Katlanmış Diploid Biberlerde Morfolojik, Sitolojik ve DNA Miktarına Göre Yapılan İnceleme Sonuçlarının Karşılaştırılması

Öz

Kapya biberlerde, tohumsuz meyve veren çeşitler geliştirmeye yönelik ıslah programlarında kullanılacak tetraploid ebeveynlerin elde edilmesi amacıyla bir dizi çalışma yapılmıştır. Bu kapsamda gerçekleştirilen kolhisin uygulamaları sonucunda gelişen bitkiler/sürgünler üzerindeki çiçeklerin kendilenmesiyle tohumlar elde edilmiştir. Olası tetraploid tohumlar ekilmiş ve bitkiler fidelikte yetiştirilmiştir. Polikarbon serada yerlerine dikilen bitkiler çeşitli morfolojik özellikleri bakımından gözlemlenmiş (yaprak alanı, çiçek çapı, meyve eni / boyu, tohum oluşumu), bu bitkilerin yapraklarında birim alandaki stoma sayıları ve boyutları kayıt altına alınmış, ayrıca genç fide döneminde alınan örneklerde flow sitometri yöntemi ile ploidi seviyesi belirlenmiştir. Böylece morfolojik, sitolojik ve DNA miktarı bakımından yapılan incelemelerin sonuçları bir arada değerlendirilebilmiştir. Üç farklı yöntemden elde edilen sonuçlar karşılaştırılarak aralarındaki ilişkiler araştırılmıştır. Çalışma sonucunda en güvenilir bulguları veren flow sitometri yöntemi esas alınarak, morfolojik ve sitolojik tahminlerin tutarlılığı yorumlanmıştır. Morfolojik ve sitolojik yollarla geniş bir bitki popülasyonu içinde ön seleksiyonun yapılabileceği, seçilen bitkilerin ploidi seviyesinin teyit edilmesi için flow sitometri yoluyla analiz yapılmasının gerekli olduğu sonucuna ulaşılmıştır.

Anahtar Kelimeler

• Capsicum annuum
• flow sitometri
• stoma
• ploidi

Comparison of The Examination Results According to Morphological, Cytological and DNA Amount in Doubled Diploid Peppers

Öz

A series of studies have been carried out to obtain tetraploid parents to be used in breeding programs to develop seedless fruiting varieties in capia peppers. In this context, the seeds obtained by the selfing of flowers on the plants / shoots developed as a result of colchicine applications were sown and the seedlings were grown in the seedling greenhouse. Seedlings planted in the polycarbonate greenhouse were observed in terms of various morphological characteristics (leaf area, flower diameter, fruit width and length, seed formation) and the number of stomas per unit area and size in the leaves of these plants were determined. Also, the ploidy level of plants was determined by flow cytometry method in the samples taken during the young seedling period. Thus, the results of the examinations made in terms of morphological, cytological and DNA amount could be evaluated together. At the end of the study, the consistency of morphological and cytological predictions was interpreted based on the flow cytometry method, which gave the most reliable result. It has been concluded that preselection can be made in a large plant population by morphological and cytological means, but analysis by flow cytometry is necessary to confirm the ploidy level of the selected plants.

Anahtar Kelimeler

• Capsicum annuum
• flow cytometry
• stoma
• ploidy

Kaynakça

1. Alsahlany, M., Zarka, D., Coombs, J., Douches, D.S. 2019. Comparison of methods to distinguish diploid and tetraploid potato in applied diploid breeding. American Journal of Potato Research 96 (3):244-254.
2. Amanah, H.A., Arumingtyas, E.L., Indriyani, S. 2016. Chromosome analysis of cayenne pepper (Capsicum frustescens L.) in colchicine induced mutation. Journal of Applied Horticulture 18(3):217-220.
3. Badu, M., Tripathy, B., Sahu, G.S., Kumar, Jena, A.K. 2017. Role of doubled haploids in vegetable crop improvement. Journal of Pharmacognosy and Phytochemistry 6(6):384-389.
4. Compton, M.E., Gray, D.J., Elmstrom, G.W. 1996. Identification of tetraploid regeneration from cotyledons of diploid watermelon cultured in vitro. Euphytica 87:165-172.
5. FAO, 2020. FAOSTAT (https://www.fao.org/ faostat/en/#data/qcl; Erişim:30.07.2022
6. Grimball, P.C. 1971. Production of seedless water melons. USDA, Technical Bulletin, No:1425.
7. Ishikawa, K., Mishiba, K., Yoshida, H., Nunomura, O. 1997. Establishment of tetraploid plants of Capsicum annuum L. by colchicines treatment with the analysis of flow cytometry. Capsicum and Eggplant Newsletter 16:44-47.
8. Ishikawa, K., Kuboki, H., Mishiba, K. 2001. Tetraploid bell pepper shows high in vitro pollen germination at 15℃. HortScience 36(7):1336.

