Pink-pigmented Facultative Methylotrophic Bacteria Research Articles

Effect of foliar spraying with Pink Pigmented Facultative Methylotrophic Bacteria as a Foliar Spraying on the Growth and Productivity of Strawberry.

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Impact of foliar fertilization of nutrients and biofertilizer on the growth and yield of hybrid cotton (Gossypium hirsutum L.)

The newly released high yielding cotton cultivars are higher boll retention rate than conventional cultivars that have higher overall nutrient demand during the flowering and boll development stages. Therefore, this field experiment was conducted at during summer, 2020 at Annamalai University experimental farm, Department of Agronomy, Faculty of Agriculture, Annamalai Nagar, Tamil Nadu, India, 608 002 to ascertain the impact of foliar application of essential nutrients and biofertilizers on growth and yield of cotton. The experiment was laid out in randomized block design with twelve treatments of foliar application macro and micro-nutrients with and without Pink pigmented facultative methylotrophic bacteria (PPFM) and replicated thrice. Foliar application of macro, micronutrients and PPFM registered the increased values of growth characters, yield attributes and yield of hybrid cotton. Among the foliar application practice tested, foliar application of 2% urea + 1% PPFM at flowering and boll development stages significantly resulted in the enhances values of growth characters viz., plant height (157.40 cm), Dry matter production (6003 kg/ha), number of monopodial branches (3.28/plant), growth analysis like Leaf area index (4.30), Crop growth rate (7.01 g/m2/day) and Relative growth rate (0.0144 g/g/day), yield attributes viz., number of sympodial branches (21.77/plant), number of squares (83.77/plant), number of bolls (45.04/plant), boll weight (5.58 g) and seed cotton yield (3185 kg/ha). This was statistically on par with the foliar application of 2% MgSO4 + 1% PPFM at flowering and boll development stages. The lower values of growth characters, yield attributes and yield were recorded under water spray treatment. Based on the results of field investigation it can be concluded that foliar application of 2% urea + 1% PPFM at flowering and boll development stages can possibly enhance the growth and yield of hybrid cotton grown under dryland condition.

Development of deficit irrigation program for improving the water use efficiency, growth, and yield of groundnut in the semiarid tropical region of India

In this study, we assessed the efficiency of a deficit irrigation program (DIP) on groundnut (<i>Arachis hypogaea</i> L.) yield and economic profits. The DIP was designed to combine frequency and volume of irrigation to develop an appropriate irrigation water management (IWM) strategy for groundnut. Two field experiments were conducted at the Agricultural College and Research Institute, Madurai, Tamil Nadu, India, with a bunch variety of groundnut, VRI 2, under a randomized block design (RBD) with three replications. The treatments included DIP in combination with a foliar application of pink-pigmented facultative methylotrophic bacteria (PPFM). The irrigation frequencies adopted for the first set of treatments were high frequency (5/5), medium frequency (7/5), and lower frequency (7/7) starting from 14 days after sowing (DAS). The effects of early water stress on groundnut pod yield and water use efficiency were investigated in a second experiment by starting irrigation at 21 DAS. The number of irrigation frequencies and irrigation volume was fixed to develop an easily readable irrigation chart for end-users. The number of irrigation frequencies was fixed as three, and the volume of irrigation was fixed as a depth of 3 cm. In the first experiment, a mini mobile pumping unit was used for scheduling irrigation frequencies; in the second experiment, the surface drip method was used. In both experiments, we also assessed the effects of foliar application of PPFM, widely used as a growth stimulant for several crops, especially under drought conditions. Overall, we observed that DIP led to up to 25.6% water saving compared to surface irrigation. Use of the DIP at the medium frequency with PPFM spray had a significantly positive influence on water use efficiency (WUE) and irrigation water use efficiency (IWUE) compared to conventional drip irrigation (CDI). Compared to surface irrigation (SI), DIP had a positive impact on growth parameters, yield attributes, and economic profitability with a higher benefit-cost ratio. Additionally, there were only minor differences between deficit irrigation (DI) results starting from 21 DAS and 14 DAS. Irrespective of the irrigation techniques, PPFM application had a positive effect on the yield attributes and WUE. Our results indicated that DI at a medium frequency (7/5) with PPFM spray, starting from 21 DAS under drip fertigation, is the best irrigation management innovation (IMI) strategy for optimum utilization of irrigation water without significant reduction in pod yield. This management had a positive influence on WUE and IWUE of summer groundnut in sandy clay loam soil. However, the exact scheduling of the irrigation may differ in different environments as well as soil types. Nevertheless, we propose that the application and further optimization of the DIP devised in this study would help the farmers attain a similar yield using a lower amount of irrigation water and lead to significant economic profits for the farmers.

