Cell-Free Supernatant (CFS) from Bacillus subtilis EB2004S and Lactobacillus helveticus EL2006H Cultured at a Range of pH Levels Modulates Potato Plant Growth under Greenhouse Conditions.

Agriculture involving industrial fertilizers is another major human made contributing factor to soil pH variation after natural factors such as soil parent rock, weathering time span, climate, and vegetation. The current study assessed the potential effect of cell-free supernatant (CFS) obtained from Bacillus subtilis EB2004S and Lactobacillus helveticus EL2006H cultured at three pH levels (5, 7, and 8) on potato (var Goldrush) growth enhancement in a greenhouse pot experiment. The results showed that CFSs obtained from B. subtilis EB2004S and L. helveticus EL2006H cultured at pH 5 significantly improved photosynthetic rates, stomatal conductance, root fresh weight, and whole plant fresh weight. interactive effects of pot pH and that of CFSs obtained from pH 5 influenced chlorophyll, plant height, and shoot and whole plant fresh weight. Moreover, treatment 52EB2004S~0.4% initiated early tuberization for potato grown at pH 7 and 8. Potato grown at pH 5, which received a 72EB2004S~0.4% CFS treatment, had greater whole plant fresh and dry weight than that treated with L. helveticus EL2006H CFS and a positive control. Taken together, the findings of this study are unique in that it probed the effect of CFS produced under differing pH conditions which revealed a new possibility to mitigate stresses in plants.

Secretome Analysis of the Plant Biostimulant Bacteria Strains Bacillus subtilis (EB2004S) and Lactobacillus helveticus (EL2006H) in Response to pH Changes.

It is well-known that there is a high frequency of plant-growth-promoting strains in Bacillus subtilis and that these can be effective under both stressful and stress-free conditions. There are very few studies of this activity in the case of Lactobacillus helveticus. In this study, the effects of pH on the secretome (proteins) in the cell-free supernatants of two bacterial strains were evaluated. The bacteria were cultured at pH 5, 7 and 8, and their secretome profiles were analyzed, with pH 7 (optimal growth pH) considered as the "control". The results showed that acidity (lower pH 5) diminishes the detectable production of most of the secretome proteins, whereas alkalinity (higher pH 8) increases the detectable protein production. At pH 5, five (5) new proteins were produced by L. helveticus, including class A sortase, fucose-binding lectin II, MucBP-domain-containing protein, SLAP-domain-containing protein and hypothetical protein LHEJCM1006_11110, whereas for B. subtilis, four (4) types of proteins were uniquely produced (p ≤ 0.05), including helicase-exonuclease AddAB subunit AddB, 5-methyltetrahydropteroyltriglutamate-homocysteine S-methyltransferase, a cluster of ABC-F family ATP-binding-cassette-domain-containing proteins and a cluster of excinuclease ABC (subunit B). At pH 8, Bacillus subtilis produced 56 unique proteins. Many of the detected proteins were involved in metabolic processes, whereas the others had unknown functions. The unique and new proteins with known and unknown functions suggest potential the acclimatization of the microbes to pH stress.

Antibiotics Resistance in Rhizobium: Type, Process, Mechanism and Benefit for Agriculture.

The use of high-quality rhizobial inoculants on agricultural legumes has contributed substantially to the N economy of farming systems through inputs from biological nitrogen fixation (BNF). Large populations of symbiotically effective rhizobia should be available in the rhizosphere for symbiotic BNF with host plants. The rhizobial populations should also be able to compete and infect host plants. However, the rhizosphere comprises large populations of different microorganisms. Some of these microorganisms naturally produce antibiotics which are lethal to susceptible rhizobial populations in the soil. Therefore, intrinsic resistance to antibiotics is a desirable trait for the rhizobial population. It increases the rhizobia's chances of growth, multiplication and persistence in the soil. With a large population of rhizobia in the soil, infectivity of host plants and the subsequent BNF efficiency can be guaranteed. This review, therefore, puts together findings by various researchers on antibiotic resistance in bacteria with the main emphasis on rhizobia. It describes the different modes of action of different antibiotics, the types of antibiotic resistance exhibited by rhizobia, the mechanisms of acquisition of antibiotic resistance in rhizobia and the levels of tolerance of different rhizobial species to different antibiotics.

Effect of NaCl stress on exoproteome profiles of Bacillus amyloliquefaciens EB2003A and Lactobacillus helveticus EL2006H.

