SNP Mapping to Locate Anthracnose Resistance in Capsicum spp. | Mongkolporn | Proceedings of The Annual International Conference, Syiah Kuala University - Life Sciences & Engineering Chapter 5789 11782 1 PB
Proceedings of The 5 th Annuual I nt ernat ional Conference Syiah Kuala Univ ersit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
SN P M a pping t o Loca t e Ant hr a cnose Re sist a nce in
Ca psicu m spp.
* 1 Orarat Mongkolporn
1
Depart m ent of Hort icult ure, Facult y of Agricult ure at Kam phaeng Saen, Kaset sart
Universit y, Kam phaeng Saen Cam pus, Nakhon Pat hom 73140, Thailand;
Cent er for Agricult ural Biot echnology, Kaset sart Universit y, Kam phaeng Saen Cam pus,
Nakhon Pat hom 73140 Thailand
* Corresponding Aut hor: orarat .m @ku.ac.t h
Abst ra ct
Two SNP m aps were const ruct ed from t wo ch ili populat ion s including Capsicum annu um x C.
chinense ‘PBC932’, and C. baccat um ‘PBC80’ x ‘CA1316’, aim ing t o ident ify QTLs for
ant hracnose resist ance. The ‘PBC9 32’- deriv ed m ap con t ain ed 12 link age groups ( LG) wit h 214
SNPs and 824 cM cov erage. The ‘PBC80’- deriv ed m ap cont ained 12 LGs wit h 403 SNPs and
1,270 cM cov erage. Based on t he ‘PBC932’ m ap, t wo QTLs corresponding t o t he ant hracnose
resist an ces in m at u re green and ripe fruit st ages were ident ified on t he sam e locat ion of LG2
bet ween t wo SNPs wit hin 14 cM. Based on t he ‘PBC80’ m ap, t hree QTLs were ident ified in t he
ripe fruit st age, which corresponded t o different resist ance t rait s t hat were assay ed by different
inoculat ion m et hods ( m icr oinj ect ion or MI and high pressure spray or HP) wit h t wo different
pat hot y pes ( PCa2 and PCa3) . All t he t hree m aj or QTLs f or t he resist ance t rait s assay ed by
PCa2/ MI , PCa3/ MI , and PCa3/ HP were locat ed on LG4 bet ween t wo SNP m ark ers wit hin 17 cM.
Ke y w ords: Cap sicu m baccat u m , Capsicu m chinen se, Collet ot richu m t runcat um ,
Collet ot r ichum acut at um , QTL
I nt r oduct ion
Ant hracnose, caused by a com plex of Colletot richum species, is a m aj or fungal disease infect ing chili
fruit in t he t ropics and subt ropics worldwide, especially Asia ( Mongkolporn and Taylor 2011). Fruit
yield losses, both pre- and post -harvested, due t o ant hracnose are severe in wet seasons, and t he
losses can be over 80% ( Mahasuk et al., 2009a) . Typical anthracnose sym ptom s appear as sunken
necrot ic t issues wit h concent ric rings of acervuli, that are oft en m oist ( Than et al., 2008; Mont ri et al.,
2009) .
Breeding for t he ant hracnose resistance has been around in Asia for over t wo decades wit h not m uch
success, due t o t he com plexit y of t he causal pathogen and t he host -pat hogen int eract ion, and t he lack
of resistance in Capsicum annuum gene pool ( Mongkolporn and Taylor 2011) . Three chili variet ies wit h
im m une resistance including C. chinense ‘PBC932’, and C. baccat um ‘PBC80’ and ‘PBC81’ were
ident ified by t he World Vegetable Cent er-AVRDC since 1998, and have been shared am ong Asian chili
breeders.