Pink Pigmented Facultative Methylotrophic Bacteria Isolated from Fermented Philippine Shrimp Paste.

Pink pigmented facultative methylotrophic (PPFM) bacteria are ecologically distributed microorganisms. They have been isolated in many types of ecosystems like soil, water, air, in association with plants and even as pathogens in humans. However, a yet unexplored area for PPFM bacteria research is in food. Hence, the objective of this study was to establish the presence of PPFM bacteria in Philippine fermented food, in particular shrimp paste, and characterise them phenotypically and genotypically. A total of 13 PPFM bacteria were obtained from Philippine shrimp paste. Sequencing of the 16S rRNA gene revealed that the PPFM bacterial isolates belong to the genus Methylobacterium. A total of 35 phenotypic characterisations were performed that included morphological, biochemical and physiological tests. Phylogenetic tree was constructed to establish the genetic relatedness of the isolates. Morphological test results showed that all 13 isolates were consistent with the established phenotypic characters of the genus such as pink colony colour, Gram negative and rod-shaped. Biochemically, the use of API® 20 NE tests showed heterogeneity of results and physiological tests exhibited that the isolates are primarily mesophilic and halotolerant, being able to grow at 2% salt. Phylogenetic analysis showed that the isolates are Methylobacterium populi, M. dankookense, M. lusitanum, M. radiotolerans and M. zatmanii. This study confirmed the presence and diversity of PPFM bacteria in Philippine shrimp paste. Further studies are needed to show the functional activity of Methylobacterium in Philippine shrimp paste production.

Potentials, Utilization, and Bioengineering of Plant Growth-Promoting Methylobacterium for Sustainable Agriculture

Plant growth-promoting bacteria (PGPB) have great potential to provide economical and sustainable solutions to current agricultural challenges. The Methylobacteria which are frequently present in the phyllosphere can promote plant growth and development. The Methylobacterium genus is composed mostly of pink-pigmented facultative methylotrophic bacteria, utilizing organic one-carbon compounds as the sole carbon and energy source for growth. Methylobacterium spp. have been isolated from diverse environments, especially from the surface of plants, because they can oxidize and assimilate methanol released by plant leaves as a byproduct of pectin formation during cell wall synthesis. Members of the Methylobacterium genus are good candidates as PGPB due to their positive impact on plant health and growth; they provide nutrients to plants, modulate phytohormone levels, and protect plants against pathogens. In this paper, interactions between Methylobacterium spp. and plants and how the bacteria promote crop growth is reviewed. Moreover, the following examples of how to engineer microbiomes of plants using plant-growth-promoting Methylobacterium are discussed in the present review: introducing external Methylobacterium spp. to plants, introducing functional genes or clusters to resident Methylobacterium spp. of crops, and enhancing the abilities of Methylobacterium spp. to promote plant growth by random mutation, acclimation, and engineering.

IMPACT OF PINK PIGMENTED FACULTATIVE METHYLOTROPHIC BACTERIA AS A BIO-STIMULANT ON GROWTH, HERB YIELD AND VOLATILE OIL ATTRIBUTES OF ROSEMARY (ROSMARINUS OFFICINALIS L.) PLANT