Salt stress can affect survival, multiplication and ability of plant growth promoting microorganisms to enhance plant growth. Changes in a microbe's proteome profile is one of the mechanisms employed by PGPM to enhance tolerance of salt stress. This study was focused on understanding changes in the exoproteome profile of Bacillus amyloliquefaciens EB2003A and Lactobacillus helveticus EL2006H when exposed to salt stress. The strains were cultured in 100 mL M13 (B. amyloliquefaciens) and 100 mL De man, Rogosa and Sharpe (MRS) (L. helveticus) media, supplemented with 200 and 0 mM NaCl (control), at pH 7.0. The strains were then incubated for 48 h (late exponential growth phase), at 120 rpm and 30 (B. amyloliquefaciens) and 37 (L. helveticus) °C. The microbial cultures were then centrifuged and filtered sterilized, to obtain cell free supernatants whose proteome profiles were studied using LC-MS/MS analysis and quantified using scaffold. Results of the study revealed that treatment with 200 mM NaCl negatively affected the quantity of identified proteins in comparison to the control, for both strains. There was upregulation and downregulation of some proteins, even up to 100%, which resulted in identification of proteins significantly unique between the control or 200 mM NaCl (p ≤ 0.05), for both microbial species. Proteins unique to 200 mM NaCl were mostly those involved in cell wall metabolism, substrate transport, oxidative stress tolerance, gene expression and DNA replication and repair. Some of the identified unique proteins have also been reported to enhance plant growth. In conclusion, based on the results of the work described here, PGPM alter their exoproteome profile when exposed to salt stress, potentially upregulating proteins that enhance their tolerance to this stress.

Lactobacillus helveticus EL2006H cell-free supernatant enhances growth variables in Zea mays (maize), Glycine max L. Merill (soybean) and Solanum tuberosum (potato) exposed to NaCl stress.

Plant growth promoting microorganisms and their derived compounds, such as cell-free supernatant (CFS), enhance plant growth under stressed and non stressed conditions. Such technology is sustainable and environmentally friendly, which is desirable amidst the climate change threat. The current study evaluated the effect of CFS obtained from Lactobacillus helveticus EL2006H on its ability to enhance mean percentage germination and mean radicle length of corn and soybean, as well as growth parameters of potato, using treatment formulations that consisted of 0.2 and 1.0% [v/v] L. helveticus EL2006H CFS concentrations and 100 mM NaCl and 150 mM NaCl levels. Results show that treatment with 100 mM NaCl lowered percentage germination of corn by 52.63%, at 72 h, and soybean by 50%, at 48 h. Treatment with 100 NaCl +0.2% EL2006H enhanced percentage germination of soybean by 44.37%, at 48 h, in comparison to that of the 100 mM NaCl control. One hundred mM NaCl lowered radicle length of corn and soybean by 38.58 and 36.43%, respectively. Treatment with 100 Mm NaCl +1.0% EL2006H significantly increased radicle length of corn by 23.04%. Treatment with 100 mM NaCl +0.2% EL2006H significantly increased photosynthetic rate, leaf greenness and fresh weight of potato. Increasing NaCl concentration to 150 NaCl lowered the effectiveness of the 0.2% EL2006H CFS on the same growth variables of potato. In general, the lower CFS concentration of 0.2% was more efficient at enhancing germination in soybean while the higher concentration of 1.0% was more efficient at enhancing radicle length of corn. There was an observed variation in the effectiveness of L. helveticus EL2006H CFS across the different CFS concentrations, NaCl levels and crop species studied. In conclusion, based on findings of this study, CFS obtained from L. helveticus can be used as a bio stimulant to enhance growth of corn, soybean and potato. However, further studies need to be conducted, for validation, especially under field conditions, for commercial application.

Microbial Derived Compounds Are a Promising Approach to Mitigating Salinity Stress in Agricultural Crops.

The use of microbial derived compounds is a technological approach currently gaining popularity among researchers, with hopes of complementing, supplementing and addressing key issues associated with use of microbial cells for enhancing plant growth. The new technology is a promising approach to mitigating effects of salinity stress in agricultural crops, given that these compounds could be less prone to effects of salt stress, are required in small quantities and are easier to store and handle than microbial cells. Microorganism derived compounds such as thuricin17, lipochitooligosaccharides, phytohormones and volatile organic compounds have been reported to mitigate the effects of salt stress in agricultural crops such as soybean and wheat. This mini-review compiles current knowledge regarding the use of microbe derived compounds in mitigating salinity stress in crops, the mechanisms they employ as well as future prospects.

