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Richard Lally , Antonio Sergio Moreira , Paul Galbally , John Culhane , Nicholas Otiento , David Ryan, Kieran Germaine and David Dowling. Development of endophytic bacterial inoculants possessing plant growth promotion traits for practical application in bio-energy plant species. - PowerPoint PPT Presentation
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Development of endophytic bacterial inoculants possessing plant growth
promotion traits for practical application in bio-energy plant species
Institute of Technology Carlow, Ireland
Richard Lally, Antonio Sergio Moreira, Paul Galbally, John Culhane, Nicholas Otiento, David Ryan, Kieran Germaine and
David Dowling
Presentation Overview
Introduction
Field experiment
Results
Conclusion
Introduction
Richard Lally•Postgraduate researcher, PhD candidate•BSc in Biotechnology with Biopharmaceuticals
EnviroCORE research department
Focus on •Environmental research techniques• Environmental issues and•Environmentally effective biotechnology innovation
My research focus Examination of the use of microorganisms for their use as live
microbial biofertilisers
Microorganisms and agriculture
Exploitation of favourable traits for agricultural benefits
How do bacterial endophytes colonise and promote growth ?
What are the microorganisms doing to make capable plant colonisation?
Are the microbes promoting growth at a field scale?
In 2012 -EnviroCORE IT Carlow was awarded SFI (Science foundation Ireland) TIDA ( Technology Innovation Development Feasibility Study)
Funding provided a platform to perform a:
1. Field trial to investigate the use of live microbial biofertilisers
2. Investigation into the bacterial gene sequences of the endophytes
Plant growth promoting traits•Siderophore production, Phosphate solubilisation, Plant hormone regulation compounds including ACC deaminase and plant hormone IAA production
Strains can colonise plant roots and rhrizoshere•Displayed numerous times in green house trials in various plant species
Have enhanced plant growth in green house trials •Shown in various experiments to enhance different aspects of plant growth
Background to our endophytes Isolated from Miscanthus giganteus
Three Pseudomonas fluorescens strains•L321, L111 and L228
Why test in the field?
Choice of crop Brassica napus – Oilseed rape
Economically important crop in Ireland
•Used for bio-fuel production and fuel substitution
•Used in culinary oil production
•Prior experiments showed that the microorganisms colonise and significantly promote plant biomass in greenhouse trials
The field trial
Three field trials1. First trial unsuccessful trial; uncompleted 2. Malones site examination of “Compass” variety; completed 3. Knockbeg site examination of “Castille” variety; completed
Treatments:L321L111L228F113Master mix (MM)Control
All treatments were applied to each trial by spray application after germination
Master mix contained the 3 endophytes including that of 7 other EnviroCORE strains
Trial 1: Uncompleted Used a seed coating incorporating our strains
Trial 2: Malones trial Three spray applications after germination
Trail 3: Knockbeg Trial Laboratory based coating formulation designed in IT Carlow followed by one spray application
Tracking microorganisms in the field An effort was made to detect the microorganisms in the field
We could not use plating techniques as we did not use genetically modified microorganisms
Attempted to track F113 using molecular methods
PCR amplification using sequence characterised amplification region markers developed by Von Felten et al., 2010
Unsuccessful as primers were amplifying regions of similar bacteria in the field
Results
The overall seed and oil yield was not impacted as a result of the biofertiliser application
The “Castille” trial found there was no positive impact as a result of the treatment biofertilisers
However
Continuous sampling from “Compass” trial showed a significant increase in stem and leaf biomass at full growth
Seed yield from “Compass” and “Castille”
F113 L228 L321 L111 MM Control0
500
1000
1500
2000
2500
Yiel
d Kg
/ha
Castille seed yield F113 L228 L111 L321 MM Control
2500
2600
2700
2800
2900
3000
3100
Yeild
Kg/
ha
Compass seed yield
No significant increase in seed yield
Error bars represent the standard error of the mean
“Compass” dry stem and leaf biomass
F113 L228 L111 L321 MM CONTROL0
5
10
15
20
25
Treatment Strain
Wei
ght
(g)
* *
*Significant increase over control
Conclusions
The bacterial inoculants F113 and master mix increased the stem and leaf biomass of Brassica napus variety “Compass”
Strain specific molecular markers are ineffective in the detection of Pseudomonas F113 wild type strain in the field
Three spray strategy may be more effective than a seed coating followed by a signal spray treatment
A variety effect can not be ruled out as a possible colonisation effect factor
The inoculants may be increasing stem development but not contributing to the overall yield
Future Work
Effect of endophytes on stem yielding crops
Stages of colonisation
Data mining using bacterial annotations
Molecular interaction between plants and bacteria
Examination of reduced chemical treatments under biofertilisers
AcknowledgementsThis research is funded by Science Foundation Ireland and Enterprise
Ireland under the Technology Innovation Development Award (TIDA), ITC Postgraduate Scholarship Scheme and IoTI (Institute of Technology
Ireland) Postgraduate Scholarship Initiative
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