Optimization of in vitro sterilization for pistachio (Pistacia vera L.) rootstocks

Öz

Pistachio (Pistacia vera L.) is one of the leading edible nut consumed all over the World due to its nutritional values. The plant is cultivated in most of the countries alongwith Turkey which is one of the the leading grower of pistachio. In Turkey, the rootstock material is currently propagated through traditional methods and there is a need of propagating plant material using modern biotechnological techniques like plant tissue culture. The provision of contaminated free explants with minimum or no phenolic compounds in the culture medium is the prerequisite of in vitro regeneration protocol. The plant material used in this study was collected at different physiological stages during different months like April- June and Sep-October. The plant material was cut into 2-3 cm long nodal segments followed by cleaning with different agents like water, soap and fungicide prior to subjected to sterilizing agents. Different sterilizing agents used in this study were HgCl2, Huwa-san (H2O2) and commercial bleach (NaOCl) for both rootstocks (UCB-1 and Buttum) with different exposure time. Sterilized explants were cultured on MS basal medium containing plant growth regulators and subcultured once a week for three weeks. Results revealed that HgCl2 as sterilizing agent was more superior than other sterilizing agents for both rootstocks. Among rootstocks, UCB-1 was more responsive than Buttum and relatively more sterilized plants were attained. On the other hand, plant material collected during June responded better and 90.0% and 50.0% sterilzied plants were attained for UCB-1 and Buttum respectively. The results revealed the significant impact of collection time, sterilizing agent type, concentration and exposure time on sterilization of P. vera rootstocks.

Anahtar Kelimeler

• In vitro
• Pistachio
• Phenolics
• Rootstocks
• Sterilization

Antep fıstığı (Pistacia vera L.) anaçlarında in vitro sterilizasyon optimizasyonu

Öz

Antep fıstığı (Pistachio vera L.), besin değerleri sebebiyle dünyada en çok tüketilen kuruyemişlerden biridir. Antep fıstığı, fıstık yetiştiriciliğinde önde olan Türkiye ile birlikte birçok ülkede yetiştirilmektedir. Türkiye’de genel olarak anaç materyali geleneksel yöntemler ile çoğaltılmaktadır ve bitki materyallerinin bitki doku kültürü gibi biyoteknolojik teknikleri kullanılarak çoğaltılmasına ihtiyaç vardır. In vitro rejenerasyon protokolünün ön koşulu, kültür ortamında kontamine olmayan, minimum veya hiç fenolik bileşik içermeyen eksplantların sağlanmasıdır. Bu çalışmada kullanılan bitki materyali, Nisan-Hazıran ile Eylül-Ekim ayları arasında değişen farklı aylarda, farklı fizyolojik aşamalardayken toplanmıştır. Bitki materyali 2-3 cm uzunluğunda nod parçaları olarak kesilmiş ardından sterilizasyon ajanlarına tabi tutulmadan önce su, sabun ve mantar ilacı gibi farklı ajanlarla temizlenmiştir. Yapılan sterilizasyon çalışmada her iki anaç (UCB-1 ve Buttum) için farklı sürelerde HgCl2, Huwa-san (H2O2) ve ticari çamaşır suyu (NaOCl) farklı sterilizasyon ajanları kullanılmıştır. Daha sonra eksplantlar, bitki büyüme düzenleyicileri içeren bazal ortamlarda kültürlendi ve üç hafta boyunca haftada bir kez kültür aktarıldı. Sonuç olarak, sterilizasyon ajanı olarak HgCl2’nin her iki anaç için de diğer sterilizasyon ajanlarından daha üstün olduğunu ortaya koymuştur. Anaçlardan UCB-1, Buttum’a göre daha duyarlı ve nispeten daha steril bitkiler elde edildi. Sonuç olarak, P. vera anaçlarının sterilizasyonunda eksplant alım zamanı, sterilizasyon ajanının türü, konsantrasyonu ve maruz kalma süresinin önemini ortaya koymuştur.

