Isolation, identification, and characterization of phosphate solubilizing bacteria, Paenibacillus sp., from the soil of Danum Valley Tropical Rainforest, Sabah, Malaysia

While phosphorus (P) is a vital element for the plant to grow, only 0.1% of the phosphate soil is directly to be uptake by plants. Consequently, P fertilizer, which is mostly taken from unrenewable resources of phosphate rock, is practically added into the croplands. Nevertheless, as the demand for this fertilizer kept increasing, the availability of resources and environmental issues are currently raising wide concerns. Alternatively, soil phosphate solubilizing bacteria (PSB) is promising to be further developed as a biofertilizer to increase the availability of P elements for plants. This study aims to screen and characterize novel PSB from the tropical rainforest soil. The soil samples were collected from the Danum Valley tropical rainforest, Sabah. Phosphatase solubilizing bacteria were then screened using the NBRIP Agar selective media. The screening results yielded five colonies, designated as PSB1, PSB2, PSB3, PSB4, and PSB5, displaying halos, with an average diameter of 10mm. Further, 16s rRNA gene sequence analysis using BLASTn suggested that PSB1, PSB2, PSB3, PSB4, and PSB5 were designated as Bacillus sp. PSB01, Pseudomonas oryzyhabitans PSB02, Staphylococcus pasteuri PSB03, Paenibacillus sp. PSB04, and Staphylococcus pasteuri PSB05, respectively. Interestingly, the Paenibacillus group is a promising biofertilizer and is currently used in the global agriculture industry. Accordingly, Paenibacillus sp. PSB04 was then selected for further characterization using Gram staining and observed under scanning electron microscope (SEM). The Gram staining revealed that Paenibacillus sp. PSB04 is a Gram-negative bacterium with a rod shape, which is in good agreement with the SEM result. The specific phosphatase activity of the extracellular fraction of this bacterium was 7378.12 U mg-1 which is the highest activity compared to previous studies. This study provides an early insight into an excellent phosphate solubilizing bacterium for the agriculture industry obtained from Danum Valley.

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Screening and Molecular Identification of Phosphate-Solubilizing Rhizobacteria from Mangrove Ecosystem of the Lombok Island

JURNAL BIOLOGI TROPIS ◽

10.29303/jbt.v20i3.1730 ◽

2020 ◽

Vol 20 (3) ◽

pp. 475

Author(s):

Lalu Zulkifli ◽

Prapti Sedijani ◽

Dewa Ayu Citra Rasmi ◽

Lalu Wira Zain Amrullah

Keyword(s):

Bacterial Species ◽

Rrna Gene ◽

Plant Growth Promoting Bacteria ◽

Phosphate Solubilizing Bacteria ◽

Genome Database ◽

Paenibacillus Sp ◽

Screening And Identification ◽

Phosphate Solubilizing ◽

Phosphate Solubilizing Rhizobacteria ◽

Lombok Island

Phosphate solubilizing rhizobacteria can be used as a component in biofertilizer formulations to increase local and national food production without causing adverse risks to the environment compared to the use of chemical fertilizers. In this regard, screening and identification of phosphate solubilizing bacteria from the rhizosphere of several mangrove species that grow on the coast of Lombok Island has been carried out. The method of isolation used is the Pikovskaya method. Screening and identification activities have obtained 5 isolates of mangrove rhizosphere bacteria (BRM) which are indicated by their ability to form clear zones on Pikovskaya media, namely isolates BRM1 and BRM4 (isolated from the rhizosphere of Avicennia marina), BRM2 and BRM3 (isolated from the rhizosphere of Rhizophora apicullata), BRM5 (isolated from the rhizosphere of R. stylosa). All rhizobacteria isolates were identified as Gram-positive bacteria. Molecular analysis based on the comparison of 16S rRNA gene sequences of isolates with the genome database at GenBank (NCBI) using Mega 10 software, showed that all BRM isolates occupied the same cluster as bacterial species from the Genus Paenibacillus in the dendrogram of the phylogenetic tree, namely Paenibacillus sp. JWLB1 strain, Paenibacillus sp. Strain NO13, P. cineris strain cu1-7, P. favisporus strain CHP14, with genetic distance ranging from 1.3 to 1.4. Many species of the Genus Paenibacillus are currently known to play an important roles as plant growth-promoting bacteria. The BRM isolates obtained in this study can be further developed as a biofertilizer component (inoculant) in saline and dryland agriculture.

