Biber ve Patlıcanda Katlanmış Haploidi Teknolojisi İle Elde Edilen Saf Hatlarda Moleküler Yöntemlerle Heterotik Grupların Oluşturulması

Öz

Anter kültürü yöntemiyle elde edilen dihaploid (DH) silindirik tipte 133 patlıcan (Solanum melongena) ve kapya tipinde 128 biber (Capsicum annuum L) genotipinde heterotik grupların oluşturulması amacıyla DNA izolasyonu yapılmıştır. İzolasyonu yapılan DNA’lar genetik benzerlik ve farklılıklarının tespit edilmesi amacıyla SRAP markırlarıyla taranmıştır. Araştırmada 32 adet primer kombinasyonu test edilmiş ve bu primerlerden polimorfik ve skorlanabilir biberde 7, patlıcanda 8 primer kombinasyonu seçilmiştir. Polimorfik primerler bütün patlıcan ve biber genotipleri ile taranmıştır. NTSYS (Numerical Taxonomy Multivariate Analysis System) modülü kullanılarak, Rohlf (2004) metoduna göre elde edilen benzerlik matrisinin UPGMA (Unweighted Pair Group Method Arithmetic Average) gruplandırması ile genetik ilişkinin seviyesi belirlenmiştir. Biberde primerlerin polimorfizm oranı %40 ile %83 arasında değişmiş, ortalama %60 olarak tespit edilmiştir. Biberde 128 genotipin 0.65 ile 1.0 düzeyinde benzer oldukları tespit edilmiştir. Patlıcanda 8 primer kombinasyonu ile toplam 71 bant elde edilmiştir ve bunların 37’si polimorfik bulunmuştur. Tüm primerlerin polimorfizm oranı %52 olarak belirlenmiştir. Genotiplerin benzerlik oranı 0.78 ile 0.99 arasında değişmiştir. Elde edilen dendrogramlarda ve iki boyutlu dağılım grafiğinde tespit edilen bulgular ışığında birbirine en uzak olan hatlar melezlenmek üzere belirlenmiştir.

Anahtar Kelimeler

• Solanum melongena
• Capsicum annuum
• DH hat
• markır
• heterotik grup
• hibrit çeşit

Construction of Heterotic Groups With Molecular Methods in Pure Lines Obtained by Doubled Haploid Technology in Pepper and Eggplant

Öz

DNA isolation was performed for heterotic grouping in DH (doubled haploid) 133 cylindrical type eggplant (Solanum melongena) and 128 pepper (Capsicum annuum L) genotypes obtained by anther culture method. The isolated DNAs were screened with SRAP markers to detect genetic similarities and differences. In the study, 32 primer combinations were tested and 7 primer combinations in polymorphic and score able pepper and 8 primer combinations in eggplant were selected from these primers. Polymorphic primers were analyzed with all eggplant and pepper genotypes. In line with the data obtained, the level of genetic relationship was determined by UPGMA (Unweighted Pair Group Method Arithmetic Average) grouping of the similarity matrix obtained according to the Rohlf (2004) method using the NTSYS (Numerical Taxonomy Multivariate Analysis System) module. The polymorphism ratio of the primers in pepper varied between 40%and 83%, with an average of 60%. It was determined that 128 genotypes were similar between 0.65 and 1.00 in pepper. In eggplant, 71 bands were obtained with 8 primer combinations and 37 of them were found to be polymorphic. Polymorphism ratio of all primers was determined as 52%. The similarity ratio of the genotypes varied between 0.78 and 0.99. In light of the findings determined in the obtained dendrograms and bi-dimensional scatterplots, the lines that are farthest from each other can be hybridized to increase the hybrid power.

