Rhizobacterial Isolates Research Articles

Prospects of iron solubilizing Bacillus species for improving growth and iron in maize (Zea mays L.) under axenic conditions

Iron (Fe) deficiency in calcareous soils is a significant agricultural challenge, affecting crop productivity and nutritional quality. This study aimed to isolate, characterize, and evaluate Fe solubilizing rhizobacterial isolates from maize rhizosphere in calcareous soils as potential biofertilizers. Forty bacterial isolates coded as SG1, SG2, …, SG40 were isolated and screened for siderophore production, with ten showing significant Fe solubilizing capabilities. These isolates were further assessed for phosphate solubilization and exopolysaccharides production. The selected bacterial isolates were also screened under axenic conditions for their ability to improve maize growth. The isolates SG8, SG13, SG24, SG30 and SG33 significantly enhanced growth parameters of maize. Notably, SG30 showed highest increment in shoot length (58%), root length (54%), root fresh and dry biomass (67% and 76%), SPAD value (67%), relative water contents (69%), root surface area (61%), and Fe concentration in shoots (79%) as compared to control. The biochemical characterization of these strains showed that all these strains have capability to solubilize insoluble phosphorus, produce indole-3-acetic acid (IAA), and ammonia with catalase, urease and protease activity. Molecular identification through 16s rRNA gene sequencing confirmed high similarity (99.7–100%) of the selected isolates to various Bacillus species, including B. pyramidoids, B. firmicutes, and B. cereus. The study provides a strong base for developing eco-friendly, cost-effective biofertilizers to address Fe deficiency in crops and promote sustainable agriculture.

Effectiveness Test of Indigenous Plant Growth Promoting Rhizobacteria (PGPR) Isolate on the Growth of Acehnese Patchouli (Pogostemon cablin Benth.) Sidikalang Variety

Plant Growth Promoting Rhizobacteria (PGPR) is a group of bacteria that live saprophytically in the root area (rhizosphere) acting as a plant growth promoter. The purpose of this study was to test the effectiveness of indigenous rhizobacterial isolates from Aceh Besar and Aceh Tengah in supporting the growth of Aceh patchouli Sidikalang variety. This research was conducted at the Laboratory of Seed Science and Technology, Faculty of Agriculture, Syiah Kuala University and patchouli planting was carried out at the Patchouli Innovation Park (Nino Park), Syiah Kuala University, which took place from December 2022 to June 2023. This study used a non-factorial Randomized Group Design (RAK) with 3 replications. The factor studied was the type of rhizobacterial isolate consisting of 15 treatment levels, namely 1 treatment without rhizobacteria, 7 treatments given indigenous rhizobacterial isolates from Lambada, Seulimeum District, Aceh Besar Regency and 7 treatments given rhizobacterial isolates from Paya Tampu village, Rusip Antara District, Central Aceh Regency. The observation parameters studied are of plant height, number of leaves, number of branches, stem diameter, area per leaf blade, root length, root volume and wet biomass weight of plants. The results showed that indigenous rhizobacterial isolates from Aceh Besar and Aceh Tengah effectively increased the average plant height of Sidikalang variety patchouli at 4 and 8 MSA (26%, and 24%), number of leaves (46%, and 58%), number of branches (78% and 50%), area per leaf blade (34%), root length (19%), root volume (60%) and plant wet biomass weight (50.61%).

Evaluation and Characterization of Rhizobacteria and Endophytes for Antagonistic and Growth Promotional Traits Against Collar Rot (Sclerotium rolfsii Sacc.) of Soybean

Background: Sclerotium rolfsii Sacc. is one of the destructive soil-borne plant pathogens due to its significant loss in terms of yield and quality in several crop plants including soybean. Use of biocontrol agents in the management of disease is an ecofriendly strategy. Therefore, the study aimed to assess in vitro antagonisticactivity of rhizobacteria and endophytes against Sclerotium rolfsii following dual culture technique. Methods: A total of 75 rhizobacterial isolates, 3 endophytes along with reference cultures were evaluated for the antagonistic activity against Sclerotium rolfsii following dual culture technique. The effective rhizobacterial isolates were tested for the various plat growth promoting rhizobacteria traits viz., indole acetic acid production, siderophore production, Phosphate solubilization, production of ammonia, hydrogen cyanide production and chitinase test following the standard procedure. Result: The two promising rhizobacterial strains i.e., AUUB 209 (Streptomyces enissocaesilis) and Streptomyces racemochromogenes (AUDT 626) were found effective with 76.79 and 64.75 percent mycelial inhibition of Sclerotium rolfsii under in vitro. The antagonistic activity of these two rhizobacteria is mainly attributed to the positive confirmation of various PGPR activities viz., production of Indole acetic acid, siderophore, phosphate solubilization and chitinase production.

