ABSTRACT

Name: Quek Chuan Rui

Student ID: 16WLU10135
Title: Bio-prospecting of antagonistic agent for controlling of Ralstonia solanacearum phylotype
4, causal agent for blood disease of banana.

Banana blood disease (BDB) have been plaguing the banana industry for a few years now,
seriously affecting the production of the food product. The causal pathogen has been recently
confirmed as Ralstonia solanacearum bacterium phylotype IV. The soil sampling was conducted
at a banana plantation managed by United Plantations located at Teluk Intan, Perak. Site
selection was made according to history of BDB occurrence. This project aims to isolate and
identify the potential bio-control agents for controlling of BDB pathogen (R. solanacearum), in a
bid to find management options for blood disease of banana. The targeted bio-control agents
from Pseudomonas sp., Bacillus sp. and Actinomycetes groups were isolated from the soil using
selective isolation agar and heat-shock isolation method. A total of 5.7x108 cfu/g Pseudomonas
sp, 2x105cfu/g Bacillus sp. and 7x106 Actinomycetes have been successfully isolated. The BDB
pathogen has been successfully isolated from an infected banana plant obtained from Bukit
Rambai, Melaka. The isolation and purification of BDB pathogen was aided by Kelman’s
Tetrazolium chloride (TZC) medium. The isolated BDB was then subjected to various challenges
tests with the potential antagonistic bacteria isolated from the soil. The challenge tests included
are Kirby-Bauer disk diffusion test and agar well diffusion test. Majority of the isolates showed
negative antagonistic effect on the BDB with the exception of one bacterium isolate. This isolate
has been subjected to identification via molecular means and the identification results will be
included in the thesis. The BDB bacterium was also sent for DNA sequencing to further confirm
the identity and results will be included in the thesis. Although the results yielded are non-
conclusive, more frequent and representative samplings could be conducted in future research
coupled with more defined agar media.

Keywords: Blood disease bacterium, Kelman’s TZC, Ralstonia solanacearum, Pseudomonas sp.,
Bacillus sp., Actinomycetes