vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJGB-23-66vavilov-3925Research ArticleГЕНЕТИЧЕСКИЕ РЕСУРСЫ И БИОКОЛЛЕКЦИИPLANT GENETICSПолучение и изучение линии с замещением хромосомы 4B пшеницы Triticum aestivumL. на хромосому 4Hmar дикого ячменя Hordeum marinum ssp. gussoneanum (4x)Development and characterization of a line with substitution of chromosome 4B of wheat Triticum aestivum L. on chromosome 4Hmar of wild barley Hordeum marinum ssp. gussoneanum (4x)https://orcid.org/0000-0002-9941-2026ПершинаЛ. А.PershinaL. A.

N

Новосибирск

ovosibirsk

Novosibirsk

pershina@bionet.nsc.ruhttps://orcid.org/0000-0002-6701-6811ТрубачееваН. В.TrubacheevaN. V.

Новосибирск

Novosibirsk

https://orcid.org/0000-0003-1939-6140ШумныйВ. К.ShumnyV. K.

Новосибирск

Novosibirsk

https://orcid.org/0000-0001-7101-9639БадаеваЕ. Д.BadaevaE. D.

Москва

Moscow

Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук; Курчатовский геномный центр ИЦиГ СО РАНРоссияInstitute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences; Kurchatov Genomics Center of ICG SB RASRussian FederationФедеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияInstitute of Cytology and Genetics of the Siberian Branch of the Russian Academy of SciencesRussian FederationИнститут общей генетики им. Н.И. Вавилова Российской академии наукРоссияVavilov Institute of General Genetics of the Russian Academy of SciencesRussian Federation202301112023276545­-552Copyright © Першина Л.А., Трубачеева Н.В., Шумный В.К., Бадаева Е.Д., 20232023Першина Л.А., Трубачеева Н.В., Шумный В.К., Бадаева Е.Д.Pershina L.A., Trubacheeva N.V., Shumny V.K., Badaeva E.D.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/3925

Интрогрессивная гибридизация является основным методом расширения генетического разнообразия мягкой пшеницы. В качестве источника новых генов для мягкой пшеницы может служить дикий ячмень Hordeum marinum ssp. gussoneanum Hudson (2n = 4x = 28), который характеризуется высокой устойчивостью к стрессовым факторам. Настоящая работа посвящена изучению возможности использования неполного амфиплоида H. marinum ssp. gussoneanum (4x)–T. aestivum (Пиротрикс 28) (2n = 54), носителя цитоплазмы дикого ячменя, в качестве источника хромосом H. marinum для их интрогрессии в геном мягкой пшеницы. С этой целью получены гибриды между линией сорта мягкой пшеницы Пиротрикс 28 (далее П28) и неполным амфиплоидом, а затем среди потомков гибридов проведен отбор цитогенетически стабильных 42­хромосомных растений с высоким уровнем фертильности. С использованием GISH­анализа обнаружено наличие пары хромосом H. marinum в геноме этих растений. По результатам С­окрашивания хромосом установлено, что у этой гибридной линии произошло замещение хромосомы 4B мягкой пшеницы на хромосому дикого ячменя 4Hmar. С помощью хромосом­специфичных SSR­маркеров Xgwm368 и Xgwm6 подтверждено отсутствие хромосомы 4B мягкой пшеницы, а с применением EST­маркеров BAWU808 и BAW112 – наличие хромосомы 4Hmar в геноме выделенной дисомной пшенично­ячменной линии. Изучение этой линии показало, что замещение хромосомы 4B мягкой пшеницы на хромосому дикого ячменя 4Hmar привело к изменению ряда признаков: сильно выраженной антоциановой окраске колеоптиле, характерной для дикого ячменя H. marinum, а также отсутствию пурпурной окраски ушек у основания листьев, которая проявляется у линии сорта пшеницы П28. По высоте растений, числу стеблей и колосьев в растении, числу колосков и зерен в главном колосе, а также числу зерен в растении линия 4Hmar(4B) превосходит родительскую линию П28, а по массе 1000 зерен ей уступает. Цитогенетическая стабильность и фертильность линии 4Hmar(4B) указывают на высокую компенса ционную способность хромосомы 4Hmar ячменя по отношению к хромосоме 4B мягкой пшеницы и гомеологию между этими хромосомами.

