Quality Of Biofertilizer Research Articles

Editorial: The role of biochar in enhancing biogas productivity and bio-fertilizer quality

Editorial: The role of biochar in enhancing biogas productivity and bio-fertilizer quality

БИОГАЗОВАЯ ТЕХНОЛОГИЯ - ЭФФЕКТИВНЫЙ СПОСОБ ПЕРЕРАБОТКИ НАВОЗА СЕЛЬСКОХОЗЯЙСТВЕННЫХ ЖИВОТНЫХ

The renewable energy sources (VIE) widespread using makes it possible the fossil fuels’ consumption that pollute the environment reducing significantly. One of the VIE types is biomass, which tends the agricultural production, especially livestock intensification growing. The main type of biomass is manure; at it processing by biogas technology using, it provides biogas, biofertilizer and electric energy, that are necessary for soil fertility and energy supply to rural consumers increasing. At the same time, biogas technology helps the manure disinfection’s problems solving, since fresh manure is an environmental pollutant by harmful gases (methane, ammonia, hydrogen sulfide, etc.) emitting. The livestock methane emissions’ amount is about 16%. The authors have developed small sized biogas manure processing installation, to the small agricultural enterprises’ conditions adapting. Distinctive features of the installation are: sawdust from reed plants for manure preparation loading into the bioreactor; of an external energy source from renewable resources (sun, wind, etc.) to heat the bioreactor in cold weather using; biofertilizer into dry residue and liquid separating; manure different types to carbon and nitrogen at C /N = 20 ratio mixing. These features make it possible the quality of biofertilizer, since reed plants contain vitamins C, carotene, starch, carbohydrates and proteins improving, as well as the crust formation to prevent; of biogas by observing the temperature regime inside the reactor and the ratio C /N = 20 output increasing; manure next fresh portion is accelerated by liquid part feeding of the biofertilizer back into the bioreactor’s process fermentation.

Shelf Life of Biofilm Producing PGPR Liquid Inoculants in Different Enriched Media Composition

Selecting a carrier material compatible with microbes is crucial for ensuring the quality of biofertilizers. This research aimed to identify a carrier material that could sustain the population of inoculants from dryland (including Delftia tsuruhatensis strain D9, Delftia sp. strain MS2As2, and Bacillus sp.) for a specified period. After conducting the study, it was found that a formulation of 3% molasses, 3% glycerol, 1% potassium sorbate, and 1% Tween-20 enriched with 1% nutrient was the most effective carrier material in maintaining the number of live inoculant cells. This formulation was able to sustain the population of inoculants up to the twelfth week, with a total population of 21.60 × 108 CFU.mL-1 with pH value of media is 5.29, and fulfilled the main criteria for biofertilizers.

Engineering Approach for Production of Arbuscular Mycorrhizal Inoculum Adapted to Saline Soil Management

(1) Background: Soil salinity impacts plant growth and productivity in steppic regions. Thus, the current study aims at producing, at low cost, an inoculum composed of native arbuscular mycorrhizal fungi (AMF) strains adapted to saline soil management. (2) Methods: The propagation of the native AMF strains was carried out in three natural saline soils presenting increasing salinity levels (4.5, 8.5 and 9.3 dS·m−1 in Boughzoul (BG), Salt Rocket (SR) and Zaafrane (ZA) sites, respectively). Three host plant species (alfalfa, clover and leek) were tested as trap cultures. AMF spore richness and diversity, the quantification of soil microbial biomass and the determination of the most probable number (MPN) were carried out before and after 24 months culture. Moreover, the mycorrhizal rates of the host plant species were assessed. (3) Results: The moderate saline soil (SR) planted with alfalfa was found to be the most suitable for AMF inoculum production with a maximum of 650 spores, 10 g−1 of soil, a mycorrhizal rate of 86%, 70 propagules·g−1 of soil and the highest microbial biomass content. (4) Conclusion: This study points out the best combination for the production of a more adapted AMF biofertilizer for saline soil management and proposes several indicators for biofertilizer quality evaluation.

