Soil Amendment in Kenya on Medium and Small Scale Farming Using Bio-Fertilizer Obtained from Cowpea and Peanut Nodules.

Methods

Summary

Molecular and microscopic studies of collected nodules will be performed at CIRAD, Montpellier in France. Field experiments will be done in Kenya. 

Nodule homogenate will be dilution-streaked on yeast extract Mannitol Agar (YMA) plates.

Re-infection assay commonly known as rhizobia trapping experiment for the purpose of nodule forming ability of the rhizobia isolates will be performed in the greenhouse using nitrogen free media and trap plants planted in leonard jars. 

Genomic DNA will be isolated from pure colonies using Qiagen kit.

DNA samples will be subjected to Sanger sequencing. Sequences obtained in this study will be analysed by BLAST algorithm for comparison of a nucleotide query sequence against nucleotide sequences from NCBI to find the closely related species in comparison with the nonredundant nucleotide BLAST database.

Phylogenetic analysis will be done on the basis of neighbor joining method. This will be done for the purpose of comparing 16S rDNA of the isolates. 

Challenges

My project challenges will be the pereption of farmers to a particular legume which in turn will influence the preference and cropping culture. 

My experimental fields within the forest also will be at risk since the forest is not protected against hervivours and rodents. 

Pre Analysis Plan

All data on SDW, NDW, RDW, tissue nitrogen concentration content per sample plant and SE from greenhouse experiments will be subjected to ANOVA using general linear procedure of SAS. 

Raw Sequences will be cleaned, edited and assembled using Bioedit 7.2 followed by BLAST algorithm for analysis of sequences.  Phylogenetic analysis will be done using maximum likelihood method in MEGA software version 11 with bootstrap significant value to determine the robustness.