Introgressive hybridization is the main method of broadening the genetic diversity of bread wheat. Wild barley Hordeum marinum ssp. gussoneanum Hudson (2n = 4x = 28) has useful agronomical traits, such as high resistance to stress factors, that could be a potential source of new genes for bread wheat improvement. This study aimed to evaluate the possibility of introgression of H. marinum chromosomes into the genome of bread wheat using an incomplete amphiploid H. marinum ssp. gussoneanum (4x)–T. aestivum (Pyrotrix 28) (2n = 54) carrying the cytoplasm of wild barley. For this purpose, we crossed the line of bread wheat variety Pyrotrix 28 with an incomplete amphiploid, and then selected cytogenetically stable 42­chromosome plants with a high level of fertility in hybrid progeny. Genomic in situ hybridization (GISH) revealed a pair of H. marinum chromosomes in the genome of these plants. C­ banding analysis confirmed that bread wheat chromosome 4B was replaced by wild barley chromosome 4Hmar. SSR markers Xgwm368 and Xgwm6 confirmed the absence of chromosome 4B, and EST markers BAWU808 and BAW112 identified chromosome 4Hmar in the genome of the isolated disomic wheat­barley substitution line. The study of this line showed that the substitution of chromosome 4B with chromosome 4Hmar resulted in a change of some morphological traits. It included intense anthocyanin coleoptile coloration, specific for H. marinum, as well as a lack of purple coloration of the ears in the leaf sheath, specific for Pyrotrix 28. Line 4Hmar(4B) showed increased performance for several traits, including plant height, number of spikes and tillers per plant, spikelet and grain number in the main spike, grain number per plant, but it had decreased values of 1000­grain weight compared to wheat. Cytogenetic stability and fertility of line 4Hmar(4B) indicated a high compensation ability of barley 4Hmar for wheat chromosome 4B and confirmed their homeology.