The viability test of phosphate solubilizing microbes on carriers during storage

Excessive use of inorganic fertilizers, especially during the green revolution, has caused pollution and contamination of the soil due to accumulated residues. The inorganic fertilizer residues then cause soil compaction, loss of available nutrients and soil organic matter, loss of soil biodiversity which indirectly reduces soil fertility and makes plants more susceptible to disease. Biofertilizers have the potential to optimize soil response to fertilization, improve soil health, and even reduce the need for inorganic fertilizers to achieve sustainable agriculture. One of the types of microbes contained in biological fertilizers that have been proven to be able to replace the need for inorganic P fertilizers and improve soil health and productivity is a group of phosphate solubilizing microbes. The quality of biofertilizers is also determined by the carrier material capable of supporting the growth of inoculants. The purpose of this study was to test the viability of phosphate solubilizing microbes on carriers during storage The microbes used were Bacillus subtilis, Pseudomonas mallei, Burkholderia sp, and Trichoderma harzianum, while the tested carrier material were hydrogel and kaolin. The results showed that hydrogel and kaolin as carriers were able to support the growth of P-solubilizing microbes inoculants during storage (until 126 day). The number of viable populations of each isolate that makes up the biofertilizer consortium was in the high population range up to 1012 cfu. g-1.

Isolation and Identification of Effective Microorganisms from Water Hyacinth Biofertilizer

The use of microorganisms to promote growth and increase crop production has begun to be studied. Microorganisms that isolated from biofertilizers vary according to the source of organic matter. One of the biofertilizer quality determinants is the type and number of populations of microorganisms. This study aims to know is the type and abundance of microorganisms, as well as nutrient content in water hyacinth biofertilizers. The experiment was conducted at the Greenhouse and Laboratory of Food Crops Cultivation, Payakumbuh State Agricultural Polytechnic, Limapuluh Kota Regency, West Sumatra from March to July 2019. The experimental stages included (1) Manufacture water hyacinth biofertilizers, the experiments using Completely Randomized Design (CRD) with four measurements of water hyacinth treatment in biofertilizer, A = 25%, B = 50%, C = 75%, and D = 100% and five replications, (2) Isolation of microorganisms from biofertilizers using pure plate and streak plate methods. Eleven bacterial species and two fungal species were identified using molecular analysis based on 16S rRNA gene fragments. Effective microorganisms that play a role in the degradation of biofertilizers are Pseudomonas aeruginosa, Bacillus subtilis, Trichoderma asperellum, and Trichoderma harzianum. Biofertilizers contain C-organic 3.61-3.92%, N 0.166-0.278%, C/N 14,10-21,75, P2O5 0.347-0.729%, K2O 1.422-2.090%, and Ca 0.140-0.259%.

Assessment of Shelf Life and Quality of Biofertilizers using Tricalcium Phosphate as an Anticaking Agent and Aluminium Silicate as the Inert Carrier

Background: Bio-fertilizers are the substances which contain living microorganisms, when applied to soil, seeds and plant root these fertilizers increases soil fertility and promote growth of the plant. Biofertilizers help plants to utilize important mineral resources, phosphorous and nitrogen. Microorganisms like Rhizobacteria, fungi and algae which provide nutrient to the soil and which are produced commercially are known as biofertilizers. The microorganisms which present in biofertilizers are Rhizobium species, Pseudomonas species and Azospirillum species etc. These biofertilizers have potential to replace conventional chemical fertilizers. The quality of biofertilizers is utmost important as they have to be used by farmers and should work well when applied to the soil. It should not form clumps after preparation. In this study, anticaking property provided by tricalcium phosphate (TCP) to individual biofertilizer containing Pseudomonas, Rhizobium and Azospirillum respectively (each separately) was studied. Methods: In our study, we have used serial dilution and direct count method (CFU) for checking viability of live microorganism for 15, 30 and 90 days duration in respective biofertilizers in our laboratory. Different percentage viz 5%, 10%, 15% and 20% of tricalcium phosphate (TCP) was used in addition to aluminium silicate as an inert carrier.Conclusion: Our study has validated that all percentage (5%, 10%, 15% and 20%) of tricalcium phosphate (TCP) is reducing clump formation as compared to control with no TCP added. On the basis of plate count method (CFU result) 10% TCP is found to be optimum to be used as an anticaking agent for biofertilizer containing Pseudomonas, Rhizobium and Azospirillum respectively.

