Impact of application of endophytic Bacillus spp

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Presentation transcript:

Impact of application of endophytic Bacillus spp Impact of application of endophytic Bacillus spp. for biocontrol of cacao diseases on native microbial communities Rachel L. Melnick Ph.D. Candidate Department of Plant Pathology The Pennsylvania State University Department of Plant Pathology

Theobroma cacao L. Tropical understory perennial Seeds processed into chocolate Home to wide range of fungal endophytes

Diseases of cacao Main limiting factor

Biological Control: Pod Diseases Cherelle wilt Thought to be physiological thinning mechanism Likely has biological component Frosty Pod Black Pod Witches’ broom Focus on biocontrol for disease management

Hypothesis Endophytic Bacillus spp. may provide sustainable control of cacao diseases, through activation of plant defense mechanisms and alteration of the microbial ecology of tree

Bacterial endophytes Bacillus spp. isolated from pods, leaves, branches, floral cushions at the INIAP station in Pichilingue, Ecuador Screened for elite qualities for biocontrol agent Identified 4 potential biological control agents

Biological Control: Witches’ Broom Bacillus pumilis ET reduced overall disease severity through dry and rainy seasons

Biological Control: Pod Diseases B. pumilis ET reduced cherelle wilt in genotype CCN-51, but not diseases See poster P-714 for more information…

What is known to date… Bacillus pumilis ET works as a biological control agent, but how? Antibiosis – Reduced growth of M. roreri, M. perniciosa, and P. capsici & chitinase producer Induced resistance – Activates expression of cacao defense genes Niche displacement?

Niche displacement Thought that most endophytes are neutral in terms of plant health Introducing beneficial endophytes likely displaces neutral microbes to increase benefits How to test?

The trees… Sprayed branches of cacao trees in cardinal directions with 1x108 CFU/ml at beginning of rainy season (Jan) & again in end of March Three replicate trees per treatment Collected leaves from branches in March and May Preserved and shipped in RNALater

ARISA 16S 23S Automated Ribosomal Intergenic Spacer Analysis tRNA Extracted genomic DNA Nested PCR with bacilli specific primers following Garbeva et al. 2003 tRNA 16S 23S Microb Ecol. 2003. 45: 302-316

Abundance & Diversity in March Applied species

Abundance in May

Witches’ Broom May Sampling Reapplication

Conclusions B. pumilis ET appears to decrease diversity of native microbes Indications that application of B. pumilis ET may displace neutral endophytes

Future work… Statistical analysis of the data presented today to get more exact measures of diversity & abundance ARISA with universal bacterial primers As soon as I get home! ARISA with universal fungal primers ARISA of pods

Acknowledgements Penn State Paul Backman Scott Geib USDA-ARS SPCL Bryan Bailey INIAP-EET Carmen Suárez Karina Solis Danilo Vera Travel Support: APS Potomac Division Travel Award The Eddie Echandi & H. David Thurston Award APS Foundation Award Penn State College of Ag. Sci. Student Travel Award Research Support: USAID IPM-CRSP USDA-ARS SPCL Penn State College of Ag. Sci. Graduate Student Competitive Grant

Questions????