Proceeding s of Th e 5th Annu ual I n t ern at ion al Co nferen ce Syiah Ku ala Univ ersit y ( AI C Unsyiah ) 20 15
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
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. 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 ionTwo 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
Proceeding s of Th e 5th Annu ual I n t ern at ion al Co nferen ce Syiah Ku ala Univ ersit y ( AI C Unsyiah ) 20 15
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
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).
Lin k ag e m ap const ru ct ion an d QTL an a 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).
Resu lts a nd D iscussion SN P m a ps an d 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) .
Fig u r 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
Proceeding s of Th e 5th Annu ual I n t ern at ion al Co nferen ce Syiah Ku ala Univ ersit y ( AI C Unsyiah ) 20 15
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
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.
Fig u r 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
Proceeding s of Th e 5th Annu ual I n t ern at ion al Co nferen ce Syiah Ku ala Univ ersit y ( AI C Unsyiah ) 20 15
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
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.
Con clusions
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 n ow led g em e n ts
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 ( AG-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.