Anahtar Kelimeler

• Anaç
• Antep fistik
• Fenolikler
• In vitro
• Sterilizasyon

Kaynakça

1. Aasim M, Hussain N, Umer EM, Zubair M, Hussain SB, Saeed S, Rafique TS, Sancak C (2010). In vitro shoot regeneration of fenugreek (Trigonella foenum-graecum L.) using different cytokinins. African Journal of Biotechnology 9(42): 7174-7179.
2. Aasim M, Karataş M, Khawar KM, Doğan M (2013). Optimization of sterilization and micropropagation of water lettuce (Pistia stratiotes L.). Journal of Applied Biological Sciences 7(3): 71-74.
3. Aasim M, Sameeullah M, Karataş M, Bakırcı Ş, Bakhsh A, Akhtar MS (2019). An insight into biotechnological approaches used for improvement and secondary metabolites for medicinal aquatic plant, water hyssop (Bacopa monnieri L.). In: Akhtar MS, Swamy MK (eds.) Natural bioactive compounds: Biotechnology, Bioengineering, and Molecular Approaches: Volume 3. Springer, Singapore. Pp. 123-152.
4. Arab MM, Yadollahi A, Hosseini-Mazinani M, Bagheri S (2014). Effects of antimicrobial activity of silver nanoparticles on in vitro establishment of G × N15 (hybrid of almond × peach) rootstock. Journal of Genetic Engineering and Biotechnology 12: 103-110.
5. Barpete S, Aasim M, Fatih ÖS, Sebahattin Ö (2019). Effect of gibberellic acid on in vitro flowering from stem node explant of Anubias barteri var. Nana. Research Journal of Biotechnology 14(9): 83-88.
6. Barpete S, Aasim M, Khawar KM, Özcan S (2021). Evaluation of toxicity levels of micronutrient strengths on regeneration of grass pea under in vitro conditions. Journal of Plant Nutrition, 44(11): 1548-1562.
7. Batti JR, Larekeng SH, Arsyad MA, Gusmiaty G Restu M (2020). In vitro growth response on three provenances of Jabon Merah based on auxin and cytokinin combinations IOP Conf. Ser. Earth and Environmental Science 486: 1-16.
8. Benmahioul B (2017). Factors affecting in vitro micropropagation of Pistachio (Pistacia vera L.). Agriculture and Forestry Jounal 1(1): 56-61.