The effect of Pink Pigmented Facultative Methylotrophic (PPFM) bacteria as a bio-stimulant on vegetative growth, macro-nutrient elements ( N,P and K ) contents , chemical constituents and volatile oil percentage and components in rosemary plant (Rosmarinus officinalis L.) were studied during 2018 and 2019 seasons. Plant height (cm), number of lateral branches/plant, herb fresh & dry weights and root fresh & dry weights of rosemary plant were all greatly affected with all applied treatments of PPFM when compared with the control. The hugest effect was recorded by 1% PPFM bacteria spraying in both studied seasons. However, no significant effect was recorded due to PPFM applied treatments on P%. Highest values of total carbohydrates were recorded by both 1 and 3%. PPFM bacteria spraying in both studied seasons. Whereas, the highest values of C/N ratios were obtained with 3% PPFM in first season and 1% PPFM in the second one. A substantial effect from PPFM bacteria spraying was observed through its effect on reducing the level of NO3 and NO2 in rosemary herb when compared with the control. In any case, 3% of PPFM bacteria spraying was more effective than other treatments in recording the least values of NO3 and NO2 in first season compared to the control. Volatile oil % was enormously increased with 3% PPFM bacteria spraying in both studied seasons than other treatments or control. The main components of volatile oil of rosemary plant were Camphor, Eucalyptol, Verbenone, Borneol, Pinene, Pinanone, Bornyl acetate and Camphene. The treatments of PPFM bacteria spraying immensely stimulated the component % in volatile oil of rosemary herb than in the control. It could be concluded that the application of PPFM bacteria as foliar spraying in rosemary plant production had many advantages especially reducing NO3 and NO2 levels and increasing volatile oil % and volatile oil fractions not to mention it increased the percent of active ingredients.

GROWTH AND PRODUCTIVITY OF EGGPLANT AS AFFECTED BY PINK PIGMENTED FACULTATIVE METHYLOTROPHIC BACTERIA

The present study was suggested as an attempt to investigate the effects of Pink Pigmented Facultative Methylotrophic Bacteria (PPFM) dipping and foliar spray on vegetative growth and yield of eggplant (Solanum melongena L.). Field experiments were carried out during the summer seasons of 2014 and 2015 at the Experimental Farm, Faculty of Agriculture, Ain Shams University. The vegetative growth of eggplant responded positively to dipping seedlings in PPFM which gave the highest values of growth parameters, i.e. number of leaves per plant, leaf area, nitrogen & potassium concentration in leaf and total protein in fruit, in the two seasons as compared with control (dipping in water). The application of PPFM as foliar spray, increased significantly plant growth (plant length, leaf number, potassium concentration in leaf, vitamin C in fruit and total yield per plant) in the two seasons as compared with the other studied foliar application treatments. The interaction between dipping seedlings in PPFM and PPFM foliar spray with all concentrations gave synergistic effects on growth parameters and yield components of eggplant, during the two growing seasons as compared with either individual foliar application or control plants.

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This work highlighted a putative link between the physiological activity and genetic diversity of Methylobacterium species and the association with sugarcane roots and rhizoplane. In total, 40 isolates previously described as pink-pigmented facultative methylotrophic bacteria (PPFMs), were evaluated for their ability to fix nitrogen and solubilize inorganic phosphate, amylase and pectinase activity. This in vitro potential was positively correlated with the community isolated from the root tissues than those from the rhizoplane. Regarding the genomic fingerprinting, the (BOX-PCR) approach revealed a low similarity among the isolates, occurring sole 7 haplotypes harboring more than 70% of similarity among band patterns. These results revealed that the genomic fingerprinting of the isolates recovery from roots is different from the rhizoplane. Besides that, these haplotypes occurred on both sugarcane varieties. Using a phylogenetic sequencing approach based on the 16S rRNA gene, we observed a high abundance of sequences similar to Methylobacterium radiotolerans colonizing both plant tissue and sugarcane varieties were observed. Hence, it was suggested that the plant should select those Methylobacterium spp. with a high biotechnological potential to promote plant growth. Therefore, the bioprospection of specific endophytic bacterial groups comprise an important source of biotechnological potential to improve sugarcane growth and production. Key words: Pink-pigmented facultative methylotrophic bacteria, plant growth promotion, BOX-PCR, 16S rDNA, Methylobacterium radiotolerans.

The abundance of pink-pigmented facultative methylotrophs in the root zone of plant species in invaded coastal sage scrub habitat.