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Seed bio-priming with phosphate solubilizing bacteria strains to improve rice (Oryza sativa L. var. FARO 44) growth under ferruginous ultisol conditions

10.1101/2021.11.23.469751 ◽

2021 ◽

Author(s):

Musa Ibrahim Saheed ◽

Beckley Ibrahim Ikhajiagbe

Keyword(s):

Bacillus Cereus ◽

Proteus Mirabilis ◽

Rice Plant ◽

Bacterial Colonization ◽

Rrna Gene ◽

Rice Seed ◽

Phosphate Solubilizing Bacteria ◽

Growth Properties ◽

Rice Seeds ◽

Phosphate Solubilizing

The research investigated the possibility of phosphate solubilizing bacteria (PSB) with plant growthpromoting (PGP) capabilities to improve growth properties of rice plant under ferruginous ultisol (FU) condition through bio-priming strategy. The PSB with PGP properties used in this research were Bacillus cereus strain GGBSU-1, Proteus mirabilis strain TL14-1 and Klebsiella variicola strain AUH-KAM-9 that were previously isolated and characterized following the 16S rRNA gene sequencing. Biosafety analysis of the PSB isolates was conducted using blood agar. The rice seeds were then bio-primed with the PSBs at 3, 12 and 24 hours priming durations and then sown in a composite FU soil sample. Differences in germination bioassay involving SEM, morphology, physiology and biomass parameters were investigated for 15 weeks after bio-priming. The composite FU soil used in the study had high pH, low bioavailable phosphorus, low water holding capacity and high iron levels which has led to a low growth properties of rice seeds without bio-priming in FU soil. Germination parameters was better in seeds bio-primed with the PSBs, especially at 12h priming duration as against seeds without priming. SEM showed more bacterial colonization in the PSB bioprimed seeds. Seed bio-priming of rice seed with Bacillus cereus strain GGBSU-1, Proteus mirabilis strain TL14-1 and Klebsiella variicola strain AUH-KAM-9 under FU soil condition significantly improved seed microbiome, rhizocolonization and soil nutrient properties, thereby enhancing growth properties of the rice plant. This suggest the ability of PSB to solubilize and mineralize soil phosphate and improve its availability and soil property for optimum plant usage in phosphate stressed and iron toxic soils.

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Isolation and Characterization of Phosphate-Solubilizing Bacteria from Mushroom Residues and their Effect on Tomato Plant Growth Promotion

Polish Journal of Microbiology ◽

10.5604/17331331.1234993 ◽

2017 ◽

Vol 66 (1) ◽

pp. 57-65 ◽

Cited By ~ 19

Author(s):

Jian Zhang ◽

Peng Cheng Wang ◽

Ling Fang ◽

Qi-An Zhang ◽

Cong Sheng Yan ◽

...

