Enhancing growth of maize seedling and plant through the integration of phosphate-solubilizing bacteria, rock phosphate, and vermicompost

1. Enhancing growth of maize seedling and plant through the
integration of phosphate-solubilizing bacteria, rock phosphate, and
vermicompost
Project code: BT456BT
Semester: 8th semester
B.Tech in Biotechnology
Date of Submission: 11/12/2079

2. Bikash Gyawali Kritima Bagale
Dewaka Poudel Sheila Khankhawas Thakuri
Rajiv Pathak
Supervisor
Team Members

3. Contents
Background
Rationale for the Study
Objectives
Methods and Methodology
Expected Outcome
Gant Chart
Conclusion
Reference
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4. Unlocking the Power of Phosphorus
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Soure: ravensdown Soure: learning chemistry

5. 5
Plant growth and development
From molecular to physiological and
biochemical processes
• Photosynthesis,
• Root development,
• Stem strengthening,
• Flower and seed formation,
• Crop maturity,
• Energy production,
• Storage and transfer reactions,
• Cell division and enlargement,
• Nitrogen fixation in legumes,
• Resistance to plant diseases,
• Conversion of sugar to starch,
• Transportation of genetic traits
• Protein synthesis,
• Biosynthesis of macromolecules,
• Respiration
Phosphorus: A crucial Macronutrient

6. History: Phosphorus fertilizers
6
History of Phosphate Fertilizer in Nepal:
History of Phosphate Fertilizer in World:

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Source:https://jabonline.in/abstract.php?article_id=784&sts=2
Phosphorus:
• Deficiency
• Roles
• Phosphorus
pool in soil
• Phosphate
solubilizing
Bacteria
• Bioavailable
Phosphorus
• Solubilization
• Enzymatic
Digestion
• Agriculture
Sustainability

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PSM – Biofertilizers

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Source: https://ib.bioninja.com.au/options/option-c-ecology-and-conser/c6-nitrogen-and-phosphorus/phosphorus-cycle.html
Source: Emami-Karvan et al.2012, 2021
Phosphorus Cycle

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Originates from the accumulation of organic remains
Significant source of phosphorus for sustainable agriculture
Produced from food waste contains a
higher level of total phosphorus
Excellent source of nutrients for plant growth
Source: https://www.ferti-organic.com/products/rock-phosphate
Source: https://www.planetnatural.com/composting-101/indoor-composting/vermicomposting/
Rock Phosphate : A Sustainable Source of Phosphorus Benefits of Vermicompost for Plant Growth and Soil Health:
for Agricultures

11. • 11
• Most cultivated crop in the world
• Animal feed
• Human consumption
• Oil production for culinary purpose
• Industrial products (biodegradable foams, plastics, and
adhesives)
• Forage
• Biofuel production
Why Maize ?

12. • Phosphorus (P) is rapidly immobilized by soil particles, creating an urgent need for sustainable
phosphorus utilization and solubilization. The application of chemical fertilizers can result in
adverse environmental effects. Nonetheless, utilizing phosphate-solubilizing bacteria may provide
a promising resolution to these challenges.
• An alternative, effective solution to tackle this problem is the integration of phosphate-solubilizing
bacteria, rock phosphate, and vermicompost.
• This study's outcomes will provide valuable information on the role of phosphate-solubilizing
bacteria, rock phosphate, and vermicompost in enhancing maize growth.
12
Rationale of the study:

13. Hypothesis:
General Hypothesis:
-Integration of phosphate- solubilizing bacteria, vermi-compost and rock phosphate enhances the
growth of maize seedlings and plants.
13
Hypothesis:

14. Objectives of the study are:
General Objective:
1. The general objectives of this research are to study the plant growth-promoting activities of
rhizospheric bacteria.
Specific objective:
The specific objectives of this research are as follows:
1. To isolate, screen and characterize the phosphate solubilizing bacteria from the rhizosphere.
2. To enhance growth of maize seedling and plant through the integration of phosphate-solubilizing
bacteria, rock phosphate, and vermicompost.
3. Analyzing the impact of phosphate-solubilizing bacteria, rock phosphate, and vermicompost on
soil.
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Objective:

15. Methodology:
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1. Site selection: -Barillot et al. in 2013
Chitwan
110m-850m
Dang
300m-1000m
Syangja
800m-1200m
Kathmandu
1200m-1400m

16. Methodology:
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2. Isolation and selection of PSB - Sharma et al., 2011
3. Soil analysis:
-phosphorous concentration, pH of the soil, acid and alkaline
phosphatase activities, phytase activity, dehydrogenase enzyme
activity , organic matter
- NAST (National Association
of Science and Technology)
4. PSB inoculum -M. A. Khan et al., 2016
5. Morphological Characterization:
- Morphology , motility
- Kit (K001-1KT, Hi Media)
-HI Media MIU medium
(SL042)
6. Phenotypic Identification of Bacterial isolates through
Biochemical test
- Indole test, Urease test, Catalase test, Voges-Proskauer (VP) test,
Methyl Red (MR) test, and Citrate test
- Tested & Control, 2016

17. Methodology:
17
7. Analysis of phosphate solubilizing activity
- qualitative and quantitative analysis
7.1.Qualitative measurement of phosphate solubilization
- P solubilization efficiency (PSE) , P solubilization index
(PSI)
- Journal & Mineralization, 2016
7.2. Quantitative measurement of phosphate
solubilization
- Pande et al., 2017
8. Seed Germination assay
- Total germination seed, seed testing, comparative
examination of germination rates of seeds and germination
capacity
- Using formula

18. Methodology:
18
9. Invitro growth promotion assay:
- Germination percentage of the seeds , vigor index
- (Gholami et al., 2009) , (Hariprasad &
Niranjana, 2009)
10. Growth chamber trial: Pot Experiment - Viruel et al., 2014 , Vashisth &
Nagarajan, 2010
11. Molecular characterization of Bacterial Isolates - (McKiernan & Danielson, 2017)
- (Weisburg et al., 1991)
- NAST
12. Phylogenetic Analysis
- ClustalW software, neighbor-joining approach,
- (Kapli et al., 2020)
- MEGA (Molecular Evolutionary
Genetics Analysis)
- Gene Bank
13. Statistical Analysis
-
- Analysis of variance (ANOVA)
method, Tukey test
- One-way factorial set up
- SPSS software

19. Expected Outcome:
The expected outcomes of this research are:
• Isolate, screen, and characterize phosphate-solubilizing bacteria from the rhizosphere.
• Enhance growth of maize seedlings and plants by integrating phosphate-solubilizing bacteria, rock
phosphate, and vermicompost.
• Analyze the impact of phosphate-solubilizing bacteria, rock phosphate, and vermicompost on soil.
19

20. Gantt Chart:
S/N Activities 2023
Januar
y
Febru
ary
March April May June July August Septem
ber
Octo
ber
Novem
ber
Decem
ber
1 Literature review
2 Sample collection
3 Isolation of PSB
4 Morphological characterization &
Biochemical Analysis
5 Quantitative estimation of
phosphate solubilizing efficiency
6 Inoculum Development
7 Testing on Maize
8 Seedling Vigor Test
9 Report Writing
10 Molecular Characterization
11 Pot Experiment
12 Statistical Analysis
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21. Budget Estimation:
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Budget estimated for our research project is around NRP 64,000/-.
Petri plates X 50 sets
8%
Laboratories Fee:
Sequencing
22%
Reagents
31%
Soil Profile Analysis
16%
Miscellaneous
23%
Budget Estimation
Petri plates X 50 sets Laboratories Fee: Sequencing Reagents Soil Profile Analysis Miscellaneous