Some Chemical and Biological Methods to Control Pepper Root Rot Disease in Iraq

Background: Pepper growing fields in the Babil and Wasit governorates in Iraq, are suffering from a decline in production due to the spread of root rot diseases. The use of biological control techniques, such as the use of beneficial bacteria in the rhizosphere and natural plant extracts, along with chemical fungicides, has recently contributed to reducing the damage of plant pathogens globally. Methods: In this research, the main cause of root rot disease in pepper plants was isolated and diagnosed using polymerase chain reaction technology using (TEF-1α) gene, antagonistic activities of 75 rhizobacterial isolates, natural extracts of garlic and nerium and the fungicides of Promess and Tecto 500 SC were tested in vitro against the pathogen, then the superior bacterial isolates were identified. The ability of biological and chemical agents to control the disease was tested under greenhouse conditions according to the completely randomized design (CRD). Result: The results of isolation showed that the fungus Fusarium solani was dominance and the isolate Fsw45 was the most virulent. 3 of 75 rhizobacterial isolates outperformed the pathogen in an antagonistic tests and identified as Pseudomonas mendocina, P. fluorescens and Staphylococcus kloosii also the fungicides and botanical extracts showed significant antagonistic activities as well in vitro. Under greenhouse conditions, the fungicide treatments excelled in disease control by 100%. As for the 3 bacterial isolates, they achieved a significant reduction in the percentage of disease incidence and disease severity index, their abilities were higher than the nerium and garlic extracts.

PLANT-GROWTH-PROMOTING AND ANTIFUNGAL ASSET OF INDIGENOUS DROUGHT-TOLERANT RHIZOBACTERIA ISOLATED FROM OLIVE (Olea europaea L.) RHIZOSPHERE

Faced with global environmental challenges, the quest for sustainable food production has gathered momentum. While abiotic stresses adversely affect plant health and productivity, Verticillium wilt causes considerable yield losses worldwide, particularly in crops such as olive. Recently, drought-tolerant bacteria have been used to alleviate both abiotic stress and pathogen pressure in crops. In this context, our work focuses on the isolation of tolerant indigenous rhizobacteria to mitigate these challenges by investigating their role in biocontrol and abiotic stress tolerance. Thus, a total of 94 rhizobacterial strains were isolated from the rhizospheres of olive trees in southeastern Morocco and characterized to identify tolerant plant growth-promoting rhizobacteria that inhibit Verticillium dahliae. 24 strains demonstrated in vitro suppression of Verticillium dahliae Klebahn, and exhibited tolerance to different abiotic stresses (drought, salinity, and high temperature). In addition, they proved xerotolerant (Aw ≤ 0.91), halotolerant (≥10% NaCl), and thermotolerant (≥ 55°C) capabilities. Beyond, these isolates showcased multifaceted plant growth-promoting traits, such as phosphate solubilization and significant synthesis of essential bioactive compounds like siderophores, indole-3-acetic acid and hydrolytic enzymes. Evaluating outcomes, three standout rhizobacterial isolates emerged due to their exceptional stress tolerance, unique plant growth-promoting qualities, and potent antagonistic potential. Molecular analysis identified them as Bacillus paranthracis (OZ-60) and Bacillus licheniformis (OZ-48 and OZ-77) through 16S rRNA sequencing. Besides enhancing plant abiotic stress resistance, these isolates hold promise in bolstering the sustainability of olive cultivation and fortifying plant defenses against pathogens.

Bacillus licheniformis and Bacillus velezensis from Rhizosphere of Clerodendrum infortunatum L. Promote Plant Growth and Resistance to Sclerotium rolfsii in Vigna unguiculata (L.) Walp.

