ECOLOGICAL DISTRIBUTION OF PHYTO-BENEFICIAL BACTERIA TOWARDS IMPROVEMENT OF MAIZE By *1, 3, 4ABIALA, M. A, 1ODEBODE, A. C, 3ADEOYE, G. O, 4HSU, F., and 4BLACKWOOD, C. B. 1Department of Botany and Microbiology, University of Ibadan, Nigeria 2Department of Biological Sciences, Ajayi Crowther University, Oyo, Nigeria 3Department of Agronomy, University of Ibadan, Nigeria 4Department of Biological Sciences, Kent State University.

Maize (Zea mays L.): Why this study? Introduction Economic importance (CIMMYT, 2010) High demand (Akande, 1994) Implication of NPK fertilizer (Gerhardson, 2002) Any alternative for NPK fertilizer? Phyto-beneficial bacteria as an alternative (Tilak et al., 2007; Hayat et al., 2010) Why this study? Objectives Phylogenetic assessment of phyto- beneficial bacteria With respect to ecological distribution in Southwestern Nigeria

Materials and Methods Field Survey Laboratory evaluation Screen house evaluation Phylogenetic analysis - Taxonomy (Wang et al., 2007) - Editing of sequences - Bioedit version 7. 0. 5 - Sequence alignment (Thompson et al., 1994) - Phylogenetic - Mega 5 (Tamura et al., 2011) Data Collection and Analysis Zone of inhibition (mm) Plumule and radicle length Phosphateand chitinase activity index Growth parameters Plant height, Stem girth, Number of leaves and Leaf area ANOVA was performed using SAS ( 9.2) - Tukey – Kramer HSD test at α = 0.05 - SNK test at α = 0.05 Figure 1: Map of Southwestern Nigeria showing Ecological zones and Study areas.

Evaluation and screening parameters Table 1: Plant growth promoting characteristics of bacterial isolates Evaluation and screening parameters Effectiveness (%) Antagonistic bioassay 20.25 Phosphate solubilization 61.20 Chitinase activity 63.30 IAA (L – Tryptophan) 100.00 IAA (D – L Tryptophan) Seed germination Radicle length 60.42 Plumule length 47.20 Plant height 81.25 Stem girth 0.00 Number of leaves 83.33 Leaf area 68.75 Disease expression at day 14 12.24 Disease expression at day 21 43.75 Bacteria load 97.92

Effect of beneficial bacteria on maize seed germination Effect of beneficial bacteria on maize root whorls Effect of beneficial bacteria on maize growth Plate 1: Diagram representing sequential effect of beneficial bacteria on maize growth

Table 2: Molecular identification in distribution of phyto-beneficial bacterial isolates Phyto-beneficial bacteria code Ecological Zone RDP Similarity match (%) Molecular identification Dendogram / phylogenetic similarity match   ILS13 Derived Savannah Bacillus sp. (100%) Bacillus sp. Bacillus niacini EPR7 Fresh water swampy forest Bacillus aeolius EPR2 Lysinibacillus sp. (100%) Lysinibacillus sp. Lysinibacillus boronitolerans IGGR11 Enterobacter sp. (93%) Enterobacter sp. Enterobacter pyrinus OSR7 Lowland rain forest Enterobacter sp. (70%) Enterobacter radicincitans IBS8 Citrobacter sp. (97%) Citrobacter sp. Citrobacter fameri UNS9 Mangrove forest Stenotrophomonas sp. (100%) Stenotrophomonas sp. Stenotrophomonas maltophilia ABS6 Azomonas sp. (26%) Azomonas sp. Azomonas macrocytogenes TDS9 Guinea Savannah Myroides sp. (100%) Myroides sp. Myroides odoratus AT-IKS IPR1 EPR4

Firmicutes (25.00%) Proteobacteria (41.67%) Bacteroidetes (33.33%) Prevalence of occurrence (%)   Phyto-beneficial bacteria Firmicutes (25.00%) Bacteroidetes (33.33%) Figure 1: Distribution of 16Sr DNA identified phyto – beneficial bacteria based on phylla taxonomy

Ecological zone Distribution (%) Figure 2: Ecological distribution of identified phyto-beneficial bacterial isolates

Fresh Water swampyforest Table 3: Identified soil bacterial isolates based on their ecological zone Bacterial isolates Guinea Savannah Derived Savannah Lowland Rainforest Fresh Water swampyforest Mangrove Forest Genus 1 (Myroides) 1 (TDS9) 1 (AT-IKS) 1 (IPR1) 1 (EPR4) Genus 2 (Enterobacter) 1 (IGGR11) 1 (OSR7) Genus 3 (Citobacter) 1 (IBS8) Genus 4 (Stenotrophomonas) 1 (UNS9) Genus 5 (Bacillus) 1 (ILS13) 1 (EPR7) Genus 6 (Azomonas) 1 (ABS6) Genus 7 (Lysinibacillus) 1 (EPR2)

EPR2 – Lysinibacillus sp. UNS9 – Stenotrophomonas sp. TDS9 ILS13 OSR7 EPR2 UNS9 TDS9 – Myroides sp. ILS13 – Bacillus sp. OSR7 – Enterobacter sp. EPR2 – Lysinibacillus sp. UNS9 – Stenotrophomonas sp. Figure 3: Map of southwestern Nigeria showing most effective phyto-beneficial bacteria per ecological zone.

CONCLUSION The less number and variation observed in beneficial bacteria across and within the ecological zones could be attributed to; (Salako et al., 2002; Eludoyin and Wokocha, 2011) A large number of physico-chemical changes taking place in the soil which occur due to agriculture. Soils of Southwestern Nigeria suffer physical degradation; Such as erosion and chemical degradation, which causes nutrient loss. These degradations can sometimes be caused by tillage, careless use of pesticides and fertilizers, and sewage slime, which eventually causes organic matter and biodiversity loss. (Ibekwe et al., 2001; Girvan et al., 2003)

THANK YOU…. Acknowledgements University of Ibadan IITA, Ibadan West Africa Res. Ass. University of Ibadan IITA, Ibadan IAR&T, Ibadan Dept. of Botany, UI Dept. of Microbiology, UI Finally, To God Almighty THANK YOU….