Ramasamy Krishnamoorthy

Arbuscular Mycorrhizal Fungi Community Structure, Abundance and Species Richness Changes in Soil by Different Levels of Heavy Metal and Metalloid Concentration

Arbuscular Mycorrhizal Fungi (AMF) play major roles in ecosystem functioning such as carbon sequestration, nutrient cycling, and plant growth promotion. It is important to know how this ecologically important soil microbial player is affected by soil abiotic factors particularly heavy metal and metalloid (HMM). The objective of this study was to understand the impact of soil HMM concentration on AMF abundance and community structure in the contaminated sites of South Korea. Soil samples were collected from the vicinity of an abandoned smelter and the samples were subjected to three complementary methods such as spore morphology, terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis (DGGE) for diversity analysis. Spore density was found to be significantly higher in highly contaminated soil compared to less contaminated soil. Spore morphological study revealed that Glomeraceae family was more abundant followed by Acaulosporaceae and Gigasporaceae in the vicinity of the smelter. T-RFLP and DGGE analysis confirmed the dominance of Funneliformis mosseae and Rhizophagus intraradices in all the study sites. Claroideoglomus claroideum, Funneliformis caledonium, Rhizophagus clarus and Funneliformis constrictum were found to be sensitive to high concentration of soil HMM. Richness and diversity of Glomeraceae family increased with significant increase in soil arsenic, cadmium and zinc concentrations. Our results revealed that the soil HMM has a vital impact on AMF community structure, especially with Glomeraceae family abundance, richness and diversity.

Alleviation of salt stress in maize plant by co-inoculation of arbuscular mycorrhizal fungi and Methylobacterium oryzae CBMB20

Arbuscular mycorrhizal fungi (AMF) and plant growth-promoting bacteria (PGPB) can have multiple impacts on phytohormone production and nutrient uptake to improve plant growth. Co-inoculation of AMF and PGPB can mitigate the effects of salinity in plants. This study was conducted to evaluate the efficacy of co-inoculation of AMF (Glomus etunicatum) and Methylobacterium oryzae CBMB20 to alleviate salt stress and improve maize growth. Single and co-inoculation of AMF and M. oryzae CBMB20 significantly increased dry biomass, AMF root colonization, and nutrient accumulation in maize plants under salt stress. The proline content and Na uptake were significantly reduced in co-inoculation of AMF and M. oryzae CBMB20. In addition, co-inoculation of AMF and M. oryzae CBMB20 (soil) showed higher AMF density and AMF colonization compared to single AMF treatment. Among all treatments, co-inoculation of AMF and M. oryzae CBMB20 (soil) was more effective to ameliorate salt stress and improve plant growth. These results demonstrate that co-inoculation of AMF and M. oryzae CBMB20 (soil application) may be an effective alternative to enhance plant growth under salinity stress.

Cold Stress Tolerance in Psychrotolerant Soil Bacteria and Their Conferred Chilling Resistance in Tomato (Solanum lycopersicum Mill.) under Low Temperatures

The present work aimed to study the culturable diversity of psychrotolerant bacteria persistent in soil under overwintering conditions, evaluate their ability to sustain plant growth and alleviate chilling stress in tomato. Psychrotolerant bacteria were isolated from agricultural field soil samples colleced during winter and then used to study chilling stress alleviation in tomato plants (Solanum lycopersicum cv Mill). Selective isolation after enrichment at 5°C yielded 40 bacterial isolates. Phylogenetic studies indicated their distribution in genera Arthrobacter, Flavimonas, Flavobacterium, Massilia, Pedobacter and Pseudomonas. Strains OS211, OB146, OB155 and OS261 consistently improved germination and plant growth when a chilling stress of 15°C was imposed and therefore were selected for pot experiments. Tomato plants treated with the selected four isolates exhibited significant tolerance to chilling as observed through reduction in membrane damage and activation of antioxidant enzymes along with proline synthesis in the leaves when exposed to chilling temperature conditions (15°C). Psychrotolerant physiology of the isolated bacteria combined with their ability to improve germination, plant growth and induce antioxidant capacity in tomato plants can be employed to protect plants against chilling stress.

Expression of an exogenous 1-aminocyclopropane-1-carboxylate deaminase gene in psychrotolerant bacteria modulates ethylene metabolism and cold induced genes in tomato under chilling stress.