In the current study, thirty bacterial strains isolated from the rhizosphere of Clerodendrum infortunatum L. were evaluated for the properties related to the plant growth promotion and disease resistance. Here, all the selected strains were screened for its antagonistic effect towards the phytopathogen Sclerotium rolfsii and also for the production of bioactive compounds known to promote the plant growth. Among these isolates, CiRb1 and CiRb16 were observed to have a broad range of plant beneficial features and were identified as Bacillus licheniformis and Bacillus velezensis respectively. Both the isolates were also demonstrated to produce the volatile organic compounds (VOCs) responsible for the growth enhancement in Brassica nigra (L.) and growth inhibition of S. rolfsii. Talc based formulations made out of both B. licheniformis and B. velezensis were further demonstrated to augment the plant growth and protection against S. rolfsii in Vigna unguiculata (L.) Walp. By the GC-MS based analysis, undecane could also be detected in the methanolic extracts prepared from both B. licheniformis and B. velezensis. Here, the selected rhizobacterial isolates were found to promote the plant growth and disease resistance through both direct and VOC mediated mechanisms. The results of the study hence reveal both B. licheniformis and B. velezensis have the potential in field application to promote the growth and control of plant diseases.

Molecular Characterization Reveals Biodiversity and Biopotential of Rhizobacterial Isolates of Bacillus Spp

Bacillus species appearas the most attractive plant growth-promoting rhizobacteria (PGPR) and alternative to synthetic chemical pesticides. The present study examined the antagonistic potential of spore forming-Bacilli isolated from organic farm soil samples of Allahabad, India. Eighty-seven Bacillus strains were isolated and characterized based on their morphological, plant growth promoting traits and molecular characteristics. The diversity analysis used 16S-rDNA, BOX-element, and enterobacterial repetitive intergenic consensus. Two strains, PR30 and PR32, later identified as Bacillus sp., exhibited potent in vitro antagonistic activity against Ralstonia solanaceorum. These isolates produced copious amounts of multiple PGP traits, such as indole-3-acetic acid (40.0 and 54.5 μg/mL), phosphate solubilization index (PSI) (4.4 and 5.3), ammonia, siderophore (3 and 4 cm), and 1-aminocyclopropane-1-carboxylate deaminase (8.1and 9.2 μM/mg//h) and hydrogen cyanide. These isolates were subjected to the antibiotic sensitivity test. The two potent isolates based on the higher antagonistic and the best plant growth-promoting ability were selected for plant growth-promoting response studies in tomatoe, broccoli, and chickpea. In the pot study, Bacillus subtilis (PR30 and PR31) showed significant improvement in seed germination (27–34%), root length (20–50%), shoot length (20–40%), vigor index (50–75%), carotenoid content (0.543–1.733), and lycopene content (2.333–2.646 mg/100 g) in tomato, broccoli, and chickpea. The present study demonstrated the production of multiple plant growth-promoting traits by the isolates and their potential as effective bioinoculants for plant growth promotion and biocontrol of phytopathogens.

Investigating the Efficacy of Dry Region Zinc Solubilizing Bacteria for Growth Promotion of Maize and Wheat under Axenic Conditions

Zinc (Zn) plays a crucial role as a micronutrient, essential for the growth, development, and proper functioning of plants. Pakistani soils are deficient of micronutrient especially zinc. Supplementation of Zn fertilizers cannot do the job because only a small fraction of Zn is available to plants and rest is fixed in soil. To enhance the Zn availability different methods are being used which are expensive. Zinc solubilizing bacteria (ZSB) can be a potential candidate for solubilization of Zn and growth promoting activities in cereals. The present study aimed to evaluate the potential of dry region bacterial strains of Bahawalpur for improving Zn uptake and growth improvement in wheat and maize. Rhizosphere samples of wheat and maize were taken from the dry regions of Bahawalpur and bacterial strains were isolated and tested for their Zn solubilizing ability and screened for urease, siderophore activity, exopolysaccharide production, phosphorus solubilization and plant growth promoting abilities. Under axenic conditions selected isolates were further screened to assessed for improving the growth of wheat and maize seedlings. The rhizobacterial isolates IUB-34, IUB-80, IUB-93 and IUB-96 performed best to improve growth, physiology, biochemical attributes and root colonization. The maximum increase in root and shoot lengths were recorded 32.2 and 35.7% in wheat under IUB-34 application and in maize 43.8 and 39.9%, respectively, under IUB-96 application as compared to uninoculated control. Furthermore, maximum root colonization was also recorded under IUB-34 in wheat and IUB-96 in maize seedlings. Moreover, the biochemical characterization of selected isolates showed IAA (Auxins) production, protease, catalase, cellulose degradation, HCN and ACC-deaminase activities by these strains. However, only IUB-34 and IUB-96 were found positive for oxidase activity. Therefore, study concluded that use of dry region ZSB can solubilize Zn and make it available for plant to improve growth of wheat and maize. So, these isolates may be utilized for coating urea as a biofertilizer to increase the biofortification in cereals.