Microbial Derived Compounds, a Step Toward Enhancing Microbial Inoculants Technology for Sustainable Agriculture.

Sustainable agriculture remains a focus for many researchers, in an effort to minimize environmental degradation and climate change. The use of plant growth promoting microorganisms (PGPM) is a hopeful approach for enhancing plant growth and yield. However, the technology faces a number of challenges, especially inconsistencies in the field. The discovery, that microbial derived compounds can independently enhance plant growth, could be a step toward minimizing shortfalls related to PGPM technology. This has led many researchers to engage in research activities involving such compounds. So far, the findings are promising as compounds have been reported to enhance plant growth under stressed and non-stressed conditions in a wide range of plant species. This review compiles current knowledge on microbial derived compounds, taking a reader through a summarized protocol of their isolation and identification, their relevance in present agricultural trends, current use and limitations, with a view to giving the reader a picture of where the technology has come from, and an insight into where it could head, with some suggestions regarding the probable best ways forward.

Nature and mechanisms of aluminium toxicity, tolerance and amelioration in symbiotic legumes and rhizobia.

Recent findings on the effect of aluminium (Al) on the functioning of legumes and their associated microsymbionts are reviewed here. Al represents 7% of solid matter in the Earth's crust and is an important abiotic factor that alters microbial and plant functioning at very early stages. The trivalent Al (Al3+) dominates at pH < 5 in soils and becomes a constraint to legume productivity through its lethal effect on rhizobia, the host plant and their interaction. Al3+ has lethal effects on many aspects of the rhizobia/legume symbiosis, which include a decrease in root elongation and root hair formation, lowered soil rhizobial population, and suppression of nitrogen metabolism involving nitrate reduction, nitrite reduction, nitrogenase activity and the functioning of uptake of hydrogenases (Hup), ultimately impairing the N2 fixation process. At the molecular level, Al is known to suppress the expression of nodulation genes in symbiotic rhizobia, as well as the induction of genes for the formation of hexokinase, phosphodiesterase, phosphooxidase and acid/alkaline phosphatase. Al toxicity can also induce the accumulation of reactive oxygen species and callose, in addition to lipoperoxidation in the legume root elongation zone. Al tolerance in plants can be achieved through over-expression of citrate synthase gene in roots and/or the synthesis and release of organic acids that reverse Al-induced changes in proteins, as well as metabolic regulation by plant-secreted microRNAs. In contrast, Al tolerance in symbiotic rhizobia is attained via the production of exopolysaccharides, the synthesis of siderophores that reduce Al uptake, induction of efflux pumps resistant to heavy metals and the expression of metal-inducible (dmeRF) gene clusters in symbiotic Rhizobiaceae. In soils, Al toxicity is usually ameliorated through liming, organic matter supply and use of Al-tolerant species. Our current understanding of crop productivity in high Al soils suggests that a much greater future accumulation of Al is likely to occur in agricultural soils globally if crop irrigation is increased under a changing climate.

Microsymbiont diversity and phylogeny of native bradyrhizobia associated with soybean (Glycine max L. Merr.) nodulation in South African soils.

The genetic diversity and identification of slow- and fast-growing soybean root nodule bacterial isolates from different agro-climatic regions in Mpumalanga, Limpopo and Gauteng Provinces of South Africa were evaluated. The 16S-rDNA-RFLP analysis of 100 rhizobial isolates and eight reference type strains placed the isolates into six major clusters, and revealed their site-dependent genomic diversity. Sequence analysis of single and concatenated housekeeping genes (atpD, glnII and gyrB), as well as the symbiotic gene nifH captured a considerably higher level of genetic diversity and indicated the dominance of Bradyrhizobium diazoefficiens and Bradyrhizobium japonicum in Mpumalanga, Limpopo and Gauteng Provinces. Gene sequence similarities of isolates with type strains of Bradyrhizobium ranged from 97.3 to 100% for the 16S rDNA, and 83.4 to 100% for the housekeeping genes. The glnII gene phylogeny showed discordance with the other genes, suggesting lateral gene transfer or recombination events. Concatenated gene sequence analysis showed that most of the isolates did not align with known type strains and might represent new species from South Africa. This underscores the high genetic variability associated with soybean Bradyrhizobium in South African soils, and the presence of an important reservoir of novel soybean-nodulating bradyrhizobia in the country. In this study, the grouping of isolates was influenced by site origin, with Group I isolates originating from Limpopo Province and Groups II and III from Mpumlanga Province in the 16S rDNA-RFLP analysis.