INTRODUCTION The common bean (Phaseolus vulgaris L.) is one of the most important legumes for direct human consumption (Lin et al. 2008). Anthracnose, caused by Colletotrichum lindemuthianum is the most widespread, recurrent and devastating disease of the common bean in Latin America and Africa (Pastor-Corrales and Tu 1989). The genetic mapping is carried out using segregating populations. Obtaining segregating populations for genetic mapping have been standard practice in the common bean research (Gepts et al. 1993; Blair et al. 2006). These populations have led to detailed studies and promising to greater efficiency in the genetic breeding of the crop worldwide, but have as limitation, the small size (Sanglard et al. 2013). Thus, the objective of this research was to phenotype the recombinant inbred lines (RIL’s) population from AND 277 × Rudá cross with the 65 and 73 races of Colletotrichum lindemuthianum

Castro, S. A. L.

Gonçalves-Vidigal, M. C.

Lacanallo, G. F.

Martins, V. S. R.

Pelembe, A. E.

Souza, T. L. P. O.

Vidigal Filho, P. S.

English

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University of Nebraska - Lincoln 
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Publications from USDA-ARS / UNL Faculty U.S. Department of Agriculture: Agricultural Research Service, Lincoln, Nebraska 
3-2017 
CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE 
COMMON BEAN TO RACES 65 AND 73 OF Colletotrichum 
lindemuthianum 
A. E. Pelembe 
Ministério da Agricultura e Segurança Alimentar 
P. S. Vidigal Filho 
Universidade Estadual de Maringá 
M. C. Gonçalves-Vidigal 
Universidade Estadual de Maringá, mcgvidigal@uem.br 
G. F. Lacanallo 
Universidade Estadual de Maringá 
V. S. R. Martins 
Universidade Estadual de Maringá 
See next page for additional authors 
Follow this and additional works at: https://digitalcommons.unl.edu/usdaarsfacpub 
Pelembe, A. E.; Vidigal Filho, P. S.; Gonçalves-Vidigal, M. C.; Lacanallo, G. F.; Martins, V. S. R.; Castro, S. A. 
L.; and Souza, T. L. P. O., "CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE COMMON BEAN 
TO RACES 65 AND 73 OF Colletotrichum lindemuthianum" (2017). Publications from USDA-ARS / UNL 
Faculty. 1707. 
https://digitalcommons.unl.edu/usdaarsfacpub/1707 
This Article is brought to you for free and open access by the U.S. Department of Agriculture: Agricultural Research 
Service, Lincoln, Nebraska at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in 
Publications from USDA-ARS / UNL Faculty by an authorized administrator of DigitalCommons@University of 
Nebraska - Lincoln. 
Authors 
A. E. Pelembe, P. S. Vidigal Filho, M. C. Gonçalves-Vidigal, G. F. Lacanallo, V. S. R. Martins, S. A. L. Castro, 
and T. L. P. O. Souza 
This article is available at DigitalCommons@University of Nebraska - Lincoln: https://digitalcommons.unl.edu/
usdaarsfacpub/1707 
5 
CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE COMMON BEAN 
TO RACES 65 AND 73 OF Colletotrichum lindemuthianum 
 
Pelembe AE1, Vidigal Filho PS2, Gonçalves-Vidigal MC2*, Lacanallo GF2, Martins VSR2, 
Castro SAL2, Souza TLPO3  
 
1Ministério da Agricultura e Segurança Alimentar, IAM, Maputo, Mozambique. 
 2 Dep. Agronomia, Universidade Estadual de Maringá, Maringá, PR, Brazil,  
3Embrapa Arroz e Feijão, Empresa Brasileira de Pesquisa Agropecuária (Embrapa) 
*E-mail: mcgvidigal@uem.br 
 
 
INTRODUCTION 
The common bean (Phaseolus vulgaris L.) is one of the most important legumes for 
direct human consumption (Lin et al. 2008). Anthracnose, caused by Colletotrichum 
lindemuthianum is the most widespread, recurrent and devastating disease of the common bean 
in Latin America and Africa (Pastor-Corrales and Tu 1989). The genetic mapping is carried out 
using segregating populations. Obtaining segregating populations for genetic mapping have been 
standard practice in the common bean research (Gepts et al. 1993; Blair et al. 2006). These 
populations have led to detailed studies and promising to greater efficiency in the genetic 
breeding of the crop worldwide, but have as limitation, the small size (Sanglard et al. 2013). 
Thus, the objective of this research was to phenotype the recombinant inbred lines (RIL’s) 
population from AND 277 × Rudá cross with the 65 and 73 races of Colletotrichum 
lindemuthianum. 
 
