Jean-Philippe Pichaut , Benoît Farinas , Marie Umber ... · Jean-Philippe Pichaut1, Benoît...

Jean-Philippe Pichaut1, Benoît Farinas1, Marie Umber1, Matthieu Chabannes2, Nathalie

Laboureau2, Pierre Olivier Duroy2, Lydiane Bonheur1, Frédéric Salmon1, Christophe Jenny1,

Marie-Line Iskra-Caruana2 & Pierre-Yves Teycheney1Marie-Line Iskra-Caruana2 & Pierre-Yves Teycheney1

1 CIRAD, UMR AGAP, F-97130 Capesterre Belle-Eau, Guadeloupe, France2 CIRAD, UMR BGPI, F-34398 Montpellier Cedex 5, France

14ème rencontres de virologie végétale, Aussois 13-17 January 2013

Horizontal gene transfer is a very important process in eukaryotic evolution and

viruses are prominent DNA donors

• Plant genomes contain sequences of viruses with dsDNA

(Caulimoviridae) and ssDNA (Geminiviridae) genomes which have

• Animal genomes contain retroviruses and LTR retrotransposons

(Caulimoviridae) and ssDNA (Geminiviridae) genomes which have

no obligatory integration step in their replication cycle.

Teycheney P-Y & Geering ADW (2011) Endogenous viral sequences in plant genomes. Recent Advances in Plant Virology, eds Caranta C, Aranda MA, Tepfer M, & Lopez-

Moya JJ (Caister Academic Press), pp 347-366.

� Some endogenous Caulimoviridae sequences of banana, petunia

and Nicotiana edwardsonii are infectious and can lead to the

release of viral particles through stress-induced activation.


Musa acuminata (A) Musa balbisiana (B)

• Fruit organoleptic qualities

• Disease resistance traits

• Rusticity and robustness

• Partial or total resistance to

biotic and abiotic stresses


Interspecific hybrid

Infectious eBSV integratedinto the genome of the M.

balbisiana parent

Activating stress

Viral particles

© J

. Vo,

QA

AF

I

Dallot S., Acuna P., Rivera C., Ramirez P., Côte F., Lockhart B.E.L., Caruana M.L. (2001). Archives of virology, 146 : 2179-2190.

Lheureux F., Carreel F., Jenny C., Lockhart B.E.L., Iskra Caruana M.L. (2003). Theoretical and applied genetics, 106 : 594-598

Côte F, Galzi S., Folliot M., Lamagnère Y., Teycheney P.-Y., Iskra-Caruana M.-L. (2010). Mol. Plant Pathol. 11: 137–144

Interspecific hybrid(AAB / AAAB)

Known activating stresses:

• interspecific crosses

• temperature differences

• micropropagation

Viral particles

Risk of outbreak


Yellow leaf streaksPseudostem

splitting

Abnormalinflorescence and bunch emergence


• Improved banana hybrid varieties with disease resistance traits are the key to

increased sustainability through the reduction of pesticide usage

• Breeding strategies rely on a narrow genetic basis, and only on 2 families of

progenitors : Musa acuminata (A) and Musa balbisiana (B)

AAw x AAcv colchicine

AAcv AAAAcv

• CIRAD’s breeding scheme:

AAcv AAAAcv

AAcv x AAcv

colchicine

AAcv x BBw ABcv AABBcv

AAcv x AAAAcv AAAcv

AAw x AAAAcv AAAcv

BBw x AAAAcv AABcv

AAcv x AABBcv AABcv

AAw x AABBcv AABcv

Tetraploid

parents

Triploid

hybrids


Tetraploid parents

AAw x AAcv colchicine

AAcv AAAAcv

AAcv x AAcv

colchicine

AAcv x BBw ABcv AABBcv

Triploid hybrids

AAcv x AAAAcv AAAcv

AAw x AAAAcv AAAcv

BBw x AAAAcv AABcv

AAcv x AABBcv AABcv

AAw x AABBcv AABcv

• Only M. acuminata progenitors are used since 2001, which narrows down the

genetic basis available for breeding new varieties

• Research efforts have been focused on the creation of M. balbisiana ressources

devoid of infectious eBSVs for breeding purposes


eBSGFV-7(BAC 71C19)

eBSGFV-9(BAC 94I16)

13.3 kb

15.6 kb

eBSOlV -1 (BAC 310O7) 22.9 kb

23.2 kbeBSOlV-2 (BAC 73B22)

ORF1

ORF2

ORF3Viral genome

✖ 1 locus / di allélic

✖ 1 locus / mono alléliceBSImV(BAC 68C24)

15.8 kb

eBSMysV-1(BAC 29H14) 11.3 kb

eBSMysV-2(BAC 86I03)

12 kb

Infectious allele

Non infectious allele

✖ 1 locus / mono allélic

✖ 2 loci / mono-allélic

15.8 kb

Gayral P, Noa-Carrazana JC, Lescot M, Lheureux F, Lockhart BE, Matsumoto T, Piffanelli P, Iskra-Caruana M-L. (2008) J Virol. 82: 6697-710.