Keyword(s):

Plant Growth ◽

Growth Promotion ◽

Wood Flour ◽

Dry Weight ◽

Rrna Gene ◽

Plant Growth Promoting Bacteria ◽

Phosphate Solubilizing Bacteria ◽

Sequence Comparisons ◽

Isolation And Characterization ◽

Phosphate Solubilizing

Phosphorus is a major essential macronutrient for plant growth, and most of the phosphorus in soil remains in insoluble form. Highly efficient phosphate-solubilizing bacteria can be used to increase phosphorus in the plant rhizosphere. In this study, 13 isolates were obtained from waste mushroom residues, which were composed of cotton seed hulls, corn cob, biogas residues, and wood flour. NBRIP solid medium was used for isolation according to the dissolved phosphorus halo. Eight isolates produced indole acetic acid (61.5%), and six isolates produced siderophores (46.2%). Three highest phosphate-dissolving bacterial isolates, namely, M01, M04, and M11, were evaluated for their beneficial effects on the early growth of tomato plants (Solanum lycopersicum L. Wanza 15). Strains M01, M04, and M11 significantly increased the shoot dry weight by 30.5%, 32.6%, and 26.2%, and root dry weight by 27.1%, 33.1%, and 25.6%, respectively. Based on 16S rRNA gene sequence comparisons and phylogenetic positions, strains M01 and M04 belonged to the genus Acinetobacter, and strain M11 belonged to the genus Ochrobactrum. The findings suggest that waste mushroom residues are a potential resource of plant growth-promoting bacteria exhibiting satisfactory phosphate-solubilizing for sustainable agriculture.

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Secretion of Gluconic Acid From Nguyenibacter sp. L1 Is Responsible for Solubilization of Aluminum Phosphate

Frontiers in Microbiology ◽

10.3389/fmicb.2021.784025 ◽

2021 ◽

Vol 12 ◽

Author(s):

Xiao Li Li ◽

Xue Qiang Zhao ◽

Xiao Ying Dong ◽

Jian Feng Ma ◽

Ren Fang Shen

Keyword(s):

Gluconic Acid ◽

Calcareous Soils ◽

Acid Soils ◽

Aluminum Phosphate ◽

Available P ◽

Rrna Gene ◽

Phosphate Solubilizing Bacteria ◽

P Deficiency ◽

Phosphate Solubilizing ◽

Major Factors

Phosphorus (P) deficiency is one of the major factors limiting plant growth in acid soils, where most P is fixed by toxic aluminum (Al). Phosphate-solubilizing bacteria (PSBs) are important for the solubilization of fixed P in soils. Many PSBs have been isolated from neutral and calcareous soils, where calcium phosphate is the main P form, whereas PSBs in acid soils have received relatively little attention. In this study, we isolated a PSB strain from the rhizosphere of Lespedeza bicolor, a plant well adapted to acid soils. On the basis of its 16S rRNA gene sequence, this strain was identified as a Nguyenibacter species and named L1. After incubation of Nguyenibacter sp. L1 for 48 h in a culture medium containing AlPO4 as the sole P source, the concentration of available P increased from 10 to 225 mg L–1, and the pH decreased from 5.5 to 2.5. Nguyenibacter sp. L1 exhibited poor FePO4 solubilization ability. When the pH of non-PSB-inoculated medium was manually adjusted from 5.5 to 2.5, the concentration of available P only increased from 6 to 65 mg L–1, which indicates that growth medium acidification was not the main contributor to the solubilization of AlPO4 by Nguyenibacter sp. L1. In the presence of glucose, but not fructose, Nguyenibacter sp. L1 released large amounts of gluconic acid to solubilize AlPO4. Furthermore, external addition of gluconic acid enhanced AlPO4 solubilization and reduced Al toxicity to plants. We conclude that secretion of gluconic acid by Nguyenibacter sp. L1, which is dependent on glucose supply, is responsible for AlPO4 solubilization as well as the alleviation of Al phytotoxicity by this bacterial strain.

