Creation of new tomato forms with fungal disease resistance genes based on marker selection
https://doi.org/10.18619/2072-9146-2021-6-16-21
Abstract
Relevance. The presented studies are aimed at obtaining new forms of tomato with a complex of genes for resistance to fungal diseases in combination with a standard type of bush and dark coloring of fruits based on marker-mediated selection.
Methodology. The biological objects of the study are varieties and hybrid forms of tomato from the collection of the Michurinsky SAU. Molecular genetic analysis was performed using the following methods. DNA extraction was carried out from young leaves using a kit for isolation of NC Sample NC manufactured by Agrodiagnostika LLC according to the manufacturer's protocol. Fermentas production kits were used for PCR. Identification of the cladosporosis resistance gene was Cf-19 performed using the DNA marker R7. The presence of a fusarious wilting resistance gene was determined by a I-2/5 marker. The amplification results were visualized by agarose gel electrophoresis.
Results. During the research, a collection of varieties and hybrid forms of tomato of the Michurinsky GAU was analyzed in order to identify genes for resistance to cladosporiosis Cf-19 and fusarium wilt I-2. A total of 52 genotypes were analyzed. It was found that most samples (41 samples) are characterized by a heterozygous state of the Cf-19 gene. All indeterminant and semi-determinant forms had both alleles. Of the 23 determinant forms presented in the collection, 10 had only one allele corresponding to recessive homozygote. Among all analyzed tomato genotypes, no dominant homozygous forms were noted. The study of the collection revealed several alleles of the I-2 gene. In total, four fragments corresponding to various alleles were amplified. A total of 50 resistant genotypes have been identified in the collection. Two alleys of the I-2 gene (633/693 bp) were identified in 42 tomato samples. Four varieties are homozygous in one allele (633 bp), which determines resistance. Three varieties have a second resistance allele (566 bp). One genotype has only an allele defining susceptibility (693 bp). On the basis of molecular analysis, as well as an assessment of the type of bush and fetal color, initial forms were selected with subsequent hybridization. 67 hybrid tomato plants were obtained. Evaluation of the presence of resistance genes showed that most of the resulting hybrids are resistant to cladosporiosis and fuzariosis. This is due to the presence of dominant alleles of Cf-19 and I-2 genes in a heterozygous state. Among the resulting hybrids, plants with a bark type of bush were identified. A total of 13 such plants were obtained.
Conclusion. Thus, the work carried out allowed to obtain hybrid forms of tomato combine the signs of resistance to two pathogens of fungal diseases and the stem type of the bush. These forms are planned to be used in further selection work.
About the Authors
I. N. ShamshinRussian Federation
Ivan N. Shamshin – Cand. Sci. (Biology), Head of the laboratory of the molecular and genetic analysis of fruit plants
Scopus ID 56708633300, Researcher ID AAZ-9047-2021
101, Internationalnaya Str., Michurinsk. Tambov region, Russian Federation, 393760
E. V. Grosheva
Russian Federation
Ekaterina V. Grosheva – Laboratory Assistant of the laboratory of molecular genetic analysis of fruit plants
Researcher ID ААА7122-2020
101, Internationalnaya Str., Michurinsk. Tambov region, Russian Federation, 393760
M. V. Maslova
Russian Federation
Marina V. Maslova – Cand. Sci. (Agriculture), Senior Researcher of Biofotonika laboratory
Researcher ID E-4506-2015
101, Internationalnaya Str., Michurinsk. Tambov region, Russian Federation, 393760
R. M. Samoilova
Russian Federation
Rufima M. Samoilova – Educational Master of the Department of horticulture, biotechnology and crop selection
101, Internationalnaya Str., Michurinsk. Tambov region, Russian Federation, 393760
References
1. Litvinov S.S. [Phytosanitary problems in modern vegetable production]. Plant protection and quarantine. 2015; 4. (in Russ.)
2. Seitbattalova A.I., Sadanov A.K., Shemshura O.N., Kaptagai R.Zh., Ismailova E.T. [The influence of pre-treatment of seeds with issop extract on the resistance of tomatoes to mushroom diseases in the field]. News of the National Academy of Sciences of the Republic of Kazakhstan series biology and medicine. 2017;5(233):222- 227. (in Russ.)
