Role of B. bassiana on Plant Defence, Biocontrol and Insect Behaviour

modification

LaboratoryofPlantPathology,DepartmentofMarineSciencesandAppliedBiology,

UniversityofAlicante,E-03080Alicante,Spain.email:lv.lopez@ua.es

http://www.fungalinteractions.org/

LuisV.Lopez-Llorca

High-Level Meetingon RedPalmWeevil ManagementRome,Italy,29-31March,2017

Palmsformoasesofhighenvironmental andeconomical value andareImportantcultural/touristic assets

RPW is major palm

pest worldwide...

RPWcandamageDatePalmValue

Tabarca Island Denia Albufereta Cartagena

Córdoba Sevilla Granada

10 µm

Biocontrol: Entomopathogenicfungi

B. bassiana: most common EntomopathogenicFungus in dry Ecosystems (SE Spain)

Mitosporic fungi

Coleopterans (Beauveria bassiana)

Thrips

Lepidopterans(Paecilomyces farinosus)

Entomophthorales(Erynia neoaphidis)Aphids

B.bassiana infects RPWunder naturalconditions

193

203

207

206

Bb203 isthemostpathogenicstraintoRPW

Ricaño, J., Güerri-Agulló, B., Serna-Sarriás, M. J., Rubio-Llorca, G., Asensio, L., Barranco, P., & Lopez-Llorca, L. V. (2013). Evaluation of the pathogenicity of multiple isolates of Beauveria bassiana (Hypocreales: Clavicipitaceae) on Rhynchophorusferrugineus (Coleoptera: Dryophthoridae) for the assessment of a solid formulation under simulated field conditions. floridaentomologist, 96(4), 1311-1324.

B.bassiana dry conidiaarecapturedbyRPW

cuticle

B.bassiana dry conidiagerminate,adhere…

1

3

…andpenetrateRPWcuticle

Güerri‐Agulló et al. (2010)

B.bassiana infectsallRPWstagesunder

fieldconditions

Güerri-Agulló et al. (2011)

Bb203increasesRPWmortalityintheField(canarypalm

trial)

% Healthy Palms

Bb203 treated 75 62 -

Non-treated 60 56 38

+ Beauveria bassiana 203 -

50% treated palms

33% treated palms

0% treated palms

B.bassiana persistsinthefield

Canarypalmtrial.Datacollected12monthsafterlastBb203treatment

97%77%

Healthy palms

Bb203reducesRPWpalminfestationintheField(date

palmtrial)

Infe

stat

ion

leve

l (%

)

Treatment

Fungi

Plants

Nematodes

Insects

Nematophagous

Fungi

B. bassiana displays a Multitrophic behaviour

Entomopathogenic

Fungi

Beauveria bassianaPochonia

chlamydosporia

Nematophagous

Fungi

Chitin Protein

B.bassiana (Bb)caninhibitfungalpathogensofpalms(Pv)

Pv

Bb

B. bassiana is a Palm Endophyte

B. bassiana is a date palm root endophyte

(MohamedMahmoud etal.2017)

SESpain

Fungal Root Endophytes helpNatural vegetation cope with stress

70% Root samples!!!

Drought, Salt and Root Pathogens

(Macia-Vicenteetal.,2008,2012)

B.bassiana inducesDatePalmdefenses

RgenecodingProteins

AntioxidantProteins

HydrolyticEnzymes

HeatshockProteins

29

3031

32

33

36

37

29

34

36

37

Control

phenolics(tannins)

Control

B.bassiana

(Gómez-Vidal,etal.2008,2009).

Chitosan: Biocontrol Fungi

enhancer

Chitin inExoskeletons/CellWalls

• Chitin main component

(flexible/resistant)

• Highly adaptative (Cambric 540MA)

• ChitinàChitosan(CDAs)

• Biodegradation

(Chitinases/Chitosanases)

Chitin

microfibril

Protein (with

cross-links)

Phenolics (DOPA,

melanin etc).

Control(nochitosan) Chitosan0.01mg/ml

500nm500nm

CW

MP

CW= cellwall

celular

PM=plasma

membrane

LB= lipidbody

C=cytoplasm

LB

C

CW

(Fusariumoxysporumf.sp.radicis-lycopersici)

Palma-Guerrero etal.(2008)

Chitosan destroys the plasma membrane of plant

pathogenic fungi

Low-fluidity Membrane makes B.bassianaresistant to chitosan

Mol.Microbiol.doi:10.1111/j.1365-2958.2010.07039.

Spots119&134:homologoustoVcP1P.chlamydosporia.`rpteaseFold=2.2and1.9

CHCl

T8s2mg/ml

CHCl

T8s2mg/ml

- - - - --Sensitive Fungi:Plant

Pathogens (Forl)

- - - - -+++++++++++++++++Chitosan

Saturated FFA

Resistant Fungi (FPI,

Biocontrol Agents):

Leakage

Chitosan

Highfluidity Lowfluidity

- - - - -Chitosan

+ +

Lowfluidity Lowfluidity

Unsaturated FFA

Palmaetal.,2010MolecularMicrobiology

Chitosan stimulates B. bassiana sporulation

Mol.Microbiol.doi:10.1111/j.1365-2958.2010.07039.

Enz.Microb.Technol.46:568–

574

Spots 119 & 134: homologous to VcP1 P. chlamydosporia. `rptease Fold = 2.2 and 1.9

Palmaetal.2010,

PATENTED

Chitosan enhances virulence of Fungal Parasites of

Invertebrates

VcP1/S8serine protease

Palma et al., 2010;

Escudero et al., 2016)PATENTED

(Lopez-Moyaetal.,2016)

Chitosanactivatesstressresponsegenesinfungi

(Jaime et al., 2012)

Model:Chitosanenhaces ROSàPromotes

Appresoria differentiationandProtease

expression inBiocontrol Fungi(Escuderoetal.2016)

P. chlamydosporia(Chitosan resistant)

NEW STRATEGY:Palm/Antagonist (eg. B.bassiana)/RPW/Chitosan-OMICS(RNAseq/Proteomics/Metabolomics)

ALLGENOMESAVAILABLE

DISCOVERYOFNEWTOOLS

(Proteins/Metabolites)FORPALM

STRESSMANAGEMENTLopez-Moyaetal.,2016

Westermann etal.,2012

Conclusions

1. B. bassiana can manage sustainably RPW

2. B. bassiana can induce palm defences

3. Chitosan enhances performance of B. bassiana

4. -omics can improve Palm stress management

THANKS FOR YOUR ATTENTION!!!

THANKS TO THE PLANT PATHOLOGY LAB

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