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Molecular mapping of genes for resistance to the bean pod weevil
                          (Apion godmani Wagner) in common bean
                                                        M.W. Blair1*, C. Muñoz1, R. Garza2, C. Cardona1
                                                   1/ CIAT - International Center for Tropical Agriculture, A. A. 6713, Cali, Colombia,
                                                          2/ INIFAP, A.P. No. 10, Chapingo, Estado de Mexico 56230, Mexico


 Justification                                                     Figures and Tables                                                                                                                                                                     Results and Discussion
The bean pod weevil (Apion godmani Wagner) is                                                                                                                                                                                                                After cloning and conversion to sequence tagged
                                                                   Table 1. Percentage seed damage by Apion godmani in recombinant
a serious insect pest of common beans                              inbred lines (RILs) and parents.
                                                                                                                                                                                                                                                         site (STS) markers, one of the markers was
(Phaseolus vulgaris L.) grown in Mexico and                              Genotype                                                         1994                     1995                           1996                        1997
                                                                                                                                                                                                                                                         polymorphic as a SCAR marker while four detected
Central America that is best controlled by host-                         Mean 10 most resistant RILs                                    5.2b ± 0.57            6.7b ± 0.53                  11.6c ± 0.84                12.5b ± 0.68
                                                                                                                                                                                                                                                         polymorphism as CAPS markers upon digestion with
                                                                                                                                                                                                                                                         restriction enzymes (Figure 2).
plant resistance available in Durango or Jalisco                         Mean 10 most suscep tible RILS s                           38.0a ± 0.96           34.2a ± 1.08                 42.0b ± 1.56                    55.6a ± 1.60

genotypes such as J-117. Given unreliable                                ‘Jamapa’ (susc eptible parent)                             44.8a ± 2.84           35.6a ± 3.19                     56.3a ± 2.80                61.2a ± 2.78                        Nine molecular markers for Apion resistance were
infestation by the insect, the use of marker-                            J 117 (resistant parent)                                       1.1b ± 0.45            4.0b ± 0.74                   9.1c ± 0.82                 6.6b ± 1.31                     mapped to loci on chromosomes 2, 3, 4 and 6 (linkage
assisted selection is desirable.                                                                                                                                                                                                                         groups b01, b08, b07and b11,respectively) based on
                                                                                                                                                                                                                                                         genetic analysis the Jamapa x J-117 population and
                                                                                                                                                                                                                                                         two reference populations (DOR364 x G19833 and
 Introduction                                                                                                                                                                                                                                            BAT93 x JaloEEP558) for which chromosome and
                                                                                                                                                                                                                                                         linkage group designations are known (Figure 3).
Apion godmani (Wagner) is a destructive insect pest of
common beans grown in highland regions of Mexico                                                                                                                                                                                                          The loci on linkage groups b01 and b11 loci may be
(states of Puebla, Durango, and Chiapas) and Central                                                                                                                                                                                                     associated with the Agr and Agm genes while
America (especially Guatemala, Honduras, El Salvador                                                                                                                                                                                                     additional resistance genes may exist on b07 and b08.
and Northern Nicaragua). Insect life cycle alternates
                                                                                                                                                                                                                                                           These are among the first specific markers
between forest soils / crop refuse where the adults over-
                                                                                                                                                                                                                                                         developed for tagging insect resistance in common
winter and fields of common bean where they
                                                                                                                                                                                                                                                         bean and are expected to be useful for evaluating the
reproduce. Bean pod weevils generally appear in bean
                                                                                                                                                                                                                                                         mechanism of resistance to A. godmani and for
crops during flowering and consume small amounts of                      Figure 1. Population distribution for 50 recombinant inbred lines
                                                                         from the Jamapa x J-117 population over four seasons.                                                                                                                           marker assisted selection of resistance in breeding
young leaves and flowers before beginning to
                                                                                                                                                                                                                                                         programs for Mexico and Central America.
reproduce. Damage occurs when the bean pod weevil
females lay their eggs in the mesocarp of 1- 4 cm long                                                                              b01 markers (I)                                                                                                                                                                             b01 markers (II)
developing common bean pods and the resulting larvae                                                                                                                                                                                                                                                                                W9-1300S-
                                                                                 OPF-10                      F10-500S                                              OPU-01                                       U1-1400R
burrow into the immature seeds inside causing yield                      M   1   2       3       4       5   6   7      8       m         bp           M       1       2    3       4         m         5       6         7         8        bp          M     1
                                                                                                                                                                                                                                                                   OPW-09
                                                                                                                                                                                                                                                                    2       3       4       m   5
                                                                                                                                                                                                                                                                                                    W9-1300S
                                                                                                                                                                                                                                                                                                        6       7       8       9
                                                                                                                                                                                                                                                                                                                                      AluI
                                                                                                                                                                                                                                                                                                                                     10    11      12   M         bp         1
                                                                                                                                                                                                                                                                                                                                                                                     OPZ-04
                                                                                                                                                                                                                                                                                                                                                                                      2       3       4       M           5
                                                                                                                                                                                                                                                                                                                                                                                                                                   Z4-800R
                                                                                                                                                                                                                                                                                                                                                                                                                                    6    7    8   bp

