PLANT BREEDING AND SEED SCIENCE Volume Czempiñ, Po land, Kraków, ul. Zawi³a 4a, Po land

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PLANT BREEDING AND SEED SCIENCE Volume 51 2005 Henryk Woœ 1, Anna Strzembicka 2 1 Plant Breed ing Com pany Strzelce, Breed ing De part ment-borowo, 64-020 Czempiñ, Po land, 2 Plant Breeding and Acclimatization Institute IHAR, Department of Cereals, 30-423 Kraków, ul. Zawi³a 4a, Po land RESISTANCE TO LEAF RUST (PUCCINIA RECONDITA F.SP.TRITICI AT THE SEEDLING STAGE AMONG SINGLE D-GENOME SUBSTITUTION LINES OF TRITICALE PRESTO (Short communication) ABSTRACT There were per formed tests on a set of twenty sin gle sub sti tu tions of the D-ge nome chro mo somes from var i ous bread wheats into win ter triticale Presto, a set of seven disomic ad di tion lines of chro mo somes from win ter rye Dankowskie Zlote to win ter wheat Grana, and all pa ren tal forms to study on the chro mo some lo ca tion of ma jor genes that are resposible for seed ling stage re sis tance to Puccinia recondita f.sp.tritici in triticale,. For each line, seed lings de vel oped to two-leaf stage were exposured to four pathotypes of the patho gen pre dom i nant in Po land to test them in the green house con di tions. There were high re sis tance among the sub sti tu tion lines of Presto, 1D(1A), 4D(4R), and 5D(5R) sug gest ing that chro mo somes in tro duced from the D-ge nome car ried re sis tance genes. Among the ad di tion lines, GH2R was highly re sis tant. Re sis tance to rust at seed lings stage of Presto lines of D- ge - nome and ad di tion lines Grana (wheat)/dankowskie Zlote (rye) shows that chro mo some 2R is re spon si ble for the re sis tance to the dis ease. Key words: leaf rust, re sis tance, sub sti tu tion, triticale, wheat INTRODUCTION The most im por tant of the three rusts oc cur ring on wheat in Po land, is the leaf rust, caused by Puccinia recondita Rob. ex Desm. f. sp. tritici appearing every year that of ten dem on strates in se vere ep i dem ics (DwuraŸna, Gajda 1980, Strzembicka 1997). Re cently this patho gen has also become a se ri ous men ace for triticale (Zamorski et al.1994, Woœ et al. 1994, 1995, Arseniuk 1996, Strzembicka et al. 1998). Se vere in fec tions caused by P. recondita f.sp.tritici re sulted sub stan tial yield losses of win ter triticale (Woœ et al. 1994). Leaf rust re sis tance of triticale, similary to that one found in wheat, may be con - di tioned by nu mer ous genes scat tered through the ge nome what make breed ing ef - forts fo cused on con cen tra tion of prof it able loci nec es sary. Some cultivars, par tic u larly the newer ones, show var i ous de gree of sus cep ti bil ity (Kociuba 1994, Communicated by Henryk J. Czembor

44 Henryk Woœ, Anna Strzembicka Woœ et al.1995, Wêgrzyn et al. 1996, Strzembicka et al.1998). It has been es tab - lished that re spon si ble for the spread of Puccinia recondita f.sp.tritici are: the phys - i o log i cal spe cial iza tion of the patho gen, its high vir u lence and in suf fi ciency of sources of re sis tance (Roelfs et al.1992, Strzembicka 1997). Tak ing un der con sid - er ation that the iden ti fi ca tion of new sources of re sis tance would be im por tant in de vel op ing of breed ing pro grams, it seems to be rea son able to search such sources. This stud ies were taken up to iden tify the chro mo somes that pos si bly carry leaf rust re sis tance among the sub sti tu tion and ad di tion lines of win ter triticale and win ter wheat. The re sults of the re sis tance eval u a tion at the seed ling stage were pre sented in this pa per. MATERIALS AND METHODS Twenty sin gle D-ge nome sub sti tu tion lines