Multifaceted Plant Growth Promoting Traits and Abiotic Stress Resistance Abilities Exhibited by Chrysanthemum Rhizobacteria

Plant growth-promoting rhizobacteria (PGPR) boost plant growth and agricultural sustainability in an ecologically friendly way. The cultivation of chrysanthemum, a globally significant flower crop, has relied heavily on substantial agrochemical inputs that have detrimental impact on the environment. To assess the potential of chrysanthemum rhizobacteria to reduce this reliance, bacterial strains were retrieved from the plant rhizosphere and subjected to an assessment of various plant growth-promoting traits. Out of the 34 rhizobacterial isolates, 21 demonstrated the production of the plant growth hormone auxin, 21 had phosphate solubilization ability, 22 were capable of nitrogen fixation, and 21 could produce ammonia. Based on these findings, seven preeminent PGPR strains, characterized by multifaceted plant growth-promoting traits, were selected for subsequent studies and identified as species belonging to Acinetobacter, Bacillus, Pantoea, Serratia and Staphylococcus. The selected strains were systematically analyzed for their capacity to endure an array of abiotic stresses. A majority of these strains demonstrated adaptation under osmotic stress ranging from -0.15 to -0.49 MPa, temperatures of 20°C and 30°C, and salt stress within the range of 3 to 7% NaCl, which suggests their potential to promote plant growth across diverse environmental conditions. Additionally, the secretion of hydrolytic enzymes such as protease, pectinase and amylase was examined, and only the Staphylococcus hominis PGPR-12 strain demonstrated the ability to produce all three extracellular hydrolases. These findings underscore the potential application of multiple isolates possessing promising plant-probiotic properties to enhance plant growth across various conditions, thereby necessitating further exploration through pot and field assays.

Drought-Tolerant Bacteria and Arbuscular Mycorrhizal Fungi Mitigate the Detrimental Effects of Drought Stress Induced by Withholding Irrigation at Critical Growth Stages of Soybean (Glycine max, L.).

Considering current global climate change, drought stress is regarded as a major problem negatively impacting the growth of soybeans, particularly at the critical stages R3 (early pod) and R5 (seed development). Microbial inoculation is regarded as an ecologically friendly and low-cost-effective strategy for helping soybean plants withstand drought stress. The present study aimed to isolate newly drought-tolerant bacteria from native soil and evaluated their potential for producing growth-promoting substances as well as understanding how these isolated bacteria along with arbuscular mycorrhizal fungi (AMF) could mitigate drought stress in soybean plants at critical growth stages in a field experiment. In this study, 30 Bradyrhizobium isolates and 30 rhizobacterial isolates were isolated from the soybean nodules and rhizosphere, respectively. Polyethylene glycol (PEG) 6000 was used for evaluating their tolerance to drought, and then the production of growth promotion substances was evaluated under both without/with PEG. The most effective isolates (DTB4 and DTR30) were identified genetically using 16S rRNA gene. A field experiment was conducted to study the impact of inoculation with DTB4 and DTR30 along with AMF (Glomus clarum, Funneliformis mosseae, and Gigaspora margarita) on the growth and yield of drought-stressed soybeans. Our results showed that the bioinoculant applications improved the growth traits (shoot length, root length, leaf area, and dry weight), chlorophyll content, nutrient content (N, P, and K), nodulation, and yield components (pods number, seeds weight, and grain yield) of soybean plants under drought stress (p ≤ 0.05). Moreover, proline contents were decreased due to the bioinoculant applications under drought when compared to uninoculated treatments. As well as the count of bacteria, mycorrhizal colonization indices, and the activity of soil enzymes (dehydrogenase and phosphatase) were enhanced in the soybean rhizosphere under drought stress. This study's findings imply that using a mixture of bioinoculants may help soybean plants withstand drought stress, particularly during critical growth stages, and that soybean growth, productivity, and soil microbial activity were improved under drought stress.

Production of Exopolysaccharides and İndole Acetic Acid (IAA) by Rhizobacteria and Their Potential against Drought Stress in Upland Rice.