3. Polyxenova V.D. [Induced resistance of plants to pathogens and abiotic stress factors: on the example of tomato]. Bulletin of the Belarusian State University. 2009;(1):48-60. (in Russ.)
4. Iida Y., van ‘t Hof P., Beenen H., Mesarich C., Kubota M., Stergiopoulos I., de Wit P. J. Novel mutations detected in avirulence genes overcoming tomato Cf resistance genes in isolates of a Japanese population of Cladosporium fulvum. PloS one. 2015;10(4). DOI 10.1371/journal.pone.0123271
5. Thomas C.M., Jones D.A., Parniske M., Harrison K., Balint-Kurti P. ., Hatzixanthis K., Jones J.D. Characterization of the tomato Cf-4 gene for resistance to Cladosporium fulvum identifies sequences that determine recognitional specificity in Cf-4 and Cf-9. The Plant Cell. 1997;9(12):2209-2224. DOI 10.1105/tpc.9.12.2209.
6. Chai X., Xu X., Wang D., Xue D., Li J. Mapping and candidate gene screening of Cladosporium fulvum resistance gene Cf-12 in tomato (Solanum lycopersicum) by high-throughput sequencing. Plant Breeding 2020;139(5):977-987. DOI 10.1111/pbr.12852
7. Zhang D., Bao Y., Sun Y., Yang H., Zhao T., Li H., Xu X. Comparative transcriptome analysis reveals the response mechanism of Cf-16-mediated resistance to Cladosporium fulvum infection in tomato. BMC plant biology. 2020;20(1):1-16. DOI 10.1186/s12870-020-2245-5
8. Liu G., Liu J., Zhang C., You X., Zhao T., Jiang J., Xu X. Physiological and RNAseq analyses provide insights into the response mechanism of the Cf-10-mediated resistance to Cladosporium fulvum infection in tomato. Plant molecular biology. 2018;96(4):403-416. DOI 10.1007/s11103-018-0706-0.
9. Kruijt M., Brandwagt B.F., De Wit P.J. Rearrangements in the Cf-9 disease resistance gene cluster of wild tomato have resulted in three genes that mediate Avr9 responsiveness. Genetics. 2004;168(3):1655-1663. DOI 10.1534/genetics.104.028985.
10. Jones D.A., Dickinson M.J., Balint-Kurti P.J., Dixon M.S., Jones J.D.G. Two complex resistance loci revealed in tomato by classical and RFLP mapping of the Cf-2, Cf-4, Cf-5, and Cf-9 genes for resistance to Cladosporium fulvum. Molecular Plant Microbe Interactions. 1993;6(3):348-357.
11. Dixon M.S., Jones D.A., Keddie J.S., Thomas C.M., Harrison K., Jones J.D. The tomato Cf-2 disease resistance locus comprises two functional genes encoding leucine-rich repeat proteins. Cell. 1996;84(3):451-459. DOI 10.1016/S0092-8674(00)81290-8.
12. Xue D.Q., Chen X.L., Zhang H., Chai X.F., Jiang J.B., Xu X.Y., Li J.F. Transcriptome analysis of the Cf-12-mediated resistance response to Cladosporium fulvum in tomato. Frontiers in plant science. 2017;(7):2012. DOI 10.3389/fpls.2016.02012.
13. Zhao T., Liu W., Zhao Z., Yang H., Bao Y., Zhang D., Xu X. Transcriptome profiling reveals the response process of tomato carrying Cf-19 and Cladosporium fulvum interaction. BMC plant biology. 2019;19(1):1-12. DOI 10.1186/s12870-019-2150-y.
14. Jordatilde D.O., de Almeida C.M.A., Malafaia C.B., da Silva M.L.R.B., dos Santos Correia M.T., de Menezes Lima V.L., da Silva, M.V. Identification of races 1, 2 and 3 of Fusarium oxysporum f. sp. lycopersici by molecular markers. African Journal of Microbiology Research. 2013;7(20):2324-2331. DOI10.5897/AJMR12.2234.
15. Gonzalez-Cendales Y., Catanzariti A.M., Baker B., Mcgrath D.J., Jones D.A.. Identification of I-7 expands the repertoire of genes for resistance to Fusarium wilt in tomato to three resistance gene classes. Molecular Plant Pathology. 2016;17(3):448-463. DOI10.1111/mpp.12294
16. Eroshevskaya A.S., Egorova A.A., Milyukova N.A., Pyrsikov A.S. Molecular-genetic analysis of tomato hybrids F1 on resistance to fuzariosis. Potatoes and vegetables. 2021;(5):37-40. (In Russ.)
