GrainGenes

ITEMS FROM SPAIN

UNIVERSIDAD POLITECNICA DE MADRID
Departamento de Biotecnologia, E.T.S. Ing. Agronomos.- C. Universitaria, 28040, Madrid, Spain.

A. Delibes, I. Lopez-Braña, M.J. Montes, M. Gomez Colmenarejo, and C. Gonzalez-Belinchon.

CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
Serrano, 115, 28006, Madrid, Spain.

D. Romero and M.F. Andres.

UNIVERSITY OF LLEIDA AND
INSTITUTE FOR FOOD AND AGRICULTURAL RESEARCH AND TECHNOLOGY (UdL-IRTA)

Center of R&D, Rovira Roure 177, 25198 Lleida, Spain.

J.A. Martín-Sánchez, E. Sin, C. Martinez, and A. Michelena.

Differential induction of defense enzymes and the chromosomal location in wheat/Aegilops introgression lines of two Heterodera avenae resistance genes of Ae. ventricosa. [p. 164-165]

Two independent introductions of genetic resistance to the CCN from Ae. ventricosa to hexaploid wheat were compared. The Cre2 (from Ae. ventricosa AP-1) and Cre5 (from Ae. ventricosa #10) genes confer a high resistance level to the CCN-Spanish pathotype Ha71 in controlled conditions and under natural infestation. However, previous studies have shown the differential behavior of the Cre2 and Cre5 genes against other CCN pathotypes (Delibes et al. 1993; Jahier et al. 2001; Ogbonnaya et al. 2001). No susceptible plants were found in the F2 progeny from a cross between the two accessions of Ae. ventricosa with the Cre2 and Cre5 genes, suggesting that their respective resistance factors were allelic. However, the Cre2 and Cre5 genes have been transferred to different chromosomal locations in the wheat introgression lines, H-93-8 and 6D/6Nv (substitution line) because some very susceptible F2 plants were found in the segregation of the resistance trait in the progeny from the cross (6D/6Nv / H-93-8). A high proportion of F2 plants appeared with null infestation, whereas other plants showed a low number of females, suggesting that the former could have both resistance genes (Cre2 and Cre5) and the latter probably only one. Pyramiding resistance genes has been used successfully in many worldwide breeding programs incorporating different resistance genes into a single cultivar to delay the adaptation of the pathogen, which needs to overcome each resistance gene simultaneously to be able to grow on the host.

The induction of several defence responses during early incompatible interaction of resistant lines with the Cre2 and Cre5 genes also has been studied. Isoelectrofocusing isozyme analysis revealed changes of peroxidase (PER), esterase (EST), and superoxide dismutase (SOD) activities in infected roots of resistant lines in comparison to their susceptible parents. Following nematode infection, changes in isozyme patterns of putative defence enzymes (PER, EST, and SOD) occurred in susceptible and resistant wheat hosts. The resistant lines (with Cre2 and Cre5 genes) differentially expressed these isozymes in timing and abundance. The highest differences between infected and uninfected roots were found for the peroxidase system, implicated in lignification, as previously described by other authors (Melillo et al. 1992; Zacheo et al. 1993; Andres et al. 2001). The results clearly confirm previous observations, that the H. avenae pathotype Ha71 was unable to overcome the resistance mechanisms conferred by the Cre2 in the H-93-8 line (Delibes et al. 1993; Andres et al. 2001). No previous study has been made with the Cre5 gene.

Molecular markers can support classical breeding in crop plants by shortening the selection time in the breeding programs. A linkage analysis of the resistance to CCN and a DNA marker, present in H-93-8 and Ae. ventricosa AP-1 (both with Cre2 gene) and absent in Cre5-carrying genotypes, was made in individual F2 plants from the cross between H-93-8 and their susceptible parent (H-10-15). The distribution of susceptibility scores for the two classes of plants (with and without markers) indicated that the linkage between the two traits could not be very tight. Evidence of linkage was found in all the clearly susceptible plants (with a female number/plant higher than 15) always without the marker. However, some plants classified as resistant did not have the DNA marker, which would be consistent with recombination between the introgressed chromosome of H-93-8, with both DNA segments, and a wheat chromosome. This result agrees with our previous work showing recombination between chromosomes of the Nv and D genomes in some H-93 lines. The partial resistance previously described in the parental line H-10-15 also could explain this fact (Delibes et al. 1993). Pathogens are known to more easily overcome resistance provided by a single gene. Durability of resistance has been increased in several crops by incorporating genetic diversity of the major resistance genes. Differences observed between the Cre2 and Cre5 genes with respect to the chromosomal location, induction of detoxifying enzymes, and behavior to different pathotypes, suggest that there are different H. avenae resistance sources for their introduction into commercial wheat cultivars. Selecting lines with Cre2 and Cre5 genes for construction of durable resistant cultivars also may be possible. These findings are significant because novel sources of resistance provide breeders with alternative genes in the event of new pathotypes emerging.