loss, reduced seed quality and lower seed viability.
Yield loss caused by the insect is variable depending on
the climatic conditions of the previouis year but can be                                                                                  1400
                                                                                                                                                                                                                                            1237

up to 90%.                                                                                                                                460                                                                                                                                                                                                                          800
                                                                                                                                                                                                                                                                                                                                                            500                                                                                   650


Objectives                                                                                                                                                                                                                                                                                                                                                  270




Given that major genes are thought to be involved in
resistance, our objective in this study was to develop
                                                                                                                                         b08 markers                                                                                                                                                                                    b11 markers
STS markers for Apion resistance using bulked
                                                                                                                                                                                                                          C1-800S-                                                                                                                                                                                        B1-1400R-
segregant analysis of recombinant inbred lines derived                                   OPK -16                     SK16                                              OPC-01                      C1–800S                  RsaI                                            OPW-06                      SW6-800R                                                  OPB-01                      B1-1400R                       TaqI
                                                                   bp        M       1       2       3       4   5          7       M            bp        M       1   2        3       4     5     6       7       8     9    10       M          bp    bp    M        1       2       3   4       5       6       7       8       M      bp           M    1         2     3   4        5       6       7       8   9       10    11   12   M   bp
from the cross Jamapa x J-117, where ‘J-117’ is a
resistant landrace source and ‘Jamapa’ is a susceptible
black-seeded cultivar from Mexico. In this study we                                                                                                                                                                                                                                                                                       1400


developed a total of nine SCAR and CAPS markers,                   890
                                                                                                                                                                                                                                                         732                                                                                                                                                                                      780

confirmed their linkage to Apion resistance genes and                                                                                            682                                                                                               682
                                                                                                                                                                                                                                                   550
                                                                                                                                                                                                                                                                                                                                                 521
                                                                                                                                                                                                                                                                                                                                                                                                                                                  600



mapped them to their genomic position based on
integrated genetic maps of common beans.
                                                                                                                                                                                                                                                   132