Peatlands are marginal agricultural lands due to highly acidic soil conditions and poor drainage systems. Drought stress is a big problem in peatlands as it can affect plants through poor root development, so technological innovations are needed to increase the productivity and sustainability of upland rice on peatlands. Rhizobacteria can overcome the effects of drought stress by altering root morphology, regulating stress-responsive genes, and producing exopolysaccharides and indole acetic acid (IAA). This study aimed to determine the ability of rhizobacteria in upland rice to produce exopolysaccharides and IAA, identify potential isolates using molecular markers, and prove the effect of rhizobacteria on viability and vigor index in upland rice. Rhizobacterial isolates were grown on yeast extract mannitol broth (YEMB) medium for exopolysaccharides production testing and Nutrient Broth (NB)+L-tryptophan medium for IAA production testing. The selected isolates identify using sequence 16S rRNA. The variables observed in testing the effect of rhizobacteria were germination ability, vigour index, and growth uniformity. EPS-1 isolate is the best production of exopolysaccharides (41.6 mg/ml) and IAA (60.83 ppm). The isolate EPS-1 was identified as Klebsiella variicola using 16S rRNA sequencing and phylogenetic analysis. The isolate EPS-1 can increase the viability and vigor of upland rice seeds. K. variicola is more adaptive and has several functional properties that can be developed as a potential bioagent or biofertilizer to improve soil nutrition, moisture and enhance plant growth. The use of rhizobacteria can reduce dependence on the use of synthetic materials with sustainable agriculture.

Influence of Soil Phosphate on Rhizobacterial Performance in Affecting Wheat Yield

As a primary nutrient in agricultural soils, phosphorus plays a crucial but growth-limiting role for plants due to its complex interactions with various soil elements. This often results in excessive phosphorus fertilizer application, posing concerns for the environment. Agri-research has therefore shifted focus to increase fertilizer-use efficiency and minimize environmental impact by leveraging plant growth-promoting rhizobacteria. This study aimed to evaluate the in-field incremental effect of inorganic phosphate concentration (up to 50 kg/ha/P) on the ability of two rhizobacterial isolates, Lysinibacillus sphaericus (T19), Paenibacillus alvei (T29), from the previous Breedt et al. (Ann Appl Biol 171:229–236, 2017) study on maize in enhancing the yield of commercially grown Duzi® cultivar wheat. Results obtained from three seasons of field trials revealed a significant relationship between soil phosphate concentration and the isolates’ effectiveness in improving wheat yield. Rhizospheric samples collected at flowering during the third season, specifically to assess phosphatase enzyme activity at the different soil phosphate levels, demonstrated a significant decrease in soil phosphatase activity when the phosphorus rate reached 75% for both isolates. Furthermore, in vitro assessments of inorganic phosphate solubilization by both isolates at five increments of tricalcium phosphate-amended Pikovskaya media found that only isolate T19 was capable of solubilizing tricalcium at concentrations exceeding 3 mg/ml. The current study demonstrates the substantial influence of inorganic phosphate on the performance of individual rhizobacterial isolates, highlighting that this is an essential consideration when optimizing these isolates to increase wheat yield in commercial cultivation.

Molecular identification and characterizations of rhizobacterial isolates collected from lentil rhizosphere of Indo-gangetic plains.

Plant growth promoting rhizobacteria (PGPR) are also known to colonize in the soil rhizosphere and prevent the development of other soil borne pathogens residing in the root surface. These microorganisms play a vital role in growth and development of the plant and also enhances the soil fertility by enriching the soil with different beneficial nutrients. This study was aimed at isolation of different rhizobacteria and their molecular characterization in search of efficient bacterial strains with multiple growth regulating activities. A total 36 bacteria were isolated from lentil root nodule as well as soil from different lentil growing fields with a view to screen/evaluate their plant growth promoting potential. Morphological characterization of isolated rhizobacterial candidates were done by observing the colonies on YEMA and nutrient agar media. Determination of CFU, Congo red test and gram staining tests were done to further screen them according to their morphology. All the isolates were then undergone molecular phylogenetic analysis using the partial sequences of the 16S rDNA. Based upon the Gram staining test, all the isolates were negative in gram reaction except six Bacillus isolates, PSB2 and AB3. Results of Ribosomal Database Project (RDP) and Basic Local Alignment Search Tool for Nucleotide Sequences (BLASTn) from 16S rDNA gene sequences showed that these isolates are genetically diverse. A total of 15 isolates of Rhizobium, 6 isolates of Bacillus, 3 isolates of Pseudomonas, 2 isolates of Phosphate Solubilizing Bacteria, 4 isolates of actinomycetes were identified by molecular sequencing of their 16S rDNA region and comparing them with the other isolates enlisted in the database of NCBI for the similarity percentage, query coverage. The purpose of the present study was to select native rhizosphere bacteria from the lentil nodule and soil of Lentil field and to evaluate their plant growth promoting potential as an alternative of chemical fertilizer for sustainable, environment friendly agriculture and assessment of their phylogenetic characterization.

