Poster for CIAT 2009 Knowledge Sharing Week

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.