The role of stress induced ethylene under low temperature stress has been controversial and hitherto remains unclear. In the present study, 1-aminocyclopropane-1-carboxylate deaminase (ACCD) gene, acdS expressing mutant strains were generated from ACCD negative psychrotolerant bacterial strains Flavobacterium sp. OR306 and Pseudomonas frederiksbergensis OS211, isolated from agricultural soil during late winter. After transformation with plasmid pRKACC which contained the acdS gene, both the strains were able to exhibit ACCD activity in vitro. The effect of this ACCD under chilling stress with regards to ethylene was studied in tomato plants inoculated with both acdS expressing and wild type bacteria. On exposing the plants to one week of chilling treatment at 12/10 °C, it was found that stress ethylene, ACC accumulation and ACO activity which are markers of ethylene stress, were significantly reduced in plants inoculated with the acdS gene transformed mutants. In case of plants inoculated with strain OS211-acdS, ethylene emission, ACC accumulation and ACO activity was significantly reduced by 52%, 75.9% and 23.2% respectively compared to uninoculated control plants. Moreover, expression of cold induced LeCBF1 and LeCBF3 genes showed that these genes were significantly induced by the acdS transformed mutants in addition to reduced expression of ethylene-responsive transcription factor 13 (ETF-13) and ACO genes. Induced expression of LeCBF1 and LeCBF3 in plants inoculated with acdS expressing mutants compared to wild type strains show that physiologically evolved stress ethylene and its transcription factors play a role in regulation of cold induced genes as reported earlier in the literature.

Genetic Diversity and Association Characters of Bacteria Isolated from Arbuscular Mycorrhizal Fungal Spore Walls.

Association between arbuscular mycorrhizal fungi (AMF) and bacteria has long been studied. However, the factors influencing their association in the natural environment is still unknown. This study aimed to isolate bacteria associated with spore walls of AMF and identify their potential characters for association. Spores collected from coastal reclamation land were differentiated based on their morphology and identified by 18S rDNA sequencing as Funneliformis caledonium, Racocetra alborosea and Funneliformis mosseae. Bacteria associated with AMF spore walls were isolated after treating them with disinfection solution at different time intervals. After 0, 10 and 20 min of spore disinfection, 86, 24 and 10 spore associated bacteria (SAB) were isolated, respectively. BOX-PCR fingerprinting analysis showed that diverse bacterial communities were associated to AMF spores. Bacteria belonging to the same genera could associate with different AMF spores. Gram positive bacteria were more closely associated with AMF spores. Isolated SAB were characterized and tested for spore association characters such as chitinase, protease, cellulase enzymes and exopolysaccharide production (EPS). Among the 120 SAB, 113 SAB were able to show one or more characters for association and seven SAB did not show any association characters. The 16S rDNA sequence of SAB revealed that bacteria belonging to the phyla Firmicutes, Proteobacteria, Actinobacteria and Bactereiodes were associated with AMF spore walls.

Mycorrhizal Symbiotic Efficiency on C3 and C4 Plants under Salinity Stress – A Meta-Analysis

A wide range of C3 and C4 plant species could acclimatize and grow under the impact of salinity stress. Symbiotic relationship between plant roots and arbuscular mycorrhizal fungi (AMF) are widespread and are well known to ameliorate the influence of salinity stress on agro-ecosystem. In the present study, we sought to understand the phenomenon of variability on AMF symbiotic relationship on saline stress amelioration in C3 and C4 plants. Thus, the objective was to compare varied mycorrhizal symbiotic relationship between C3 and C4 plants in saline conditions. To accomplish the above mentioned objective, we conducted a random effects models meta-analysis across 60 published studies. An effect size was calculated as the difference in mycorrhizal responses between the AMF inoculated plants and its corresponding control under saline conditions. Responses were compared between (i) identity of AMF species and AMF inoculation, (ii) identity of host plants (C3 vs. C4) and plant functional groups, (iii) soil texture and level of salinity and (iv) experimental condition (greenhouse vs. field). Results indicate that both C3 and C4 plants under saline condition responded positively to AMF inoculation, thereby overcoming the predicted effects of symbiotic efficiency. Although C3 and C4 plants showed positive effects under low (EC < 4 ds/m) and high (>8 ds/m) saline conditions, C3 plants showed significant effects for mycorrhizal inoculation over C4 plants. Among the plant types, C4 annual and perennial plants, C4 herbs and C4 dicot had a significant effect over other counterparts. Between single and mixed AMF inoculants, single inoculants Rhizophagus irregularis had a positive effect on C3 plants whereas Funneliformis mosseae had a positive effect on C4 plants than other species. In all of the observed studies, mycorrhizal inoculation showed positive effects on shoot, root and total biomass, and in nitrogen, phosphorous and potassium (K) uptake. However, it showed negative effects in sodium (Na) uptake in both C3 and C4 plants. This influence, owing to mycorrhizal inoculation, was significantly higher in K uptake in C4 plants. For our analysis, we concluded that AMF-inoculated C4 plants showed more competitive K+ ions uptake than C3 plants. Therefore, maintenance of high cytosolic K+/Na+ ratio is a key feature of plant salt tolerance. Studies on the detailed mechanism for the selective transport of K in C3 and C4 mycorrhizal plants under salt stress is lacking, and this needs to be explored.