Isolation, screening, identification, and characterization of arsenic resistant plant growth promoting rhizobacteria isolated from Pennisetum purpureum

Microbes that can progress plant productivity and health are recognized as PGPR. Rhizobacteria perform a main role in the phytoremediation process by increasing phytoremediation efficacy, thus the need to identify the superior rhizobacteria has been gaining serious attention. In the present study, Pennisetum purpureum were exposed to a series of As-polluted sand including control (0, 5, 20, 40, 60, and 80 mg kg−1) in plastic crates in a greenhouse environment. Four weeks after exposure, the plants could survive with toxicity of As pollution up to 40 mg kg−1. A total of 22 groups of bacterial isolates were confirmed based on the morphological characteristics, biochemical characteristics, and Gram staining. Preliminary and secondary As screening tests were also conducted. Identification was directed utilizing a molecular identification system. After As screening tests, 17 rhizobacterial isolates presented possible tolerance to As. Based on the molecular identification, 9 bacterial isolates were confirmed which were two [28N A UPM (Bacillus subtilis) and 28N U UPM (Bacillus australimaris)] gram-positive and seven [28N H UPM (Burkholderia seminalis), 28N G UPM (Enterobacter cloacae), 28N D UPM (Pseudomonas stutzeri), 28N I UPM (Sphingobacterium thalpophilum), 28N M UPM (Proteus mirabilis), 28N K UPM (Neisseria perflava) and 28N S UPM (Pseudomonas boreopolis)] gram-negative bacteria. All the nine identified rhizobacteria showed excellent results in terms of PGP characters. Among them, 28N U UPM (Bacillus australimaris) performed the highest PGP traits. The findings revealed that these highly As-resistant rhizobacteria have the potential to produce PGP traits and can help P. purpureum growth and productivity.

Antifungal Potential of Bacillus subtilis subsp. inaquosorum RLS76 for Management of Fusarium Wilt Disease of Bt- Cotton

Fusarium wilt is a destructive disease affecting Bt cotton that is caused by Fusarium oxysporum f. sp. vasinfectum. This seed and soilborne disease pose a significant risk to Bt cotton production worldwide, including India. Therefore, to cope with this problem in an eco-friendly manner, we explored the potentiality of the rhizobacterial isolate possessing antifungal activity as a natural arsenal against the fungal pathogen fusarium wilt of Bt cotton. The rhizobacterial isolate RLS76 inhibited 85.39% of the fungal phytopathogen in the dual culture method. The RLS76 isolate's 16S rRNA partial gene sequence was determined to be Bacillus subtilis subsp. inaquosorum RLS76, with a closest phylogenetic affiliation of 99.53%. The crude extract of Bacillus subtilis subsp. inaquosorum RLS76 demonstrated 34.38% inhibition of the growth of the pathogen after a week of incubation by using an agar well diffusion method. Bacillus subtilis subsp. inaquosorum RLS76 produced volatile organic compounds that inhibited the fungal growth of phytopathogens by 54.84% in a closed environment. Also, GCMS analysis of the crude extract of Bacillus subtilis subsp. inaquosorum RLS76 identified four compounds, namely Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-; Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl); 2,5-cyclohexadiene-1,4-dione, 2,5-Dihydroxy-3-methoxy-6-methyl-; 9,12-Octadecadienoic acid (Z,Z)-,phenylmethyl ester. Out of four compounds, three exhibit antimicrobial activity except 2,5-Dihydroxy-3-methoxy-6-methyl. The RLS76 strain of Bacillus subtilis subsp. inaquosorum displayed a transition in hue, shifting from blue to orange, surrounding the colony on CAS medium by producing siderophore qualitatively. The quantitative siderophore produced in the crude extract was 86.06 SU after 60-72 h of incubation. The RLS76 strain of Bacillus subtilis subsp. inaquosorum also showed the ability to produce a proteolytic enzyme outside the cell. This enzyme caused a noticeable clear zone around the colony when the bacteria were grown on a skim-milk agar plate. The Bacillus subtilis subsp. inaquosorum RLS76 also exhibited phosphate solubilization activity with the clear halo around the colonies on NBRIP medium. All these results suggest that Bacillus subtilis subsp. inaquosorum RLS76 exhibits good biocontrol as well as a plant growth promoting candidate against the phytopathogen of Fusarium wilt of Bt cotton.