of win ter triticale Presto (sub sti tu tion 7D(7R) miss ing), seven disomic ad di tion lines of chro mo somes of win ter rye Dankowskie Zlote to win ter wheat Grana (GHR1-GHR7) and the pa ren tal forms were tested with re spect to re sis tance (Ta ble 1).The win ter triticale sub sti tu tion lines and wheat-rye ad di tion lines were ob tained from A.J. Lukaszewski, Uni ver - sity of Cal i for nia, Riv er side, USA. Ex per i ments were car ried out un der green house con di tions within the years 2000-2002. Seedlings de vel oped to two-leaf stage were in oc u lated with the same four pathotypes of Puccinia recondita f.sp.triticiin ev ery year, i.e. 4c, 40b, 83c, and 95b. These four pathotypes were found as pre dom i nant ones in the Pol ish pop u la - tion of the wheat leaf rust (Strzembicka 1997) and were highly vir u lent to the dif - fer ent set of fif teen isogenic lines car ry ing the fol low ing re sis tance genes Lr1,Lr2a, Lr2b, Lr2c, Lr3a, Lr9, Lr11, Lr15, Lr17, Lr19, Lr21, Lr23, Lr24, Lr26 and Lrr28, re spec tively. This set of dif fer en tials is used in all of Eu rope (Mesterhazy et al.2000). The virulence characteristics of the pathotypes of Puccinia recondita f.sp.triticiused in this study: PathotypeVirulent on the Lr genes 4c2c, 3, 11, 15, 17, 21, 26, 28 40b 3, 11, 15, 17, 21, 26 83c 3, 11, 15, 17, 21, 26, 28 95b 2b, 2c, 3, 11, 15, 17, 21, 26, 28 The in oc u la tion of seed lings was per formed by spray ing the leaves with a wa ter sus pen sion of spores sup ple mented Tween 20 used as a wet ting agent. Fol low ing in oc u la tion, the seed ling were placed in a mois ture cham ber for 24 h and then trans - ferred to the room with 18-20 C and 10-12 h day/night pe riod. The vir u lence of patho gen against leaves was eval u ated ac cord ing to the scale sug gested by Roelfs et al.(1992), where: 0-im mune (no vis i ble uredia), 0; - very re sis tant (hy per sen si - tive flecks), 1- re sis tant (small uredia with ne cro sis), 2 re sis tant to mod er ately re - sis tant (small to me dium uredia of ten sur rounded by chlorosis or ne cro sis), 3 mod er ately sus cep ti ble (me dium sized uredia with or with out chlorosis), 4 sus - cep ti ble (large uredia with out chlorosis), x re sis tant, het er o ge neous.

Re sis tance to leaf rust (Puccinia recondita f.sp.tritici at the seed ling stage among sin gle d ge nome 45 Re sis tance at the seed ling stage to Puccinia recondita f.sp.tritici of some sub sti tu tion lines of triticale Presto, euploids and ad di tion lines of Grana (wheat)/dankowskie Zlote (rye) Table 1 Substitution, Pathotypes of Puccinia recondita f.sp.tritici Addition, translocation 4c 40b 83c 95b 4c 40b 83c 95b Presto 1D(1A) 0; 0; 0; 0; 0; 0; 0; 0;1 Presto 1D(1B) 2 2 2 1,2 0; 0; 1,2 1 Presto 1D(1R) 1,2 x x 0; 0; 0; 0; 0; Presto 2D(2A) 0;2 2,x 2 2 0; 1 0; 0;1 Presto 2D(2B) 1,2 2 2 0;2 0; 1 1 0; Presto 2D(2R) 2 4 4 2 2 3 3 2 Presto 3D(3A) 2 x 0; 2 0;1 0; 0; 1 Presto 3D(3B) 0; 0; 2,x 0; 1 0; 0; 0; Presto 3D(3R) 1,2 2,x 1,2 x 0;1 2,x 2 2;x Presto 4D(4A) 2 2 x 0; 0;1 0;1 0; 0; Presto 4D(4B) 0; 2,x 2 0; 0; 0; 0;1 0;1 Presto 4D(4R) 0; 0; 0;1 0;1 0; 0; 1 1 Presto 5D(5A) 0;1 1,2 0;1 1 0; 0; 0; 0; Presto 5D(5B) 1 2 2 1 0; 0;2 0;1 0;1 Presto 5D(5R) 0; 0;1 0; 1 0; 0; 0; 0;1 Presto 6D(6A) 1 2 x 0; 0; 0; 0;1 0; Presto 6D(6B) 2 1,2 1,2 0; 0; 1 0; 0;1 Presto 6D(6R) 1,2 1,2 1,2 2 0; 0; 0; 1,2 Presto 7D(7A) 0;1 x 2 1,2 0; 0; 0; 0;1 Presto 7D'N(7B) 1,2 0;2 2 1 0; 0; 1 0; GH 1R 4 4 4 4 4 4 4 4 GH 2R 0; 0;1 0; 1 0; 1 0; 1 GH 3R 4 4 4 4 4 4 4 4 GH 4R 4 4 4 4 4 4 4 4 GH 5R 4 4 4 4 4 4 4 4 GH 6R 4 4 4 4 4 4 4 4 GH 7R 4 4 4 4 4 4 4 4 Grana (wheat) 4 4 4 4 4 4 4 Dañk.Z³ote (rye) 0 0 0 0 0 0 0 0 Presto (triticale) 0;2 0;1 1 2 0;1 0; 0; 0; (Valdy- 1) Presto 1R.1D 5+10-2/ WR4 - - - - 0; 2 0;x 0; Valdy - 2 Presto 1R.1D 5+10-2/ WR4 - - - - 0; 0; 0; 0; Check- Kolibri 4 4 4 4 4 4 4 4 Leaf rust re ac tion scale (Roelfs et al.1992): 0 -immune, 0;-very resistant, 4 - susceptible, x - heterogeneous reaction