Hordeum marinum ssp. gussoneanumмягкая пшеницапшенично­ячменная замещенная линия 4Hmar(4B)Hordeum marinum ssp. gussoneanumbread wheatwheat­barley substitution line 4Hmar(4B)ReferencesBadaeva E.D., Badaev N.S., Gill B.S., Filatenko A.A. Intraspecific karyotype divergence in Triticum araraticum (Poaceae). Plant Syst. Evol. 1994;192:117-145. DOI: 10.1007/BF00985912.Badaeva E.D., Badaev N.S., Gill B.S., Filatenko A.A. Intraspecific karyotype divergence in Triticum araraticum (Poaceae). Plant Syst. Evol. 1994;192:117-145. DOI: 10.1007/BF00985912.Bothmer R.V., Jacobsen N., Baden C., Jorgensen R.B., Linde-Laursen I. An Ecogeographical Study of the Genus Hordeum. Rome: IBPGR, 1991.Bothmer R.V., Jacobsen N., Baden C., Jorgensen R.B., Linde-Laursen I. An Ecogeographical Study of the Genus Hordeum. Rome: IBPGR, 1991.Davoyan R.O., Bebyakina I.V., Davoyan O.R., Zinchenko A.N., Davoyan E.R., Kravchenko A.M., Zubanova Y.S. The use of synthetic forms in the preservation and exploitation of the gene pool of wild common wheat relatives. Russ. J. Genet. Appl. Res. 2012;2:480-485. DOI: 10.1134/S2079059712060044.Davoyan R.O., Bebyakina I.V., Davoyan O.R., Zinchenko A.N., Davoyan E.R., Kravchenko A.M., Zubanova Y.S. The use of synthetic forms in the preservation and exploitation of the gene pool of wild common wheat relatives. Russ. J. Genet. Appl. Res. 2012;2:480-485. DOI: 10.1134/S2079059712060044.Dospekhov B.A. Methodology of Field Experiments (With the Basics of Statistical Processing of Research Results. Moscow: Agropromizdat Publ., 1985. (in Russian)Dospekhov B.A. Methodology of Field Experiments (With the Basics of Statistical Processing of Research Results. Moscow: Agropromizdat Publ., 1985. (in Russian)Fang Y., Yuan J., Wang Z., Wang H., Xiao J., Yang Z. Development of T. aestivum L.–H. californicum alien chromosome lines and assignment of homoeologous groups of Hordeum californicum chromosomes. J. Genet. Genomics. 2014;41(8):439-477. DOI: 10.1016/j.jgg.2014.06.004.Fang Y., Yuan J., Wang Z., Wang H., Xiao J., Yang Z. Development of T. aestivum L.–H. californicum alien chromosome lines and assignment of homoeologous groups of Hordeum californicum chromosomes. J. Genet. Genomics. 2014;41(8):439-477. DOI: 10.1016/j.jgg.2014.06.004.Garthwaite A.J., von Bothmer R., Colmer T.D. Salt tolerance in wild Hordeum species is associated with restricted entry of Na+ and Cl–Garthwaite A.J., von Bothmer R., Colmer T.D. Salt tolerance in wild Hordeum species is associated with restricted entry of Na+ and Cl–into the shoots. J. Exp. Bot. 2005;56(419):2365-2378. DOI: 10.1093/jxb/eri229.into the shoots. J. Exp. Bot. 2005;56(419):2365-2378. DOI: 10.1093/jxb/eri229.Hao M., Zhang L., Ning S., Huang L., Yuan Z., Wu B., Yan Z., Dai S., Jiang B., Zheng Y., Liu D. The resurgence of introgression breeding, as exemplified in wheat improvement. Front. Plant Sci. 2020;11: 252. DOI: 10.3389/fpls.2020.00252.Hao M., Zhang L., Ning S., Huang L., Yuan Z., Wu B., Yan Z., Dai S., Jiang B., Zheng Y., Liu D. The resurgence of introgression breeding, as exemplified in wheat improvement. Front. Plant Sci. 2020;11: 252. DOI: 10.3389/fpls.2020.00252.Islam S., Malik A.I., Islam A.K.M.R., Colmer T.D. Salt tolerance in a Hordeum marinum–Triticum aestivum amphiploid, and its parents. J. Exp. Bot. 2007;58(5):1219-1229. DOI: 10.1093/jxb/erl293.Islam S., Malik A.I., Islam A.K.M.R., Colmer T.D. Salt tolerance in a Hordeum marinum–Triticum aestivum amphiploid, and its parents. J. Exp. Bot. 2007;58(5):1219-1229. DOI: 10.1093/jxb/erl293.Khlestkina E.K., Antonova E.V., Pershina L.A., Soloviev A.A., Badaeva E.D., Borner A.A., Salina E.A. Variability of Rc (red coleoptile) alleles in wheat and wheat-alien genetic stock collections. Cereal Res. Commun. 2011;39(4):465-474. DOI: 10.1556/CRC.39.2011.4.1.Khlestkina E.K., Antonova E.V., Pershina L.A., Soloviev A.A., Badaeva E.D., Borner A.A., Salina E.A. Variability of Rc (red coleoptile) alleles in wheat and wheat-alien genetic stock collections. Cereal Res. Commun. 2011;39(4):465-474. DOI: 10.1556/CRC.39.2011.4.1.Kobylyanskiy V.D. Biological characters of wild barley species in relation to aims of breeding. Biologicheskiy Zhurnal Armenii = Armenian Journal of Biology. 1967;20(10):41-51. (in Russian)Kobylyanskiy V.D. Biological characters of wild barley species in relation to aims of breeding. Biologicheskiy Zhurnal Armenii = Armenian Journal of Biology. 