Materials and Methods
  Plant Material:    Recombinant inbred line population (50                                                                                                                                                                                                        Figure 2. (above). SCAR and CAPS markers developed from
genotypes) was developed from a cross of Jamapa (susceptible)                                                                                                                                                                                                      polymorphic RAPD bands found in bulked segregant analysis of
and J-117 (resistant) through single seed descent to the F5                                                                                                                                                                                                        Apion godmani resistant and susceptible parents and individuals
generation and evaluated for Apion resistance over four                                                                                                                                                                                                            of the Jamapa x J-117 population. SCAR.
consecutive seasons (1994 – 1997) at the Santa Lucía de Prías
Experiment Station (INIFAP) near Texcoco. Resistance was                                                                                                                                                                                                           Figure 3. (left). Linkage relationships of Apion resistance RAPD,
measured as damage level on a random sample of 30 pods per                                                                                                                                                                                                         SCAR and CAPS markers in recombinant inbred line populations
                                                                                                                                                                                                                                                                   Jamapa x J-117 (JJ), DOR364 x G19833 (DG) and BAT93 x Jalo
plot taken at crop maturity (Table 1).
                                                                                                                                                                                                                                                                   EEP 558 (BJ) for linkage groups b01, b07, b08 and b11.
  Bulked Segregant Analysis: DNAs from the four most resistant
and four most susceptible recombinant inbred lines were
combined into their respective bulks. A total of 131 decamer                                                                                                                                                                                                        References
primers were used to discover random amplified (RAPD)
polymorphisms among the parents that were also observed in                                                                                                                                                                                                         Blair MW, Muñoz C, Garza R, Cardona C (2006) Molecular
                                                                                                                                                                                                                                                                   mapping of genes for resistance to the bean pod weevil (Apion
the resistant and susceptible bulks. Both repulsion and coupling                                                                                                                                                                                                   godmani Wagner) in common bean. Theor Appl Genet 112:
markers were considered as long as the presence or absence of                                                                                                                                                                                                      913–923.
the bands was consistent between the bulks and the parents.

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Poster62: Molecular mapping of genes for resistance to the bean pod weevil (Apion godmani Wagner) in common bean