Pink-pigmented facultative methylotrophic bacteria (PPFMs) are associated with the roots, leaves and seeds of most terrestrial plants and utilize volatile C1 compounds such as methanol generated by growing plants during cell division. PPFMs have been well studied in agricultural systems due to their importance in crop seed germination, yield, pathogen resistance and drought stress tolerance. In contrast, little is known about the PPFM abundance and diversity in natural ecosystems, let alone their interactions with non-crop species. Here we surveyed PPFM abundance in the root zone soil of 5 native and 5 invasive plant species along ten invasion gradients in Southern California coastal sage scrub habitat. PPFMs were present in every soil sample and ranged in abundance from 102 to 105 CFU/g dry soil. This abundance varied significantly among plant species. PPFM abundance was 50% higher in the root zones of annual or biennial species (many invasives) than perennial species (all natives). Further, PPFM abundance appears to be influenced by the plant community beyond the root zone; pure stands of either native or invasive species had 50% more PPFMs than mixed species stands. In sum, PPFM abundance in the root zone of coastal sage scrub plants is influenced by both the immediate and surrounding plant communities. The results also suggest that PPFMs are a good target for future work on plant-microorganism feedbacks in natural ecosystems.

Evaluation of pink-pigmented facultative methylotrophic bacteria for phosphate solubilization

Thirteen pink-pigmented facultative methylotrophic (PPFM) strains isolated from Adyar and Cooum rivers in Chennai and forest soil samples in Tamil Nadu, India, along with Methylobacterium extorquens, M. organophilum, M. gregans, and M. komagatae were screened for phosphate solubilization in plates. P-solubilization index of the PPFMs grown on NBRIP-BPB plates for 7days ranged from 1.1 to 2.7. The growth of PPFMs in tricalcium phosphate amended media was found directly proportional to the glucose concentration. Higher phosphate solubilization was observed in four strains MSF 32 (415mgl(-l)), MDW 80 (301mgl(-l)), M. komagatae (279mgl(-l)), and MSF 34 (202mgl(-l)), after 7days of incubation. A drop in the media pH from 6.6 to 3.4 was associated with an increase in titratable acidity. Acid phosphatase activity was more pronounced in the culture filtrate than alkaline phosphatase activity. Adherence of phosphate to densely grown bacterial surface was observed under scanning electron microscope after 7-day-old cultures. Biochemical characterization and screening for methanol dehydrogenase gene (mxaF) confirmed the strains as methylotrophs. The mxaF gene sequence from MSF 32 clustered towards M. lusitanum sp. with 99% similarity. This study forms the first detailed report on phosphate solubilization by the PPFMs.

Influence of Fertilizer Levels and Growth Substances on Yield and Economics of Hybrid Maize

A field experiment was conducted at Tamil Nadu Agricultural University, Coimbatore during kharif 2008 to study the influence of fertilizer levels and foliar spray of plant growth substances on yield and economics of hybrid maize under irrigated condition. The experiment was laid out in a split plot design replicated thrice. Three fertilizer levels viz., 150:75:75, 200:100:100 and 250:125:125 NPK kg ha-1 constituted the main plot treatments. Foliar spray of growth substances viz., control (no spray), salicylic acid 100 ppm, boric acid 0.3%, PGR consortia 1.5%, TNAU Panchagavya 3% and Pink-pigmented facultative methylotrophic bacteria (PPFM) 106 dilution were assigned to sub plot. The results revealed that yield, gross return and net return were higher under the treatment combination 250:125:125 NPK kg ha-1 along with the foliar spray of PGR consortia @ 1.5% twice at 45 DAS and 55 DAS. However, the highest benefit cost ratio was recorded under the treatment combination 200:100:100 NPK kg ha-1 along with the foliar spray of PGR consortia @ 1.5%.

Influence of Fertilizer Levels and Growth Regulating Substances on Growth, Nutrient Use Efficiency and Yield of Hybrid Maize

A field experiment was conducted at Tamil Nadu Agricultural University, Coimbatore during kharif 2008 to study the influence of fertilizer levels and foliar spray of plant growth substances on growth, nutrient use efficiency and yield of hybrid maize under irrigated condition. The experiment was laid out in a split plot design replicated thrice. Three fertilizer levels viz., 150:75:75, 200:100:100 and 250:125:125 NPK kg ha-1 constituted the main plot treatments. Foliar spray of growth substances viz., control (no spray), salicylic acid 100 ppm, boric acid 0.3%, PGR consortia 1.5%, TNAU Panchagavya 3% and Pink-pigmented facultative methylotrophic bacteria (PPFM) 106 dilution were assigned to sub plot. The results of the experiment revealed that among the fertilizer levels, 250:125:125 NPK kg ha-1 recorded taller plants, higher LAI, DMP, nutrient uptake and yield. However, the yield was comparable with 200:100:100 NPK kg ha-1. Regarding the growth substances, PGR consortia 1.5% recorded better growth parameters, higher nutrient uptake and yield than the other growth substances. The highest agronomic efficiency (AE) and apparent recovery (AR) for nitrogen, phosphorus and potassium were recorded under the treatment combination 200:100:100 NPK kg ha-1 along with the foliar spray of PGR consortia @ 1.5%.