1967;20(10):41-51. (in Russian)Kruse A. Hordeum × Triticum hybrids. Hereditas. 1973;73(1):157-161. DOI: 10.1111/j.1601-5223.1973.tb01078.x.Kruse A. Hordeum × Triticum hybrids. Hereditas. 1973;73(1):157-161. DOI: 10.1111/j.1601-5223.1973.tb01078.x.Li A., Lui D., Yang W., Kishii M. Synthetic hexaploid wheat: yesterday, today, and tomorrow. Engineering. 2018;4(4):552-558. DOI: 10.1016/j.eng.2018.07.001.Li A., Lui D., Yang W., Kishii M. Synthetic hexaploid wheat: yesterday, today, and tomorrow. Engineering. 2018;4(4):552-558. DOI: 10.1016/j.eng.2018.07.001.Malik A.I., English J.P., Colmer T.D. Tolerance of Hordeum marinum accessions to O2 deficiency, salinity and these stresses combined. Ann. Bot. 2009;103(2):237-248. DOI: 10.1093/aob/mcn142.Malik A.I., English J.P., Colmer T.D. Tolerance of Hordeum marinum accessions to O2 deficiency, salinity and these stresses combined. Ann. Bot. 2009;103(2):237-248. DOI: 10.1093/aob/mcn142.Martín A. Tritordeum: a man-made cereal. In: 4th Conference of Cereal Biotechnology and Breeding Jointly Organized by EUCARPIA Cereal Section November 6–9, 2017, Budapest, Hungary. Budapest, 2017;5-6.Martín A. Tritordeum: a man-made cereal. In: 4th Conference of Cereal Biotechnology and Breeding Jointly Organized by EUCARPIA Cereal Section November 6–9, 2017, Budapest, Hungary. Budapest, 2017;5-6.Martín A., Álvarez J.B., Martín L.M., Barro F., Ballesteros J. The development of tritordeum: a novel cereal for food processing. J. Cereal Sci. 1999;30(2):85-95. DOI: 10.1006/jcrs.1998.0235.Martín A., Álvarez J.B., Martín L.M., Barro F., Ballesteros J. The development of tritordeum: a novel cereal for food processing. J. Cereal Sci. 1999;30(2):85-95. DOI: 10.1006/jcrs.1998.0235.Melz G., Thiele V. Chromosome locations of genes controlling ‘purple leaf base’ in rye and wheat. Euphytica. 1990;49:155-159. DOI: 10.1007/BF00027265.Melz G., Thiele V. Chromosome locations of genes controlling ‘purple leaf base’ in rye and wheat. Euphytica. 1990;49:155-159. DOI: 10.1007/BF00027265.Molnár-Láng M., Linc G. Wheat-barley hybrids and introgression lines. In: Molnár-Láng M., Ceoloni C., Dolezel J. (Eds.). Alien Introgression in Wheat. Cytogenetics, Molecular Biology and Genomics. Cham: Springer, 2015;315-346. DOI: 10.1007/978-3-319-234946_12.Molnár-Láng M., Linc G. Wheat-barley hybrids and introgression lines. In: Molnár-Láng M., Ceoloni C., Dolezel J. (Eds.). Alien Introgression in Wheat. Cytogenetics, Molecular Biology and Genomics. Cham: Springer, 2015;315-346. DOI: 10.1007/978-3-319-234946_12.Molnár I., Linc G., Dulai S., Nagy E.D., Molnár-Láng M. Ability of chromosome 4H to compensate for 4D in response to drought stress in a newly developed and identified wheat-barley 4H(4D) disomic substitution line. Plant Breed. 2007;126(4):369-374. DOI: 10.1111/j.1439-0523.2007.01300.Molnár I., Linc G., Dulai S., Nagy E.D., Molnár-Láng M. Ability of chromosome 4H to compensate for 4D in response to drought stress in a newly developed and identified wheat-barley 4H(4D) disomic substitution line. Plant Breed. 2007;126(4):369-374. DOI: 10.1111/j.1439-0523.2007.01300.Molnár-Láng M., Linc G., Szakács É. Wheat-barley hybridization: the last 40 years. Euphytica. 2014;195:315-329. DOI: 10.1007/s10681013-1009-9.Molnár-Láng M., Linc G., Szakács É. Wheat-barley hybridization: the last 40 years. Euphytica. 2014;195:315-329. DOI: 10.1007/s10681013-1009-9.Pershina L.A., Numerova O.M., Belova L.I., Devyatkina E.P., Rakovtseva T.S., Shumny V.K. Expression of fertility during morphogenesis in self-pollinated backcrossed progenies of barley-wheat amphiploids [Hordeum geniculatum All. (2n = 28) × Triticum aestivum L. (2n = 42)] (2n = 70). Russ. J. Genet. 2004;40:510-514. DOI: 10.1023/B:RUGE.0000029153.61243.c2.Pershina L.A., Numerova O.M., Belova L.I., Devyatkina E.P., Rakovtseva T.S., Shumny V.K. Expression of fertility during morphogenesis in self-pollinated backcrossed progenies of barley-wheat amphiploids [Hordeum geniculatum All. (2n = 28) × Triticum aestivum L. (2n = 42)] (2n = 70). Russ. J. Genet. 2004;40:510-514. DOI: 10.1023/B:RUGE.0000029153.61243.c2.Pershina L.A., Devyatkina E.P., Belova L.I., Trubacheeva N.V., Arbuzova V.S., Kravtsova L.A. Features of alloplasmic wheat–bar ley substitution and addition lines (Hordeum marinum subsp. gusso neanum)-Triticum aestivum. Russ. J. Genet. 