterial cells were arranged linearly and laterally on the growing root axis. This phenomenon was more noticeable in the seedlings dipped in the bacterial culture on the 3rd day after germination. The results indicate that attachment to the seed coat and the rhizoplane by the plant growth-promoting rhizobacterium (PGPR) is an important factor in the successful colonization of the rhizoplane. The significance of the work is to ascertain that the inoculated Bacillus spp. adhered to and established in the rhizoplane of maize. It can therefore be used as a PGPR and as a biocontrol agent.

Bacillus↗

[Research progress on PGPR/AMF interactions].

As one of the rhizospheric microorganisms PGPR(Plant Growth Promoting Rhizobacteria) and AMF(Arbuscular Mycorrhizal Fungi) play an important role in promoting plant growth. It is of significance to further study and elucidate the interactions between them to utilize and regulate the interactions among rhizospheric microorganisms, and promote and protecte plant growth. Many research results show that on one hand, there exists synergism between PGPR and AMF. AMF can transfer PGPR or act as a media in the process of spread of PGPR along roots, where PGPR create many beneficial conditions for the infection of AMF. Both of them can indirectly enhance the other side's colonization or infection ability through their own promoting role on plant growth. On the other hand, they compete with each other for nutrients and niches, and probably produce some secondary metabolites which cause detrimental effects on the other. However, whether these interactions are synergistic or competitive depends upon the AM fungal or PGPR species involved. So far, the research work is extensive, even in molecular level in some aspects, but not systematic and deep. It is believed however, with the development of techniques in molecular biology and the increasing application of advanced testing methods, the new breakthroughs will be gained in the study and understanding on the interactions.

Bacterial Physiological Phenomena↗

Gibberellins-producing rhizobacteria increase endogenous gibberellins content and promote growth of red peppers.

The growth of red pepper plants was enhanced by treatment with the rhizobacterium, Bacillus cereus MJ-1. Red pepper shoots showed a 1.38-fold increase in fresh weight (fw) and roots showed a 1.28-fold fw gain. This plant growth-promoting rhizobacterium (PGPR) has been reported to produce gibberellins (GAs). Other GAs-producing rhizobacteria, Bacillus macroides CJ-29 and Bacillus pumilus CJ-69, also enhanced the fw of the plants. They were less effective than B. cereus MJ-1, though. The endogenous GAs content of pepper shoots inoculated with MJ-1 was also higher than in shoots inoculated with CJ-29 or CJ-69. When inoculated with MJ-1, bacterial colonization rate of the roots was higher than that of roots inoculated with CJ-29 or CJ-69. These results support the idea that the plant growth-promoting effect of the bacteria also positively related with the efficiency of root colonization by the bacteria. In addition, we identified the major endogenous GAs of the red pepper as originating from both the early C-13 hydroxylation and the early non C-13 hydroxylation pathways, with the latter being the predominant pathway of GA biosynthesis in red pepper shoots.

Bacillus↗

Type III secretion in plant growth-promoting Pseudomonas fluorescens SBW25.

In vivo expression technology (IVET) analysis of rhizosphere-induced genes in the plant growth-promoting rhizobacterium (PGPR) Pseudomonas fluorescens SBW25 identified a homologue of the type III secretion system (TTSS) gene hrcC. The hrcC homologue resides within a 20-kb gene cluster that resembles the type III (Hrp) gene cluster of Pseudomonas syringae. The type III (Rsp) gene cluster in P. fluorescens SBW25 is flanked by a homologue of the P. syringae TTSS-secreted protein AvrE. P. fluorescens SBW25 is non-pathogenic and does not elicit the hypersensitive response (HR) in any host plant tested. However, strains constitutively expressing the rsp-specific sigma factor RspL elicit an AvrB-dependent HR in Arabidopsis thaliana ecotype Col-0, and a host-specific HR in Nicotiana clevelandii. The inability of wild-type P. fluorescens SBW25 to elicit a visible HR is therefore partly attributable to low expression of rsp genes in the leaf apoplast. DNA hybridization analysis indicates that rsp genes are present in many plant-colonizing Pseudomonas and PGPR, suggesting that TTSSs may have a significant role in the biology of PGPR. However, rsp and rsc mutants retain the ability to reach high population levels in the rhizosphere. While functionality of the TTSS has been demonstrated, the ecological significance of the rhizosphere-expressed TTSS of P. fluorescens SBW25 remains unclear.

Bacterial Proteins↗

Growth promotion and yield enhancement of peanut (Arachis hypogaea L.) by application of plant growth-promoting rhizobacteria.