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The Application of Phosphate Solubilizing Microbes Biofertilizer to Increase Soil P and Yield of Maize on Ultisols Jatinangor

KnE Life Sciences ◽

10.18502/kls.v2i6.1037 ◽

2017 ◽

Vol 2 (6) ◽

pp. 179

Author(s):

Betty Natalie Fitriatin ◽

Pujawati Suryatmana ◽

Anny Yuniarti ◽

Noor Istifadah

Keyword(s):

Soil Phosphorus ◽

Block Design ◽

Fertilizer Efficiency ◽

Phosphate Solubilizing Bacteria ◽

Soil P ◽

Soil Phosphate ◽

Randomized Block Design ◽

P Fertilizer ◽

Aspergillus Sp ◽

Phosphate Solubilizing

Ultisols has problems of low availability of nutrients, especially phosphorus. To improve soil phosphate and P fertilizer efficiency, it is necessary to develop biofertilizer such as phosphate solubilizing microbes. Phosphate solubilizing microbes (PSM) have the capability of dissolving soil phosphorus which have been adsorbed and can mineralize organic P to become inorganic P, hence increasing the avalibility of P in the soil. Phosphate solubilizing bacteria (Pseudomonas mallei and Pseudomonas cepacea) and phosphate solubilizing fungi (Penicillium sp. and Aspergillus sp) were selected based on their ability to dissolve P. The experiment was conducted at Jatinangor, West Java Indonesia to study the application of PSM biofertilizer to increase soil P and yield of maize. Experiment used a Randomized Block Design (RBD) in factorial pattern, consisting of two factors with three replications. The first factor consisted of PSM biofertilizer, which were; without PSM, 5 L ha-1 of PSM and 50 kg ha-1 of PSM. The second factor was P fertilizer with five levels (0%, 25%, 50%, 75% and 100% dosage of recommendation). The results showed that the application of PSM biofertilizer increased soil phosphate and yield of maize on Ultisol Jatinangor. The dosage of P inorganic fertilizers was reduced by 50%.Keywords: ultisol, maize, biofertillizer, phospate-solubilizing bacteria.

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Isolasi Rhizobakteri dan Pengaruh Aplikasinya dengan Pupuk N-P terhadap Mutu Benih dan Pertumbuhan Bibit Tetua Betina Jagung

Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) ◽

10.24831/jai.v45i3.13138 ◽

2018 ◽

Vol 45 (3) ◽

pp. 255

Author(s):

Pitri Ratna Asih ◽

Memen Surahman ◽

Dan Giyanto

Keyword(s):

Field Experiments ◽

Seed Quality ◽

Female Parent ◽

Dry Weight ◽

Hybrid Seed ◽

Phosphate Solubilizing Bacteria ◽

Isolation And Identification ◽

Seed Field ◽

P Fertilizer ◽

Phosphate Solubilizing

Increased productivity of maize can be done with the use of high quality seeds from improved varieties such as hybrid seed. The objectives of this study were Increasing productivity of maize female parent is important in order to reduce the price of hybrid seed. The objectives of this study were to determine the nitrogen fixing bacteria compatible with phosphate solubilizing bacteria (PSB), and using those bacteria to increase physiological seed quality and seedling growth of maize female parent. The research consisted of laboratory and field experiments. Laboratory experiment for the isolation and identification of rhizobacteria resulted in 25 Azotobacter and 29 Actinomycetes non-pathogenic isolates capable of fixing nitrogen and PSB selected for compatibility tests were AB3, B28, P12, P14, P24, and P31. The compatibility test showed 25 pairs of BPF with Azotobacter and 16 pairs of BPF with Actinomycetes were mutually compatible. The BPF pair with Azotobacter or Actinomycetes P24-AzL7, P24-AzL9, B28-AcCKB4, P24-AcCKB9, P24-AcCKB20, and P24-AcCKW5 were able to increase the vigor index of hybrid maize female parent seed. Field experiment was arranged in a split plot design with three replications. The main plot was dosage of N-P fertilizer (0%, 25%, 50%, 75%, and 100% of recommendation dosage), and the subplot was 12 rhizobacteria treatments selected from 25 compatible pairs of BPF with Azotobacter and 16 pairs of BPF with Actinomycetes and 1 control. The application of compatible pairs of bacteria had a significant effect on plant height, the number of leaves at 3 and 4 weeks after planting and plant dry weight. However, the best treatment i.e. B28-AcCKB4 was not significantly different with the nutrient broth treatment (as control).