22. Conclusion:
• Given the cultural and practical importance of maize in Nepal, addressing
fertilizer issues using this research can boost productivity in a sustainable manner
and improve soil health and fertility.
• This study has the potential to break in new oppurtunity for farmers and
researchers, exploring possibilities and leading to outstanding results in
agricultural sustainability and productivity.
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23. References:
23
Barillot, C.D.C., Sarde, CO., Bert, V. et al. A standardized method for the sampling of rhizosphere and rhizoplan soil
bacteria associated to a herbaceous root system. Ann Microbiol 63, 471–476 (2013).
Sharma, S., Kumar, V., & Tripathi, R. B. (2011). Isolation of Phosphate Solubilizing Microorganism (PSMs) From
Soil. Journal of Microbiology and Biotechnology Research Scholars Research Library J. Microbiol. Biotech. Res,
1(2), 90–95. http://scholarsresearchlibrary.com/archive.html
Khan, M. A., Marwat, K. B., & Khan, S. A. (2016). INOCULATION OF ARBUSCULAR MYCORRHIZAL FUNGI
AND PHOSPHATE SOLUBILIZING BACTERIA IN THE PRESENCE OF ROCK PHOSPHATE IMPROVES
PHOSPHORUS UPTAKE AND GROWTH OF MAIZE. 48(2), 739–747.
Tested, M., & Control, Q. (2016). Biochemical Tests for the Identification of Aerobic Bacteria. In Clinical
Microbiology Procedures Handbook. https://doi.org/10.1128/9781555818814.ch3.17.1
Journal, G., & Mineralization, P. S. (2016). Isolation of Phosphate Solubilizing Bacteria from Maize Rhizosphere
and Their Potential for Rock Growth Promotion. 0451(February). https://doi.org/10.1080/01490451.2016.1146373

24. References:
24
Pande, A., Pandey, P., Mehra, S., Singh, M., & Kaushik, S. (2017). Phenotypic and genotypic characterization of
phosphate solubilizing bacteria and their efficiency on the growth of maize. Journal of Genetic Engineering and
Biotechnology, 15(2), 379–391. https://doi.org/10.1016/j.jgeb.2017.06.005
Hariprasad, P., & Niranjana, S. R. (2009). Isolation and characterization of phosphate solubilizing rhizobacteria to
improve plant health of tomato. Plant and Soil, 316(1–2), 13–24. https://doi.org/10.1007/s11104-008-9754-6
Gholami, A., Shahsavani, S., & Nezarat, S. (2009). Effect of Pgpr on Germination,Seedling Growth and Yeild of Maize.
World Academy of Science, Engineering and Technology, 19–24.
Vashisth, A., & Nagarajan, S. (2010). Effect on germination and early growth characteristics in sunflower (Helianthus
annuus) seeds exposed to static magnetic field. Journal of Plant Physiology, 167(2), 149–156.
https://doi.org/10.1016/j.jplph.2009.08.011

25. References:
25
McKiernan, H. E., & Danielson, P. B. (2017). Molecular Diagnostic Applications in Forensic Science. In Molecular
Diagnostics: Third Edition. Elsevier Ltd. https://doi.org/10.1016/B978-0-12-802971-8.00021-3
Weisburg, W. G., Barns, S. M., Pelletier, D. A., & Lane, D. J. (1991). 6S Ribosomal DNAAmplification for
Phylogenetic Study. In JOURNAL OF BACTERIOLOGY (Vol. 173, Issue 2). http://jb.asm.org/
Kapli, P., Yang, Z., & Telford, M. J. (2020). Phylogenetic tree building in the genomic age. Nature Reviews Genetics,
21(7), 428–444. https://doi.org/10.1038/s41576-020-0233-0
MoALD, 2021. (2021). Statistical Information On Nepalese Agriculture (2077/78 ). Publicatons of the Nepal in Data
Portal, 73, 274. https://nepalindata.com/resource/statistical-information-nepalese-agriculture-207374-201617/

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