The ‘PBC932’ and ‘PBC80’ have been t he m aj or sources of ant hracnose resistance in Thailand, and
were used as donor parents producing t hree chili populat ions t o previously st udy t he genet ics of
ant hracnose resistance ( Pakdeevaraporn et al., 2005, Mahasuk et al., 2009a, Mahasuk et al., 2009b,
Mahasuk et al., 2013) . A key genet ic discovery in all t hree populat ions was t hat the resist ances at
different fruit m aturit y st ages were cont rolled by different genes. Of t he t hree populat ions, t wo were
used to m ap QTLs conferring t he resistance t o ant hracnose in this st udy, including an interspecific C.
annuum ‘Bangchang’ x C. chinense ‘PBC932’, and an int raspecific C. baccatum ‘PBC80’ x ‘CA1316’.
Single-nucleot ide polym orphism s (SNPs) are t he m ost abundant and st able form of genet ic variat ion
in m ost genom es, t herefore high m ap resolut ion can be fast achieved ( Ganal et al. 2009) . Several
high t hroughput SNP detect ion system s have been developed. Com pet it ive allele- specific PCR
( current ly called Kom pet it ive Allele Specific PCR or KASPTM) from KBioscience offers a user- friendly
high- t hroughput assay. Wit h t he advantage of t he high t hroughput KASP t echnology, two SNP m aps
were const ruct ed aim ing t o ident ify QTLs for ant hracnose resist ances derived from t wo chili variet ies
ie. C. chinense ‘PBC932’ and C. baccat um ‘PBC80’.
M a t e r ia ls a nd M e t hods
M a pping popula t ions and phe notyping
Two F2 single crossed populat ions segregat ing for ant hracnose resistance, i.e. interspecific Capsicum
annuum cv. ‘Bangchang’ x C. chinense ‘PBC932’, and int raspecific C. baccat um ‘PBC80’ x ‘CA1316’,
were used t o m ap wit h t he SNP m arkers. The ant hracnose resist ance was evaluat ed at m ature green
and ripe fruit m aturit y stages. The ‘PBC932’-derived populat ion was assayed wit h Collet ot richum
143
Proceedings of The 5 th Annuual I nt ernat ional Conference Syiah Kuala Univ ersit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
t runcat um ( form er nam e C. capsici) isolate ‘158ci’ by m icroinject ion ( MI ) following Mont ri et al. ( 2009)
. The ‘PBC80’- derived populat ion was assayed wit h two Collet ot richum acut atum pathot ypes including
PCa2 (Ca313) and PCa3 (CaMJ5) as ident ified by Mongkolporn et al. ( 2010) by MI and high pressure
spray ( HP) (Mahasuk et al., 2013).
Link age m ap const ruct ion and QTL ana lysis
The SNP m arkers obt ained from each chili populat ion were m apped using JoinMap 3.0 ( van Ooijen and
Voorrips 2001) . QTL analysis of t he anthracnose resist ance was perform ed using MapQTL 4.0 with
int erval m apping at LOD 3.0 ( van Ooijen et al., 2002).
Results a nd D iscussion
SN P m a ps and QTL loca t ions
PBC932-derived m ap: Approxim at ely 20% ( 214) of t he t otal 1,024 SNPs developed from the C.
annuum genom e were m apped. The m ap contained 12 linkage groups ( LG) covering 824 cM ( Fig.1) .
Figur e 1 . A SNP m ap of ‘Bangchang’ x ‘PBC932’ populat ion, form ing 12 linkage groups wit h 824 cM
t ot al coverage, const ruct ed by JoinMap 3.0, LOD 6.0- 10.0. The side bars indicate t he locat ions of t he
QTLs for ant hracnose resist ance
Two m aj or QTLs corresponding t o t he resistances t o ant hracnose in m ature green and ripe fruit , were
ident ified on the sam e locat ion of t he LG2 wit hin t wo SNP m arkers at 14 cM. ( Fig. 1) .