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Isolation and characterization of phosphate solubilizing bacteria from corn rhizosfer

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/911/1/012063 ◽

2021 ◽

Vol 911 (1) ◽

pp. 012063

Author(s):

Haswania ◽

H Karim ◽

A.A. Azis ◽

N Iriany ◽

O Jumadi

Keyword(s):

Soil Samples ◽

Phosphate Solubilizing Bacteria ◽

Methyl Red ◽

Biochemical Tests ◽

Gram Staining ◽

Isolation And Characterization ◽

Sucrose Fermentation ◽

Phosphate Solubilizing ◽

Sample Dilution

Abstract The aim of this study was to isolate and characterize the Phosphate solubilizing bacteria from the rhizosphere of Zea mays L., Jeneponto Regency. This research was conducted in several stages; i.e, sampling, medium preparation, sample dilution, isolation, characterization in the form of gram staining, biochemical tests, and quantitative tests of phosphate solubility. Soil samples were diluted in 0.9% NaCl and soil containing microbes was isolated on the Picovskaya medium. Three isolates were obtained which could dissolve phosphate, namely J2KN1, J3KR2, and J3TG3 isolates. The isolates were generally round in shape with raised elevations, white, slimy, smooth, shiny surface, milky white, shape like coccus and bacillus, and gram-negative. Some of the isolates had positive motility, indole, voges, methyl red, glucose, and sucrose fermentation in the biochemical test. The quantitative tests of the ability to dissolve phosphate showed that J2KN1 isolate had the highest concentration of 51.1 μM, and the J3KR1 and J3TG3 isolates had a concentration of 45.2 μM and 37.6 μM, respectively.

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Isolation and Characterization of a Phosphate-Solubilizing Halophilic BacteriumKushneriasp. YCWA18 from Daqiao Saltern on the Coast of Yellow Sea of China

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2011/615032 ◽

2011 ◽

Vol 2011 ◽

pp. 1-6 ◽

Cited By ~ 44

Author(s):

Fengling Zhu ◽

Lingyun Qu ◽

Xuguang Hong ◽

Xiuqin Sun

Keyword(s):

Liquid Medium ◽

Soil Phosphorus ◽

Halophilic Bacterium ◽

Eastern Coast ◽

Rrna Gene ◽

Phosphate Solubilizing Bacteria ◽

Phosphorus Cycle ◽

Isolation And Characterization ◽

Moderately Halophilic Bacterium ◽

Phosphate Solubilizing

Phosphate-solubilizing bacteria (PSB) function in soil phosphorus cycle, increasing the bioavailability of soil phosphorus for plants. Isolation and application of salt-tolerant or halophilic PSB will facilitate the development of saline-alkali soil-based agriculture. A moderately halophilic bacterium was isolated from the sediment of Daqiao saltern on the eastern coast of China, which also performs phosphate-solubilizing ability. The bacterium was assigned to genusKushneriaaccording to its 16S rRNA gene sequence, and accordingly named asKushneriasp. YCWA18. The fastest growth was observed when the culturing temperature was 28∘C and the concentration of NaCl was 6% (w/v). It was founds that the bacterium can survive at a concentration of NaCl up to 20%. At the optimum condition, the bacterium solubilized 283.16 μg/mL phosphorus in 11 days after being inoculated in 200 mL Ca3(PO4)2containing liquid medium, and 47.52 μg/mL phosphorus in 8 days after being inoculated in 200 mL lecithin-containing liquid medium. The growth of the bacterium was concomitant with a significant decrease of acidity of the medium.