Anahtar Kelimeler

• Solanum melongena
• Capsicum annuum
• DH lines
• markers
• heterotic groups
• hybrid species

Kaynakça

1. Adetula, O.A. 2006. Genetic diversity of Capsicum using random amplified polymorphic DNAs. Afr. J. Biotech. 5:120-122.
2. Alghamdi, S.S., Al-Faifi, S.A., Migdadi, H.M., Khan M.Al., EL-Harty, EH., Ammar, M.H. 2012. Molecular diversity assessment using sequence related amplified polymorphism (SRAP) markers in Vicia faba L. International Journal of Molecular Science 13:16457-16471 (doi:10.3390/ijms1312 16457).
3. Ali, Z., Xu, Z.L., Zhang, D.Y., He, X.L., Bahadur, S., Yi, J.X. 2011. Molecular diversity analysis of eggplant (Solanum melongena) Genetic Resources, Genetics and Molecular Research, 10(2):1141-1155.
4. Behera, T., Sharma, P., Singh, B., Kumar, G., Kumar, R., Mohapatra, T., Singh, N. 2006. Assessment of genetic diversity and species relationships in eggplant (Solanum melongena L.) using STMS markers. Scientia Horticulturae, 107:352-357.
5. Bhadragoudar, M.R., Patil, C.G. 2011. Assessment of genetic diversity among Capsicum annuum L. genotypes using RAPD markers. Afr. J. Biotechnol 10:17477-17483.
6. Bozokalfa, M.K., Aşcıoğlu, T.K., Eşiyok, D. 2017. Biber genotiplerinin genetik çeşitliliklerinin SRAP markörleri kullanılarak belirlenmesi. Anadolu Tarım Bilimleri Dergisi, 32(3):321-329.
7. Bozokalfa, M.K., Eşiyok, D., Turhan, K. 2009. Patterns of phenotypic variation in a germplasm collection of pepper (Capsicum annuum L.) from Turkey. Spanish Journal of Agricultural Research, 7:83-95.
8. Comlekcioglu, N., Ellialtioglu, S.S. 2018. Review on the research carried out on in vitro androgenesis of peppers (Capsicum annuum L.) in Turkey. Research Journal of Biotechnology, 13(6), 75-84.