The 12 LGs in bot h m aps well corresponded t o the assigned Capsicum chrom osom es ( P1- P12)
( Capsicum annuum genom e dat abase; version 1.5, ht t p: / / peppergenom e.snu.ac.kr/ ) , except for t he
LG3, LG8 and LG9 in the ‘PBC80’ m ap. The LG3 and LG8 were fused wit h t he m arkers from P3, P5
and P9; and LG9 was fused wit h the m arkers from P3 and P5. Three m aj or QTLs ident ified in the
‘PBC80’ were on t he sam e locat ion of LG4. Previous genet ic analysis revealed t hat t he resist ance t rait s
by different inoculat ion m ethods, MI and HP, appeared t o be cont rolled by 2- linked genes, while t he
144
Proceedings of The 5 th Annuual I nt ernat ional Conference Syiah Kuala Univ ersit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
resist ance assayed wit h different Collet ot richum pathot ypes, PCa2 and PCa3, appeared t o be t he sam e
gene ( Mahasuk et al., 2013) .
PBC80- derived m ap: Approxim at ely 35% ( 403) of t he t otal 1,165 SNPs developed from t he C.
baccat um genom e were m apped. The m ap cont ained 12 LGs covering 1,270 cM ( Fig.2) . Three m ajor
QTLs corresponding to t he ant hracnose resistance t rait s at ripe fruit st age, including t he resistance to
PCa2 by m icroinject ion ( PCa2/ MI ) , resistance t o PCa3 by m icroinj ect ion ( PCa3/ MI ) and resistance t o
PCa3 by high- pressure spray ( PCa3/ HP) were ident ified on t he sam e locat ion of LG4 flanked by t wo
SNP m arkers 17 cM (Fig. 2).
I nt erest ingly, P4 was also resided by ot her disease resistance genes, including t ospoviruses and
pot yvirus ( Dj ian- Caporalino et al., 2006) . Previously, t wo QTLs for ant hracnose resist ance from
different donor parent, Capsicum baccat um ‘PBC81’ were m apped on P9 and P12 ( Lee et al., 2010;
Lee et al., 2011). Our t wo m inor QTLs were also on P9 and P12 (data is not shown) . The SNPs t hat
flanked all t he ident ified QTLs in bot h m aps will be great ly beneficial t o select for t he anthracnose
resist ance in chili breeding program s.
Figur e 2 . A SNP m ap of ‘PBC80’ x ‘CA1316’ populat ion, form ing 12 linkage groups wit h 1,270 cM t ot al
coverage, const ructed by JoinMap 3.0, LOD 6.0-10.0. The side bars indicate t he locat ions of t he QTLs
for ant hracnose resistance
All t he QTLs for ant hracnose resist ance ident ified in t his st udy were derived from t he sam e
populat ions t hat had been genet ically st udied ( Mahasuk et al., 2009a; 2013). The t wo m aj or QTLs
ident ified in t he ‘PBC932’ responsible for t he resist ance at m at ure green and ripe fruit st ages were on
145
Proceedings of The 5 th Annuual I nt ernat ional Conference Syiah Kuala Univ ersit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
t he sam e locat ion of LG2 or P2 (Fig 3, data is not shown) . Genet ically t he genes responsible for t he
resist ance t rait s on m at ure green and ripe fruit, co1 and co2, were linked ( Mahasuk et al., 2009a) .
Therefore the co1 and co2 convincingly resided in t he ident ified QTL area.
Conclusions
Two QTLs for ant hracnose resist ance in ‘PBC932’ were located on P2, while t hree QTLs ident ified in
‘PBC80’ were located on P4.
Ack now ledgem e nts
The project was co- funded by t he Nat ional Center for Genet ic Engineering and Biotechnology, Nat ional
Science and Technology Developm ent Agency, t he East West Seed Com pany ( Thailand), and t he
Center of Excellence on Agricult ural Biot echnology, Science and Technology Postgraduate Educat ion
and Research Developm ent , Office of Higher Educat ion Com m ission, Minist ry of Educat ion ( AGBI O/ PERDO-CHE) . The Royal Golden Jubilee Scholarship, t he Thailand Research Fund, granted a PhD
scholarship. Field experim ent s were supported by t he Tropical Veget able Research Center, Kasetsart
Universit y, Kam phaeng Saen Cam pus.