Iskra-Caruana M-L, Baurens FC, Gayral P, Chabannes M. (2010). Mol. Plant Micr. Interact 23: 1394-402

Gayral P, Blondin L, Guidolin O, Carreel F, Hippolyte I, Perrier X, Iskra-Caruana M-L (2010) J. Virol 84: 7346-59


�PCR Markers

OB

INO

L’E

WA

I

eBSOlV-1

eBSOlV-2

GO

LDF

ING

ER

eBSGFV-7

eBSGFV-9

eBSImV-1

IMO

VE

eBSImV-2

- Structures

- Integration

- Allèles

eBSMysV-1

IMO

VE

eBSImV-2

eBSMysV-2

MY

SO

RE

eBSV specific signature for each BSV species

Internal structure definition is ongoing


GF OL Im

PKW BBw GF7/GF9 OL1/OL2 Im

B. Honduras BBw GF7/GF9 OL1/OL2 n

Pisang Batu BBw GF7/GF9 OL1/OL2 Im

B. Cameroun BBw GF? OL? Im

Pisang Klutuk BBw GF7/GF9 OL1/OL2 Im

Porp (Popoulou) AABcv n OL? Im?

Langka AABcv n OL? n

Safet Velchi ABcv GF? OL1 n

Ney Poovan ABcv GF? OL1 n

Figue Pomme (Ekona) ABcv GF? OL1 n

Accessions with M. balbisiana genetic

background were screened by PCR, using

primer pairs differentiating infectious and

non infectious eBSVs

OL1, GF7, Im : infectious

OL2, GF9: non infectious

n : no eBSV

Figue Pomme (Ekona) ABcv GF? OL1 n

FlhorBan 902 AAAcv n n n

IDN 110T AAAA n n n

Kunnan D ABcv n OL? Im?

Kunnan T AABB n OL? Im?

FlhorBan 914 AABhyb GF9 OL2 Im

FlhorBan 910 AABhyb GF7 OL2 Im

French Clair AABcv GF7 OL1 n

Kelong Mekintu AABcv GF? OL1 n

Burro (CEMSA) ABBcv GF7/GF9 OL2/OL2 Im

Pelipita ABBcv GF7/GF9 OL1/OL2 Im

Bunga Oisan AABcv GF7 OL1 n

� Several accessions harbour both

infectious and non infectious eBSOLV

and eBSGFV alleles

� Modified alleles are present in some

cultivars (Cameroun, Kunnan, Kelong

Mekintu…)

� cv Honduras is free of eBSImV


Breeding♂ X ♀

Seeds No seeds

Embryo rescue Failed

Plantlets

Plants weaning

eBSV signatures


B. Honduras BBw GF7/GF9 OL1/OL2 n B. Honduras BBw GF7/GF9 OL1/OL2 nX

• Progeny of 70 screened by PCR

OL1/OL2 OL1/OL1 OL2/OL2

GF7/GF9 30 13 7

Allelic signatures (%)

GF7/GF7 7 6 3

GF9/GF9 10 3 4

• eBSV signature results fit expected segregation ratios

• 4 % of plants are devoid of all infectious alleles


B. Cameroun BBw GF ? OL? ImB. Honduras BBw GF7/GF9 OL1/OL2 n X


OL1/OL2 OL1 OL2 ND

Allelic signatures (%)

GF7/GF9 7 13 7 2

GF7 3 14 4 2

GF9 1 14 14 4

ND 0 3 2 11

• 24 % harbour modified eBSOLV and eBSGFV alleles


B. Cameroun BBw GF ? OL? ImB. Honduras BBw GF7/GF9 OL1/OL2 n X


OL1/OL2 OL1 OL2 ND

GF7/GF9 - - - -

Allelic signatures (%) Imové -

GF7 - - - -

GF9 - - 1.3 1.3

ND - - - 7.2


• 1.3 % are devoid of all infectious alleles

��Molecular screening tools allow:

�� the study of eBSV alleles segregation

�� the search for individuals devoid of infectious

eBSV alleles

��Molecular screening tools allow:

�� the study of eBSV alleles segregation

�� the search for individuals devoid of infectious

eBSV alleles

M.balbisiana genitors devoid of infectious eBSV alleles

were created and are being used for breeding improved

banana interspecific hybrids


�� Unravel the structure of modified eBSV alleles

��Southern-blot

�� Sequencing

�� Unravel the structure of modified eBSV alleles

��Southern-blot

�� Sequencing

�� Study the infectious status of modified alleles�� Study the infectious status of modified alleles

�� Optimize molecular screening tools specific of modified

eBSV alleles

�� Optimize molecular screening tools specific of modified

eBSV alleles

�� Study the infectious status of modified alleles

�� CMR X IDN110 (AAB) progeny is being assessed

for the expression of infectious eBSVs

�� Study the infectious status of modified alleles

�� CMR X IDN110 (AAB) progeny is being assessed

for the expression of infectious eBSVs


Lydiane BonheurIsabelle Acina

MamboleMarie Umber

Jean-Philippe

Pichaut

Pierre-Yves

Teycheney

Jean-Philippe Pichaut , Benoît Farinas , Marie Umber ... · Jean-Philippe Pichaut1, Benoît...

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