Improved biofertilizer properties of digestate from codigestion of brewer\u2019s spent grain and palm oil mill effluent by manure supplementation

The biofertilizer quality of anaerobically codigested agroindustrial residues from brewer’s spent grain and palm oil mill effluent was evaluated after supplementation with different livestock manure in order to validate its use as organic fertilizer. Manure supplementation assay was performed using different animal manure-inoculum to demonstrate the influence of inoculum-type on the nutrient status, plant growth promoting bacteria (PGPB) and other plant growth promoting attributes of the resultant digestate. In addition to elevated nutrient levels (K > P > Ca > Mg > S > N), the plethora of essential microbial groups (phosphate solubilizers > diazotrophs > auxin producers) that enhance nutrition and promote plant growth was evinced in the supplemented digestate compared to the control. On the other hand, environmental risk assessment revealed a notable yet inadequate reduction in indicator bacteria and putative pathogens (> 3.0 log CFU mL− 1) with potentially toxic elements within publicly available requirements. The preponderance of PGPB with excellent biofertilizing attributes observed in this study could be leveraged upon by plants thus substantiating its potential for use as organic fertilizer. However, the presence of pathogens highlights the importance of post-treatment hygienization to eliminate its biosafety risk.

Enhanced Biogas and Biofertilizer Production from Anaerobic Codigestion of Harvest Residues and Goat Manure

Comparative assays were conducted to assess the biogas and biofertilizer production efficiency from anaerobically codigested goat manure (GM) and harvest residues: corn stover (CS) and rice straw (RS). All digesters were operated simultaneously under mesophilic temperature of 40°C and notable phosphate solubilizing and nitrogen fixing bacterial populations indicated qualitative biofertilizer quality of the digestates. Codigestion of the substrates significantly increased biogas yield (p < 0.05) compared to monodigestion, and the highest cumulative yield of 573 ml/g VS was obtained from codigested rice straw (RS) and goat manure (GM). With a significant decimation in number of pathogens (p < 0.05), a 2 – 3 fold increase in populations of plant growth promoting bacteria (Bacillus and Pseudomonas species) was observed in digestate from codigestion assays when compared to monodigestion (control) and were identified as Clostridium sp., Bacillus subtilis, Bacillus megaterium, Lactobacillus sp., Pseudomonas fluorescens including methanogens: Methanothrix sp., Methanobacterium sp. and Methanosarcina sp. On the average, codigestion assays resulted in enhanced biogas yield and biofertilizer quality that was 2.3 – 4.1 times higher than single substrate digestion and generally improved the efficiency of biogas and biofertilizer production.

Anaerobic co-digestion of swine manure with sweet potato or cassava in different C/N ratios

ABSTRACT: Anaerobic co-digestion (AcoD) of waste is a method of increasing methane (CH4) yield and improving biofertilizer quality. This study aimed to evaluate the best AcoD conditions for swine manure (SM) with sweet potato (SP) or cassava (C) in different amounts in semi-continuous biodigesters. Initially, using batch biodigesters, an AcoD test of the SM with SP or C was performed, adopting carbon/nitrogen (C/N) ratios of 10/1, 13/1, 17/1, and 22/1. Based on the results, a C/N ratio of 10/1 was chosen, which was the proportion that resulted in the highest reduction of volatile solids (VS) and specific biogas production. From these results, the experiment was carried out in semi-continuous biodigesters, consisting of three treatments (control (SM), SP 10/1, and C 10/1) with five replicates and a hydraulic retention time (HRT) of 30 days. Total solid (TS) and volatile solid (VS) reductions, biogas and CH4 yields, alkalinity, and volatile acidity were measured. The control treatment differed from the others and resulted in decreased biogas yield (752 LN.kgVSadded -1), CH4 (449 LN.kgVSadded -1), and CH4 content (59.7%). The AcoD treatments (SP and C) did not differ significantly for biogas yield (respectively, 901 and 883 LN.kgVSadded -1) and CH4 (respectively, 590 and 547 LN.kgVSadded -1); however, they differed in CH4 content (65.5% and 61.9% respectively). The treatments showed general reduction averages of 76.1% and 85.9% for TS and VS, respectively, with no statistical difference found between them. The AcoD of the SM with SP or C increased the production and quality of the biogas, increasing the concentration of CH4 therein.

In vitro test for compatibility of biofertilizers containing phosphate solubilizers and nitrogen-fixing bacteria

The quality of biofertilizers is determined by the content of isolates and synergism between isolates. The experiment was aimed to investigate the compatibillty of biofertilizers containing P-solubilizers and N-fixing bacteria. In vitro test using isolates of PSB and N-fixing bacteria was done to study the synergystic character by using streak method on nutrient agar for the bacteria and potato dextrose agar for the fungi. Species of P-solubilizers were Pseudomonas mallei, P. cepaceae, Aspergillus niger, and Penicillium sp., and species of N-fixing bacteria were Azotobacter chroococum and Azospirillum sp. The results revealed that the P-soluilizers microbes and N-fixing bacteria were synergistic and did not antagonistic to each other. Biofertilizers which contain P-solubilizers and N-fixing bacteria can be used for agricultural purposes.