Re fe re n ce s
Dj ian - Caporalin o, C., Lefebv re, V., Sage- Daubèze, A.M., Palloix , A. ( 200 6) . Capsicum . I n: Singh RJ ( ed) Genet ic
resources, ch rom osom e engineering and cr op im prov em en t : Veget able Crops, v ol 3, 185 - 243. CRC Press,
Boca Rat on, FL, USA.
Ganal, M.W., Alt m ann, T., Roeder, M.S. ( 200 9) . SNP ident ificat ion in crop plant s. Cu rrent Opin ions in Plant Biology ,
12: 211–21 7.
Lee, J., Do, J., Yoon, J. ( 2011) . Dev elopm ent of STS m ark ers link ed t o t he m aj or QTLs for resist ance t o t he pepper
ant hracnose caused by Collet ot richum acut at um and C. capsici. Hort icult ure, Env ir onm ent , and Biot echnology ,
52: 596- 601 .
Lee, J., Hong, J., Do, J., Yoon, J., Lee, J.D. , Hong, J.H., Do, J.W., Yoon , J.B. ( 201 0) . I dent ificat ion of QTLs for
resist an ce t o ant hracn ose t o t wo Collet ot richu m species in p epper. Journal of Crop Science and Biot echnology ,
13: 227–23 3.
Mahasuk , P., Chint haisong, J., Mongk olporn, O. ( 2013) . Different ial resist ances t o ant hracn ose in Capsicu m
baccat um as responding t o t wo Collet ot richum pat h ot y pes and inoculat ion m et hods. Breeding Science, 63 :
333–338.
Mahasuk , P., Khum peng, N., Wasee, S., Tay lor, P.W.J. , Mongk olporn, O. ( 2009a) . I nherit an ce of resist ance t o
ant hracnose ( Collet ot richum capsici) at seedling and fruit ing st ages in ch ili pepper ( Capsicum spp.) . Plant
Breeding, 128: 701- 706.
Mahasuk , P., Tay lor, P.W.J., Mongk olporn , O. ( 2009b) . I dent ificat ion of t wo new genes conferring resist an ce t o
Collet ot r ichum acut at um in Cap sicu m baccat um . Phy t opat hology , 99: 1100- 11 04.
Mongk olporn, O., Mont ri, P., Supak aew, T., Tay lor, P.W.J. ( 2010) . Different ial react ions on m at u re green and ripe
chili fruit infect ed by t hree Collet ot richu m spp. Plant Disease, 94: 306- 310.
Mongk olporn, O., Tay lor, P.W.J. ( 2011) . Capsicum . I n: Kole C ( ed) Wild crop relat iv es: Genom ic and breeding
resources, v ol 5, 43- 57. Springer, New York , USA.
Mont ri, P., Tay lor, P.W.J., Mongk olpor n, O. ( 20 09) . Pat h ot y pes of Collet ot richum capsici, t he causal agent of chili
ant hracnose, in Thailand. Plant Disease, 93: 17- 20.
Pak deev araporn, P., Wasee, S., Tay lor, P.W.J., Mongk olpor n, O. ( 200 5) . I nherit an ce of resist ance t o ant hracnose
caused by Collet ot richu m capsici in Capsicu m . Plant Breedin g, 124: 206- 208.
Than, P.P., Raj esh, J., Hy de, K.D., Pongsupasam it , S., Mongk olporn, O., Tay lor, P.W. J. ( 2008 ) . Charact erizat ion
and pat hogenicit y of Collet ot richum species associat ed wit h ant hracnose infect ion on chili ( Capsicu m spp.) .
Plant Pat hology , 57: 562- 572.
v an Ooij en, J.W., Boer, M.P., Jansen, R.C., Maliepaard, C. ( 2002) . MapQTL® 4.0 , Soft ware for t he calculat ion of
QTL posit ion s on genet ic m aps. Plant Resear ch I nt ernat ional, Wageningen, t he Net herlands.
v an Ooij en, J.W., Voorrips, R.E. ( 2 001) . JoinMap® v ersion 3 .0: soft ware for t he calculat ion of genet ic link age m aps.