2009;45(10):1223-1229. DOI: 10.1134/S102279540910010X.Pershina L.A., Devyatkina E.P., Belova L.I., Trubacheeva N.V., Arbuzova V.S., Kravtsova L.A. Features of alloplasmic wheat–bar ley substitution and addition lines (Hordeum marinum subsp. gusso neanum)-Triticum aestivum. Russ. J. Genet. 2009;45(10):1223-1229. DOI: 10.1134/S102279540910010X.Pershina L.A., Belova L.I., Trubacheeva N.V., Osadсhaya T.S., Shumny V.K., Belan I.A., Rosseeva L.P., Nemchenko V.V., Abakumov S.N. Alloplasmic recombinant lines (H. vulgare)-T. aestivum with 1RS.1BL translocation: initial genotypes for production of common wheat varieties. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2018;22(5):544-552. DOI: 10.18699/VJ18.393. (in Russian)Pershina L.A., Belova L.I., Trubacheeva N.V., Osadсhaya T.S., Shumny V.K., Belan I.A., Rosseeva L.P., Nemchenko V.V., Abakumov S.N. Alloplasmic recombinant lines (H. vulgare)-T. aestivum with 1RS.1BL translocation: initial genotypes for production of common wheat varieties. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2018;22(5):544-552. DOI: 10.18699/VJ18.393. (in Russian)Rahman S., Islam S., Yu Z., She M., Nevo E., Ma W. Current progress in understanding and recovering the wheat genes lost in evolution and domestication. Int. J. Mol. Sci. 2020;21(16):5836. DOI: 10.3390/ijms21165836.Rahman S., Islam S., Yu Z., She M., Nevo E., Ma W. Current progress in understanding and recovering the wheat genes lost in evolution and domestication. Int. J. Mol. Sci. 2020;21(16):5836. DOI: 10.3390/ijms21165836.Rey M.-D., Calderón M.-C., Rodrigo M.J., Zacarías L., Alós E., Prieto P. Novel bread wheat lines enriched in carotenoids carrying Hordeum chilense chromosome arms in the ph1b background. PLoS One. 2015;10(8):e0134598. DOI: 10.1371/journal.pone.0134598.Rey M.-D., Calderón M.-C., Rodrigo M.J., Zacarías L., Alós E., Prieto P. Novel bread wheat lines enriched in carotenoids carrying Hordeum chilense chromosome arms in the ph1b background. PLoS One. 2015;10(8):e0134598. DOI: 10.1371/journal.pone.0134598.Röder M.S., Korzun V., Wendehake K., Plaschke J., Tixier M.H., Leroy P., Ganal M.W. A microsatellite map of wheat. Genetics. 1998; 149(4):2007-2023. DOI: 10.1093/genetics/149.4.2007.Röder M.S., Korzun V., Wendehake K., Plaschke J., Tixier M.H., Leroy P., Ganal M.W. A microsatellite map of wheat. Genetics. 1998; 149(4):2007-2023. DOI: 10.1093/genetics/149.4.2007.Rubiales D., Moral A. Resistance of Hordeum chilense against loose smuts of wheat and barley (Ustilago tritici and U. nuda) and its expression in amphiploids with wheat. Plant Breed. 2011;130(1): 101-103. DOI: 10.1111/j.1439-0523.2010.01818.x.Rubiales D., Moral A. Resistance of Hordeum chilense against loose smuts of wheat and barley (Ustilago tritici and U. nuda) and its expression in amphiploids with wheat. Plant Breed. 2011;130(1): 101-103. DOI: 10.1111/j.1439-0523.2010.01818.x.Said M., Cabrera A. A physical map of chromosome 4Hch from H. chilense containing SSR, STS and EST-SSR molecular markers. Euphytica. 2009;167:253-259. DOI: 10.1007/s10681-009-9895-6.Said M., Cabrera A. A physical map of chromosome 4Hch from H. chilense containing SSR, STS and EST-SSR molecular markers. Euphytica. 2009;167:253-259. DOI: 10.1007/s10681-009-9895-6.Shchapova A.I., Kravtsova L.A. Cytogenetics of Wheat-rye Hybrids. Novosibirsk: Nauka Publ., 1990. (in Russian) Taketa S., Takeda K. Production and characterization of a complete set of wheat-wild barley (Hordeum vulgare ssp. spontaneum) chromosome addition lines. Breed Sci. 2001;51(3):199-206. DOI: 10.1270/jsbbs.51.199.Shchapova A.I., Kravtsova L.A. Cytogenetics of Wheat-rye Hybrids. Novosibirsk: Nauka Publ., 1990. (in Russian) Taketa S., Takeda K. Production and characterization of a complete set of wheat-wild barley (Hordeum vulgare ssp. spontaneum) chromosome addition lines. Breed Sci. 2001;51(3):199-206. DOI: 10.1270/jsbbs.51.199.Trubacheeva N.V., Badaeva E.D., Osadchaya T.S., Pershina L.A. Use of H. vulgare EST markers, GISH and C-banding to study bread wheat–H. marinum subsp. gussoneanum (2n = 28) introgression lines. Cereal Res. Commun. 2019;47(4):593-603. DOI: 10.1556/0806.47.2019.37.Trubacheeva N.V., Badaeva E.D., Osadchaya T.S., Pershina L.A. Use of H. vulgare EST markers, GISH and C-banding to study bread wheat–H. marinum subsp. gussoneanum (2n = 28) introgression lines. Cereal Res. Commun. 2019;47(4):593-603. DOI: 10.1556/0806.47.2019.37.

The authors declare that there are no conflicts of interest present.