Cooperation with other institutions. We are cooperating with Drs T. Bleve-Zacheo and M. T. Melillo (CNR-Bari, Italy) in the histochemical localization of enzymes related to H. avenae resistance in wheat.

Financial support. This work was supported by grants AGF98-1057-CO4 and AGL2001-3824-CO4 from the Comision Interministerial de Ciencia y Tecnologia of Spain.

References.

• Delibes A, Romero D, Aguaded S, Duce A, Mena M, Lopez-Braña I, Andres MF, Martín-Sánchez JA, and Garcia-Olmedo F. 1993. Resistance to the cereal cyst nematode (Heterodera avenae Woll.) transferred from the wild grass Aegilops ventricosa to hexaploid wheat by a "stepping stone" procedure. Theor Appl Genet 87:402-408.
• Jahier J, Abelard P, Tanguy AM, Dedryver F, Rivoal R, Khatkar S, and Bariana HS. 2001 . The Aegilops ventricosa segment on chromosome 2AS of the wheat cultivar VPM1 carries the cereal cyst nematode gene Cre5. Plant Breed 120:125-128.
• Melillo MT, Bleve-Zacheo T, and Zacheo G. 1992. Role of peroxidase and esterase isoenzymes in pea roots infected with Heterodera goettingiana. Nemat Medit 20:171-179.
• Zacheo G, Orlando C, and Bleve-Zacheo T. 1993. Characterization of anionic peroxidases in tomato isolines infected by Meloidogyne incognita. J Nemat 25:249-256.
  

Publications. [p. 166-167]

• Andres MF, Pinochet J, Hernandez-Dorrego A, and Delibes A. 2000. Detection and analysis of inter- and intraspecific diversity of Pratylenchus spp. using isozyme markers. Plant Pathol 49:640-649.
• Andres MF, Romero D, Montes MJ, and Delibes A. 2001. Genetic relationships and isozyme variability in Heterodera avenae complex determined by isoelectrofocusing. Plant Pathol 50:270-279.
• Andres MF, Melillo MT, Delibes A, Romero MD, and Bleve-Zacheo T. 2001. Changes in wheat root enzymes correlated with resistance to cereal cyst nematodes. New Phytol 152:343-354.
• Andres MF, Melillo MT, Delibes A, Romero MD, and Bleve-Zacheo T. 2001. Biochemical and cytochemical changes in a resistant wheat/Aegilops ventricosa introgression line during cereal cyst nematode infection. Phytopath Medit (in press).
• Andres MF, Melillo MT, Delibes A, Romero MD, and Bleve-Zacheo T. 2000. Histopatologia y cambios enzimaticos producidos por la infeccion de nematodos del complejo Heterodera avenae en una linea de introgresion Trigo / Aegilops ventricosa. In: Proc X Cong Soc Esp Fitopatol, Valencia, Spain, 3-6 October, 2000.
• Andres MF, Melillo MT, Delibes A, Romero MD, and Bleve-Zacheo T. 2001. Biochemical and cytochemical changes in a resistant wheat/Aegilops ventricosa introgression line during cereal cyst nematode infection. In: Proc 11th Cong Medit Phytopath Union and the 3rd Cong Soc Port Fitopatol Evora (Portugal), 17-20 September, 2001.
• Delibes A, Lopez Braña I, Montes MJ, Gomez-Colmenarejo M, Romero MD, Andres MF, Martin-Sanchez JA, Sin E, Martinez C, Michelena A, del Moral J, and Mejias A. 2001. Transfer of resistance genes to Hessian fly and Cereal Cyst nematode from Aegilops triuncialis to hexaploid wheat and its use in breeding programs. Ann Wheat Newslet 47:198-200.
• Montes MJ, Romero MD, Sin E, Lopez-Braña I, Andres MF, Martín-Sánchez JA, Benavente E., Gomez-Colmenarejo M. and Delibes A. 2000. Caracterizacion de la linea de introgresion trigo/Aegilops triuncialis portadora del gen de resistencia a Heterodera avenae woll. (CreAet). Estudio de su relacion con otros genes de resistencia en trigo. In: Proc X Cong Soc Esp Fitopatol, Valencia, Spain, 3-6 October, 2000.
• Montes MJ, Lopez-Braña I, Gomez-Colmenarejo M, and Delibes A. 2001. Enzimas relacionadas con el estres oxidativo inducidas en raices de trigos resistentes Heterodera avenae, en respuesta a la infestacion por el nematodo. In: Proc Cong Soc Esp Genet, Sevilla, Spain, 19-22 September, 2001.
• Nombela G and Romero MD. 2000. Host response to Pratylenchus thornei of a wheat line carrying the cre2 gene for resistance to Heterodera avenae. Nematology 1(4):381-388 (2000).
• Ogbonnaya S, Seah I, Delibes A, Jahier J, Lopez-Braña I, Eastwood RF, and Lagudah ES. 2001. Molecular-genetic characterisation of nematode resistance from Aegilops ventricosa and its derivatives in wheat. Theor Appl Genet 102:263-269.
• Sin E, Montes MJ, Romero MD, Lopez-Braña I, Andres MF, Martín-Sánchez JA, Benavente E, Gomez-Colmenarejo, M, and Delibes A. 2001. Transferencia del gen de resistencia a Heterodera avenae (Cre7) desde Aegilops triuncialis a trigo hexaploide. In: Proc Cong Soc Esp Genet, Sevilla, Spain, 19-22 September, 2001.
  