Wageningen: Plant Research I n t ernat ional, Wageningen, t he Net herlands.
146
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
SN P M a pping t o Loca t e Ant hr a cnose Re sist a nce in
Ca psicu m spp.
* 1 Orarat Mongkolporn
1
Depart m ent of Hort icult ure, Facult y of Agricult ure at Kam phaeng Saen, Kaset sart
Universit y, Kam phaeng Saen Cam pus, Nakhon Pat hom 73140, Thailand;
Cent er for Agricult ural Biot echnology, Kaset sart Universit y, Kam phaeng Saen Cam pus,
Nakhon Pat hom 73140 Thailand
* Corresponding Aut hor: orarat .m @ku.ac.t h
Abst ra ct
Two SNP m aps were const ruct ed from t wo ch ili populat ion s including Capsicum annu um x C.
chinense ‘PBC932’, and C. baccat um ‘PBC80’ x ‘CA1316’, aim ing t o ident ify QTLs for
ant hracnose resist ance. The ‘PBC9 32’- deriv ed m ap con t ain ed 12 link age groups ( LG) wit h 214
SNPs and 824 cM cov erage. The ‘PBC80’- deriv ed m ap cont ained 12 LGs wit h 403 SNPs and
1,270 cM cov erage. Based on t he ‘PBC932’ m ap, t wo QTLs corresponding t o t he ant hracnose
resist an ces in m at u re green and ripe fruit st ages were ident ified on t he sam e locat ion of LG2
bet ween t wo SNPs wit hin 14 cM. Based on t he ‘PBC80’ m ap, t hree QTLs were ident ified in t he
ripe fruit st age, which corresponded t o different resist ance t rait s t hat were assay ed by different
inoculat ion m et hods ( m icr oinj ect ion or MI and high pressure spray or HP) wit h t wo different
pat hot y pes ( PCa2 and PCa3) . All t he t hree m aj or QTLs f or t he resist ance t rait s assay ed by
PCa2/ MI , PCa3/ MI , and PCa3/ HP were locat ed on LG4 bet ween t wo SNP m ark ers wit hin 17 cM.
Ke y w ords: Cap sicu m baccat u m , Capsicu m chinen se, Collet ot richu m t runcat um ,
Collet ot r ichum acut at um , QTL
I nt r oduct ion
Ant hracnose, caused by a com plex of Colletot richum species, is a m aj or fungal disease infect ing chili
fruit in t he t ropics and subt ropics worldwide, especially Asia ( Mongkolporn and Taylor 2011). Fruit
yield losses, both pre- and post -harvested, due t o ant hracnose are severe in wet seasons, and t he
losses can be over 80% ( Mahasuk et al., 2009a) . Typical anthracnose sym ptom s appear as sunken
necrot ic t issues wit h concent ric rings of acervuli, that are oft en m oist ( Than et al., 2008; Mont ri et al.,
2009) .
Breeding for t he ant hracnose resistance has been around in Asia for over t wo decades wit h not m uch
success, due t o t he com plexit y of t he causal pathogen and t he host -pat hogen int eract ion, and t he lack
of resistance in Capsicum annuum gene pool ( Mongkolporn and Taylor 2011) . Three chili variet ies wit h
im m une resistance including C. chinense ‘PBC932’, and C. baccat um ‘PBC80’ and ‘PBC81’ were
ident ified by t he World Vegetable Cent er-AVRDC since 1998, and have been shared am ong Asian chili
breeders.
The ‘PBC932’ and ‘PBC80’ have been t he m aj or sources of ant hracnose resistance in Thailand, and
were used as donor parents producing t hree chili populat ions t o previously st udy t he genet ics of
ant hracnose resistance ( Pakdeevaraporn et al., 2005, Mahasuk et al., 2009a, Mahasuk et al., 2009b,
Mahasuk et al., 2013) . A key genet ic discovery in all t hree populat ions was t hat the resist ances at
different fruit m aturit y st ages were cont rolled by different genes. Of t he t hree populat ions, t wo were
used to m ap QTLs conferring t he resistance t o ant hracnose in this st udy, including an interspecific C.
annuum ‘Bangchang’ x C. chinense ‘PBC932’, and an int raspecific C. baccatum ‘PBC80’ x ‘CA1316’.