9. Cravero, V., Martin, E., Cointry, E. 2007. Genetic diversity in Cynara cardunculus determined by sequence-related amplified polymorphism markers. Section Title:Plant Biochemistry, 132(2):208-212.
10. Dhaliwal, M.S., Jindal, S.K., Gaikwad, A.K. Singh, K. 2013. Genetic diversity analysis and DNA fingerprinting of elite Chili pepper lines using SSR markers. International Journal of Vegetable Science, 19 (3):207-216.
11. Dice, LR. 1945. Measures of the amount of ecologic association between species. Ecology 26(3):297-302.
12. Doyle, J.J., Doyle, J.L. 1990. Isolation of plant DNA from fresh tissue. Focus, V.12.
13. Escribano, M.R., Santalla, M., Casquero, P.A., De Ron, A.R. 1998. Patterns of genetic diversity in landraces of common bean (Phaseolus vulgaris L.) from Galicia. Plant Breeding, 117:49-56.
14. Fang, C., Yang, Z., Liu, D., Liu, X., Liang, G., Yang, H., Li, Y. 2010. Analysis of eggplant linkage map using SRAP molecular markers. Southwest China Journal of Agricultural Sciences, 23(5):1591-1594.
15. FAO, 2020. FAOSTAT Production Databases. Available online at:(http://www.faostat.fao.org; Erişim:25 Ağustos 2022).
16. Fidan, H., Sarıkaya, P. 2020. Antalya ili patlıcan (Solanum melongena) yetiştiriciliğinde sorun olan virüs hastalıkları. Mediterranean Agricultural Sciences, 33(1):27-35.
17. Gaikwad, A.B., Archak, S., Gautam, D. 2013. DNA profiling of Capsicum annuum L. cultivars based on AFLP and ISSR markers. Geneconserve:12(49):4-12.
18. Geleta, L.F., Labuschagne, M.T., Viljoen, C.D. 2005. Genetic variability in pepper (Capsicum annuum L.) estimated by morphological data and amplified fragment length polymorphism markers. Biodiversity & Conservation, 14:2361-2375.
19. Guo, S.G., Zhang, J.G., Sun, H.H., Salse, J., Lucas, W.J., Zhang, H.Y., Zheng, Y., Mao, L.Y., Ren, Y., Wang, Z.W., Min, J.M., Guo, X.S., Murat, F., Ham, B.K., Zhang, Z.L., Gao, S., Huang, M.Y., Xu, Y.M., Zhong, S.L., Bombarely, A., Mueller, L.A., Zhao, H., He, H.J., Zhang, Y., Zhang, Z.H., Huang, S.W., Tan, T., Pang, E.L., Lin, K., Hu, Q., Kuang, H.H., Ni, P.X., Wang, B., Liu, J.A., Kou, Q.H., Hou, W.J., Zou, X.H., Jiang, J., Gong, G.Y., Klee, K., Schoof, H., Huang, Y., Hu, X.S., Dong, S.S., Liang, D.Q., Wang, J., Wu, K., Xia, Y., Zhao, X., Zheng, Z.Q., Xing, M., Liang, X.M., Huang, B.Q., Lv, T., Wang, J.Y., Yin, Y., Yi, H.P., Li, R.Q., Wu, M.Z., Levi, A., Zhang, X.P., Giovannoni, J.J., Wang, J., Li, Y.F., Fei, Z.J., Xu, Y. 2013. The draft genome of watermelon (Citrullus lanatus) and resequencing of 20 diverse accessions. Nature Genetics, 45(1):51-58.
20. Haussmann, B.I.G, Parzies, H.K., Presterl, T., Susic, Z., Miedaner, T. 2004. Plant genetic resources in crop improvement. Plant Genetic Resources, 2(1):3-21.
21. Hayati, B.A.T., Shidfar, M., Çömlekçioğlu, N., Ellialtıoğlu, Ş.Ş. 2020. In vitro androgenesis in pepper and the affecting factors on success:I. Carbon source and concentrations. Biotech Studies, 29(2):62-68.
22. Isik, N., Doganlar, S., Frary, A., 2011. Genetic diversity of Turkish olive varieties assessed by simple sequence repeat and sequence-related amplified polymorphism Markers. Crop Science, 51:1646-1654.
23. Keleş, D. 2007. Farklı biber genotiplerinin karakterizasyonu ve düşük sıcaklığa tolerans. Çukurova Üniversitesi Fen Bilimleri Enstitüsü Bahçe Bitkileri Anabilim Dalı (Doktora Tezi). Adana.
24. Lantos, C., Juhasz, A.G., Vagi, P., Mihaly, R., Kristof, Z., Pauk, J. 2012. Androgenesis induction in microspore culture of sweet pepper (Capsicum annuum L.). Plant Biotechnology Rep, 6:123-132.
25. Li, H., Chen, H., Zhuang, T., Chen, J. 2010. Analysis of genetic variation in eggplant and related Solanum species using sequence-related amplified polymorphism markers. Scientia Horticulturae, 125(1):19-24.
26. Munoz-Falcón, J.E., Vilanova, S., Plazas, M., Prohens, J. 2011. Diversity, relationships, and genetic fingerprinting of the Listada de Gandía eggplant landrace using genomic SSRs and EST-SSRs. Scientia Horticulturae 129:238-246.
27. Mutlu, N., Boyacı, F.H., Göçmen, M., Abak, K. 2008. Development of SRAP, SRAP-RGA, RAPD and SCAR markers linked with a Fusarium wilt resistance gene in eggplant. Theor Appl Genet. 117:1303-1312.
28. Nacar Ç., 2014. Yazlık kabaklarda (Cucurbita pepo L.) morfolojik ve moleküler yöntemlerle heterotik grupların tespit edilmesi ile verim, erkencilik ve meyve özelliklerinde heterozis. Çukurova Üniversitesi Fen Bilimleri Enstitüsü Bahçe Bitkileri Anabilim Dalı, Adana, 481s.