UNIVERSIDAD POLITÉCNICA DE MADRID

Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos, Ciudad Universitaria, 28040 Madrid, Spain.

A. Delibes, I. Lopez Braña, M.J. Montes, M. Gomez Colmenarejo, and C. Gonzalez-Belinchon.

UNIVERSITY OF LLEIDA AND
INSTITUTE FOR FOOD AND AGRICULTURAL RESEARCH AND TECHNOLOGY (UdL-IRTA)

Center of R&D, Rovira Roure 177, 25198 Lleida, Spain.

J.A. Martín-Sánchez, E. Sin, C. Martinez, and A. Michelena.

JUNTA DE EXTREMADURA

Servicio de Investigacion Agraria, Finca La Orden, 06187 Guadajira, Badajoz, Spain.

J. del Moral, F. Perez Rojas, and F.J. Espinal.

Resistance to Hessian fly conferred by the gene H27. Relationships with other sources of resistance and its effect on some agronomic traits. [p. 167-169]

Hessian fly is one of the most destructive pest of wheat. A Hessian fly resistance gene from Ae. ventricosa and its transfer to hexaploid wheat via interspecific hybridization had been described (Delibes et al. 1997). Transfer line H-93-33, which has 42 chromosomes and has been derived from the cross (T. turgidum/Ae. ventricosa//T. aestivum) was highly resistant to the Spanish Hessian fly population tested. Resistance in Ae. ventricosa 10, Ae. ventricosa AP1, and H-93-33 (DS 4D/4N^v^ ) was located on chromosome 4Nv and is inherited as a single dominant factor (H27), linked to a isozyme marker (Acph-N^v^1), in the introgression line.

Lines derived from the backcross of H-93-33 line, as donor parent, and T. aestivum cultivars Astral, Adalid, and Cargifaro, as alternative recurrent parents, showed a high level of resistance to M. destructor. These lines, which were selected for resistance using Acph-N^v^1 marker, were morphologically similar to bread wheat. Isogenic lines, with and without the H27 gene, have been compared by different characters affecting to seed production and bread quality. As average value, plants with and without the marker showed no difference for number of spikelets/spike and kernel weight/spike, but plants with the marker showed lower spike compactness and a lower number of kernels/spikelet and higher 1,000-kernel weight. However, some lines with the marker show no difference that those lacking the marker. No significant differences were observed in protein contents and SDS-sedimentation value. The same HMW-glutenin pattern as the recurrent progenitor was found in the resistant lines analyzed, which is an acceptable composition for bread-making performance.

Preliminary results in agronomic trials show a superior yield of some lines in relation with checks, both in normal conditions and with insect attack. Table 1 gives the results of some lines obtained during last season at three different locations.