46 Henryk Woœ, Anna Strzembicka RESULTS AND DISCUSSION In all ex per i ments, a re sis tant or mod er ately re sis tant re ac tions were typ i cal for all sub sti tu tion lines of tested triticale; Presto it self had a good level of re sis - tance. De pend ing on the pathotypes used, the sub sti tu tion lines showed var i ous host re sponses, with in fec tions of types of 0, 1, 2, X. Among the D-ge nome sub - sti tu tion lines of Presto, lines 1D(1A), 4D(4R), 5D(5R) showed higher re sis - tance to Puccinia recondita f.sp. tritici than the euploid Presto and it was found for both years of test ing. It in di cates that chro mo somes 1D, 4D and 5D pres ent in the set can be iden ti fied as ones car ry ing the re sis tance genes. Less likely, chro mo somes 1A, 4R and 5R can be thought as car ry ing genes of suppressors of re sis tance. It may be pointed out that in a dif fer ent study (Woœ et al., this vol - ume) sub sti tu tion 1D(1A) had the most pos i tive ef fect on breadmaking qual ity of all sub sti tu tions tested. This study shows that ad di tion ally, it im proves dis - ease re sis tance. Among the disomic wheat-rye ad di tion lines, only GH2R showed high seed - ling re sis tance to all four used pathotypes (Ta ble 1). Clearly, this chro mo some car ries a ma jor re sis tance lo cus al though its pres ence was not ap par ent in the set of Presto sub sti tu tions. This lo cus could be ex ploited in wheat breed ing as there is avail able a set of 120 2B-2R recombinants and it could be eas ily used to en gi - neer the proper trans fer (A.J. Lukaszewski, pers. comm). While this study gave no in di ca tion with re gard to the iden tity of the re sis tance gene (it may be also Lr25), known to be on the seg ment of 2RL in the Transec translocation (Driscoll and An der son 1967). The patho gen iso lated from rust-in fected triticale fields is usu ally iden ti fied as Puccinia recondita f.sp.tritici (McIntosh et al. 1995, Arseniuk 1996).This im plies that like wheat, triticale is gen er ally re sis tant to f.sp secalis. Be cause f.sp.secalis is avirulent on wheat, Qui nones et al.(1972) sur mised that re sis - tance in triticale was con trolled only by genes de riv ing from the wheat par ent com po nent. If it ap peared true in at least some in stances, rye re sis tance genes clearly would op er ate in other cases. Singh and McIntosh (1990) iden ti fied gene LrSatu, lo cated in a rye chro mo - some, that oc curred at high fre quency in CIMMYT triticale pop u la tion. The rye-de rived genes Lr 25 and Lr26 that showed to be ef fec tive in hexaploid wheat back grounds, in some coun tries they ought to be ef fec tive in triticale (McIntosh et al.1995). Re sis tance to dis eases in triticale has been con sid ered as one of its most im - por tant and du ra ble ad van tages (Arseniuk et al.2000). Re sis tance to rust at seed lings stage of Presto lines of D- ge nome and ad di tion lines Grana (wheat)/dankowskie Zlote (rye) shows that chro mo some 2R is re - spon si ble for the re sis tance to the dis ease. The re sults of seed ling tests on leaf rust re sis tance sug gest that some of an a - lyzed sub sti tu tion lines of triticale and ad di tion line GH2R may be the valu able source of re sis tance. They will be eval u ated in the field con di tions, in adult plant stage and used for fur ther in ves ti ga tions.

