Molecular Marker Application to Incorporate Salinity Tolerance to West Africa Rice Varieties Isaac Kofi Bimpong1, Baboucarr Manneh1, Bathe Diop1 and Glenn B. Gregorio2 1Africa Rice Centre, Sahel station, B.P 96, Saint Louis, Senegal, 2 International Rice Research Institute (IRRI), DAPO Box 7777, Manila, Philippines Salinity is a serious constraint for rice production in West Africa. Molecular markers are now routinely being used to identify and transfer useful genes/QTLs for salinity tolerance. One of the major QTL (Saltol) derived from the salt-tolerant Indian landrace Pokkali, has been mapped on chromosome 1.The QTL for salt tolerance (SKC1), maintains K+ homoeostasis in the salttolerant cultivars under salt stress, and the SKC1 gene encodes a member of HKT-type transporters. Fine-mapping of the Saltol QTL and the development of indel markers based on candidate genes in this region is being pursued at IRRI. Marker-assisted backcrossing (MABC) has been used to transfer the Saltol QTL into popular varieties such as IR64, BR28, and BR11. A diverse set of germplasm including the NERICAs (New Rice for Africa), collection from Cassamance regions in Senegal and Gambia are being characterized for physiological traits underlying salinity tolerance to identify new sources of tolerance for use as donors in breeding and identification of novel QTLs. Besides, MABC is currently being used to introgress Saltol into few varieties popular in West Africa. The long term goal is to identify and combine genes/QTLs controlling different physiological mechanisms to achieve a higher level of salt tolerance in high yielding West African rice varieties. 4 Nericas-L9, -L23, -L24 & L27: tolerant & different genes from Saltol NL25 Genotype Mean salt injury at EC 12 dSm-1 NERICA-L9 NERICA-L23 3 3 NL23 NL34 NL28 NL27 NL30 NL13 NL29 NL20NL31 NL21 NL15 NL24 NL22 NL16 NL14 NL7 NL8 NL9 NL4 NL1 NL2 NL33 NL32 NL5 NL3 NERICA_L24 5 NERICA_L27 FL 478(Tolerant check) IR 29 (Sensitive check) 3 NL10 NL26 NL12 NL18 NL11 NL19 5 FL478 NL17 NL6 9 IR64 CG14 0 Objectives 0.2 Allelic diversity for Saltol QTL with 34 lowland NERICAs To use MABC approach to introgress Saltol into several African varieties that will subsequently be deployed for commercial cultivation Identify new source of suitable donors of major QTLs/genes other than Saltol Identify new salt tolerance breeding lines using the participatory varietal selection (PVS) approach Comparison of 15 major QTL/genes published as tolerant to salinity other than Saltol with identified tolerant germplasm qNa+/ Trait qNa+/ qDWSchrm Saltol qNa+S K+SV- qNa+L based K+ SV 9/ qSTRQSst2 QPh4 chrm6 QSds8 10 (control) V-1-1 2.2 V-3.1 QTL 8-1 9 Materials and methods Evaluation of physiological traits for salt tolerance RM 10843 Name Jarmissa Camaro A total of 348 germplasm comprising of NERICAs, O. glaberrimas and O. sativas were evaluated for salinity tolerance under controlled conditions. Salt stress was imposed 5 d after germination by adding NaCl to an EC of 12 dS m-1 in Yoshida nutrient solution. IR29 (sensitive) and FL 478 (highly tolerant) were used as checks. Entries were scored based on visual symptoms using IRRI’s Standard evaluation system (SES) scores. RM 572 RM 208 RM 145 RM 563 RM 551 RM 273 RM 336 RM 281 RM 38 RM 215 n.a Codeh mano n.a Madina koyo Gold coast fingo RAM121 TOG6224 TOG7230 NERICA L9 MABC to introgress Saltol in African Varieties African variety/FL478 // African variety RM 311 n.a n.a BC1F1 NERICA L23 n..a. Selected BC1F1 NERICA L24 n.a BC1F1 / African variety BC2F1 Selected BC2F1 BC2F1 / African variety BC3F1 Selected BC3F1 BC3F1 X Recipient varieties: – IR64, Sahel 108, Sahel 201, BG90-2, Kogoni 91-1, Bouake189, Rassi, NERICA L-19, ITA212 Donors: – Pokkali, FL478 (salt tolerance) • Routine use of MAB to transfer Saltol : using flanking SSRs: 11-13 Mb on chrom1 – Foreground (36 markers) – & recombinant markers used for selection after each cross – SSRs for background selection (179 markers) NERICA L27 HASAWI Pokkali FL478 n.a n.a n.a Blue = tolerance in germplasm is different from the already identified & published QTLs/gene. Yellow = alleles is similar to the already identified & published QTLs/gene n.a= not clear or poor amplification PVS for Salt Tolerant in 3 countries Gambia Mali Senegal 1 WAR 1 WAS 201-B-2 IR 66946-3R-178-1-1 2 IR 1907-3R-2-1-2-B IR 59418-7-B-27-3 IR 72593-B-3-2-1-2-B 3 IR 63275-B-1-1-1-3-3-2 WAS 202-B-1 4 5 WAS 73-B-B-84-2 BC3F2 Results Current status of MABC Cross combination Mapping popn used Rassi x FL478 BC2F1 Lines with Seeds produced Saltol identified with Saltol background 7 out of 29 BC2F2 & BC3F1 Rassi x FL478 BC3F2 5 out of 23 BC3F3 & BC4F2 Sahel 108 x POKKALI BC2F1 6 out of 37 BC2F2 & BC3F1 To genotype for Saltol (10 markers used; 3 each at the upper & lower flanking region 4 Saltol specific markers Background genotyping using 151 polymorphic markers WAS 197-B-8-2 IR 1829-3R-89-1-1 CT6163-8-9-5-2-M-134M IR 76393-2B-7-1-13-1 WAS 73-B-B-231-4 & WAS 236 After 2 phases of PVS 4-6 promising lines have been selected by farmers Selected lines being validated in final round of PVS Main selection criteria :Earliness, plant height, tillering ability, panicle length and weight, resitance to birds, Conclusions New Germplasm Tolerant to Salinity 8 Salt tolerant landraces from 188 identified from hydroponic and field screening Molecular analysis show 5 of 8 appear to have different genes from Saltol 4 out of 104 accession of O. glaberrima; (TOG7230, RAM26, RAM121) tolerant 6 IR 59418-7B-21-3 Variety Tolerance to Salinity Country of Origin Jarmissa Many Fingo Camaro Condeh Mano Madina Koyo 5 3 5 5 5 Gambia Gambia Gambia Gambia Senegal Gold Coast Fingo 3 Gambia Nafisatu FL 478 (Tolerant Ck) 5 4 Gambia IRRI IR 29 (Sensitive Ck) 9 IRRI New source of tolerance have been identified in both O. sativa & O. glaberrima Saltol gene/QTL have been introgressed into the background of 3 African mega varieties More than 5 breeding & mapping populations at different stages have been developed 10 salt tolerant lines/varieties identified by NARs after 2 season of PVS