Single-nucleot ide polym orphism s (SNPs) are t he m ost abundant and st able form of genet ic variat ion
in m ost genom es, t herefore high m ap resolut ion can be fast achieved ( Ganal et al. 2009) . Several
high t hroughput SNP detect ion system s have been developed. Com pet it ive allele- specific PCR
( current ly called Kom pet it ive Allele Specific PCR or KASPTM) from KBioscience offers a user- friendly
high- t hroughput assay. Wit h t he advantage of t he high t hroughput KASP t echnology, two SNP m aps
were const ruct ed aim ing t o ident ify QTLs for ant hracnose resist ances derived from t wo chili variet ies
ie. C. chinense ‘PBC932’ and C. baccat um ‘PBC80’.
M a t e r ia ls a nd M e t hods
M a pping popula t ions and phe notyping
Two F2 single crossed populat ions segregat ing for ant hracnose resistance, i.e. interspecific Capsicum
annuum cv. ‘Bangchang’ x C. chinense ‘PBC932’, and int raspecific C. baccat um ‘PBC80’ x ‘CA1316’,
were used t o m ap wit h t he SNP m arkers. The ant hracnose resist ance was evaluat ed at m ature green
and ripe fruit m aturit y stages. The ‘PBC932’-derived populat ion was assayed wit h Collet ot richum
143
Proceedings of The 5 th Annuual I nt ernat ional Conference Syiah Kuala Univ ersit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
t runcat um ( form er nam e C. capsici) isolate ‘158ci’ by m icroinject ion ( MI ) following Mont ri et al. ( 2009)
. The ‘PBC80’- derived populat ion was assayed wit h two Collet ot richum acut atum pathot ypes including
PCa2 (Ca313) and PCa3 (CaMJ5) as ident ified by Mongkolporn et al. ( 2010) by MI and high pressure
spray ( HP) (Mahasuk et al., 2013).
Link age m ap const ruct ion and QTL ana lysis
The SNP m arkers obt ained from each chili populat ion were m apped using JoinMap 3.0 ( van Ooijen and
Voorrips 2001) . QTL analysis of t he anthracnose resist ance was perform ed using MapQTL 4.0 with
int erval m apping at LOD 3.0 ( van Ooijen et al., 2002).
Results a nd D iscussion
SN P m a ps and QTL loca t ions
PBC932-derived m ap: Approxim at ely 20% ( 214) of t he t otal 1,024 SNPs developed from the C.
annuum genom e were m apped. The m ap contained 12 linkage groups ( LG) covering 824 cM ( Fig.1) .
Figur e 1 . A SNP m ap of ‘Bangchang’ x ‘PBC932’ populat ion, form ing 12 linkage groups wit h 824 cM
t ot al coverage, const ruct ed by JoinMap 3.0, LOD 6.0- 10.0. The side bars indicate t he locat ions of t he
QTLs for ant hracnose resist ance
Two m aj or QTLs corresponding t o t he resistances t o ant hracnose in m ature green and ripe fruit , were
ident ified on the sam e locat ion of t he LG2 wit hin t wo SNP m arkers at 14 cM. ( Fig. 1) .
The 12 LGs in bot h m aps well corresponded t o the assigned Capsicum chrom osom es ( P1- P12)
( Capsicum annuum genom e dat abase; version 1.5, ht t p: / / peppergenom e.snu.ac.kr/ ) , except for t he
LG3, LG8 and LG9 in the ‘PBC80’ m ap. The LG3 and LG8 were fused wit h t he m arkers from P3, P5
and P9; and LG9 was fused wit h the m arkers from P3 and P5. Three m aj or QTLs ident ified in the
‘PBC80’ were on t he sam e locat ion of LG4. Previous genet ic analysis revealed t hat t he resist ance t rait s
by different inoculat ion m ethods, MI and HP, appeared t o be cont rolled by 2- linked genes, while t he
144
Proceedings of The 5 th Annuual I nt ernat ional Conference Syiah Kuala Univ ersit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
resist ance assayed wit h different Collet ot richum pathot ypes, PCa2 and PCa3, appeared t o be t he sam e
gene ( Mahasuk et al., 2013) .