Re sis tance to leaf rust (Puccinia recondita f.sp.tritici at the seed ling stage among sin gle d ge nome 47 REFERENCES Arseniuk E. 1996. Triticale dis eases a re view. De vel op ments in plant breed ing. Triticale: to day and to mor - row. Eds. H. Guedes-Pinto, N.Darvey, V.Carnide, Kluwer Ac a demic Pub lish ers: 499-525. Arseniuk E., Woœ H., WoŸniak-Strzembicka A. 2000. As pects of triticale Dis ease re search in Po land. Vortr. Pflanzenzuchtg. 49. 63-72. Driscoll C.J. and L.M. An der son. 1967. Cytogenetic stud ies of Transec - a wheat-rye translocation line. Can J. Genet. and Cytol 9:375-380. DwuraŸna M., Gajda Z. 1990. Zagadnienie rdzy zbo owej w hodowli pszenicy. Biul. IHAR. 173-174, 79-81. Kociuba W. 1994. Field es ti ma tion of re sis tance to fun gal dis eases in col lec tion of spring triticale (X Triticosecale Wittmack).Genetica Polonica 35B, 187-191. Mesterhazy A., Bartos P.,Goyeau H., Niks R.E., Csosz M., Andersen O., Casulli F., Ittu M.,Jones E.,Manisterski J.,Manninger K.,Pasquini M., Rubiales D.,Schachermayr G.,Strzembicka A.,Szunics L.,Todorova M. Unger O., Vanco B., Vida G., Wal ter U. 2000. Eu ro pean vir u lence sur vey for leaf rust in wheat. Agronomie 20. 793-804. McIntosh R.A., Wellings C.R., Park R.F. 1995. Wheat Rusts: An at las of re sis tance genes, CSIRO. Kluwer Acad.Publ. Dordrecht. Qui nones M.A., Llarter E.N., Samborski D.J. 1972. The in her i tance of re sis tance to Puccinia recondita in hexaploid triticale. Canad. Jour nal of Genet. and Cyt. 14, 495-505. Roelfs A.P.Singh R.P. Saari E.E.1992. Rust Dis ease of Wheat. Con cepts and meth ods of dis ease man age - ment. Mex ico D.F. CIMMYT. 81 pp. Singh S.J., McIntosh R.A. 1990. Link age and ex pres sion of genes for re sis tance to leaf rust and stem rust in triticale. Ge nome 33,115-118. Strzembicka A.1997.Vir u lence of Puccinia recondita f.sp.tritici in Po land. J. Appl. Genet. 38B. 101-105. Strzembicka A.,Wêgrzyn S.,Grzesik H.1998.Ocena rodów hodowlanych pszen yta pod wzglêdem odpornoœci na rdzê brunatn¹. Biuletyn IHAR 205/206. 273-277. Wêgrzyn S., Strzembicka A., Grzesik H., Gajda Z. 1996. Eval u a tion of the triticale breed ing ma te ri als with re spect to their re ac tion to fun gal dis eases. Triticale: To day and Tommorow. De vel op ments in Plant Breed ing, Kluwer Ac a demic Pub lish ers, 315-320. Woœ H., Maækowiak W., Cichy H. 1994. Sus cep ti bil ity of win ter triticale to glume blotch, leaf rust and scald. Hod. Roœl. Akl. i Nas.T.38.Z.1-4, 223-227. Woœ H., Maækowiak W., Apolinarska B. 1995. Wstêpne wyniki nowej metody okreœlania odpornoœci pszen yta ozimego na Puccinia recondita. Biuletyn IHAR 195/196, 191-196. Zamorski C., Schollenberger M., Sieklucka M. 1994. The role of win ter triticale crop in overwintering of Puccinia recondita f.sp.tritici and Puccinia striiformis. Zeszyty Naukowe Akad. Rol. Szczecin. 527-532