Application of arbuscular mycorrhizal fungi in combination with nitrogen fixing bacteria and other potential soil microbes as biofertilizer for soybean plant

The use of mycorrhizae as phosphate solubilizing fungus has been known since decade. Mycorrhizae could promote the growth of almost all species of plants. Biofertilizer product based on mycorrhizae fungus has been produced commercialization and easily found in the market. This study was aimed to understand the positive effect of mycorrhizae inoculation when it mixed with other microbes especially nitrogen fixing bacteria and growth hormone production bacteria. An experiment was set up at green house to run the activities of microbes inoculation on soybean plants. Soybean was mainly used for testing the potential of soil microbes since the target of the experiment is increasing the production of soybean seed and decreasing the used of chemical fertilizer. The result of this study showed that multi strains inoculation was better that single inoculation. Single mycorrhizae inoculation produced slightly higher number of pods. However when the mycorrhizae was combining with other potential microbes the number of pods produced by the plants even significantly higher than only single inoculation. Updating the quality of biofertilizer is needed by including the biodiversity of soil microbes to gain the promotion of green farming program.

FACTORIAL ANALYSIS ON PROCESSING FACTORS FOR NITROGEN, PHOSPHORUS AND POTASSIUM CONTENTS IN MUSHROOM WASTE

The agriculture industry relies heavily on the use of bio-fertilizer and the main components in bio-fertilizer are nitrogen (N), phosphorous (P), and potassium (K). Thus, a study was conducted to identify the N, P, and K contents in mushroom waste (MW). These components are numerous building blocks that plants need for healthy growth. Therefore, by increasing the N, P and K contents in MW, it can be utilized to produce high and better quality of bio-fertilizer. Five independent factors, i.e., aging of waste (fresh 0 day & aged 14 days), waste pH (7 & 8.5), composition [MW only & mixture of MW & spent medium (SM)], technique of drying (oven 50 °C & sunlight), and MW size (powder & cut) were the affecting factors on N, P, and K contents in MW. A 25-1 fractional factorial design was used to investigate the effect of the independent factors as well as the interaction factors on the N, P, and K contents. The N, P, and K contents were measured using HACH spectrophotometer. The objective of this research is to identify the best combination of processing factors. Some of the independent factors were shown to have significant effects on the N, P and K contents. The results showed that the most significant factor in N content are MW size and aging of waste, while for P and K contents are technique of drying and MW size. The best condition was identified to maximize the amount of N, P, and K contents in MW. The identified conditions were the MW aged for 7 days, MW size at powder form, waste pH at 7, drying under sun light and the composition MW only. Based on the proposed best condition the N (12.08 mg/L), P (3.04 mg/L), and K (8.09 mg/L) contents were achieved. The results show that fractional factorial design was suitable in investigating the effect of a large number of factors with a minimum number of experiments.

CHARACTERIZATION AND QUALITY ASSESSMENT OF INDONESIAN COMMERCIAL BIOFERTILIZERS

Biofertilizers currently sold in the market are not labeled with a distinct quality standard. As such, farmers may buy low quality biofertilizers, which can reduce their profit and trust on the benefit of biofertilizers. This paper presents the characteristics of various products of commercial biofertilizers as well as farmers’ knowledge and experience on the products. The study was carried out in 2004-2006 by collecting and analyzing data on registered commercial biofertilizers, checking their availability at the market (39 agricultural shops), and interviewing farmers on their knowledge and experience on the use of biofertilizers on various farming systems in Bandung District, West Java (86 respondents) and Semarang District, Central Java (77 respondents). The quality of biofertilizers was tested in the laboratory based on microbial density and its functional (phenotypic) traits. The study showed that amongst various brands of biofertilizers commercialized, 41 brands of them have been officially registered as commercial products. Two brands of other biofertilizers found in agricultural shops were registered as organic or inorganic fertilizers. In general, each biofertilizer contained two or more microbial strains and was claimed to have multiple functions. However, most of them (>90%) were not labeled with expiry date information. Macronutrient contents (NPK) of some microbial carriers were almost equal to those of organic fertilizers. Around 38% of respondents in Bandung knew biofertilizers and less than 10% have ever used them. In Semarang, however, familiarity and personal experience of the respondents were much lower, i.e. 10% and 3%, respectively. About 67% and 50% of agricultural shops in Bandung and Semarang sold biofertilizers, respectively. Laboratory analyses showed that microbial density of five biofertilizers tested was lower than that of product specification, although most of them were positive for N-fixing and P-solubilizing traits. Some microbial strains contained less than 103 cfu based on the dilution level testing. These figures imply the urgent need to improve the existing quality standard system of biofertilizers including its control mechanisms.