9. Benmahioul B, Kaid-Harche M, Daguin F (2016). In vitro regeneration of Pistacia vera L. from nodal explants. Journal of Forest Science 62(5): 198-203.
10. Çağlar A, Tomar O, Vatansever H, Ekmekçi E (2017). Antepfıstığı (Pistacia vera L.) ve insan sağlığı üzerine etkileri. Akademik Gıda 15(4): 436-447.
11. Da Silva JAT, Kulus T, Zhang D, Zeng X, Ma S, Piqueras A (2016a). Disinfection of explants for saffron (Crocus sativus) tissue culture. Environmental and experimental botany 14: 183-198.
12. Da Silva JAT, Winarto B, Dobránszki J, Cardoso JC, Zeng S (2016b). Tissue disinfection for preparation of Dendrobium in vitro culture. Folia Horticulturae 28: 57-75.
13. Dogan M, Karatas M, Aasim M (2016). In vitro shoot regeneration from shoot tip and nodal segment explants of Pogostemon erectus (Dalzell) Kuntze, a multipurpose ornamental aquatic plant. Fresenius Environmental Bulletin. 25(11): 4777-4782.
14. Ertürk YE, Geçer MK, Gülsoy E, Yalçın S (2015). Production and marketing of pistachio. Journal of the Institute of Science and Technology 5(2): 43-62.
15. FAO (2018). Pistachio production in 2017. UN Food and Agriculture Organization, Corporate Statistical Database (FAOSTAT).
16. Hesami M, Daneshvar MH (2016). Development of a regeneration protocol through indirect organogenesis in Chenopodium quinoa willd. Indo-American Journal of Agricultural and Veterinary Sciences 4: 25-32.
17. Hesami M, Daneshvar MH (2018). In vitro adventitious shoot regeneration through direct and indirect organogenesis from seedling-derived hypocotyl segments of Ficus religiosa L.: an important medicinal plant. HortScience 53: 55-61.
18. Hesami M, Daneshvar MH, Yoosefzadeh-Najafabadi M (2018a). Establishment of a protocol for in vitro seed germination and callus formation of Ficus religiosa L., an important medicinal plant. Jundishapur Journal of Natural Pharmaceutical Products 13(4): e62682.
19. Hesami M, Daneshvar MH, Lotfi-Jalalabadi A (2017). Effect of sodium hypochlorite on control of in vitro contamination and seed germination of Ficus religiosa. Iranian Journal of Plant Physiology 7: 2157-2162.
20. Hesami M, Daneshvar MH, Yoosefzadeh-Najafabadi M, Alizadeh M (2018b). Effect of plant growth regulators on indirect shoot organogenesis of Ficus religiosa through seedling derived petiole segments. Journal of Genetic Engineering and Biotechnology 16: 175–180.
21. Hesami M, Naderi R, Tohidfar M (2019). Modeling and optimizing in vitro sterilization of chrysanthemum via multilayer perceptron-non-dominated sorting genetic algorithm-II (MLP-NSGAII). Frontiers in plant science 10: 282.
22. Hesami M, Naderi R, Yoosefzadeh-Najafabadi M (2018c). Optimizing sterilization conditions and growth regulator effects on in vitro shoot regeneration through direct organogenesis in Chenopodium quinoa. BioTechnologia 99: 49–57.
23. Jafari M, Daneshvar MH, Lotfi-Jalalabadi A (2016). Control of in vitro contamination of Passiflora caerulea by using of sodium hypocholorite. Indo-American Journal of Agricultural and Veterinary Sciences 4: 8–15.
24. Karataş M, Aasim M (2014). Efficient in vitro regeneration of medicinal aquatic plant water hyssop (Bacopa monnieri L. Pennell). Pakistan Journal of Agricultural Sciences 51(3): 667-672.
25. Kılınç FM¸ Süzerer V, Özden Çiftçi Y, Onay A, Yıldırım H, Uncuoğlu AA, Tilkat E, Koç I, Akdemir OF, Metin OK (2015). Clonal micropropagation of Pistacia lentiscus L. and assessment of genetic stability using IRAP markers. Plant Growth Regulation 75: 75–88.
26. Leng L., Su S., Wei F., Yu F. and Duan Y. 2009. Correlation between browning, total phenolic content, polyphenol oxidase and several antioxidation enzymes during pistachio tissue culture. Acta Horticulturae 829: 127-131.
27. Marín JA, García E, Lorente P, Arbeloa A, Andreu P (2017). Propagation of pistachio applying in vitro culture techniques. Acta horticulturae 1155: 321-326.
28. Mascarello C, Fascella G, Zizzo GV, Mantovani E, Ruffoni B (2007). In vivo and in vitro propagation of Pistacia lentiscus L. Acta Horticulturae 764: 299–306.
29. Mihaljević I, Dugalić K, Tomaš V, Viljevac M, Pranjić A, Cmelik Z, Puskar B, Jurkovic Z (2013). In vitro sterilization procedures for micropropagation of 'Oblačinska' sour cherry. Journal of Agriculture Science 58: 117-126.
30. Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15: 473-497.
31. Nongalleima K, Dikash Singh T, Amitabha D, Deb L, Sunitibala Devi H (2014). Optimization of surface sterilization protocol, induction of axillary shoots regeneration in Zingiber zerumbet (L.) Sm. as affected by season. Biological Rhythm Research 45: 317-324.
32. Onay A, Pirinç V, Yıldırım H, Basaran D (2004). In vitro micrografting of mature pistachio (Pistacia vera var. Siirt). Plant Cell, Tissue and organ culture 77(2): 215-219.
33. Ozden-Tokatli Y, Ozudogru EA, Akcin A (2005). In vitro response of pistachio nodal explants to silver nitrate. Scientia Horticulturae 106: 415-426.
34. Snedecor GW, Cochran WG (1967). Statistical Methods, The Iowa State University Press, Iowa, USA, 327-330.
35. Tilkat E, Akdemir H (2013). A rapid and effective protocol for surface sterilization and in vitro culture initiation of adult male pistachio (Pistacia vera L . cv . “Atlı”). Academia Journal of Scientific Research 1:134-141.
36. Tilkat E, Onay A, Yıldırım H, Ayaz E (2009). Direct plant regeneration from mature leaf explants of pistachio, Pistacia vera L. Scientia Horticulturae 121(3): 361-365.