Screening for broad spectrum antagonistic Bacillus spp. with plant growth promotion capability from cassava stem and vermi-compost for control black stem rot disease of cassava
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Abstract
This research article had main objective for screening Bacillus spp. which being as broad-spectrum antagonistic bacteria and plant growth promotion as well. The Bacillus spp. isolates were isolated from healthy stem of cassava varieties and vermi-compost using NA medium. The Gram-positive bacteria which colony morphology resemble to Bacillus spp. were collected. The ability to inhibit growth of selected plant pathogenic bacteria Xanthomonas euvesicatoria, Pectobacterium carotovorum and Ralstonia solanacearum were tested by dual culture bioassay. The ability to control the major plant pathogenic fungi, e.g., Sclerotium rolfsii, Lasiodiplodia spp. and Colletotrichum capsici were also tested by dual culture bioassay. Ability for plant growth promotion was evaluated using 3 criteria including the ability to produce indole-3-acetic acid (IAA), phosphate solubilization and siderophore production. Total 106 isolates were obtained, and 12 Bacillus isolates were good candidates for further testing to control L. theobromae, the causal agent of black stem rot disease of cassava. The ability of 12 selected Bacillus isolates to inhibit the growth of Lasiodiplodia-LS02 and -LS08 were tested by dual culture bioassay compared with the broad-spectrum antagonistic Bacillus-BS and Bacillus-NTS3 obtained from Assoc. Prof. Petcharat Thummabenjapone and control (agar disc) treatment. The Lasiodiplodia-LS02 and Lasidiplodia-LS08 were the most aggressive isolates. The species determination using the nucleotide sequence of internal transcribed spacer of ITS1 -5.8s-ITS2 region using primer ITS5f and ITS4r showed that the Lasiodiplodia-LS02 and Lasiodiplodia-LS08 were designated as L. theobromae. Results revealed that the 3 top isolates were CS1, CS22 and CS99, which strongly inhibited at 48.72-56.41 inhibition percentage, significantly better than the ability of Bacillus-BS and Bacillus-NTS3 isolates. They produced IAA at a rate of 2.91-7.41 µg/ml, phosphate solubilization index was 1.17-1.22 and produced siderophore with an index at 1.70-2.40. Based on the nucleotide sequence of 16S rDNA,
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References
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