Breeding with the aid of Molecular Markers

Breeding with the aid of Molecular Markers

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  DEVELOPMENT OF MULTIPLEDISEASE RESISTANT BREEDING PARENTS WITH THE AID DISEASE RESISTANT DEVELOPMENT OF MULTIPLE OF MOLECULAR MARKERS BREEDING PARENTS WITHA. Buruchara, A. MOLECULAR MARKERS C.M. Mukankusi, R. THE AID OF Male, S. Sebuliba C.M. Mukankusi, R. A. Buruchara, A. Male, S. Sebuliba currently being utilized for Multiple Table 1. Molecular markers Background: disease resistance breeding in PABRA Table 1. Molecular markers currently being utilized for Multiple Background: disease resistance Markers Trait breeding in PABRA Source Common bean (Phaseolus vulgaris L.) plays an essential role in sustaining live- lihoods of smallholder farmers and their families in Africa. At farm level, beans Trait Markers 709 Source 54 (Mahuku et al., 2004) are attacked by aPhaseolus vulgaris L.) plays an essentialviral diseases leading to Common bean ( combination of fungal, bacterial and role in sustaining live- ALS OPE4 MEX poor yields (Fig. 1). The farmers and their families in Africa. At farm(MDR)beans lihoods of smallholder deployment of multiple disease resistant level, varie- ALS PF9250 OPE4709 G10474 and G10909 MEX 54 (Mahuku et al., 2004) ties is probably the cheapest method of managing and problem. The use of to are attacked by a combination of fungal, bacterial this viral diseases leading Pythium root rot PYAA19 RWR719 (Buruchara et al) poor yields (Fig. 1). The deploymentcould speed up the development ofvarie- MDR parents in breeding programs of multiple disease resistant (MDR) MDR PF9250 G10474 and G10909 ties is probably the cheapest method of managing this problem. The use of PYB08 RWR719 719 RWR (Buruchara et al) varieties. Marker Assisted Selection (MAS) o ers advantages within NARS Pythium root rot PYAA19 MDR parents in breeding programs could speed up the development of MDR breeding programs in targeting challenging breeding objectives requiring com- Anthracnose SAB-3 PYB08 RWR 719 (Vallejo and Kelly, 2001) G2333 varieties. Marker Assisted Selection (MAS) o ers advantages within NARS plex breeding schemes. breeding programs in targeting challenging breeding objectives requiring com- Anthracnose SAS-13 SAB-3 G2333 (Vallejo and Kelly,1998) G2333 (Young et al., 2001) plex breeding schemes. SBB-14 G2333 (Young et al., 1998) and Kelly, 2001) G2333 , AB 136 (Awale However, the national programs lack the facilities and capacity to use these SAS-13 techniques. Taking advantage of lackbiotechnology facility at Kawanda in However, the national programs the the facilities and capacity to use these SBB-14 SH-18 G2333 , AB 136 (Awale and Kelly, 2001) G2333 (Awale and Kelly, 2001) Uganda, andTaking advantage of the biotechnologyutilizing molecular markers techniques. the available markers, CIAT-PABRA is facility at Kawanda in BCMV SH-18 ROC11 G2333 (Awale and Kelly, 2001) Various Uganda, and the available markers, CIAT-PABRA is (Angular leaf Spot - ALS - linked to resistance genes to three fungal diseases utilizing molecular markers Pythium root rots and Anthracnose)fungal diseases (Angular leaf Spot - ALSMo- linked to resistance genes to three and one viral disease (Bean Common - BCMV BCMNV ROC11 SW13 Various Various saic Virus root rots and its necrotic strain one viral disease (Bean CommonVirus Pythium (BCMV) and Anthracnose) and Bean Common Mosaic Necrotic Mo- BCMNV CBB SW13 820 SAP Various et al., 2000, Deidre et al., 2007 Miklas (BCMNV) (Table 1) .and its necroticpyramid the resistance genes Necrotic Virus saic Virus (BCMV) The aim is to strain Bean Common Mosaic in common CBB SAP820 Miklas et al., 2000, Deidre et al., 2007 background in an e . ort of developing MDR parents that can bein common (BCMNV) (Table 1) The aim is to pyramid the resistance genes utilized by NARS breeding programs. developing MDR parents that can be utilized by background in an e ort of Conclusions and Way Forw ard NARS breeding programs. Conclusions and Way Forward • Crosses between Pythium/ ALS resistance plants and Anth/ BCMV/ • Crosses between have been conducted plants and Anth/ BCMV/ BCMNV plants Pythium/ ALS resistance BCMNV plants have been conducted • MDR parents developed will be made available to NARS for utilization in • MDR parents developed will be made available to NARS for utilization in own breeding programs. own breeding programs. • We envisage a mechanism where the capacity of NARS in using MAS will • We envisage a mechanism where the capacity of NARS in using MAS contact be enhanced through this process with back up from continuous will be enhanced through this facility in Kawanda. from continuous contact with the biotechnology process with back up with the biotechnology facility in Kawanda. • Some partners have been trained (MAS and Disease Phenotyping) • Some partners have been trained (MAS and Disease Phenotyping) Fig 1. Some of the most common diseases of beans in Africa Fig 1. Some of the most common diseases of beans in Africa Marker Assisted Selection (MAS) Marker Assisted Selection (MAS) Figure 2: Part analysis