Cytogenetic and flow cytometric analysis of new allotetraploid forms derived from wild cotton species with A1 and G genomes

Sevara Arslanova; Dilrabo Ernazarova; Feruza Rafiyeva; Abdulqahhor Toshpulatov; Madina Kholova; Barno B. Oripova; Faxriddin Kushanov

doi:10.66432/kdhxfr21

Sevara K. Arslanova * Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan , Dilrabo K. Ernazarova Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan , Samarkand State University named after Sharof Rashidov , Feruza U. Rafiyeva Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan , Abdulqahhor Kh. Toshpulatov Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan , Madina D. Kholova Chirchik Branch of Tashkent State Medical University , Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan , Barno B. Oripova Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan , Tashkent State Medical University , Faxriddin N. Kushanov Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan , National University of Uzbekistan

* Corresponding author: arslanovasevara87@gmail.com

DOI:

https://doi.org/10.66432/kdhxfr21

Keywords:

Cotton, Gossypium L., G. australe, hybridization, allopolyploidy, cytogenetics, chromosome, meiosis, mitosis.

Abstract

Wild Australian and Afro-Asian cotton species are characterized by their natural resistance to diseases and pests, serving as valuable genetic resources for the introgression of these beneficial traits into cultivated cotton varieties. This study presents the results of research aimed at determining the ploidy level of novel allopolyploid hybrid forms derived from wild Australian (Gossypium nelsonii) and Afro-Asian (G. herbaceum) cotton species using interspecific hybridization and experimental polyploidy methods. Cytogenetic analyses demonstrated that the interspecific F₁ hybrids possess a diploid chromosome set (2n=2x=26), whereas the synthetic F₁C hybrids developed via experimental polyploidy exhibited a multiplied chromosome set (2n=4x=52). Furthermore, flow cytometry analysis conclusively confirmed this chromosome doubling, showing that the nuclear DNA content doubled from an average value of - 12.8 in the diploid parental forms to 25.492 in the F₁C allotetraploids. The study evaluated the interspecific crossability of species belonging to different genomic groups and successfully generated genetically stable synthetic allotetraploids. This demonstrates the feasibility of utilizing wild diploid species in genetic and breeding research.

Author Biographies

Feruza U. Rafiyeva, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan

Doctor of Biological Sciences, Leading Researcher at the Laboratory of Plant Genomic Studies, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan.
Abdulqahhor Kh. Toshpulatov, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan

Junior Researcher at the Laboratory of Plant Genomic Studies, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan.
Madina D. Kholova, Chirchik Branch of Tashkent State Medical University, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan

PhD in Biological Sciences, Senior Researcher at the Laboratory of Plant Genomic Studies, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan.
Faxriddin N. Kushanov, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan, Tashkent Region, Kibray District, 102122, Uzbekistan, National University of Uzbekistan

Doctor of Biological Sciences, Professor at the Laboratory of Plant Genomic Studies, Institute of Genetics and Plant Experimental Biology, Academy of Sciences of the Republic of Uzbekistan.

References

1. Mammadov, J., Buyyarapu, R., Guttikonda, S.K., Parliament, K., Abdurakhmonov, I.Y. and Kumpatla, S.P. (2018) Wild relatives of maize, rice, cotton, and soybean: treasure troves for tolerance to biotic and abiotic stresses. Front. Plant. Sci. 9, 886.

2. Mehetre SS, Patil SC, Pawar SV, Pardedhi SU, Shinde GC, Aher AR (2004) Ovulo embryo cultured hybrid between amphi-diploid (Gossypium arboreum × Gossypium anomalum) and Gossypium hirsutum. Curr Sci 87:286–289

3. Kunbo, W. and Jonathan, W. (2018) Designations for individual genomes and chromosomes in Gossypium. J. Cotton Res. 1, 3.

4. Wendel, J.F., Brubaker, C.L. and Seelanan, T. (2010) The Origin and Evolution of Gossypium. In Physiology of Cotton (Stewart, J.M., Oosterhuis, D.M., Heitholt, J.J. & Mauney, J.R. eds), pp. 1–18. Netherlands, Dordrecht: Springer.