Although plant growth-promoting rhizobacteria (PGPR) have been reported to influence plant growth, yield and nutrient uptake by an array of mechanisms, the specific traits by which PGPR promote plant growth, yield and nutrient uptake were limited to the expression of one or more of the traits expressed at a given environment of plant-microbe interaction. We selected nine different isolates of PGPR from a pool of 233 rhizobacterial isolates obtained from the peanut rhizosphere on the basis of ACC-deaminase activity. The nine isolates were selected, initially, on the basis of germinating seed bioassay in which the root length of the seedling was enhanced significantly over the untreated control. All the nine isolates were identified as Pseudomonas spp. Four of these isolates, viz. PGPR1, PGPR2, PGPR4 and PGPR7 (all fluorescent pseudomonads), were the best in producing siderophore and indole acetic acid (IAA). In addition to IAA and siderophore-producing attributes, Pseudomonas fluorescens PGPR1 also possessed the characters like tri-calcium phosphate solubilization, ammonification and inhibited Aspergillus niger and A. flavus in vitro. P. fluorescens PGPR2 differed from PGPR1 in the sense that it did not show ammonification. In addition to the traits exhibited by PGPR1, PGPR4 showed strong in vitro inhibition to Sclerotium rolfsii. The performances of these selected plant growth-promoting rhizobacterial isolates were repeatedly evaluated for 3 years in pot and field trials. Seed inoculation of these three isolates, viz. PGPR1, PGPR2 and PGPR4, resulted in a significantly higher pod yield than the control, in pots, during rainy and post-rainy seasons. The contents of nitrogen and phosphorus in soil, shoot and kernel were also enhanced significantly in treatments inoculated with these rhizobacterial isolates in pots during both the seasons. In the field trials, however, there was wide variation in the performance of the PGPR isolates in enhancing the growth and yield of peanut in different years. Plant growth-promoting fluorescent pseudomonad isolates, viz. PGPR1, PGPR2 and PGPR4, significantly enhanced pod yield (23-26%, 24-28% and 18-24%, respectively), haulm yield and nodule dry weight over the control in 3 years. Other attributes like root length, pod number, 100-kernel mass, shelling out-turn and nodule number were also enhanced. Seed bacterization with plant growth-promoting P. fluorescens isolates, viz. PGPR1, PGPR2 and PGPR4, suppressed the soil-borne fungal diseases like collar rot of peanut caused by A. niger and PGPR4 also suppressed stem rot caused by S. rolfsii. Studies on the growth patterns of PGPR isolates utilizing the seed leachate as the sole source of C and N indicated that PGPR4 isolate was the best in utilizing the seed leachate of peanut, cultivar JL24. Studies on the rhizosphere competence of the PGPR isolates, evaluated on the basis of spontaneous rifampicin resistance, indicated that PGPR7 was the best rhizoplane colonizer and PGPR1 was the best rhizosphere colonizer. Although the presence of growth-promoting traits in vitro does not guarantee that an isolate will be plant growth promoting in nature, results suggested that besides ACC-deaminase activity of the PGPR isolates, expression of one or more of the traits like suppression of phytopathogens, solubilization of tri-calcium phosphate, production of siderophore and/or nodulation promotion might have contributed to the enhancement of growth, yield and nutrient uptake of peanut.

Arachis↗

[Research advances in plant growth-promoting rhizobacteria and its application prospects].

The study of plant growth-promoting rhizobacteria will provide possible methods to overcome the difficulties in controlling soil borne plant diseases. PGPR can colonize in rhizosphere at high population density, inhibit plant pathogens and deleterious microorganisms there, and promote crop growth and its yield. More importantly, some PGPR strains applied as biocontrol agents can make treated plant produce induced systemic resistance (ISR), and thus, increases plant overall health. In recent two decades, the researches of PGPR in abroad are very active, and many PGPR products have been applied successfully. In our contry, more attention should be paid to the study of the basic theory and application of GPPR and the industrialization of PGPR products.

Biodiversity↗

Phylloplane bacteria increase seedling emergence, growth and yield of field-grown groundnut (Arachis hypogaea L.).

AIM: To isolate and characterize groundnut-associated bacterial isolates for growth promotion of groundnut in field. METHODS AND RESULTS: Three hundred and ninety-three groundnut-associated bacteria, representing the geocarposphere, phylloplane and rhizosphere, and endophytes were applied as seed treatment in greenhouse. Maximum increase in plant biomass (up to 26%) was observed following treatment with a rhizosphere isolate identified as Bacillus firmis GRS 123, and two phylloplane isolates Bacillus megaterium GPS 55 and Pseudomonas aeruginosa GPS 21. There was no correlation between the production of L-tryptophan-derived auxins and growth promotion by the test isolates. Actively growing cells and peat formulations of GRS 123 and GPS 55, and actively growing cells of GPS 21, significantly increased the plant growth and pod yield (up to 19%) in field. Rifampicin-resistant mutants of GRS 123 and GPS 21 colonized the ecto- and endorhizospheres of groundnut, respectively, up to 100 days after sowing (DAS), whereas GPS 55 was recovered from both the habitats at 100 DAS. CONCLUSION: Seed bacterization with phylloplane isolates promoted groundnut growth indicating the possibility of isolating rhizosphere beneficial bacteria from different habitats. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of phylloplane bacteria as effective plant growth-promoting rhizobacteria (PGPR) broadens the spectrum of PGPR available for field application.

Anti-Bacterial Agents↗