ITEMS FROM ITALY

ISTITUTO DI GENETICA VEGETAL - CNR
via Amendola 165/a, 70126 Bari, Italy.

Capillary electrophoretic analysis of gliadins in wheat and related species. [p. 67]

A.R. Piergiovanni, N. Volpe, and G. Taranto.

The narrow genetic basis of modern wheat cultivars is the result of the genetic erosion that occurred in the last century. In recent years, many studies have focused on the utilization of wheat germ plasm and of some related Triticum species in wheat breeding. The main objective of this research is the broadening of the genetic bases of modern cultivars. The achievement of wheat-improvement programs is related strictly to the use of powerful techniques to analyze in detail the protein profile of germ plasm accessions of wheat and related Triticum species. This approach allows us to identify the subunits related to end-use quality traits that are present in each accession analyzed.

In the last decade, capillary electrophoretic analysis (CE) of monomeric gliadins and glutenins has assumed increasing importance. Capillary electrophoresis has proven to be a method alternative to PAGE and complementary to HPLC. Moreover, this technique allows rapid and high-resolution separation of proteins with full automation.

We are starting to analyze by CE some related Triticum species, such as the tetraploid the cultivated emmer T. turgidum subsp. dicoccum, hexaploid spelt, and Ae. tauschii, in order to evaluate the genetic variability for gliadins and glutenins of these species.

Publications.

• Laghetti G, Piergiovanni AR, Volpe N, and Perrino P. 1999. Agronomic performance of Triticum dicoccon Schrank and T. spelta L. accessions under southern Italian conditions. Agr Med 129:199-211.
• Piergiovanni AR and Blanco A. 1999. Variation of HMW glutenin and g-gliadin subunits in selected accessions of Triticum dicoccon (Schrank) and T. spelta (L.). Cereal Res Commun 27:205-211.
• Piergiovanni AR and Volpe N. 2003. Capillary electrophoresis of gliadins a tool in the discrimination and characterisation of hulled wheats (T. dicoccon Schrank and T. spelta L.) lines. Cereal Chem 80:269-273.

EXPERIMENTAL INSITUTUE FOR CEREAL RESEARCH

via Mulino, 3 - 26866 S. Angelo Lodigiano (LO), Italy.

Allelic variability at the waxy loci in Italian wheat germ plasm. [p. 68]

G. Boggini, M. Cattaneo, C. Basone, and P. Vaccino.

Waxy mutations in bread wheat are derived from the loss of functionality of the granule-bound starch synthase I (GBSSI), the key enzyme in amylose synthesis, whose coding loci, Wx-A1, Wx-D1, and Wx-B1, occurr on the short arms of chromosome 7A, 7D, and the long arm of chromosome 4A, respectively. Wheat lines with one or two GBSS null alleles produces starch with reduced amylose content and are designated 'partial-waxy', whereas the loss of all three GBSS isoforms gives rise to amylose-free starch, termed waxy. Waxy wheats may find useful application in the production of modified food starch and for the improvement of the shelf-life of baked products; moreover, they strongly influence Asian noodle quality.

A breeding program was set up to develop waxy wheat lines adapted to Italian environments from partial-waxy cultivars previously identified from a screening of Italian germ plasm. Nine crosses were made and the F1 and F2 generations were grown in the greenhouse without any selection. F3 generations were grown in the field and selection was performed on the basis of agronomical characters. Single F3 plants were harvested, and a sample of seeds/plant was analyzed by means of colorimetric analyses with a solution of KI/I2 followed by electrophoretic fractionation of the GBSS isoforms. A study involving PCR analysis is in progress in order to develop a relatively simple method, possibly automated, to screen rapidly each partial waxy wheat line. A couple of primers were able to detect the null allele at the Wx-D1 locus of Bai-Huo, but did not recognize the same locus in the Italian landrace Cologna Lunga, suggesting the presence of a new null allele is probable. The same primers seem to detect polymorphism for the Wx-A1 locus among cultivars, whereas another set of primers tested seems to be specific for the Wx-B1 locus.

Strampelli cultivars: biochemical, technological, and agronomical characterization. [p. 68]

G. Boggini, A. Brandolini, M. Perenzin, S. Empilli, P. Vaccino, and M. Cattaneo.

Nazareno Strampelli was one of the most important breeders in the world. His work preceded that of Norman Borlaug and the "Green revolution" by nearly 30 years. During his long career, he released more than 70 bread wheat cultivars, largely grown in Italy and other countries and that also appear in the pedigree of significant recent cultivars. The aim of this work combines the description of these cultivar, their comparison with modern wheats, and their rescue from loss.