9. İnan, S. 2007. Karpuz (Citrullus lanatus (Thunb.) Matsum ve Nakai)’da in vivo ve in vitro yöntemlerle tetraploid bitki elde edilmesi (Yüksek Lisans Tezi). Çukurova Üniversitesi Fen Bilimleri Enstitüsü, Adana, 80s.
10. Jaskani, M.J., Kwon, S.W., Koh, G.C., Huh, Y.C., Ko, B.R. 2004-a. Induction and characterization of tetraploid watermelon. Journal of The Korean Society for Horticultural Science 45:60-65.
11. Jaskani, M.J., Kwon, S.W., Kim, E.J., Ko, B.R. 2004-b. Polyploidy affects fruit characteristics, seed morphology and germination in watermelon (Citrullus lanatus). Journal of The Korean Society for Horticultural Science 45(5):233-237.
12. Jaskani, M.J., Kwon, S.W., Dae, H.K. 2005. Flow cytometry of DNA contents of colchicine treated watermelon as a ploidy screening method at M1 stage. Pakistan Journal of Botany 37(3):685-696.
13. Khan, M.R., Hasnunnahar, M., Isshiki, S. 2013. Production of amphidiploids of the hybrids between Solanum macrocarpon and eggplant. HortScience 48(4):422-424.
14. Kim I.S. Okubo, H., Fujieda, K. 1992. Endogenous levels of IAA in relation to parthenocarpy in cucumber (Cucumis sativus L.). Scientia Horticulturae 52:1-8.
15. Koh, G. 2002. Tetraploid production of Moodeungsan watermelon. Korean Society for Horticultural Science 43(6):671-676.
16. Kulkarni, M., Borse, T., Chaphalkar, S. 2008. Mining anatomical traits:a novel modeling approach for increased water use efficiency under drought condition in plants. Czech Journal of Genetics. Plant Breeding 44:11-21.
17. Kulkarni, M., Borse, T. 2010. Induced polyploidy with gigas expression for root traits in Capsicum annuum (L.). Plant Breeding 129(4):461-464.
18. Mears, J.A. 1980. Chemistry of polyploids:a summary with comments on Parthenium (Asteraceae-Ambrosiinae). In:Lewis WH (ed.) Polyploidy:biological relevance. New York:Plenum Press 13:77-102 (https://doi.org/10.1007/ 978-1-4613-3069-1_5).
19. Praça, M.M., Carvalho, C.R., Clarindo, W.R. 2009. A practical and reliable procedure for in vitro induction of tetraploid tomato. Scientia Horticulturae 122(3):501-505.
20. Rey, H.Y., Sansberro, P.A., Collavino, M.M., Daviña, J.R., González, A.M., Mroginski, L.A. 2002. Colchicine, trifluralin and oryzalin promoted development of somatic embryos in Ilex paraguariensis (Aquifoliaceae). Euphytica 123:49-56.
21. Sarı, N., Abak, K., Pitrat, M. 1999. Comparison of ploidy level screening methods in watermelon:Citrullus lanatus (Thunb.) Matsum. and Nakai. Scientia Horticulturae 82:265-277.
22. Sattler, M.C., Carvalho, C.R., Clarindo, W.R. 2016. The polyploidy and its key role in plant breeding. Planta 243(2):281-296.
23. Shao, J., Chen, C., Deng, X. 2003. In vitro induction of tetraploid in pomegranate (Punica granatum). Plant Cell Tissue Organ Culture 75:241-246.
24. Sjut, V., Bangerth, F. 1982. Induced parthenocarpy:a way of changing the levels of endogenous hormones in tomato fruits (Lycopersicon esculentum Mill.):1. Extractable hormones. Plant Growth Regulatuors 1:243-251.
25. Sliwinska, E. 2018. Flow cytometry-a modern method for exploring genome size and nuclear DNA synthesis in horticultural and medicinal plant species. Folia Horticulturae 30(1):103-128.
26. Takizawa, K., Ishikawa, K., Nunomura, O., Ito, T. 2008. Ploidy level effect on physiology of pepper plant as affected by fruit loading. ISHS Acta Horticulturae 779:International Symposium on Growing Media. Acta Horticulturae 779:689-697.
27. Tal, M. 1980. Physiology of polyploids. In:Lewis WH (ed.). Polyploidy:biological relevance. New York:Plenum Press. 13:61-76. PMID:550845.
28. Tanaka, M. 1950. Studies on artificial polyploid egg plants. I. The production of tetra-ploid eggplants by means of colchicine. Reports Kihara Institute of Biological Research 4:59-65.
29. Tuna, M. 2014. Flow sitometri ve tarımsal araştırmalarda kullanımı. II. flow sitometri ve tarımsal araştırmalarda kullanımı eğitim programı notları, 16-17 Ocak 2014, Tekirdağ.
30. Xiong, Y.C., Li, F.M, Zhang, T. 2006. Performance of wheat crops with different chromosome ploidy:root-sourced signals, drought tolerance, and yield performance. Planta 224:710-718 (doi:10.1007/s00425-006-0252-x).