MATERIAL AND METHODS 
This work was conducted at the Núcleo de Pesquisa Aplicada a Agricultura (Nupagri) of 
the Universidade Estadual de Maringá (UEM), Paraná state, Brazil. The genetic material 
consisted of 393 Recombinant Inbred Lines (RIL’s), and two parent (AND 277 and Rudá). Seed 
samples of the RILs population were provided by Dr. Thiago Lívio Pessoa Oliveira de Souza of 
the CNPAF/Embrapa. These seeds were sown in plastic trays containing peat base substrates. In 
each tray were sown 10 RIL’s to further inoculation with the races 65 and 73 of C. 
lindemuthianum, both obtained from the mycology collection of Nupagri. The inoculum 
preparation was performed according to the methodology proposed by Cárdenas et al. (1964). 
The counting of spores in each pathogen race was made with the aid of hemacytometer 
(Chamber of Neubauer- Preciss) and trinocular biological microscope, Motic® brand - mod 
BA210. After counting, the spore suspension was adjusted to a concentration of 1.2 x 106 spores 
mL-1. The visual evaluation of symptoms in each plant was performed 7 to 10 days after 
inoculation. Severity scale proposed by Pastor-Corrales et al. (1995) was considered, with score 
ranging from 1 to 9. Plants with scores from 1 to 3 were considered resistant, whereas those 
having scores from 4 to 9 were considered susceptible.  
 
  
6 
RESULTS AND DISCUSSION 
A total of 393 RIL’s from cross Rudá × AND 277 were evaluated and it was observed a 
segregation of 202 resistant lines and 191 susceptible lines to race 65 (2 = 0.307; p = 0.579) and 
203 resistant lines and 190 susceptible lines to the race 73 (2 = 0.430; p = 0.512), which set the 
monogenic ratio of 1:1. This results shows that the RIL’s co-segregated for both inoculated 
races, indicating that a same gene confer resistance two races 65 and 73 of C. lindemuthianum 
(Table 1).  
Table 1. Inheritance test in Recombinants Inbred Lines derived from the cross Rudá × AND 277 
inoculated with races 65 and 73 of C. lindemuthianum 
Races Score  Observed ratio 
 Expected ratio  
2 p value AND 277 Rudá  Ra Sb  R S  
Race 65 1 5  202 191  1 1  0.307 0.579 
Race 73 1 6  203 190  1 1  0.430 0.512 
a = Resistant; b = Susceptible 
A segregation fitted a 1:1 (R:S) ratio in 393 RIL's population for races 65 and 73,  
indicating that there was no distortion of segregation (changing of genetic structure) for this 
character, which highlights the potential of these lines in genetic mapping. It was noted that the 
393 RIL's showed similarity in the phenotypic behavior. Except one, all the lines resistant to the 
race 65 also were resistance to race 73, and susceptible lines to the race 65 were also susceptible 
to the race 73. This fact highlights the co-segregation of RIL's in relation to resistance to the both 
races (65 and 73). Genetic analysis revealed that the recombinant inbred lines co-segregated for 
65 and 73 races of Colletotrichum lindemuthianum, showing that the Co-14 gene present in AND 
277 confers resistance to both races.  
 
The reaction of RIL's to the races 65 and 73 of Colletotrichum lindemuthianum presented 
monogenic inheritance, being governed by a single gene. The recombinant inbred lines derived 
from the cross AND 277 × Rudá due to the large size, can be used in mapping genes that control 
quantitative and qualitative traits and molecular markers, hence they present the potential for the 
development of a consensus map to common bean. 
 
ACKNOWLEDGEMENTS 
This research was financially supported by CNPq and CAPES. This research was 
financially supported by the CNPq and Capes. AE Pelembe was supported by a scholarship from 
CNPq. PS Vidigal Filho and MC Gonçalves-Vidigal are grateful for grants from CNPq.  
 