According to the coefficient of parentage based on the pedigrees described by Strampelli, the cultivars were clustered in different groups that shared the local population Rieti as common parent. The narrow genetic background of the Strampelli germ plasm was confirmed by the limited variability observed in the gliadin and glutenin composition. An AFLP analysis also was used to assess the molecular genetic diversity. The high variability found appeared related to the incomplete homogeneity and stability of the material released by Strampelli, but also to the large use of heterogeneous local populations in the crosses.

Agronomic traits varied significantly among the cultivars, whereas rather low differences were shown for the most important quality traits. Exploitation of this germ plasm for breeding purposes is in progress.

Puroindoline and kernel hardness in Triticum aestivum and Triticum monococcum. [p. 68-69]

N.E. Pogna, L. Gazza, G. Boggini, M. Corbellini, and P. Vaccino.

Registered Italian cultivars were classified according to their hardness by means of Single Kernel Characterization System (SKCS) and puroindoline composition. Three groups were identified: very soft, with SKCS indices lower than 14; soft, with SKCS indices ranging from 20 to 43; and medium hard, with SKCS indices higher than 55. Only the cultivar Enesco was present in the range 44-55. A bulk sample of this genotype showing a bimodal distribution for SKCS index was grown as spaced plants, and the grain was characterized by SKCS, A-PAGE fractionation, and PCR amplification. According to previous results, very soft grain contained Pin A and Pin B glycine-type, whereas medium-hard grain texture was associated with either the absence of Pin A or the presence of a Pin B serine type. Studies are in progress on a plant heterozygous for the alleles Pin B serine and glicine type.

Breeding for resistance to powdery mildew. [p. 69]

A. Brandolini, M. Corbellini, and G. Reffo.

The breeding program for bread wheat cultivars based on backcrossing and MAS selection and aimed at the introgression of powdery mildew resistance gene Pm13, derived from Ae. longissima, is currently at the BC5 stage. To fix the gene, a cycle of selfing was made, and top crosses to assess homozygosity of the lines are in progress. Line evaluation for similarity to the recurrent parents also is under way.

Genetic analysis of einkorn wheat quality traits. [p. 69]

A. Brandolini, P. Vaccino, P. Ramelli, M. Corbellini, and G. Boggini.

Gliadin and glutenin storage protein fingerprinting of 668 samples of domesticated einkorn was completed. Correlations with SDS-sedimentation test results from 2 years were performed. The total number of HMW-glutenin bands scored was 39; six belong to the x HMW-subunit group, eight to the gamma HMW-subunit group, and 25 to the LMW group. The total number of gliadin bands recorded were 44, 20 in the omega region, 10 in the gamma region, and 14 in the a/b region. Eight glutenin bands and eight gliadin bands were correlated significantly with an increase in bread-making quality. Progenies of two-, three- and four-way crosses of lines with good agronomic and quality traits (earlyness, free-threshing, short straw, gluten quality, and large kernel) are undergoing different stages of evaluation.

Publications. [p. 69-70]

• Balconi C, Lanzanova C, Baldoni E, Conti E, Cattaneo M, and Lupotto E. 2003. In vivo and in vitro bioassays for testing resistance to Fusarium ssp. in wheat genotypes. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 3:1102-1104.
• Boggini G, Vaccino P, Brandolini A, and Cattaneo M. 2003. Genetic variability of Strampelli bread wheat realisations detected by storage protein composition and by AFLP. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 1:101-104.
• Brandolini A, Vaccino P, and Bruschi G. 2003. Technological properties of einkorn flour: the role of storage proteins and starch. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 1:427-430.
• Cattaneo M, Corbellini M, Empilli S, and Boggini G. 2003. Agronomical and qualitative traits in some Strampelli bread wheat cultivars. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 1:121-124.
• Gazza L, Niglio A, Vaccino P, and Pogna NE. 2003. The long arm of chromosome 5D of bread wheat contains a Pina-D1a-like sequence. In: Proc10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 3:1330-1332.
• Raciti CN, Doust MA, Lombardo GM, Boggini G, and Pecetti L. 2003. Characterization of durum wheat mediterranean germplasm for high and low molecular weight glutenin subunits in relation with quality. Eur J Agron 19:373-382.
• Reffo G, Corbellini M, Bruschi G, and Brandolini A. 2003. Marker-assisted introgression of the Pm13 powdery mildew resistance gene in Italian bread wheat cultivars. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 2:801-803.
• Salamini F and Brandolini A. 2003. Wild relatives of cultivated wheats. In: Abstr Unpubl Contrib Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. Pp. 3-4 (Abstract).
• Sgrulletta D, De Stefanis E, Cammerata A, and Corbellini M. 2003. Near infrared transmittance spectroscopy for grain hardness determination in bread wheat (T. aestivum L.). Tecnica Molitoria Internat 2:111-116.
• Spina A, Palumbo M, and Boggini G. 2003. Bread-making quality of new durum wheat cultivars. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 3:1394-1396.
• Upelniek VP, Nikolaev AA, Boggini G, and Kudryavtsev AM. 2003. Genetic polymorphism of Strampelli bread wheat varieties evaluated by gliadin alleles. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 2:646-648.
• Vaccino P, Perenzin M, and Concaro C. 2003. Identification of waxy mutants in T. aestivum and their use in a breeding program. In: Proc 10th Internat Wheat Genet Symp (Pogna NE, Romano M, Pogna EA, and Galterio G eds). Istituto Sperimentale per la Cerealicoltura, Roma, Italy. 2:846-848.
  

