1. Lila N. Salhi, David Bernard-Perron, Suha Jabaji, B. Franz Lang
Microorganisms with biofertilizing and biocontrol properties in cranberries
Lila N. Salhi, David Bernard-Perron, Suha Jabaji, B. Franz Lang

2. Overview What are plant endophytes? Benefits of endophytes for plants
Only few endophytes are known
Are there more? what are they good for?
Result: endophyte diversity is completely underestimated
Which ones are beneficial?
Conclusions : Exiting future opportunities

3. Why microorganisms? Every child knows (?):
bacteria dissolve minerals (P, K) and make them available via plant roots.
Microbial community is involved in:
Stimulation of plant growth
Improvement of soil fertility
Holguin et al., 1999). Ciencia y Desarrollo 25 (No.144): 26-35

4. Plant growth promoting microorganisms
Colonize plant roots (rhizosphere and endophytes).
Dominate in competition with other microbes.
Promote plant growth by:
Wide spectrum of hormonal stimulation factors
Biofertilization (nutrient solubilization, nitrogen fixation)
Pathogen-control (antibiotics, fungicides, insecticides …)
Rothballer et al., 2003; Gamalero et al., 2004, Persello-Cartieauxet al., 2003

5. Are endophytes always plant growth promoting?
No!
Potential interaction types
(fluid, sometimes transitional):
Neutral : No effect
Mutualistic : beneficial
Pathogenic : harmful
Zhu et al, 2012

6. Fungal endophytes of cranberries: little is known
Ericoid mycorrhiza
Dark, septate endophytes
Growth inside plant cells.
Hyphae don't extend into soil
Heterogeneous group
Darkly-pigmented
Septate hyphae
Rhizoscyphus ericae
Oidiodendron maius
Phialocephala fortinii
Smith and Read, 2008; Jumpponen and Trappe, 1998

7. … and bacterial endophytes: even less is known
Pseudomonas syringae
Gram negative with polar flagella
Usually a pathogen, but …
Successful in controlling Cottonball disease (Monilinia oxycocci)
Pseudomonas sp
voland et al, 1999

8. Questions and Hypotheses
A much more diverse universe of endophytes?
Which isolates in our microbial collection are beneficial?
How to implant them sustainably with plants ?

9. 1. Isolation from plant tissue 3. DNA Sequencing and Blast (NCBI)
Molecular identification of microorganisms
1. Isolation from plant tissue
ITS
ITS1-F
BMBC-F
ITS4-R
5.8S
ITS1
ITS2
28S
18S
600 nt
750 nt
IGS
ETS
rRNA gene in fungi
Leave
Stem
Root
2. DNA Extraction and PCR
tRNA
534-R
27-F
507 nt 5S
23S
16S
rRNA gene in bacteria
3. DNA Sequencing and Blast (NCBI)
BLAST
Muyzer et al. (1993) ; White et al ; blast.ncbi.nlm.nih.gov

10. Halo representing mineral solubilization
Checking of microorganism properties
Phosphate
solubilization
Nitrogen
fixation
Biocontrol
NBRIP medium
Insoluble phosphate
Nitrogen
free medium
PDA medium
Halo representing mineral solubilization
Bacterial growth
Fungal or bacterial
growth inhibition

11. 2 – Beneficial Proprieties
Results
1 – Microbial Diversity
2 – Beneficial Proprieties

12. 137 microorganisms isolated
Unexpected endophyte diversity: the tip of an iceberg?
Roots
65 Fungi, 36 Bacteria
137 microorganisms isolated
Trimmatostroma betulinum
Not identified
Bacteria :
Bacillus sp.
Paenibacillus sp
Pantoea sp
Fungi
Others
Stems
Leaves
Roots

13. Stems 16 Fungi, 8 Bacteria Bacteria: Fungi: Others Stems Leaves
Alternaria sp.
Nemania serpens
Fungi:
Paenibacillus sp.
Bacteria:
Others
Stems
Leaves
Roots

14. Leaves 8 Bacteria Bacteria: Others Stems Leaves Roots
Bacillus subtilis
Bacteria:
Others
Stems
Leaves
Roots

15. Phosphate solubilization
Which endophytes may be beneficial?
Phosphate solubilization
Nitrogen fixation
Bacillus subtilis
Burkholderia sp.
Rahnella aquatilis
Rhodococcus qingshengii Burkholderia sp.
Rahnella aquatilis
Panteoa sp.
Cohnella ginsengisoli
Rhizobium sp.

16. Penicillium janthinellum
Biocontrol
Pseudomonas sp.
Penicillium janthinellum
Fungal-fungal
interaction
Bacterial-fungal
interaction

17. Conclusions
Unexpectedly large endophyte diversity in cranberry tissues
Different types of microbes in roots and aerial plant parts.
Some endophytes have plant growth promoting properties.
Others have strong fungicide properties.
What Next?

18. Thanks ! This work was possible thanks to financial support by Le ministère de l'Agriculture, des Pêcheries et de l'Alimentation du Québec