29. Nsabiyera, V., Logose, M., Ochwo-Ssemakula, M., Sseruwagi, P., Gibson, P., Ojiewo, C. 2013. Morphological characterization of local and exotic hot pepper (Capsicum annuum L.) collections in Uganda. Bioremediation Biodiversity and Bioavailability, 7(1):22-32.
30. Pacheco-Olvera, A., Hernández-Verdugo, S., Rocha-Ramírez, V., González-Rodríguez, A., Oyama, K. 2012. Genetic diversity and structure of pepper (Capsicum annuum L.) from Northwestern Mexico analyzed by microsatellite markers. Crop Sci. 52:231-241.
31. Pınar, H., Coşkun, Ö.F., Uysal, E., Gülşen, O., Yetişir, H. 2017. Yöresel cırgalan biberi genotiplerinin ISSR markırları ile karakterizasyonu. Akademik Ziraat Dergisi, 6:145-150.
32. Pınar, H., Coşkun, Ö.F., Uysal, E., Gülşen, O., Yetişir, H. 2017. Farklı yamula patlıcanı genotiplerinin genetik benzerliklerinin ISSR moleküler markır yardımıyla belirlenmesi. Akademik Ziraat Dergisi, 6:157-162.
33. Prohens, J., Blanca, J.M., Nuez, F. 2005. Morphological and molecular variation in a collection of eggplants from a secondary center of diversity:implications for conservation and breeding. Journal of the American Society for Horticultural Science, 130:54-64.
34. Rai, V.P., Kumar, R., Kumar, S., Rai, A., Kumar, S., Singh, M., Singh, S.P., Rai, A.B., Rajneesh, P. 2013. Genetic diversity in Capsicum germplasm based on microsatellite and random amplified microsatellite polymorphism markers. Physiol. Mol. Biol. Plants 19(4):575-586.
35. Rana, M., Sharma, R., Sharma, P., Bhardwaj, S.V., Sharma, M. 2014. Estimation of Genetic Diversity in Capsicum annuum L. Germplasm Using PCR-Based Molecular Markers. National Academy Science Letters. 37(3):295-301.
36. Rohami, M., Mohammadi, A., Khosroshahli, M., Ahmadi, H., Darandeh, D. 2010. Karyotype Analysis of several Ecotypes of Capsicum annuum L. in Iran. Not. Bot. Hort. Agrobot. Cluj. 38(3):177-180.
37. Rohlf, F.J. 2004. NTSYS-pc numerical taxonomy and multivariate analysis system Version 2.11V Exeter software. Setauket, New York.
38. Singh, A., Singh, M., Singh, R., Kumar, S., Kaloo, G. 2006. Genetic diversity within the genus (Solanecea) as revealed by RAPD markers. Current Science, 90:711-716.
39. Stagel, A., Portis, E., Toppino, L., Rotino, G.L., Lanteri, S. 2008. Gene-based microsatellite development for mapping and phylogeny studies in eggplant. BMC genomics, 357-370.
40. Thomas, W.T.B., Forster, B., Gertsson, B. 2003. Doubled haploids in breeding. In:Maluszynski M., Kasha K.J., Forster B.P., Szarejko I (eds) Doubled Haploid Production in Crop Plants. Kluwer Academic Publishers, Dordrecht, pp:337-349.
41. Tiwari, S.K., Karihaloo, J.L., Nowsheen, H., Gaikwad, A.B. 2009. Molecular Characterization of Brinjal (Solanum melongena L.) Cultivars Using RAPD and ISSR Markers, Journal of Plant Biochemistry and Biotechnology, 18(2):189-195.
42. Toquica, S.P., Rodriguez, F., Martinez, E., Duque, M.C., Tohme, J. 2003. Molecular characterization by AFLPs of Capsicum germplasm from the amazon department in Colombia, characterization by AFLPs of Capsicum. Genetic Resources and Crop Evolution, 50:639-647.
43. Ulutürk, Z.I. 2009. Determination of Genetic Diversity in Watermelon (Citrullus lanatus (Thunb.) Matsum & Nakai) Germplasms. İzmir Institute of Technology, Graduate School of Engineering and Sciences, Master of Science, İzmir, 51p.
44. Uysal, E., Pinar, H., Uzun, A. 2019. SRAP Marker based compariation -with Yamula eggplant genotypes and some other eggplant varieties. Current Trends in Natural Sciences 8(15):95-100.
45. Uzun, A., Coskun, O.F., Yaman, M., Pinar, H., Paris, K., 2017. Identification of genetic similarities among walnut (Juglans regia L) genotypes selected from central Anatolia region of Turkey with SRAP markers. Alatarım 16(1):26-34.
46. Yu, S., Deng, Y., Yao, J., Li, S., Xin, X., Duan, D. 2012. Population genetics of wild Hizikia fusiformis (Sargassaceae, Phaeophyta) along China’s coast. Journal of Applied Phycology, 24:1287-1294.
47. Zhang, X.M., Zhang, Z.H., Gu, X.Z., Mao, S.L., Li, X.X., Chadœuf, J., Palloix, A., Wang, L.H., Zhang, B.X. 2016. Genetic diversity of pepper (Capsicum spp.) germplasm resources in China reflects selection for cultivar types and spatial distribution. Journal of Integrative Agriculture, 15(9):1991-2001.