ISTITUTO SPERIMENTALE PER LA CEREALICOLTURA - EXPERIMENTAL INSTITUTE FOR CEREAL RESEARCH
Via Cassia 176, 00191 Roma, Italy.

 

Reaction to wheat soilborne mosaic virus of 85 cultivars of durum wheat during six seasons in northern and central Italy. [p. 70-71]

V. Vallega, and C. Rubies-Autonell and C. Ratti (Dipartimento di Scienze e Tecnologie Agroalimentari (DiSTA), Area di Patologia Vegetale, Università di Bologna Via Fanin 44, 40127 Bologna, Italy).

Beginning in the 1995-96 season onward, about 30 of the durum wheat cultivars marketed in Italy are tested each season for resistance to WSBMV in a field near Bologna with WSBMV only and at a site near Rome with both WSBMV and WSSMV. Resistance is evaluated on the basis of symptom severity, virus concentration in leaves (with DAS-ELISA), and agronomic performance (i.e., grain yield, test weight, 1,000-kernel weight, plant height, and heading date). A total of 85 cultivars were assayed in six seasons characterized by severe disease pressure (Bologna 1995-96, 1996-97, 2000-01, 2001-02, and 2002-03; Rome 1998-99). The cultivars tested are listed in Table 1. Each cultivar is classified for resistance to WSBMV on the basis of DAS-ELISA values recorded in one or more seasons. Only seven cultivars among all those tested showed ELISA values close to zero and very mild symptoms. Three of these cultivars were assayed in only one season, indicating that their seemingly high level of resistance needs confirmation.

Simple correlations between symptom severity scores and grain yields were highly significant in all these trials. The estimated yield loss attributable to SBWMV or to a mixed WSBMV and WSSSMV infection (Rome 1998-99) are summarized in Table 2. Interestingly, even cultivars with relatively mild symptoms suffered severe losses; from a minimum of 17 to 39 %, depending on the season. Correlations between symptom severity scores and DAS-ELISA readings also were highly significant in all trials. Indeed, the results obtained for cultivars tested in various seasons indicate that breeders may achieve substantial progress towards WSBMV resistance by selecting on the basis of visible symptoms alone. On the other hand, results also showed that highly resistant cultivars may be distinguished from moderately resistant ones only by the use of ELISA or other techiques which estimate virus concentration.

Reaction of 81 cultivars of common wheat in northern Italy to wheat soilborne mosiac virus during five seasons. [p. 71-72]

C. Rubies-Autonell, C. Ratti, and V. Vallega.

Eighty-one common wheat cultivars currently marketed in Italy were assayed in a field near Bologna for resistance to WSBMV during five seasons characterized by severe disease pressure (1995-96, 1996-97, 2000-01, 2001-02, and 2002-03). Each cultivar was assayed for 1-5 seasons. Resistance was evaluated on the basis of symptom severity using ELISA values from extracts of leaves and agronomic performance. The cultivars tested are listed in Table 3 and classified for resistance to WSBMV on the basis of DAS-ELISA values only. Twenty-two cultivars among those tested showed ELISA values close to zero and very mild symptoms. None of the lines showed symptoms as severe as those displayed by the most susceptible durum wheats.