5. Chen, Y., Wang, Y., Wang, K., Zhu, X., Guo, W., Zhang, T. and Zhou, B. (2014) Construction of a complete set of alien chromosome addition lines from Gossypium australe in Gossypium hirsutum: morphological, cytological, and genotypic characterization. Theor. Appl. Genet. 127, 1105–1121.

6. Liu, Q., Chen, Y., Chen, Y., Wang, Y., Chen, J., Zhang, T. and Zhou, B. (2015) A new synthetic allotetraploid (A1A1G2G2) between Gossypium herbaceum and G. australe: bridging for simultaneously transferring favorable genes from these two diploid species into upland cotton. PLoS ONE, 10, e0123209.

7. Wang, Y., Feng, S., Li, S., Tang, D., Chen, Y., Chen, Y. and Zhou, B. (2018c) Inducement and identification of chromosome introgression and translocation of Gossypium australe on Gossypium hirsutum. BMC Genom.19, 15.

8. Benkang, G. and Cun, M. (1996) China Cotton Breeding resistant to Disease. Nanjing: Jiangsu Science and Technology Publishing Press.

9. Gu, B., Li, J., Gu, P., Qian, S., Huang, J., Zhou, B., Peng, Y. et al. (1993) The identification of resistance to Verticillium and Fusarium wlt in Gossypium genus wild species. Jiangsu Agricul. Sci. 5, 36–37.

10. Wang, C., Ulloa, M., Duong, T. and Roberts, P.A. (2018a) Quantitative trait loci mapping of multiple independent loci for resistance to Fusarium oxysporum f. sp. vasinfectum races 1 and 4 in an interspecific cotton population. Phytopathology, 108, 759–767.

11. Benbouza, H., Lognay, G., Scheffler, J., Baudoin, J.P. and Mergeai, G. (2009) Expression of the ‘glanded-plant and glandless-seed’ trait of Australian diploid cottons in different genetic backgrounds. Euphytica, 165, 211–221.

12. Wendel J F, Cronn R C, 2003. poliploidy and the evolutionary history of cotton. Adv. Agron. 78(78): 139–186.

13. Wendel J.F., Brubaker C.L., and Seelanan T. The origin and evolution of Gossypium // Physiology of cotton. Springer, Dordrecht, The Netherlands. 2010. -P. 1–18.

14. Gai Huang, Jin-Quan Huang, Xiao-Ya Chen, Yu-Xian Zhu. Recent Advances and Future Perspectives in Cotton Research. Annual Review of Plant Biology 2021 72:1, 437-462 https://doi.org/10.1146/annurev-arplant-080720-113241

15. Zhou T, Wang N, Wang Y, Zhang XL, Li BG, Li W, Su JJ, Wang CX, Zhang A, Ma XF, Li ZH. Nucleotide Evolution, Domestication Selection, and Genetic Relationships of Chloroplast Genomes in the Economically Important Crop Genus Gossypium. Front Plant Sci. 2022 Apr 15; 13:873788. doi: 10.3389/fpls.2022.873788.pp. 1-15.

16. Kulkarni VN, Khadi BM, Maralappanavar MS, Deshapande LA, Narayanan S (2009). The worldwide gene pools of Gossypium arboreum L. and G. herbaceum L., and their improvement. In: Paterson AH, editor. Genetics and Genomics of Cotton. Springer Science Business Media, LLC. pp. 69–97.

17. Ranjan A., Nigam D., Asif M.H. et al., Genome wide expression profiling of two accession of G. herbaceum L. in response to drought. BMC Genomics 13, 94 (2012). https://doi.org/10.1186/1471-2164-13-94 P. 1-22.

18. Liu G., Mei, H., Wang, S. et al. Association mapping of seed oil and protein contents in upland cotton. Euphytica 205, 637–645 (2015). https://doi.org/10.1007/s10681-015-1450-z.