of the plants from the cross G2333 X MCM1015 using the ROC 11 Figure 2: Part analysis of the plants recessive bc-3G2333 Sample 5 is positive control 11 marker to detect presence of the from the cross gene. X MCM1015 using the ROC variety marker to detect presence of the recessive bc-3 gene. Sample 5 is positive control variety MCM 5001, Sample 1 is negative control RAB 487. Absence of a band indicates presence of MCM 5001, Sample 1 is negativeband indicates absence of the gene. indicatesbp molecular the gene while presence of a control RAB 487. Absence of a band L is 100 presence of •• MAS refers to the use of DNA markers that are tightly-linked to target MAS refers to the use of DNA markers that are tightly-linked to target the gene while presence of a band indicates absence of the gene. L is 100 bp molecular marker. marker. loci as aasubstitute for or to assist phenotypicscreening loci as substitute for or to assist phenotypic screening •• MAS is the most promising application of DNA markers for cultivar de- MAS is the most promising application of DNA markers for cultivar de- velopment velopment Materials and Methods: Materials and Methods: •• Single crosses between sources of resistance, aim of screening up to Single crosses between sources resistance, aim of screening up to 1500 FF2plants per cross 1500 2 plants per cross •• MCM5001and MCM 1015 as sources of IIand bc- 3 genes for MCM5001and MCM sources of and bc- 3 genes for BCMV/ BCMNV BCMV/ BCMNV •• G2333—Co-4,,Co-5 and Co-7 for resistance to anthracnose, G2333— Co-4 Co-5 and resistance to anthracnose, •• RWR719 and MLB-49-89A-Pythiumroot rot, RWR719 and MLB-49-89A-Pythium root rot, •• MEX54- phg for resistance MEX54- phg for resistance to ALS. •• DNA extracted from leaves of 2 week old F2 plants DNA extracted from leaves of 2 plants •• Two mm discs used as templates in PCR reactions using speci cc mo- Two mm discs used as templates in PCR reactions using speci mo- lecular markers. lecular markers. Figure 3: 3: Breeding schemedevelop Multiple Disease resistance parents to Angular leaf spot, spot, Anthracnose, Figure Breeding scheme to to develop Multiple Disease resistance parents to Angular leaf Anthracnose, Pythium root rot rot and BCMV/BCMNV Pythium root and BCMV/BCMNV •• Plants positive for 2-3 gene combination selected and double crosses Plants positive for 2-3 gene combination selected and double crosses conducted. conducted. References References Awale, H.E., and J.D. Kelly. 2001. Development of SCAR markers linked to Co-42 gene in common bean. Ann. Awale, H.E., and J.D. Kelly. 2001. Development of SCAR markers linked to Co-42 gene in common bean. Ann. Results Rept. Bean Improv. Coop. 44:119-120. Results Rept. Bean Improv. Coop. 44:119-120. Buruchara et al. Buruchara et al. Mahuku, G., Montoya, C., Henríquez, M.A., Jara, C., T eran, H., and Beebe, S. 2004. Inheritance and charac- To date 10 out of 340 plants have been found to possess four gene combina- Mahuku, G., Montoya, C., Henríquez, M.A., Jara, C., T in the and Beebe, S. 2004. Inheritance and charac- terization of the angular leaf spot resistance gene H., common bean accession, G 10474 and eran, To date 10 out of 340 plants have been found to possess four gene combina- identi cation of anangular leaf spot resistance gene in the common bean accession, G 10474 and terization of the AFLP marker linkedto the resistance gene. Crop Science 44: 1817-1824. tion; “ Co-4”, Co-5, I and bc3” resistance genes for anthracnose and BCMV/ Miklas, P.N., J.R. Smith, R.of an AFLP Grafton,linkedto the resistance D. P. Coyne.Science 44: 1817-1824. identi cation Riley, K.F. marker S.P Singh, G. Jung, and gene. Crop 2000. Marker-assisted tion; “ Co-4”, Co-5, I and bc3” resistance genes for anthracnose and BCMV/ Miklas,breeding for pyramided resistance to common bacterial blight in D. P. Coyne. 2000. Marker-assisted P.N., J.R. Smith, R. Riley, K.F. Grafton, S.P Singh, G. Jung, and common bean. Annu. Rep. Bean BCMNV (Fig 2 and 3). BCMNV (Fig 2 and 3). Improv. Coop. 43:39-40 resistance to common bacterial blight in common bean. Annu. Rep. Bean breeding for pyramided Improv. Coop. 43:39-40 Vallejo, V., and J.D. Kelly. 2001. Development of a SCAR marker linked to Co-5 gene in common bean. Ann. Sixty two out of 103 F2 plants have been found to posses both the Pythium and Vallejo,Rept. Bean Improv.2001. Development of a SCAR marker linked to Co-5 gene in common bean. Ann. V., and J.D. Kelly. Coop. 44:121-122. Sixty two out of 103 F2 plants have been found to posses both the Pythium and Young, R.A.,Rept. Bean Improv. Coop. 44:121-122. Angular Leaf spot resistance genes (Fig 3). M. Melotto, R.O., Nodari and J.D. Kelly. 1998. Marker assisted dissection of the oligogenic Angular Leaf spot resistance genes (Fig 3). Young, R.A., M. Melotto, R.O., Nodaricommon bean cultivar G2333. assisted dissection96:87-94 anthracnose resistance in the and J.D. Kelly. 1998. Marker Theor. Appl. Genet. of the oligogenic anthracnose resistance in the common bean cultivar G2333. Theor. Appl. Genet. 96:87-94 For more information: [email protected] For more information: [email protected]