CHARACTERIZATION AND QUALITY ASSESSMENT OF INDONESIAN COMMERCIAL BIOFERTILIZERS

Biofertilizers currently sold in the market are not labeled with a distinct quality standard. As such, farmers may buy low quality biofertilizers, which can reduce their profit and trust on the benefit of biofertilizers. This paper presents the characteristics of various products of commercial biofertilizers as well as farmers’ knowledge and experience on the products. The study was carried out in 2004-2006 by collecting and analyzing data on registered commercial biofertilizers, checking their availability at the market (39 agricultural shops), and interviewing farmers on their knowledge and experience on the use of biofertilizers on various farming systems in Bandung District, West Java (86 respondents) and Semarang District, Central Java (77 respondents). The quality of biofertilizers was tested in the laboratory based on microbial density and its functional (phenotypic) traits. The study showed that amongst various brands of biofertilizers commercialized, 41 brands of them have been officially registered as commercial products. Two brands of other biofertilizers found in agricultural shops were registered as organic or inorganic fertilizers. In general, each biofertilizer contained two or more microbial strains and was claimed to have multiple functions. However, most of them (>90%) were not labeled with expiry date information. Macronutrient contents (NPK) of some microbial carriers were almost equal to those of organic fertilizers. Around 38% of respondents in Bandung knew biofertilizers and less than 10% have ever used them. In Semarang, however, familiarity and personal experience of the respondents were much lower, i.e. 10% and 3%, respectively. About 67% and 50% of agricultural shops in Bandung and Semarang sold biofertilizers, respectively. Laboratory analyses showed that microbial density of five biofertilizers tested was lower than that of product specification, although most of them were positive for N-fixing and P-solubilizing traits. Some microbial strains contained less than 103 cfu based on the dilution level testing. These figures imply the urgent need to improve the existing quality standard system of biofertilizers including its control mechanisms.

Spent mushroom substrate biochar as a potential amendment in pig manure and rice straw composting processes

ABSTRACTSpent mushroom substrate (SMS) is a bulky waste byproduct of commercial mushroom production, which can cause serious environmental problems and, therefore, poses a significant barrier to future expansion of the mushroom industry. In the present study, we explored the use of SMS as a biochar to improve the quality of bio-fertilizer. Specifically, we performed a series of experiments using composting reactors to investigate the effects of SMS biochar on the physio-chemical properties of bio-fertilizer. Biochar was derived from dry SMS pyrolysed at 500°C and mixed with pig manure and rice straw. Results from this study demonstrate that the addition of biochar significantly reduced electrical conductivity and loss of organic matter in compost material. Nutrient analysis revealed that the SMS-derived biochar is rich in fertilizer nutrients such as P, K, Na, and N. All of these findings suggest that SMS biochar could be an excellent medium for compost.

Effectiveness of rabbit manure biofertilizer in barley crop yield.

The quality of biofertilizers is usually assessed only in terms of the amount of nutrients that they supply to the crops and their lack of viable pathogens and phytotoxicity. The goal of this study was to determine the effectiveness of a liquid biofertilizer obtained from rabbit manure in terms of presence of pathogens, phytotoxicity, and its effect on the grain yield and other agronomic traits of barley (Hordeum vulgare L.). Environmental effects of the biofertilizer were also evaluated by following its influence on selected soil parameters. We applied the biofertilizer at five combinations of doses and timings each and in two application modes (foliar or direct soil application) within a randomized complete block design with three replicates and using a chemical fertilizer as control. The agronomic traits evaluated were plant height, root length, dry weight, and number of leaves and stems at three growth stages: tillering, jointing, and flowering. The effectiveness of the biofertilizer was significantly modified by the mode of application, the growth stage of the crop, and the dose of biofertilizer applied. The results showed that the foliar application of the biofertilizer at the tillering stage produced the highest increase in grain yield (59.7%, p < 0.10). The use of the biofertilizer caused significant changes in soil, particularly concerning pH, EC, Ca, Zn, Mg, and Mn. It is our view that the production and use of biofertilizers are a reliable alternative to deal with a solid waste problem while food security is increased.