PBC80- derived m ap: Approxim at ely 35% ( 403) of t he t otal 1,165 SNPs developed from t he C.
baccat um genom e were m apped. The m ap cont ained 12 LGs covering 1,270 cM ( Fig.2) . Three m ajor
QTLs corresponding to t he ant hracnose resistance t rait s at ripe fruit st age, including t he resistance to
PCa2 by m icroinject ion ( PCa2/ MI ) , resistance t o PCa3 by m icroinj ect ion ( PCa3/ MI ) and resistance t o
PCa3 by high- pressure spray ( PCa3/ HP) were ident ified on t he sam e locat ion of LG4 flanked by t wo
SNP m arkers 17 cM (Fig. 2).
I nt erest ingly, P4 was also resided by ot her disease resistance genes, including t ospoviruses and
pot yvirus ( Dj ian- Caporalino et al., 2006) . Previously, t wo QTLs for ant hracnose resist ance from
different donor parent, Capsicum baccat um ‘PBC81’ were m apped on P9 and P12 ( Lee et al., 2010;
Lee et al., 2011). Our t wo m inor QTLs were also on P9 and P12 (data is not shown) . The SNPs t hat
flanked all t he ident ified QTLs in bot h m aps will be great ly beneficial t o select for t he anthracnose
resist ance in chili breeding program s.
Figur e 2 . A SNP m ap of ‘PBC80’ x ‘CA1316’ populat ion, form ing 12 linkage groups wit h 1,270 cM t ot al
coverage, const ructed by JoinMap 3.0, LOD 6.0-10.0. The side bars indicate t he locat ions of t he QTLs
for ant hracnose resistance
All t he QTLs for ant hracnose resist ance ident ified in t his st udy were derived from t he sam e
populat ions t hat had been genet ically st udied ( Mahasuk et al., 2009a; 2013). The t wo m aj or QTLs
ident ified in t he ‘PBC932’ responsible for t he resist ance at m at ure green and ripe fruit st ages were on
145
Proceedings of The 5 th Annuual I nt ernat ional Conference Syiah Kuala Univ ersit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
t he sam e locat ion of LG2 or P2 (Fig 3, data is not shown) . Genet ically t he genes responsible for t he
resist ance t rait s on m at ure green and ripe fruit, co1 and co2, were linked ( Mahasuk et al., 2009a) .
Therefore the co1 and co2 convincingly resided in t he ident ified QTL area.
Conclusions
Two QTLs for ant hracnose resist ance in ‘PBC932’ were located on P2, while t hree QTLs ident ified in
‘PBC80’ were located on P4.
Ack now ledgem e nts
The project was co- funded by t he Nat ional Center for Genet ic Engineering and Biotechnology, Nat ional
Science and Technology Developm ent Agency, t he East West Seed Com pany ( Thailand), and t he
Center of Excellence on Agricult ural Biot echnology, Science and Technology Postgraduate Educat ion
and Research Developm ent , Office of Higher Educat ion Com m ission, Minist ry of Educat ion ( AGBI O/ PERDO-CHE) . The Royal Golden Jubilee Scholarship, t he Thailand Research Fund, granted a PhD
scholarship. Field experim ent s were supported by t he Tropical Veget able Research Center, Kasetsart
Universit y, Kam phaeng Saen Cam pus.
Re fe re n ce s
Dj ian - Caporalin o, C., Lefebv re, V., Sage- Daubèze, A.M., Palloix , A. ( 200 6) . Capsicum . I n: Singh RJ ( ed) Genet ic
resources, ch rom osom e engineering and cr op im prov em en t : Veget able Crops, v ol 3, 185 - 243. CRC Press,
Boca Rat on, FL, USA.
