International Journal of Life Science and Agriculture Research

Study aims to evaluate the effect of two rhizobacterial isolates on the germination and vigor of rice seeds and to identify the strains through molecular characterization. A factorial completely randomized design was used, involving concentrations of rhizobacterial (0%, 25%, 50%, 75%, and 100%) and soaking durations (1 and 4 hours), with three replications, totaling 30 experimental units. Variables included seed viability and seed vigor.  Diversity is assessed by Shannon and Simpson Diversity Index. Molecular identification used 16S rRNA gene sequences. Phylogenetic trees were constructed using the NeighborJoining method, and evolutionary distances were calculated with the Kimura 2-parameter model. The results showed that soaking duration of 4 hours significantly enhanced seed vigor, the average vigor index from 76.80% to 92.07%, and growth rate from 51.88% to 72.53%, regardless of rhizobacterial concentration. The highest maximum growth potential (100%) was recorded 25% concentration and 4-hour soaking. Shannon Diversity Index for two species is: 0.693 and Simpson Diversity Index 0,5. Molecular analysis identified Isolate 1 related to Bacillus cereus strain ATCC 14579 with 95% identity. Isolate 2 showed 88% similarity, suggesting potential novelty. These findings support rhizobacteria as bioenhancement, increasing ecologically important diversity and strengthening their relevance in sustainable rice cultivation. 

Rhizobacteria, seed bioenhancement, Bacillus cereus, microbial diversity, rice

1. Al-Azmiya NU, Khumairah FH, Setiawati MR. 2021. Uji Viabilitas Isolat Bakteri Penambat Nitrogen Halotoleran pada Komposisi Bahan Pembawa yang Berbeda. Jurnal Agroekotek 13: 97–104. DOI: 10.33512/jur.agroekotetek.v13i1  
2. Alyousif, N. 2022. Distribution, occurrence and molecular characterization of bacillus related species isolated from different soil in basrah province, iraq. Biodiversitas Journal of Biological Diversity, 23(2). DOI: 10.13057/biodiv/d230209 
3. Bhadani N, Sarmah H, Prasad K, Gupta N, & Sengupta T. 2024. Salt-induced reduction of hyperswarming motility inbacillus cereusmhs is associated with reduced flagellation, reduced nanotube formation and reduction in expression of quorum sensing regulator. BioRxiv 1: 1-24. DOI: 10.1101/2024.12.17.628887 
4. Cardoso C, Bazzo J, Marinho J, & Zucareli C. 2021. Effect of seed vigor and sowing densities on the yield and physiological potential of wheat seeds. Journal of Seed Science 43: 1-11. DOI: 10.1590/2317-1545v43241586 
5. Castan D, Gomes F, & Marcos Filho J. 2018. Vigor-s, a new system for evaluating the physiological potential of maize seeds. Scientia Agricola 75 : 167-172. DOI: 10.1590/1678-992x-2016-0401 
6. Cheng H, Ye , Wu T, & Ma H. 2023. Evaluation and heritability analysis of the seed vigor of soybean strains tested in the huanghuaihai regional test of china. Plants 12 : 1-11. DOI: 10.3390/plants12061347 
7. Chiboub B, Maâtougui A, Aboukhalid K, Otouya S, Zarqi F, Nazih A, & Baghour M. 2024. Effect of gibberellic acid (ga3) and temperature on seed germination of capparis spinosa l. Journal of Applied and Natural Science 16 :12-16. DOI: 10.31018/jans.v16i1.5287 
8. Dong W, Liu H, Ning Z, Bian, Zeng L, & Xie D. 2023. Inoculation with bacillus cereus dw019 modulates growth, yield and rhizospheric microbial community of cherry tomato. Agronomy 13: 1-15. DOI: 10.3390/agronomy13061458 
9. Fu Y, Ma L, Li J, Hou D, Zeng B, Zhang L & Luo L. 2024. Factors influencing seed dormancy and germination and advances in seed priming technology. Plants 13 : 1-19. DOI: 10.3390/plants13101319 
10. Forti C, Shankar A, Singh A, Balestrazzi A, Prasad V & Macovei A. 2020. Hydropriming and biopriming improve medicago truncatula seed germination and upregulate DNA repair and antioxidant genes. Genes 11: 1-15. DOI: 10.3390/genes11030242 
11. Hiroyuki Nonogaki Hiroyuki., George W. Bassel  J., Derek Bewley. 2010. Germination Still a mystery. Plant Science 179: 574-581. 