In vitro Evaluation of Rhizobacteria Isolated from Different Oilseed Growing Areas against Fusarium oxysporum f.sp. ricini and Rotylenchulus reniformis Infesting Castor

A study was conducted to evaluate the efficacy of 108 rhizobacterial isolates collected from different Oilseed rhizospheres against the wilt pathogen, Fusarium oxysporum f.sp. ricini and reniform nematode, Rotylenchulus reniformis under in vitro conditions. Among the 108 isolates tested, four isolates were reported to inhibit the radial growth of Fusarium oxysporum f.sp. ricini by more than 50% and caused more than 80% of juvenile (J4) mortality of reniform nematode. The effective rhizobacterial isolates were subjected to morphological and biochemical characterization for the assessment of plant growth promoting traits and bio-control properties.

Peribacillus frigoritolerans T7-IITJ, a potential biofertilizer, induces plant growth-promoting genes of Arabidopsis thaliana.

This study aimed to isolate plant growth and drought tolerance-promoting bacteria from the nutrient-poor rhizosphere soil of Thar desert plants and unravel their molecular mechanisms of plant growth promotion. Among our rhizobacterial isolates, Enterobacter cloacae C1P-IITJ, Kalamiella piersonii J4-IITJ, and Peribacillus frigoritolerans T7-IITJ, significantly enhanced root and shoot growth (4-5-fold) in Arabidopsis thaliana under PEG-induced drought stress. Whole genome sequencing and biochemical analyses of the non-pathogenic bacterium T7-IITJ revealed its plant growth-promoting traits, viz., solubilization of phosphate (40-73µg/ml), iron (24±0.58mm halo on chrome azurol S media), and nitrate (1.58±0.01µg/ml nitrite), along with production of exopolysaccharides (125±20µg/ml) and auxin-like compounds (42.6±0.05µg/ml). Transcriptome analysis of A. thaliana inoculated with T7-IITJ and exposure to drought revealed the induction of 445 plant genes (log2fold-change>1, FDR<0.05) for photosynthesis, auxin and jasmonate signalling, nutrient uptake, redox homeostasis, and secondary metabolite biosynthesis pathways related to beneficial bacteria-plant interaction, but repression of 503 genes (log2fold-change < -1) including many stress-responsive genes. T7-IITJ enhanced proline 2.5-fold, chlorophyll 2.5-2.8-fold, iron 2-fold, phosphate 1.6-fold, and nitrogen 4-fold, and reduced reactive oxygen species 2-4.7-fold in plant tissues under drought. T7-IITJ also improved the germination and seedling growth of Tephrosia purpurea, Triticum aestivum, and Setaria italica under drought and inhibited the growth of two plant pathogenic fungi, Fusarium oxysporum, and Rhizoctonia solani. P. frigoritolerans T7-IITJ is a potent biofertilizer that regulates plant genes to promote growth and drought tolerance.

Use of ginger extract and bacterial inoculants for the suppression of Alternaria solani causing early blight disease in Tomato

Early blight (EB), caused by Alternaria solani, is a serious problem in tomato production. Plant growth-promoting rhizobacteria promote plant growth and inhibit plant disease. The present study explored the bio-efficacy of synergistic effect of rhizobacterial isolates and ginger powder extract (GPE) against tomato EB disease, singly and in combination. Six fungal isolates from symptomatic tomato plants were identified as A. solani on the basis of morphological features i.e., horizontal septation (6.96 to 7.93 µm), vertical septation (1.50 to 2.22 µm), conidia length (174.2 to 187.6 µm), conidial width (14.09 to 16.52 µm), beak length (93.06 to 102.26 µm), and sporulation. Five of the twenty-three bacterial isolates recovered from tomato rhizosphere soil were nonpathogenic to tomato seedlings and were compatible with each other and with GPE. Out of five isolates tested individually, three isolates (St-149D, Hyd-13Z, and Gb-T23) showed maximum inhibition (56.3%, 48.3%, and 42.0% respectively) against mycelial growth of A. solani. Among combinations, St-149D + GPE had the highest mycelial growth inhibition (76.9%) over the untreated control. Bacterial strains molecularly characterized as Pseudomonas putida, Bacillus subtilis, and Bacillus cereus and were further tested in pot trials through seed bacterization for disease control. Seeds treated with bacterial consortia + GPE had the highest disease suppression percentage (78.1%), followed by St-149D + GPE (72.2%) and Hyd-13Z + GPE (67.5%). Maximum seed germination was obtained in the bacterial consortia + GPE (95.0 ± 2.04) followed by St-149D + GPE (92.5 ± 1.44) and Hyd-13Z + GPE (90.0 ± 2.04) over control (73.8 ± 2.39) and chemical control as standard treatment (90.0 ± 2). Ginger powder extracts also induce the activation of defence-related enzymes (TPC, PO, PPO, PAL, and CAT) activity in tomato plants. These were highly significant in the testing bacterial inoculants against A. solani infection in tomato crops.