REFERENCES 
Blair et al. (2006). Theor Appl Genet 112:1149-1163 
Cárdenas et al. (1964). Euphytica 13:178-186 
Gepts et al. (1993). Ann Rep Bean Improv Coop 36:24-38 
Lin et al. (2008). Food Chemistry 107:399–410 
Pastor-Corrales and Tu (1989). CIAT 77-104  
Pastor-Corrales et al. (1995). Plant Disease 79:63-67 
Sanglard et al. (2013). Ann Rep Bean Improv Coop 56:23-24. 
  


CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE COMMON BEAN TO RACES 65 AND 73 OF \u3ci\u3eColletotrichum lindemuthianum\u3c/i\u3e

DigitalCommons@University of Nebraska

University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Publications from USDA-ARS / UNL Faculty U.S. Department of Agriculture: Agricultural Research Service, Lincoln, Nebraska 3-2017 CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE COMMON BEAN TO RACES 65 AND 73 OF Colletotrichum lindemuthianum A. E. Pelembe Ministério da Agricultura e Segurança Alimentar P. S. Vidigal Filho Universidade Estadual de Maringá M. C. Gonçalves-Vidigal Universidade Estadual de Maringá, mcgvidigal@uem.br G. F. Lacanallo Universidade Estadual de Maringá V. S. R. Martins Universidade Estadual de Maringá See next page for additional authors Follow this and additional works at: https://digitalcommons.unl.edu/usdaarsfacpub Pelembe, A. E.; Vidigal Filho, P. S.; Gonçalves-Vidigal, M. C.; Lacanallo, G. F.; Martins, V. S. R.; Castro, S. A. L.; and Souza, T. L. P. O., "CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE COMMON BEAN TO RACES 65 AND 73 OF Colletotrichum lindemuthianum" (2017). Publications from USDA-ARS / UNL Faculty. 1707. https://digitalcommons.unl.edu/usdaarsfacpub/1707 This Article is brought to you for free and open access by the U.S. Department of Agriculture: Agricultural Research Service, Lincoln, Nebraska at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Publications from USDA-ARS / UNL Faculty by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Authors A. E. Pelembe, P. S. Vidigal Filho, M. C. Gonçalves-Vidigal, G. F. Lacanallo, V. S. R. Martins, S. A. L. Castro, and T. L. P. O. Souza This article is available at DigitalCommons@University of Nebraska - Lincoln: https://digitalcommons.unl.edu/usdaarsfacpub/1707 5 CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE COMMON BEAN TO RACES 65 AND 73 OF Colletotrichum lindemuthianum  Pelembe AE1, Vidigal Filho PS2, Gonçalves-Vidigal MC2*, Lacanallo GF2, Martins VSR2, Castro SAL2, Souza TLPO3   1Ministério da Agricultura e Segurança Alimentar, IAM, Maputo, Mozambique.  2 Dep. Agronomia, Universidade Estadual de Maringá, Maringá, PR, Brazil,  3Embrapa Arroz e Feijão, Empresa Brasileira de Pesquisa Agropecuária (Embrapa) *E-mail: mcgvidigal@uem.br   INTRODUCTION The common bean (Phaseolus vulgaris L.) is one of the most important legumes for direct human consumption (Lin et al. 2008). Anthracnose, caused by Colletotrichum lindemuthianum is the most widespread, recurrent and devastating disease of the common bean in Latin America and Africa (Pastor-Corrales and Tu 1989). The genetic mapping is carried out using segregating populations. Obtaining segregating populations for genetic mapping have been standard practice in the common bean research (Gepts et al. 1993; Blair et al. 2006). These populations have led to detailed studies and promising to greater efficiency in the genetic breeding of the crop worldwide, but have as limitation, the small size (Sanglard et al. 2013). Thus, the objective of this research was to phenotype the recombinant inbred lines (RIL’s) population from AND 277 × Rudá cross with the 65 and 73 races of Colletotrichum lindemuthianum.  