Methylobacterium Genome Sequences: A Reference Blueprint to Investigate Microbial Metabolism of C1 Compounds from Natural and Industrial Sources

BackgroundMethylotrophy describes the ability of organisms to grow on reduced organic compounds without carbon-carbon bonds. The genomes of two pink-pigmented facultative methylotrophic bacteria of the Alpha-proteobacterial genus Methylobacterium, the reference species Methylobacterium extorquens strain AM1 and the dichloromethane-degrading strain DM4, were compared.Methodology/Principal FindingsThe 6.88 Mb genome of strain AM1 comprises a 5.51 Mb chromosome, a 1.26 Mb megaplasmid and three plasmids, while the 6.12 Mb genome of strain DM4 features a 5.94 Mb chromosome and two plasmids. The chromosomes are highly syntenic and share a large majority of genes, while plasmids are mostly strain-specific, with the exception of a 130 kb region of the strain AM1 megaplasmid which is syntenic to a chromosomal region of strain DM4. Both genomes contain large sets of insertion elements, many of them strain-specific, suggesting an important potential for genomic plasticity. Most of the genomic determinants associated with methylotrophy are nearly identical, with two exceptions that illustrate the metabolic and genomic versatility of Methylobacterium. A 126 kb dichloromethane utilization (dcm) gene cluster is essential for the ability of strain DM4 to use DCM as the sole carbon and energy source for growth and is unique to strain DM4. The methylamine utilization (mau) gene cluster is only found in strain AM1, indicating that strain DM4 employs an alternative system for growth with methylamine. The dcm and mau clusters represent two of the chromosomal genomic islands (AM1: 28; DM4: 17) that were defined. The mau cluster is flanked by mobile elements, but the dcm cluster disrupts a gene annotated as chelatase and for which we propose the name “island integration determinant” (iid).Conclusion/SignificanceThese two genome sequences provide a platform for intra- and interspecies genomic comparisons in the genus Methylobacterium, and for investigations of the adaptive mechanisms which allow bacterial lineages to acquire methylotrophic lifestyles.

Phenotypic Characterization of Pink Pigmented Facultative Methylotrophic Bacteria from Soil Exposed to Vehicular Soot

Phenotypic Characterization of Pink Pigmented Facultative Methylotrophic Bacteria from Soil Exposed to Vehicular Soot

Phenotypic characterization of air-borne pink pigmented facultative methylotrophic bacteria from high vehicular traffic density environment in Manila, Philippines

Phenotypic characterization of air-borne pink pigmented facultative methylotrophic bacteria from high vehicular traffic density environment in Manila, Philippines

Genetic diversity and phylogeny of pink-pigmented facultative methylotrophic bacteria isolated from the phyllosphere of tropical crop plants

Genetic diversity of pink-pigmented facultative methylotrophic bacteria belonging to the genus, Methylobacterium, was assessed using 16S ribosomal RNA (rRNA) gene sequencing, amplified ribosomal DNA restriction analysis (ARDRA), and differential carbon-substrate utilization profile in the phyllosphere of cotton, maize, sunflower, soybean, and mentha plants. Methylobacterium populi, Methylobacterium thiocyanatum, Methylobacterium suomiense, M. aminovorans, and Methylobacterium fujisawaense were identified to colonize the phyllosphere of these crop plants. Among these, M. populi found to be the dominating species followed by M. aminovorans. The diversity indices like Shannon index of diversity, Pielou index of evenness, and Margalef index of richness calculated based on combined data of ARDRA, 16S rRNA gene sequence, and differential carbon-substrate utilization revealed that sunflower leaves showed richest methylobacterial diversity, followed by soybean, while cotton and mentha leaves recorded the lowest diversity. The outcome of this study indicates that diversity of Methylobacterium in phyllosphere depends upon the host plant species colonized.