Ganal, M.W., Alt m ann, T., Roeder, M.S. ( 200 9) . SNP ident ificat ion in crop plant s. Cu rrent Opin ions in Plant Biology ,
12: 211–21 7.
Lee, J., Do, J., Yoon, J. ( 2011) . Dev elopm ent of STS m ark ers link ed t o t he m aj or QTLs for resist ance t o t he pepper
ant hracnose caused by Collet ot richum acut at um and C. capsici. Hort icult ure, Env ir onm ent , and Biot echnology ,
52: 596- 601 .
Lee, J., Hong, J., Do, J., Yoon, J., Lee, J.D. , Hong, J.H., Do, J.W., Yoon , J.B. ( 201 0) . I dent ificat ion of QTLs for
resist an ce t o ant hracn ose t o t wo Collet ot richu m species in p epper. Journal of Crop Science and Biot echnology ,
13: 227–23 3.
Mahasuk , P., Chint haisong, J., Mongk olporn, O. ( 2013) . Different ial resist ances t o ant hracn ose in Capsicu m
baccat um as responding t o t wo Collet ot richum pat h ot y pes and inoculat ion m et hods. Breeding Science, 63 :
333–338.
Mahasuk , P., Khum peng, N., Wasee, S., Tay lor, P.W.J. , Mongk olporn, O. ( 2009a) . I nherit an ce of resist ance t o
ant hracnose ( Collet ot richum capsici) at seedling and fruit ing st ages in ch ili pepper ( Capsicum spp.) . Plant
Breeding, 128: 701- 706.
Mahasuk , P., Tay lor, P.W.J., Mongk olporn , O. ( 2009b) . I dent ificat ion of t wo new genes conferring resist an ce t o
Collet ot r ichum acut at um in Cap sicu m baccat um . Phy t opat hology , 99: 1100- 11 04.
Mongk olporn, O., Mont ri, P., Supak aew, T., Tay lor, P.W.J. ( 2010) . Different ial react ions on m at u re green and ripe
chili fruit infect ed by t hree Collet ot richu m spp. Plant Disease, 94: 306- 310.
Mongk olporn, O., Tay lor, P.W.J. ( 2011) . Capsicum . I n: Kole C ( ed) Wild crop relat iv es: Genom ic and breeding
resources, v ol 5, 43- 57. Springer, New York , USA.
Mont ri, P., Tay lor, P.W.J., Mongk olpor n, O. ( 20 09) . Pat h ot y pes of Collet ot richum capsici, t he causal agent of chili
ant hracnose, in Thailand. Plant Disease, 93: 17- 20.
Pak deev araporn, P., Wasee, S., Tay lor, P.W.J., Mongk olpor n, O. ( 200 5) . I nherit an ce of resist ance t o ant hracnose
caused by Collet ot richu m capsici in Capsicu m . Plant Breedin g, 124: 206- 208.
Than, P.P., Raj esh, J., Hy de, K.D., Pongsupasam it , S., Mongk olporn, O., Tay lor, P.W. J. ( 2008 ) . Charact erizat ion
and pat hogenicit y of Collet ot richum species associat ed wit h ant hracnose infect ion on chili ( Capsicu m spp.) .
Plant Pat hology , 57: 562- 572.
v an Ooij en, J.W., Boer, M.P., Jansen, R.C., Maliepaard, C. ( 2002) . MapQTL® 4.0 , Soft ware for t he calculat ion of
QTL posit ion s on genet ic m aps. Plant Resear ch I nt ernat ional, Wageningen, t he Net herlands.
v an Ooij en, J.W., Voorrips, R.E. ( 2 001) . JoinMap® v ersion 3 .0: soft ware for t he calculat ion of genet ic link age m aps.
Wageningen: Plant Research I n t ernat ional, Wageningen, t he Net herlands.
146