19. Wang Y, Feng S, Li S, Tang D, Chen Y, Chen Y, Zhou B. Inducement and identification of chromosome introgression and translocation of Gossypium australe on Gossypium hirsutum. BMC Genomics. 2018 Jan 4;19(1):15. doi: 10.1186/s12864-017-4398-7.

20. Yin X, Zhan R, He Y, Song S, Wang L, Ge Y, et al. (2020) Morphological description of a novel synthetic allotetraploid (A1A1G3G3) of Gossypium herbaceum L. and G. nelsonii Fryx. suitable for disease-resistant breeding applications. PloS ONE15(12): e0242620. https://doi.org/ 10.1371/journal.pone.0242620.

21. 24 Maru, B., Parihar, A., Kulshrestha, K. et al. Induction of poliploidy through colchicine in cotton (G. herbaceum) and its conformity by cytology and flow cytometry analyses. J Genet 100, 52 (2021). doi.org/10.1007/s12041-021-07-z.

22. Namazov SHE, Mamarahimov BI, Matyoqubov SK, Sodiqova OH, Karimov SH, Bobokhujayev SHU, Sanamyan MF, Darmanov MM (2024). Cytogenetic analysis of cotton hybrids derived from introgressive lines. SABRAO J. Breed. Genet. 56(3): 906-917. http://doi.org/10.54910/sabrao2024.56.3.1.

23. Khidirov, M.T.; Ernazarova, D.K.; Rafieva, F.U.; Ernazarova, Z.A.; Toshpulatov, A.K.; Umarov, R.F.; Kholova, M.D.; Oripova, B.B.; Kudratova, M.K.; Gapparov, B.M.; et al. Genomic and Cytogenetic Analysis of Synthetic poliploids between Diploid and Tetraploid Cotton (Gossypium) Species. Plants 2023, 12, 4184. https://doi.org/10.3390/plants12244184.

24. Abdurakhmonov IY. 2014 -Editor of “World Cotton Germpasm Resources” book, Intech, Croatia (http://www.intechopen.com).

25. Abdurakhmonov IY, Ayubov MS, Ubaydullaeva KA, Buriev ZT, Shermatov SE, Ruziboev HS, Shapulatov UM, Ulloa M, Yu JZ, Percy RG, Saha S, Sharma GC, Devor EJ, Sripathi VR, Kumpatla SP, Van Der Krol A, Kater HD, Khamidov K, Salikhov SI, Jenkins J, Abdukarimov A and Pepper AE. 2016. RNA interference for functional genomics and improvement of cotton (Gossypium spp.). Front. Plant Sci. 7:202. doi: 10.3389/fpls.2016.00202.

26. Kushanov FN, Buriev ZT, Shermatov SE, Turaev OS, Norov TM, Pepper AE, et al. (2017). QTL mapping for flowering-time and photoperiod insensitivity of cotton Gossypium darwinii Watt. PLoS ONE 12(10): e0186240. https://doi.org/10.1371/journal.pone.0186240.

27. Dospexov B.A. Metodika polevogo opita. Moskva, Agropromizdat, 1985. 351 p.

28. Pausheva Z.P. Praktikum po sitologii. -M., Kolos, 1988. -287 p.

29. Galán Sauco, V., Grajal Martin, M. J., Fernández-Galvan, D., Coello-Torres, A., Juárez, J., Navarro, L. Occurrence of spontaneous tetraploid nucellar mango plants //HortScience. – 2001. – T. 36. – №. 4. – P. 755-757.

30. Doležel J., Greilhuber J., Suda J. Estimation of nuclear DNA content in plants using flow cytometry //Nature protocols. – 2007. – Т. 2. – №. 9. – P. 2233-2244.

31. Dhooghe, E., K.Van Laere, T.Eeckhaut, L.Leus and J.Van Huylenbroeck (2011) Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tissue Organ Cult. 104: 359–373.).