EVALUATION OF MICROBIAL POTENTIAL OF RHIZOBACTERIAL ISOLATES ASSOCIATED WITH SPENT MUSHROOM COMPOST AGAINST BACTERIAL WILT OF TOMATO

Bacterial wilt, caused by the soil-borne pathogen Ralstonia solanacearum, poses a significant threat to tomato crops worldwide. This study aims to assess the microbial potential of rhizobacterial isolates obtained from spent mushroom compost in suppressing bacterial wilt of tomatoes. Spent mushroom compost is a byproduct of mushroom cultivation and is known to harbor diverse microbial communities with potential plant-beneficial properties. Tomato leaves that were contaminated were collected from a number of places in the Rawalpindi Area, Punjab, Pakistan. Rhizobacterial isolates were collected from the rhizosphere of tomato plants grown in the presence of spent mushroom compost. These isolates were then subjected to laboratory evaluations for their antagonistic activity against R. solanacearum. Selected rhizobacterial isolates were further characterized for their plant growth-promoting traits. The potential of these isolates to enhance tomato plant growth and confer resistance against bacterial wilt was evaluated through greenhouse experiments. Results indicated that certain rhizobacterial isolates exhibited substantial antagonistic activity against R. solanacearum. Additionally, these isolates demonstrated multiple plant growth-promoting traits, suggesting a potential dual role in both pathogen suppression and plant enhancement. Greenhouse experiments revealed a significant reduction in the incidence of bacterial wilt in tomato plants and increase in the growth promotion traits were observed while treated with the selected rhizobacterial isolates alone and in combination compared to control groups. The findings from this study highlight the promising role of rhizobacterial isolates associated with spent mushroom compost in managing bacterial wilt in tomatoes.

Biofilm producing plant growth promoting bacteria in combination with glycine betaine uplift drought stress tolerance of maize plant.

The escalating threat of drought poses a significant challenge to sustainable food production and human health, as water scarcity adversely impacts various aspects of plant physiology. Maize, a cornerstone in staple cereal crops, faces the formidable challenge of drought stress that triggers a series of transformative responses in the plant. The present study was carried out in two sets of experiments. In first experiment, drought stress was applied after maintaining growth for 45 days and then irrigation was skipped, and plant samples were collected at 1st, 3rd and 6th day of drought interval for evaluation of changes in plant growth, water relation (relative water content) and antioxidants activity by inoculating indigenously isolated drought tolerant biofilm producing rhizobacterial isolates (Bacillus subtilis SRJ4, Curtobacterium citreum MJ1). In the second experiment, glycine betaine was applied as osmoregulator in addition to drought tolerant PGPR to perceive modulation in photosynthetic pigments (Chlorophyll a and b) and plant growth under varying moisture stress levels (100, 75 and 50% FC). Results of the study revealed upsurge in root and shoot length, fresh and dry biomass of root and shoot besides increasing chlorophyll contents in water stressed inoculated plants compared to uninoculated plants. Glycine betaine application resulted in an additional boost to plant growth and photosynthetic pigments, when applied in combination with bacterial inoculants. However, both bacterial inoculants behaved differently under drought stress as evident from their biochemical and physiological attributes. Isolate SRJ4 proved to be superior for its potential to express antioxidant activity, leaf water potential and relative water contents and drought responsive gene expression while isolate MJ1 showed exclusive increase in root dry biomass and plant P contents. Though it is quite difficult to isolate the bacterial isolates having both plant growth promoting traits and drought tolerance together yet, such biological resources could be an exceptional option to be applied for improving crop productivity and sustainable agriculture under abiotic stresses. By exploring the combined application of PGPR and glycine betaine, the study seeks to provide insights into potential strategies for developing sustainable agricultural practices aimed at improving crop resilience under challenging environmental conditions.