The anaerobic co-digestion of sheep bedding and ⩾50% cattle manure increases biogas production and improves biofertilizer quality

Sheep manure pellets are peculiarly shaped as small ‘capsules’ of limited permeability and thus are difficult to degrade. Fragmentation of manure pellets into a homogeneous mass is important for decomposition by microorganisms, and occurs naturally by physical shearing due to animal trampling, when sheep bedding is used. However, the high lignocellulose content of sheep bedding may limit decomposition of sheep manure. Here, we evaluated if co-digestion of sheep bedding with cattle manure would improve the yield and quality of the useful products of anaerobic digestion of sheep bedding—biogas and biofertilizer—by providing a source of nutrients and readily available carbon. Mixtures of sheep bedding and cattle manure in varying proportions (0%, 25%, 50%, 75%, or 100% cattle manure) were added to 6-L digesters, used in a batch system, and analyzed by uni and multivariate statistical tools. PC1, which explained 64.96% of data variability, can be referred to as ‘organic fraction/productivity’, because higher rates of organic fraction consumption (COD, cellulose and hemicellulose contents) led to higher digester productivity (biogas production, nutrient concentration, and sample stability changes). Therefore, productivity and organic fraction variables were most influenced by manure mixtures with higher (⩾50%) or lower (⩽25%) ratios of cattle manure, respectively. Increasing the amount of cattle manure up to 50% enhanced the biogas potential production from 142Lkg−1TS (0% of cattle manure) to 165, 171, 160Lbiogaskg−1TS for the mixtures containing 100%, 75% and 50% of cattle manure, respectively. Our results show that the addition of ⩾50% cattle manure to the mixture increases biogas production and improves the quality of the final biofertilizer.

Cryogel-supported titanate nanotubes for waste treatment: Impact on methane production and bio-fertilizer quality

By reducing the cadmium (Cd2+) content in biomass used for bio-based products such as biogas, a less toxic bio-based fertilizer can be obtained. In this work, we demonstrate how a macroporous polymer can support titanate nanotubes, and we take advantage of its known selective adsorption behavior towards Cd2+ in an adsorption process from real nutrient-rich process water from hydrolysis of seaweed, a pollutant-rich biomass. We show that pretreatment steps involving alteration in area-to-volume ratio performed in aerated and acidic conditions release the most Cd2+ from the solid material. By integrating an adsorption step between hydrolysis and the biomethane, we show that it was possible to obtain high Cd2+ removal (ca. 94%) despite molar excess (between 100 and 500) of co-present ions (e.g., Mg2+, Ca2+, Na+, K+) and with maintained total phosphorous content. The bio-methane potential did not significantly decrease as compared to a process without cadmium removal and the yielded bio-fertilizer followed Swedish guideline values. This study provides a sound and promising alternative for a novel remediation step, enabling higher use of otherwise tricky and to some extent overlooked biomass sources.

Identification and Diagnostics of Plant-Symbiotic and Phytopathogenic Bacteria

Rapid and reliable diagnostics and identification of pathogenic and symbiotic bacteria are at the top of the agenda. In the first case, they are important to control and prevent crop damages, and thus reduce economic losses. In the second, it's necessary to design and monitor quality of biofertilizer to raise its effectiveness and crop capacity. Development of accurately, rapidly, technically and commercially accessible methods remains a critical problem for the bacteria with comprehensive phylogenetic structure. In this work, we investigated pathogenic Xanthomonas and Ralstonia and symbiotic Sinorhizobium. The aim of this investigation was to examine the applicability of the novel methods for phylogenetic study, identification and diagnostics of closely related species of these genera. The conventional phenotypic and genotypic (16S rRNA, gyrB) methods were applied as referents. Novel polymerase chain reaction (PCR)-based approaches, single-adapter amplified fragment length polymorphism (saAFLP) and comparative analyses of hin-region and Xcc0006-0007 sequences, were first employed for the investigations. Phenotypic tests, 16S rRNA and gyrB analysis distinguished bacteria at the genus level, but failed to identify them to the species robustly. The new methods identified bacteria at the inter-species level more precisely. This identification agreed with the accepted genera's classifications. The only exceptions were X. fuscans & X. citri and X. perforance & X. euvesicatoria which clustered together. The further outcome of this study was achieved hin-region-based genus-specific PCR primers for the express-diagnostics of the genera. Summary, these new methods can be applied for genome-based phylogeny investigations and as convenient and accurate tools for identification and routine laboratory diagnostics of these comprehensive genera.