Simple correlations between symptom severity scores and grain yield were highly significant in all trials, except in the 2000-01 season, which was characterized by the presence of severe epidemics of other wheat diseases as well. The estimated yield losses attributable to WSBMV infection are summarized in Table 4. Like the durum wheats, even cultivars with relatively mild symptoms suffered severe losses, from a minimum of 13 % to a maximum of 29 %, depending on the season. Correlations between symptom severity scores and DAS-ELISA readings were highly significant in all trials. Few, if any, of the common wheats assayed suffered grain yield losses as severe as those recorded for the most susceptible durum wheats.

Publications. [p. 72]

• Ratti C, Rubies-Autonell C, Canova A, Montanari O, and Vallega V. 2002. Molecular characterization of Italian isolates of soilborne wheat mosaic virus. Atti Giornate Fitopatologiche, Baselga del Pine, Trento, 7-11 April, 2002. 2:585-592 (In Italian).
• Rubies-Autonell C, Ratti C, Fornara M, and Vallega V. 2003. Viral diseases, resistant cultivars. Terra e Vita 54(36):58-61 (In Italian).
• Rubies-Autonell C, Vallega V, and Ratti C. 2003. Reactions of cultivars of common wheat (Triticum aestivum L.) to soilborne wheat mosaic virus in northern Italy during 1996-97. J Plant Dis Protect 110(4):332-336.
• Rubies-Autonell C, Vallega V, Ratti C, Canestrale R, and Sarti A. 2004. Soil-borne wheat mosaic virus: behaviour of durum and common wheat cultivars. Results of the trials carried out in 2002-2003. L'Informatore Agrario 5:49-54 (In Italian).
• Vallega V, Rubies-Autonell C, and Ratti C. 2002. Reactions to soilborne wheat mosaic of durum wheat cultivars grown during 2000-2001 in Northern Italy. In: Proc 5th Symp Internat Working Group of Plant Viruses with Fungal Vectors (IWGPVFV), Zurich, Switzerland, 22-25 July, 2002. Pp. 52-55.
• Vallega V, Rubies-Autonell C, and Ratti C. 2003. Reaction of durum wheat cultivars to mixed SBWMV and WSSMV infection in central Italy. Phytopath Medit 42:177-182.
• Vallega V, Rubies-Autonell C, Ratti C, Canestrale R, and Sarti A. 2002. Reaction of wheat cultivars to soilborne wheat mosaic virus: results of the trials carried out in 2000-2001. L'Informatore Agrario 58:69-73 (In Italian).

UNIVERSITY OF BOLOGNA, COLLEGE OF AGRICULTURE
Dipartimento di Scienze e Tecnologie Agroalimentari (DiSTA), Area di Patologia Vegetale, Via Filippo Re 8, 40126 Bologna, Italy.

Reaction of thirty cultivars of durum wheat to SBWMV during 2001-02. [p. 72-73]

C. Rubies-Autonell and C. Ratti, and V. Vallega (Istituto Sperimentale per la Cerealicoltura, Via Cassia 176, 00191 Rome, Italy).

Wheat soilborne mosaic virus is widespread in Italy, especially in the northern and central regions, where it causes severe losses on both common and durum wheat crops. Yield losses of up to 50-70 % have been recorded on susceptible cultivars. Fields with WSBMV have been identified also in southern Italy and Sicily. During 2001-02, 30 durum wheat cultivars were assayed in a severely WSBMV-infested field situated near Minerbio (Bologna) to study their response to the virus. The cultivars were grown in 10-m2 plots distributed in the field according to a randomized block design with three replicates. As in previous years, reactions to WSBMV were evaluated on the basis of DAS-ELISA readings, symptom severity, and agronomic performance. Samples for DAS-ELISA were collected 13 March and 3 April. Symptoms were scored 13 March, 3 April, and 22 April using a 0-4 scale. Disease pressure was severe and resulted in highly diverse reactions among cultivars (Table 1). Mean symptom severity ranged from 0.1 to 3,2; mean ELISA values from 0 to 0.991, and grain yield from 1.08 to 5.91 t/ha. Most cultivars proved susceptible in terms of both symptom expression and ELISA values, and this was clearly reflected by their relatively low grain yields.