12. Homthong M, Kaewpuk W, Yamsuan S, Thongsima A, Mokkapan K & Pikulthong V. 2022. Screening of indole-3-acetic acid pgpb from three agricultural systems at nakhon pathom, thailand. Biodiversitas Journal of Biological Diversity, 23(11). DOI: 10.13057/biodiv/d231147 
13. 1Ilangathilaka K, Rupasena L, & Fernando S. 2021. Factors affecting rice farmers’ choice of formal seed. Applied Economics & Business  5 : 51-60. DOI: 10.4038/aeb.v5i1.28 
14. 1Kandasami S, Weerasuriya N, Gritsiouk D, Patterson G, Saldıas S, Ali S & Lazarovits G. 2020. Size variability in seed ́ lot impact seed nutritional balance, seedling vigor, microbial composition and plant performance of common corn hybrids. Agronomy 10: 1-17 . DOI: 10.3390/agronomy10020157 
15. Kanjanasopa D, Wanthakarn Aiedhet, Suraphon Thitithanakul, Chanyarat Paungfoo Lonhienne.2021. Plant Growth Promoting Rhizobacteria as Biological Control Agent in Rice. Agricultural Sciences 12: 1-8.  DOI: 10.4236/as.2021.121001 
16. 1Kaushik N. 2024. Enhancing germination percentage and seed vigor in horticultural crops through biopriming techniques. Bio Web of Conferences  110: 1-9. DOI: 10.1051/bioconf/202411001012 
17. Marwan H, Mulyati S, Sarman S & Hayati I. 2020. Fluorescent pseudomonads from plant rhizosphere as biological  agent to control white root disease and growth-promoting on rubber plants. Biodiversitas Journal of Biological Diversity 21(11). DOI: 10.13057/biodiv/d211139 
18. Miljaković D, Marinković J, Tamindžić G, Đorđević V, Tintor B, Milošević D & Nikolić, Z. 2022. Bio-priming of soybean with Bradyrhizobium japonicum and Bacillus megaterium: strategy to improve seed germination and the initial seedling growth. Plants 11 : 1-15. DOI: 10.3390/plants11151927 
19. Mhada M, Zvinavashe A, Hazzoumi Z, Zeroual Y, Marelli B & Kouisni L. 2021. Bioformulation of silk-based coating to preserve and deliver rhizobium tropici to phaseolus vulgaris under saline environments. Frontiers in Plant Science 12:1-11. DOI: 10.3389/fpls.2021.700273 
20. Moreno Gavíra, A, Diánez F, Sánchez Montesinos B & Santos M. 2020. Paecilomyces variotii as a plant-growth promoter in horticulture. Agronomy 10: 1-14. DOI: 10.3390/agronomy10040597 
21. Mushtaq N, Iqbal A, Batool S, Janiad S, Ahmed M, Al‐Asmari F, & Zongo E. 2025. Alginate beads encapsulated auxinproducing pgpr as a biofertilizer promotes triticum aestivum growth. Acs Omega 10:4303-4314. DOI:10.1021/acsomega.4c06066 
22. Nithyadevi Nithyadevi G, Jerlin R, Thirusendura Selvi D &Thiyagarajan R. 2022. Effect of seed soaking duration on germination and other physiological parameters of Castor (Ricinus communis L.) Ecology Environment and Conservation 28: S565-S570. DOI: 10.53550/EEC.2022.v28i01s.077 
23. Nivetha N, Asha A, Krishna G. Chinnusamy V, & Paul S. 2024. Rhizobacteria bacillus spp. mitigate osmotic stress and improve seed germination in mustard by regulating osmolyte and plant hormone signaling. Physiologia Plantarum 176: e14202.  DOI:10.1111/ppl.14202 
24. Nontji M, Ayu KP, Saida Saida, Suriyanti Suriyanti, Muliaty Galib, Anwar Robbo, Ida Suryani, Lailatul Qadar. 2023. Increacing Plant Health using Plant Growth Regulator from Rice Rhizobacteria. Online Journal Biological Science 23: 50-56. DOI: 10.3844/ojbsci.2023.50.56 
25. Ornellas FRS, Aurisangela, ODS, Carlos PP, Marcelo dNA, Araujo, Denis SDC, Barbara Fd, Rafael, MB. 2020. Gene expression, biochemical and physiological activities in evaluating melon seed vigor through ethanol release. Scientia Horticulturae 261: 01-09. DOI: 10.1016/j.scienta.2019.108884 
