Liming and Nitrogen Fertilization Effects on Soil Microbial Community in Long Term No-till

Abstract Soil management influences organic matter decomposition rates as well soil microbial community functional behavior. No-till (NT) is the most used management system by farmers due to its conservation practices and high productivity. The main objective of this study was to evaluate the impact of surface-applied lime, nitrogen (N) application, and black oat residues on soil microbial community of a Typic Hapludox under continuous NT. Therefore, soil chemical attributes, microbial biomass carbon, basal respiration, metabolic quotient, most probable number of diazotrophs, as well as bacterial functional analysis were performed. The effect of liming and N fertilization amendments inputs were saw in soil respiration and metabolic quotient measurements, showing them to be good indicators of soil quality. Further studies should be carried out in order to molecularly identify microbial communities present in soils with different liming and N fertilization management to evaluate the behavior of specific bacterial taxa under such conditions.

Plant growth-promoting properties of free-living diazotrophic rhizobacteria from Tangerine (Citrus reticulate L.) var Batu 55

Aims@#Microbial cultures with multi-biological activities in increasing plant growth were developed to be an alternative way to reduce dependency on chemical fertilizer and to support plants productivity. The aims of this study were to characterize the potency of Tangerine var. Batu 55 diazotroph rhizobacteria as Plant Growth Promoting Rhizobacteria (PGPR) agents and to identify diazotroph rhizobacteria with multi-biological activities especially the activity of nitrogen fixation, ammonia (NH3) production, phosphate solubilizing, and Indole Acetic Acid (IAA) production.@*Methodology and results@#A total of 21 nitrogen-fixing bacteria (diazotroph) were isolated from Tangerine rhizosphere soil. Screening of PGPR isolates candidates were performed by in vitro assays consist of phytohormone Indole Acetic Acid (IAA) production, ammonia production, and phosphate-solubilizing assay. Candidates of PGPR isolates were identified based on 16S rDNA sequences. The result revealed that three isolates (Dbs 1, Dbs 2, and Dbm 3) had multibiological activities. Isolates of Dbs 1, Dbs 2, and Dbm 3 capable producing ammonia up to 10 μg/mL; 9.1 μg/mL; and 3.8 μg/mL, activity of IAA production were 30.08 μg/mL; 24.68 μg/mL; and 190.77 μg/mL, activity of phosphate solubilizing were 11.3 μg/mL; 8.6 μg/mL; and 2.2 μg/mL, respectively. Based on 16S rDNA, Dbs 1, Dbs 2, and Dbm 3 were identified as Acinetobacter schindleri, Pseudomonas syncyanea, and P. moraviensis, respectively. Based on our knowledge, this is the first report P. syncyanea was exhibited plant growth-promoting properties.@*Conclusion, significance and impact of study@#Candidates of PGPR isolates could be alternative PGPR agents, but still need to evaluate the effect of three PGPR isolates application on citrus plant growth.

Effect of biofertilizer on the diversity of nitrogen - fixing bacteria and total bacterial community in lowland paddy fields in Sukabumi West Java, Indonesia

Aims: Some of methanotrophic bacteria and nitrous oxide (N2O) reducing bacteria have been proven able to support the plant growth and increase productivity of paddy. However, the effect of application of the methanotrophics and N2O reducing bacteria as a biofertilizer to indigenous nitrogen-fixing bacteria and total bacterial community are still not well known yet. The aim of the study was to analyze the diversity of nitrogen-fixing bacteria and total bacterial communty in lowland paddy soils. Methodology and results: Soil samples were taken from lowland paddy fields in Pelabuhan Ratu, Sukabumi, West Java, Indonesia. There were two treatments applied to the paddy field i.e biofertilizer-treated field (biofertilizer with 50 kg/ha NPK) and control (250 kg/ha NPK fertilizer). There were nine different nifH bands which were successfully sequenced and most of them were identified as unculturable bacteria and three of them were closely related to Sphingomonas sp., Magnetospirillum sp. and Ideonella dechloratans respectively. In addition, there were 20 different 16S rDNA bands which were successfully sequenced. Phylogenetic analysis of the sequence showed that there were 5 phyla of bacteria, i.e. Proteobacteria (Alphaproteobacteria and Gammaproteobacteria), Chlorofexi, Gemmatimonadetes, Clostridia, and Bacteroidetes respectively. Alphaproteobacteria was the most dominant group in lowland paddy field. Microbial diversities in the biofertilizer-treated field were lower than that of 100% fertilizer-treated field either based on nifH and 16S rDNA genes. Conclusion, significance and impact study: Biofertilizer treatment has lower microbial diversity than control, either based on nifH and 16S rDNA genes.

Estimation of Nitrogenase Enzyme Activities and Plant Growth of Legume and Non-legume Inoculated with Diazotrophic Bacteria

Biological Nitrogen Fixation (BNF) process benefits the agriculture sector especially for reducing cost of nitrogen fertilizer. In the process, the diazotrophs convert N2 into ammonia (NH3) which is useable by plants. The BNF process is catalysed by nitrogenase enzyme that involved protons and electrons together with evolution of H2 therefore, the assessment of N2 fixation is also available via H2 production and electron allocation analysis. Thus, the aims of this experiment were to estimate the nitrogenase enzyme activities and observe the influence of diazothrophs on growth of legume (soybean) and non legume (rice) plants. Host plants were inoculated with respective inocula; Bradyrhizobium japonicum (strain 532C) for soybean while Azospirillum brasilense (Sp7) and locally isolated diazotroph (isolate 5) for rice. At harvest, the plants were observed for plant growth parameters, H2 evolution, N2 fixation and electron allocation coefficient (EAC) values. The experiment recorded N2 fixation activities of inoculated soybean plants at 141.2 μmol N2 h-1 g-1 dry weight nodule, and the evolution of H2 at 144.4 μmol H2 h-1 g-1 dry weight nodule. The electron allocation coefficient (EAC) of soybean was recorded at 0.982. For inoculated rice plants, none of the observations was successfully recorded. However, results for chlorophyll contents and plant dry weight of both plants inoculated with respective inocula were similar to the control treatments supplied with full nitrogen fertilization (+N). The experiment clearly showed that inoculation of diazotrophic bacteria could enhance growth of the host plants similar to plants treated with nitrogenous fertilizer due to efficient N2 fixation process

Early colonization pattern of maize (Zea mays L. Poales, Poaceae) roots by Herbaspirillum seropedicae (Burkholderiales, Oxalobacteraceae)

The bacterium Herbaspirillum seropedicae is an endophytic diazotroph found in several plants, including economically important poaceous species. However, the mechanisms involved in the interaction between H. seropedicae and these plants are not completely characterized. We investigated the attachment of Herbaspirillum to maize roots and the invasion of the roots by this bacterium using H. seropedicae strain SMR1 transformed with the suicide plasmid pUTKandsRed, which carries a mini-Tn5 transposon containing the gene for the Discosoma red fluorescent protein (Dsred) constitutively expressed together with the kanamycin resistance gene. Integration of the mini-Tn5 into the bacterial chromosome yielded the mutant H. seropedicae strain RAM4 which was capable of expressing Dsred and could be observed on and inside fresh maize root samples. Confocal microscopy of maize roots inoculated with H. seropedicae three days after germination showed that H. seropedicae cell were attached to the root surface 30 min after inoculation, were visible in the internal tissues after twenty-four hours and in the endodermis, the central cylinder and xylem after three days.