The cultivars Solex, Neodur, San Carlo, Gianni, Meridiano, and Verdi were highly resistant, even if none were symptomless throughout the season. The cultivars Cannizzo, Carioca, Claudio, Colosseo, Marco, Orobel, Vesuvio, and Vetrodur, with yields between 1.08 and 3.95 t/ha, proved particularly susceptible. Symptom severity scores were highly correlated with ELISA values (0.868**), grain yield (-0.668**), and plant height (-0.868**), but not with test weight, 1,000-kernel weight, and heading date. Cultivars with symptom scores ranging from 2.1 to 3.2 had an estimated loss in grain yield of approximately 43 % and a plant height reduction of about 25 %. Cultivars showing only relatively mild symptoms (1.12.0) also were negatively affected in terms of grain yield (-17 %) and plant height (-7 %).

Reaction of 33 cultivars of common wheat to wheat soilborne mosaic virus in 2001-02. [p. 73-74]

V. Vallega (Istituto Sperimentale per la Cerealicoltura, Via Cassia 176, 00191 Rome, Italy), and C. Ratti and C. Rubies-Autonell.

During the 2001-02 season, 33 cultivars of common wheat were assayed in a severely WSBMV-infested field situated near Minerbio (Bologna) to study their response to the virus. Entries were grown in 10-m^2^ plots distributed in the field according to a randomized block design with three replicates. Resistance to WSBMV was evaluated on the basis of DAS-ELISA readings, symptom severity, and agronomic performance. Samples for DAS-ELISA were collected 13 March and 3 April. Symptoms were scored on 26 February, 13 March, and 3 April using a 0-4 scale. As in all our previous trials, Italian common wheats showed much higher levels of resistance to WSBMV than durum wheat cultivars assayed in an adjacent field. Symptom severity scores among common wheats ranged from 0 to 1.3, ELISA values from 0 to 1.102, and grain yield from 4.77 to 7.33 t/ha (Table 2). The cultivars Belfiore, Enesco, Levis, Sirmione, Artico, Colfiorito, Tremie, and Centro showed slight symptoms and null or low ELISA values. The cultivar Centro produced the highest yields. Symptom severity scores highly correlated with ELISA values (0.716**) and grain yield (-0.447**) but not with test weight, 1000-kernel weight, plant height, or heading date. Cultivars with symptom scores ranging from 1.1 to 1.3 had an estimated mean grain yield loss of approximately 17 %.

Reaction of 114 cultivars of durum wheat to wheat soilborne misaic virus in northern Italy.[p. 74-75]

C. Rubies-Autonell, C. Ratti, and A. Perez de Anucita; M.C. Sanguineti and M. Maccaferri (DiSTA Area di Miglioramento Genetico,Università di Bologna, Via Fanin 44, 40127 Bologna); and V. Vallega (Istituto Sperimentale per la Cerealicoltura, Via Cassia 176, 00191 Rome).

During the 2002-03 season, 114 cultivars of durum wheat of various origins were grown near Bologna in a field with natural inoculum sources of WSBMV and evaluated for resistance to this pathogen on the basis of symptom severity and virus concentration in leaves. Our main objectives were to provide breeders with information on the resistance of cultivars widely used as parents in their programs and to enable virologists around the world to investigate the existence of differential reactions to WSBMV. The latter aspect, in fact, has acquired renewed importance in that several authors, based on the results of nucleotide-sequence analyses, have recently suggested that the WSBMV found in Italy and other European countries is different from that present in North America. The experiment included cultivars obtained by national breeding programs in Australia, Austria, Canada, France, Italy, Spain, the U.S., ICARDA, and CIMMYT. The cultivars were grown in 122-cm, solid-seeded rows with two replications. Symptom severity was scored three times during the season (13 March, 2 April, and 16 April) using a 0 to 4 scale, where 0-1 = slight or no symptoms; 1.1-2 = mild mottling and stunting; 2.1-3 = mottling and stunting; and 3.1-4 = severe mottling and stunting, with virus-killed plants. Virus concentration was determined by DAS-ELISA on leaf extracts collected 13 March, 2 April, and 16 April. Extracts were from a bulk of the apical half of the second youngest leaf of 10 plants/plot. The results of the experiment are summarized in Table 3 (p. 75). About one-half of the cultivars assayed had very low ELISA values and mild symptoms (Table 3). A number of these cultivars had moderately high ELISA values and relatively more severe WSBMV in previous trials conducted at the same site, but none of those previously found resistant to WSBMV showed high ELISA values and/or severe symptoms. Evidently, disease pressure during the 2002-03 growing season was sufficient to uncover very susceptible cultivars but not to distinguish resistant types from moderately susceptible ones. The experiment will be repeated in 2003-04.