The harvest index of experimental lines was superior to that of the checks (data not shown). Quality parameters (sedimentation with SDS and protein content) of experimental lines rank between Astral (low bread-making quality) and Adalid (good bread-making quality).

A high resistance level to the Hessian fly biotype prevailing in Azuaga (Badajoz, Spain) was found in cultivars with different resistance genes; including Abe (H5), Kay (H11), Ella (H9), Howell (H9), 841453H (H12), Brule (H18), 86925RAI-16 (H13), KS89WGRC6 (H24) and KS86HF (as yet unnamed); located on chromosomes 1A (H5 and H11), 5A (H9 and H12), 6DL (H13 and H24), and 2BS (KS86HF).

The segregation of resistance in the F1 and F2 populations from the crosses 'Kay/H-93-33', 'Ella/H-93-33', 'Howell/H-93-33', '841453H/H-93-33', and 'Brule/H-93-33' has been studied. About 200 plants F2 were recorded from each cross.

All plants of the F1 generation showed a similar resistance level than their resistant parents. Most of F2 plants were resistant, and a few plans were susceptible. These results support the hypothesis of two different loci, with two alleles in each cross, being the resistance genes dominant. Thus, the resistance to M. destructor in H-93-33 line would be determined by one different locus with respect to the genes H9, H11, H12, and H18, and the unnamed gene from KS86HF.

Cultivar diversification, cultivar mixtures, multilines, and pyramiding of resistance genes have been used successfully in many worldwide breeding programs. The last strategy incorporates different resistance genes into a single cultivar to delay the adaptation of the pathogen. Plants with two resistance genes to M. destructor could be obtained in the F2 generation from the crosses previously mentioned. A high proportion of F2 plants with appeared to have no attack, whereas the other showed low number of pupae, which suggests the existence of resistant plants carrying two genes (H27 and that of the other resistant parents) and others with only.

Cooperation with other institutions. We are cooperating with Acorex (a cooperative of Extremadure farmers).

Financial support. This work was supported by grants AGF98-1057-CO4, PTR 95-0496-OP CO 1021001 from CICYT "Comision Interministerial de Ciencia y Tecnologia" of Spain.

References.

• Delibes A, Del Moral J, Martín-Sánchez JA, Mejias A, Gallego M, Casado D, Sin E, and Lopez-Braña I. 1997. Hessian fly-resistance gene transferred from chromosome 4Mv of Aegilops ventricosa to Triticum aestivum. Theor Appl Genet 94:858-864 .
• Delibes A, Lopez-Braña I, Martín-Sánchez JA, Sin E, Martinez C, Michelena A, Del Moral J, and Mejias A. 1997. Transfer of one gene for resistance to Hessian fly (Mayetiola destructor) from Aegilops ventricosa to cultivars of wheat. Ann Wheat Newslet 43:214-215
  

Publications.

• Del Moral J, Delibes A, Martín-Sánchez JA, Mejias A, Lopez-Braña I, Sin E, Martinez C, Montes MJ, and Jimenez J. 1999. Caracteristicas genotipicas y fenotipicas de una coleccion de lineas de trigo resistentes al mosquito (Mayetiola destructor Say). In: Proc VII Jornadas Cientificas de la Soc Esp Entomologia Aplicada. Almeria, Spain, November, 1999.
• Del Moral J, Delibes A, Martín-Sánchez JA, Perez Rojas F, Rubiales D, Merino M, Sin E, Martinez C, Montes MJ, Senero M, and Campos L. 2001. Seleccion de una coleccion de lineas avanzadas de trigo con resistencia a Mayetiola destructor Say. por su comportamiento frente a los patogenos y accidentes fisiologicos mas importantes de este cultivo en la Campiña Sur de Extremadura. In: II Cong Nacional Entomologia Aplicada. VIII Jornadas Cientificas de la S.E.E.A. Pamplona, Spain, November, 2001.
• Sin E, Martín-Sánchez JA, Martinez C, Zarco J, Del Moral J, Mejias A, Lopez-Braña I, Montes MJ, Benavente E, and Delibes A. 2000. Resistance to Hessian fly in T. aestivum transferred from Ae. triuncialis and Ae. ventricosa. Relationships with other resistance sources and its effects in agronomic characters. In: Wheat in a global environment. Proc 6th Internat Wheat Conf (Bedö Z ed). Agric Res Inst Hung Acad Sci, Budapest, Hungary. p. 185.