Plant Growth–Promoting Methylobacterium Induces Defense Responses in Groundnut (Arachis hypogaea L.) Compared with Rot Pathogens

This study, framed in two different phases, studied the plant-growth promotion and the induction of systemic resistance in groundnut by Methylobacterium. Seed imbibition with Methylobacterium sp. increased germination by 19.5% compared with controls. Combined inoculation of Methylobacterium sp. with Rhizobium sp. also significantly increased plant growth, nodulation, and yield attributes in groundnut compared with individual inoculation of Rhizobium sp. Methylobacterium sp. challenge-inoculated with Aspergillus niger/Sclerotium rolfsii in groundnut significantly enhanced germination percentage and seedling vigour and showed increased phenylalanine ammonia lyase (PAL), beta-1,3-glucanase, and peroxidase (PO) activities. Under pot-culture conditions, in Methylobacterium sp. seed-treated groundnut plants challenge-inoculated with A. niger/S. rolfsii through foliar sprays on day 30, the activities of enzymes PO, PAL, and beta-1,3-glucanase increased constantly from 24 to 72 hours, after which decreased activity was noted. Five isozymes of polyphenol oxidase and PO could be detected in Methylobacterium-treated plants challenged with A. niger/S. rolfsii. Induced systemic resistance activity in groundnut against rot pathogens in response to methylotrophic bacteria suggests the possibility that pink-pigmented facultative methylotrophic bacteria might be used as a means of biologic disease control.

Pink-pigmented facultative methylotrophic bacteria accelerate germination, growth and yield of sugarcane clone Co86032 (Saccharum officinarum L.)

The existence of Methylobacterium as a symbiont with sugarcane and its influence on crop growth at various stages was examined. Pink-pigmented facultative methylotrophic bacteria (PPFMs) strains isolated from different parts of the sugarcane clone Co86032 showed growth on methanol, and were further confirmed based on the mxaF gene encoding the α-subunit of the methanol dehydrogenase by polymerase chain reaction amplification using specific primers. True seeds inoculated with PPFMs had a higher germination percent and rate of germination than the control. A combined treatment of seed imbibition, soil application and phyllosphere spray increased specific leaf area, plant height, number of internodes, and cane yield. Immunological determination of cytokinin in young and mature leaves significantly increased when the epiphytic population on the leaf surface increased. Trends in sugar qualities in the form of Pol (sucrose) % in cane, Brix % in cane, and commercial cane sugar were similar to that of cane yield. These effects might be mediated by the production or synthesis of plant hormones.

Indole-3-acetic acid production by pink-pigmented facultative methylotrophic bacteria

In examining the presence of indole-3-acetic acid (IAA) in supernatants of pink-pigmented facultativemethylotrophic (PPFMs) bacterial cultures, three out of the 16 isolates tested showed a positive reaction ina colorimetric assay. The presence was further unambiguously con?rmed by high-performance liquidchromatography in combination with NMR. The IAA production was signi?cantly stimulated byL-tryptophan. These results prove that PPFM bacteria are able to produce the plant hormone IAA.

Plant colonization by pink-pigmented facultative methylotrophic bacteria (PPFMs)

Bacteria belonging to the genus Methylobacterium are characterized by being able to rely on methanol as a sole carbon and energy source and by presenting a more or less intense pink reddish pigmentation. These bacteria, also referred to as pink-pigmented methylotrophic bacteria (PPFMs), are common inhabitants of the phyllosphere and are found in many other environmental samples. Since they grow slowly they are often overlooked and their impact on phyllosphere microbial communities and on the plants harboring them is not well studied nor has their ecology been elucidated. In a survey of PPFM colonization in three different agricultural sites, PPFM populations were identified on both red clover and winter wheat, but red clover was more consistently colonized. Isolations from collected leaves showed PPFM populations to decrease from spring towards summer, but they increased again towards the end of the cropping season. Isolates from red clover readily colonized winter wheat leaves and vice versa in greenhouse experiments, but population sizes were dependent on the application procedure. Tested isolates had also good potential to colonize the rhizosphere, especially after seed inoculations. Confocal scanning laser microscopy showed gfp-tagged isolates to colonize the surface of clover leaves by forming large aggregates.