Exploration of Rice Husk Compost as an Alternate Organic Manure to Enhance the Productivity of Blackgram in Typic Haplustalf and Typic Rhodustalf

The present study was aimed at using cellulolytic bacterium Enhydrobacter and fungi Aspergillus sp. for preparing compost from rice husk (RH). Further, the prepared compost was tested for their effect on blackgram growth promotion along with different levels of recommended dose of fertilizer (RDF) in black soil (typic Haplustalf) and red soil (typic Rhodustalf) soil. The results revealed that, inoculation with lignocellulolytic fungus (LCF) Aspergillus sp. @ 2% was considered as the most efficient method of composting within a short period. Characterization of composted rice husk (CRH) was examined through scanning electron microscope (SEM) for identifying significant structural changes. At the end of composting, N, P and K content increased with decrease in CO2 evolution, C:N and C:P ratios. In comparison to inorganic fertilization, an increase in grain yield of 16% in typic Haplustalf and 17% in typic Rhodustalf soil over 100% RDF was obtained from the integrated application of CRH@ 5 t ha−1 with 50% RDF and biofertilizers. The crude protein content was maximum with the combined application of CRH, 50% RDF and biofertilizers of 20% and 21% in typic Haplustalf and typic Rhodustalf soils, respectively. Nutrient rich CRH has proved its efficiency on crop growth and soil fertility.

ACC Deaminase Producing Arsenic Tolerant Bacterial Effect on Mitigation of Stress Ethylene Emission in Maize Grown in an Arsenic Polluted Soil

Arsenic is a known hazardous metalloid not only to the animals but also to plants. With high concentrations, it can impede normal plant growth and cause even death of plants at extremely high levels. A known plant response to stress conditions such as toxic levels of metal (loids) is the production of stress ethylene, causing inhibitory effect on root growth in plants. When the effect of various arsenic concentrations was tested to maize plant, the stress ethylene emission proportionately increased with increasing concentration of As(V). The inoculation of two arsenic tolerant bacteria; Pseudomonas grimonti JS126 and Pseudomonas taiwanensis JS238 having respective high and low 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity reduced stress ethylene emission by 59% and 30% in maize grown in arsenic polluted soils. The result suggested the possible use of Pseudomonas grimonti JS126 for phytoremediation of arsenic polluted soils.

Effect of Dispersed and Proximate Inoculation Methods of Glomus etunicatum on Root Colonization of Sorghum-Sudangrass Hybrid

Information on the effective application method of arbuscular mycorrhizal fungi (AMF) inoculum is still inadequate. This work was performed to assess two AMF inoculation methods (dispersed and proximate) on root colonization of sorghum- sudangrass hybrid (Sorghum bicolor L.). In dispersed inoculation method, spores were inoculated in 2 kg pots of soil in which 5 day-old seedlings were transplanted and maintained for 50 days. In the proximate inoculation method, spores were first introduced in 500 mL pots where seeds were sown. After 10 days, the seedlings with the 500 mL soil were transferred to 2 kg pots without disturbing the contents. After 50 days of growth, root colonization and arbuscule abundance significantly increased (over 100%) in proximate method of inoculation. Moreover, sorghum-sudangrass hybrid had higher shoot growth (182.5 cm) and Glomalin related soil protein (GRSP) production in proximate method. Nutrient accumulation, particularly total nitrogen (82.61 mg plant-1), was also found to be higher in proximate method of inoculation. Our results demonstrate that the proximate method of inoculation may improve the early stage mycorrhizal symbiosis and inoculum performance in Saemangeum reclaimed soil.