MATERIAL AND METHODS This work was conducted at the Núcleo de Pesquisa Aplicada a Agricultura (Nupagri) of the Universidade Estadual de Maringá (UEM), Paraná state, Brazil. The genetic material consisted of 393 Recombinant Inbred Lines (RIL’s), and two parent (AND 277 and Rudá). Seed samples of the RILs population were provided by Dr. Thiago Lívio Pessoa Oliveira de Souza of the CNPAF/Embrapa. These seeds were sown in plastic trays containing peat base substrates. In each tray were sown 10 RIL’s to further inoculation with the races 65 and 73 of C. lindemuthianum, both obtained from the mycology collection of Nupagri. The inoculum preparation was performed according to the methodology proposed by Cárdenas et al. (1964). The counting of spores in each pathogen race was made with the aid of hemacytometer (Chamber of Neubauer- Preciss) and trinocular biological microscope, Motic® brand - mod BA210. After counting, the spore suspension was adjusted to a concentration of 1.2 x 106 spores mL-1. The visual evaluation of symptoms in each plant was performed 7 to 10 days after inoculation. Severity scale proposed by Pastor-Corrales et al. (1995) was considered, with score ranging from 1 to 9. Plants with scores from 1 to 3 were considered resistant, whereas those having scores from 4 to 9 were considered susceptible.     6 RESULTS AND DISCUSSION A total of 393 RIL’s from cross Rudá × AND 277 were evaluated and it was observed a segregation of 202 resistant lines and 191 susceptible lines to race 65 (2 = 0.307; p = 0.579) and 203 resistant lines and 190 susceptible lines to the race 73 (2 = 0.430; p = 0.512), which set the monogenic ratio of 1:1. This results shows that the RIL’s co-segregated for both inoculated races, indicating that a same gene confer resistance two races 65 and 73 of C. lindemuthianum (Table 1).  Table 1. Inheritance test in Recombinants Inbred Lines derived from the cross Rudá × AND 277 inoculated with races 65 and 73 of C. lindemuthianum Races Score  Observed ratio  Expected ratio  2 p value AND 277 Rudá  Ra Sb  R S  Race 65 1 5  202 191  1 1  0.307 0.579 Race 73 1 6  203 190  1 1  0.430 0.512 a = Resistant; b = Susceptible A segregation fitted a 1:1 (R:S) ratio in 393 RIL's population for races 65 and 73,  indicating that there was no distortion of segregation (changing of genetic structure) for this character, which highlights the potential of these lines in genetic mapping. It was noted that the 393 RIL's showed similarity in the phenotypic behavior. Except one, all the lines resistant to the race 65 also were resistance to race 73, and susceptible lines to the race 65 were also susceptible to the race 73. This fact highlights the co-segregation of RIL's in relation to resistance to the both races (65 and 73). Genetic analysis revealed that the recombinant inbred lines co-segregated for 65 and 73 races of Colletotrichum lindemuthianum, showing that the Co-14 gene present in AND 277 confers resistance to both races.   The reaction of RIL's to the races 65 and 73 of Colletotrichum lindemuthianum presented monogenic inheritance, being governed by a single gene. The recombinant inbred lines derived from the cross AND 277 × Rudá due to the large size, can be used in mapping genes that control quantitative and qualitative traits and molecular markers, hence they present the potential for the development of a consensus map to common bean.  ACKNOWLEDGEMENTS This research was financially supported by CNPq and CAPES. This research was financially supported by the CNPq and Capes. AE Pelembe was supported by a scholarship from CNPq. PS Vidigal Filho and MC Gonçalves-Vidigal are grateful for grants from CNPq.   REFERENCES Blair et al. (2006). Theor Appl Genet 112:1149-1163 Cárdenas et al. (1964). Euphytica 13:178-186 Gepts et al. (1993). Ann Rep Bean Improv Coop 36:24-38 Lin et al. (2008). Food Chemistry 107:399–410 Pastor-Corrales and Tu (1989). CIAT 77-104  Pastor-Corrales et al. (1995). Plant Disease 79:63-67 Sanglard et al. (2013). Ann Rep Bean Improv Coop 56:23-24.   

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CO-SEGREGATION OF RECOMBINANT INBRED LINES OF THE COMMON BEAN TO RACES 65 AND 73 OF \u3ci\u3eColletotrichum lindemuthianum\u3c/i\u3e

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