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Mineral Phosphate Solubilizing Bacteria Isolated from Various Plant Rhizosphere under Different Aluminum Content

Indonesian Journal of Biotechnology ◽

10.22146/ijbiotech.7558 ◽

2015 ◽

Vol 10 (2) ◽

Author(s):

Dolly Iriani Damarjaya ◽

Jaka Widada ◽

Keishi Senoo ◽

Masaya Nishiyama ◽

Shigeto Otsuka

Keyword(s):

Dry Weight ◽

Rrna Gene ◽

Plant Growth Promoting Bacteria ◽

Phosphate Solubilizing Bacteria ◽

Gene Sequence Analysis ◽

Mineral Phosphate ◽

Plant Growth Promoting ◽

The Media ◽

Formation Method ◽

Phosphate Solubilizing

The objectives of this study was to isolate and characterize the mineral phosphate solubilizing bacteriafrom rhizosphere and evaluate their potential as plant growth promoting bacteria in Al-toxic soils. The halozone formation method was used to isolate PSB using the media containing insoluble phosphates (Ca-P or Al-P)as a source of phosphate. Eight of acid and Al-tolerant PSB isolates that were able to solubilize Ca-P wereobtained from rhizosphere of clover, wheat, corn, and sunflower grown in Al-toxic soil. Identification of theisolates based on the 16S rRNA gene sequence analysis demonstrated that the isolates were strains of Burkholderia(5 strains), Pseudomonas (1 strain), Ralstonia (1 strain), and unidentified bacterium (1 strains). All PSB isolatesshowed the capability to dissolve Ca-P, and only 1 strain (Ralstonia strain) was able to dissolve Al-P in agar platemedium. The P-solubilization by these isolates was correlated with pH of medium. Inoculation of the bacterialstrains on clover on Al-toxic medium showed that all isolates increased the plant dry weight compared withuninoculated treatment. Our results showed that those PSB isolates have potential to be developed as a biofertilizerto increase the efficiency of P-inorganic fertilizer used in Al-toxic soils.

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Application of Empty Fruit Bunches Compost and Types of P Fertilizer on the Growth and Phosphorus Uptake in Oil Palm Seedlings

Agrotechnology Research Journal ◽

10.20961/agrotechresj.v4i2.40784 ◽

2020 ◽

Vol 4 (2) ◽

pp. 59

Author(s):

Pauliz Budi Hastuti ◽

Sri Manu Rohmiyati

Keyword(s):

Oil Palm ◽

Phosphorus Uptake ◽

P Uptake ◽

Phosphate Solubilizing Bacteria ◽

Empty Fruit Bunches ◽

P Fertilizer ◽

Number Of Leaves ◽

Phosphate Solubilizing ◽

Growth Of Seedlings ◽

Randomized Method

This research aimed to evaluate the effect of empty fruit bunches (efb) compost, phosphate-solubilizing bacteria (PSB), lime, and types of P fertilizer on the growth and availability of Phosphorus and P uptake in the pre-nursery (PN) oil palm seedlings in latosols. The pot experiments were arranged in a Factorial Completely Randomized method. The first factor was the application of efb compost: without efb compost, efb compost, without efb compost + PSB, and without efb compost + lime. The second factor was the types of P fertilizer: SP-36, RP, guano, and NPKMg + Urea as a control. The results showed that the combination of without efb compost +lime and RP fertilizer produces the highest number of leaves. The influence of a single factor showed that the application of without efb, efb compost, without efb compost +PSB, without efb compost + lime, resulted in the same growth of seedlings, except on plant height and the length of the leaf. The various types of P fertilizer led to the same growth of seedlings except for stem diameter. The highest available P was obtained in the combination of without efb compost + PSB with SP-36 fertilizer, which was 631.1% compared to control (without efb compost and NPKMg, Urea) and the lowest in the combination of without efb compost + lime with (NPKMg, Urea). The treatment without efb compost + lime with SP-36 fertilizer increased P uptake of leaves by 55.6% and stem by 47.1% compared to control.

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