  • 1. Molecular mapping of genes for resistance to the bean pod weevil (Apion godmani Wagner) in common bean M.W. Blair1*, C. Muñoz1, R. Garza2, C. Cardona1 1/ CIAT - International Center for Tropical Agriculture, A. A. 6713, Cali, Colombia, 2/ INIFAP, A.P. No. 10, Chapingo, Estado de Mexico 56230, Mexico Justification Figures and Tables Results and Discussion The bean pod weevil (Apion godmani Wagner) is After cloning and conversion to sequence tagged Table 1. Percentage seed damage by Apion godmani in recombinant a serious insect pest of common beans inbred lines (RILs) and parents. site (STS) markers, one of the markers was (Phaseolus vulgaris L.) grown in Mexico and Genotype 1994 1995 1996 1997 polymorphic as a SCAR marker while four detected Central America that is best controlled by host- Mean 10 most resistant RILs 5.2b ± 0.57 6.7b ± 0.53 11.6c ± 0.84 12.5b ± 0.68 polymorphism as CAPS markers upon digestion with restriction enzymes (Figure 2). plant resistance available in Durango or Jalisco Mean 10 most suscep tible RILS s 38.0a ± 0.96 34.2a ± 1.08 42.0b ± 1.56 55.6a ± 1.60 genotypes such as J-117. Given unreliable ‘Jamapa’ (susc eptible parent) 44.8a ± 2.84 35.6a ± 3.19 56.3a ± 2.80 61.2a ± 2.78 Nine molecular markers for Apion resistance were infestation by the insect, the use of marker- J 117 (resistant parent) 1.1b ± 0.45 4.0b ± 0.74 9.1c ± 0.82 6.6b ± 1.31 mapped to loci on chromosomes 2, 3, 4 and 6 (linkage assisted selection is desirable. groups b01, b08, b07and b11,respectively) based on genetic analysis the Jamapa x J-117 population and two reference populations (DOR364 x G19833 and Introduction BAT93 x JaloEEP558) for which chromosome and linkage group designations are known (Figure 3). Apion godmani (Wagner) is a destructive insect pest of common beans grown in highland regions of Mexico The loci on linkage groups b01 and b11 loci may be (states of Puebla, Durango, and Chiapas) and Central associated with the Agr and Agm genes while America (especially Guatemala, Honduras, El Salvador additional resistance genes may exist on b07 and b08. and Northern Nicaragua). Insect life cycle alternates These are among the first specific markers between forest soils / crop refuse where the adults over- developed for tagging insect resistance in common winter and fields of common bean where they bean and are expected to be useful for evaluating the reproduce. Bean pod weevils generally appear in bean mechanism of resistance to A. godmani and for crops during flowering and consume small amounts of Figure 1. Population distribution for 50 recombinant inbred lines from the Jamapa x J-117 population over four seasons. marker assisted selection of resistance in breeding young leaves and flowers before beginning to programs for Mexico and Central America. reproduce. Damage occurs when the bean pod weevil females lay their eggs in the mesocarp of 1- 4 cm long b01 markers (I) b01 markers (II) developing common bean pods and the resulting larvae W9-1300S- OPF-10 F10-500S OPU-01 U1-1400R burrow into the immature seeds inside causing yield M 1 2 3 4 5 6 7 8 m bp M 1 2 3 4 m 5 6 7 8 bp M 1 OPW-09 2 3 4 m 5 W9-1300S 6 7 8 9 AluI 10 11 12 M bp 1 OPZ-04 2 3 4 M 5 Z4-800R 6 7 8 bp loss, reduced seed quality and lower seed viability. Yield loss caused by the insect is variable depending on the climatic conditions of the previouis year but can be 1400 1237 up to 90%. 460 800 500 650 Objectives 270 Given that major genes are thought to be involved in resistance, our objective in this study was to develop b08 markers b11 markers STS markers for Apion resistance using bulked C1-800S- B1-1400R- segregant analysis of recombinant inbred lines derived OPK -16 SK16 OPC-01 C1–800S RsaI OPW-06 SW6-800R OPB-01 B1-1400R TaqI bp M 1 2 3 4 5 7 M bp M 1 2 3 4 5 6 7 8 9 10 M bp bp M 1 2 3 4 5 6 7 8 M bp M 1 2 3 4 5 6 7 8 9 10 11 12 M bp from the cross Jamapa x J-117, where ‘J-117’ is a resistant landrace source and ‘Jamapa’ is a susceptible black-seeded cultivar from Mexico. In this study we 1400 developed a total of nine SCAR and CAPS markers, 890 732 780 confirmed their linkage to Apion resistance genes and 682 682 550 521 600 mapped them to their genomic position based on integrated genetic maps of common beans. 132 Materials and Methods Plant Material: Recombinant inbred line population (50 Figure 2. (above). SCAR and CAPS markers developed from genotypes) was developed from a cross of Jamapa (susceptible) polymorphic RAPD bands found in bulked segregant analysis of and J-117 (resistant) through single seed descent to the F5 Apion godmani resistant and susceptible parents and individuals generation and evaluated for Apion resistance over four of the Jamapa x J-117 population. SCAR. consecutive seasons (1994 – 1997) at the Santa Lucía de Prías Experiment Station (INIFAP) near Texcoco. Resistance was Figure 3. (left). Linkage relationships of Apion resistance RAPD, measured as damage level on a random sample of 30 pods per SCAR and CAPS markers in recombinant inbred line populations Jamapa x J-117 (JJ), DOR364 x G19833 (DG) and BAT93 x Jalo plot taken at crop maturity (Table 1). EEP 558 (BJ) for linkage groups b01, b07, b08 and b11. Bulked Segregant Analysis: DNAs from the four most resistant and four most susceptible recombinant inbred lines were combined into their respective bulks. A total of 131 decamer References primers were used to discover random amplified (RAPD) polymorphisms among the parents that were also observed in Blair MW, Muñoz C, Garza R, Cardona C (2006) Molecular mapping of genes for resistance to the bean pod weevil (Apion the resistant and susceptible bulks. Both repulsion and coupling godmani Wagner) in common bean. Theor Appl Genet 112: markers were considered as long as the presence or absence of 913–923. the bands was consistent between the bulks and the parents.