17. Takken F., Rep M. The arms race between tomato and Fusarium oxysporum. Molecular plant pathology. 2010;11(2):309-314.
18. Catanzariti A.M., Lim G.T., Jones D.A. The tomato I-3 gene: a novel gene for resistance to Fusarium wilt disease. New Phytologist. 2015;207(1):106-118. DOI 10.1111/nph.13348.
19. Gardner R.G. Mountain Spring'tomato; NC8276 and NC84173 tomato breeding lines. HortScience. 1992;27(11);1233-1234.
20. El Mohtar C.A., Atamia H.S., Dagher R.B., Abou-Jawdah Y., Salus M.S., Maxwell D.P. Marker-assisted selection of tomato genotypes with the I-2 gene for resistance to Fusarium oxysporum f. sp. lycopersici race 2. Plant disease. 2007;91(6):758-762. DOI 10.1094/PDIS-91-6-0758.
21. Simsek D., Pinar H., Mutlu N. Development of Fusarium oxysporum f. Sp. Lycopersici (FOL) and Fusarium oxysporum f. Sp. Radicis lycopersici (FORL) resistant tomato lines with the aid of marker assisted selection. Current Trends in Natural Sciences. 2018;7(13):281-285.
22. Pidigam S., Thuraga V., Munnam S.B., Amarapalli G., Kuraba G., Pandravada S.R., Sudini H.K. Genetic diversity, population structure and validation of SSR markers linked to Sw-5 and I-2 genes in tomato germplasm. Physiology and Molecular Biology of Plants. 2021;1-16. DOI 10.1007/s12298-021-01037-8.
23. Adzhieva V.F., Grushetskaya Z.E., Malyshev S.V., Nekrashevich N.A., Babak O.G., Kilchevsky A.V. [Creation of a complex of DNA markers for tomato genes that determine the content of carotenoids and resistance to diseases and pests]. II International. научн. - практ. conf. "Modern trends in the selection and seed production of vegetable crops. Traditions and perspectives. " Moscow, Russia. August 2-4, 2010; p. 47. (in Russ.)
24. Yu S.C., Zou Y.M. A co-dominant molecular marker of Fusarium wilt resistance gene I-2 derived from gene sequence in tomato. Yi Chuan= Hereditas. 2008;30(7):926-932. DOI 10.3724/sp.j.1005.2008.00926.
25. Arens P., Mansilla C., Deinum D., Cavellini L., Moretti A., Rolland S., Vosman B. Development and evaluation of robust molecular markers linked to disease resistance in tomato for distinctness, uniformity and stability testing. Theoretical and applied genetics. 2010;120(3):655-664. DOI 10.1007/s00122-009-1183-2.
26. Zhao T., Jiang J., Liu G., He S., Zhang H., Chen X., Xu X. Mapping and candidate gene screening of tomato Cladosporium fulvum-resistant gene Cf-19, based on highthroughput sequencing technology. BMC plant biology. 2016;16(1):1-10. DOI 10.1186/s12870-016-0737-0.
27. Nevamea A. Y. M., Xiaa L., Wentinga Z., Nchongbohc C. G., Wenhua L., Hasand M. M., Longtinga S. Validation of some disease-resistance molecular markers associated with multiple diseases in tomato for marker-assisted selection program. Scienceasia. 2020;46(1):19-29. DOI 10.2306/scienceasia1513-1874.2020.006.
28. Shamshin I.N., Maslova M.V., Gryazneva Yu.V. [Analysis of genetic collection of tomato varieties and hybrid forms by resistance to cladosporiosis using DNA markers]. Proceedings on applied botany, genetics and breeding. 2019;180(3):63-70. DOI: 10.30901/2227-8834-2019-3-63-70 (in Russ.)
Review
For citations:
Shamshin I.N., Grosheva E.V., Maslova M.V., Samoilova R.M. Creation of new tomato forms with fungal disease resistance genes based on marker selection. Vegetable crops of Russia. 2021;(6):16-21. (In Russ.) https://doi.org/10.18619/2072-9146-2021-6-16-21