26. 2Sui J, Yang J, Li C, Zhang L & Hua X. 2023. Effects of a microbial restoration substrate on plant growth and rhizosphere microbial community in a continuous cropping poplar. Microorganisms 11: 1-14 DOI: 10.3390/microorganisms11020486 
27. Sulistiyani TR, Mia Kusmiati, Gita Azizah Putri. 2021 . Analisis 16S rRNA dan Skrening Enzim Bacillus dari Tanah Rhizosfer Pulau Lombok. Jurnal Ilmu Petanian. 26: 582-590. DOI: 10.18343/jipi.26.4.582 
28. Rodrigues Lenia, Amaia Nogales, Joao Nunes, Leonardo Rdrigues, Lee D Hansen, Helia cardoso. 2023. Germination of Pisum sativus Lseeds is assosiated with Alternativere Respiratory Pathway. Biology 12:1-22. DOI: 10.3390/biology12101318 
29. Santan C, Freitas L, Oliveira T, Crevelari J, Cruz D, Gravina L & Gravina G. 2019. Evaluation of physiological quality of seeds of improved snap bean lines under different storage. Journal of Experimental Agriculture International 36: 1-10. DOI:10.9734/jeai/2019/v36i430242
30. Savvas D, Magkana P, Yfantopoulos D, Kalozoumis P & Ntatsi G. 2024. Growth and nutritional responses of zucchini squash to a novel consortium of six bacillus sp. strains used as a biostimulant. Agronomy 14: 1-16. DOI: 10.3390/agronomy14020362 
31. Suriani S, Baharuddin B, Muis A, Junaid M, Mirsam H & Azrai M. 2023. Morpho-physiological and molecular characteristics of bacteria causing stalk rot disease on corn in gorontalo, indonesia. Biodiversitas Journal of Biological Diversity, 24(3). DOI: 10.13057/biodiv/d240349 
32. Stinke Kat, Omkar, Tilman Weber, Akos T Kovacs. 2021. Phylogenetic Distribution of Secondary Metabolites in the Bacillus subtilis Species Complex. American Societi For Microbiology 6:2-9. DOI:10.1128/msystems.00057-21 
33. Tania S, Hossain M & Hossain M. 2019. Effects of hydropriming on seed germination, seedling growth and yield of bitter gourd. Journal of the Bangladesh Agricultural University 17 : 281-287. DOI: 10.3329/jbau.v17i3.43197 
34. Verma P, Hiremani N, Gawande S, Sain S, Nagrale D, Narkhedkar N & Prasad Y. 2022. Modulation of plant growth and antioxidative defense system through endophyte biopriming in cotton (Gossypium spp.) and non-host crops. Heliyon, 8 : 1-9. DOI: 10.1016/j.heliyon.2022.e09487 
35. Wahyudi A, Fithriansyah N, Amri M, Priyanto J & Nawangsih A. 2021. Screening of chitinase-producing rhizosphere actinomycetes and their genetic diversity. Biodiversitas Journal of Biological Diversity, 22(10). DOI: 10.13057/biodiv/d221008 
36. Wang, Y. 2020. Feasibility analysis of NIR for detecting sweet corn seeds vigor. Journal of Cereal Science 93 DOI:10.1016/j.jcs.2020.102977 
37. Xie Y, Lihui Wei, Yinghua Ji, Shuo Li. 2022. "Seed Treatment with Iron Chlorine E6 Enhances Germination and Seedling Growth of Rice. Agriculture 12: 218.  DOI: 10.3390/agriculture12020218 
38. Yan H, Zhang Z, Lv Y & Nie Y. 2024. Integrated multispectral imaging, germination phenotype, and transcriptomic analysis provide insights into seed vigor responsive mechanisms in quinoa under artificial accelerated aging. Frontiers in Plant Science 15: 1-8. DOI: 10.3389/fpls.2024.1435154 
39. Zulaika E, Solikhah F, Utomo M, Endah N & Assavalapsakul W. 2022. Cellulose degrading bacteria isolated from palangkaraya, central kalimantan, indonesia as peat fiber decomposer to accelerate peat soil compression. Biodiversitas Journal of Biological Diversity, 23(3). DOI: 10.13057/biodiv/d230356