Influence of in vitro Micropropagation Growth Conditions on Stomatal and Morphological Characteristics of Mature Pistacia vera L.

Year 2020, , 799 - 807, 01.06.2020

Emine Ayaz Tilkat , Hülya Hoşgören , Alevcan Kaplan , Engin Tilkat

https://doi.org/10.21597/jist.658085

Abstract

This research was conducted to reveal the stomatal anatomy, stomatal index and water loss (%) of mature pistachio leaves as well as the leaves of different phases (multiplication, rooting, hardening and regenerated plant) of micropropagation of mature pistachio trees obtained from the in vitro. Microscopic observations on surfaces of these leaves showed variety from elliptical to ovate stomata with length of 0.81-2.02 μm and width of 1.58-3.80 μm. An increase in stomatal index (SI) in the leaves of plants grown in vitro was observed most specifically in the hardening phase. (17.49±0.04). The stomatal index declined in the leaves of plantlets transferred to in vivo conditions subsequent to the hardening phase. In order to measure water loss, leaves obtained from all types of samples were dried in the oven between 30 minutes and 2 hours and weighed. The percent water loss of in vitro leaves of multiplication phase was greater than the other phases. The stomatal differentation was found to be influenced by the different hardening regimes applied. Hardening by covering the pots with polyethylene bags improved the survival rate. This study indicates that optmization of in vitro micropropagation stages is necessary to avoid transplantation stress.

Keywords

Hardening, in vitro Culture, Stomatal Characteristics, Water Loss, Stomatal Index, P. vera L.

Supporting Institution

TUBITAK and Dicle University

Project Number

TUBITAK ( Project No: TOVAG 3355) & DUAPK (Dicle University Research Fund Project No: 05-FF- 23 & 05- FF-61)

Thanks

This study was financially assisted by The Scientific and Technological Research Council of Turkey (TUBITAK Project No: TOVAG 3355) and Dicle University Research Fund (DUAPK Project No: 05-FF- 23 & 05- FF-61), we also thank to the director of the Pistachio Research Institute, Gaziantep, Turkey, for assisting in the supply of plant samples used in the experiments. The authors also wish to thank Linda Thain Ali and anonymous reviewers for their valuable comments.

Olgun Antepfıstığı’nın (Pistacia vera L.) Stomatal ve Morfolojik Özellikleri Üzerine In vitro Mikropropagasyon Büyüme Koşullarının Etkisi

Abstract

Bu çalışma, olgun antepfıstığı ağaçlarının in vitro olarak mikroçoğaltımının farklı evrelerinden (çoğaltma, köklendirme, alıştırma ve rejenere bitki) elde edilen yaprakların stoma anatomisini, stoma indeksini ve su kaybını (%) ortaya çıkarmak için yapılmıştır. Yaprak yüzeyinde yapılan mikroskobik gözlemler, 0.81-2.02 μm uzunluğunda ve 1.58-3.80 μm genişliğinde eliptikten ovat stomaya kadar çeşitlilik göstermiştir. In vitro yetiştirilen bitkilerin özellikle alıştırma aşamasında yapraklarında stoma indeksinde (SI) bir artış gözlenmiştir (17.49±0.04). Alıştırma aşamasından sonra in vivo koşullara transfer edilen bitki yapraklarının stoma indeksi azalmıştır. Su kaybını ölçmek için, her çeşit numuneden elde edilen yapraklar, 30 dakika ile 2 saat arasında fırında kurutularak tartılmıştır. Çoğaltma aşamasındaki su kaybı yüzdesinin diğer aşamalara göre daha büyük olduğu tespit edilmiştir. Stoma farklılaşmasının, uygulanan doğal şartlara aktarım metodu tarafından da etkilendiği tespit edilmiştir. Saksıların polietilen poşetlerle kapatılması yoluyla gerçekleştirilen aklimatizasyon yönteminin yaşama oranını arttırdığı tespit edilmiştir. Bu çalışma, transplantasyon stresinden kaçınmak için in vitro mikro-çoğaltma aşamalarının optimizasyonunun gerekli olduğunu göstermektedir.

Keywords

Alıştırma, in vitro kültür, stoma özellikleri, su kaybı, stoma indeksi, P. vera L.

Project Number

TUBITAK ( Project No: TOVAG 3355) & DUAPK (Dicle University Research Fund Project No: 05-FF- 23 & 05- FF-61)

References

• Abousalim A, 1990. Micropropagation and Micrografting of Pistachio (P. vera L. and Pistacia atlantica Desf.). Wye College, University of London, UK, Department of Horticulture, Ph.D. Thesis (Printed).
• Al Ramadhani RMA, 1985. The effect of some growth regulator on the initiation and growth of Pistacia vera L. Mosul University, Mosul (Iraq) College of Science.
• Assmann SM, 1993. Signal transduction in guard cells. Annual Review of Cell and Developmental Biology, 9: 345- 375.
• Balakrishnan V, Ram Latha M, Ravindran KC and Robinson JP, 2009. Clonal Propagation of Morus alba L. through Nodal and Axillary Bud Explants. Botany Research International, 2 (1): 42-49.
• Barghchi M, 1982. In vitro propagation of Pistacia species. University of Nottingham, UK, Ph.D. Thesis (Printed).
• Blanke MM, Belcher AR, 1989. Stomata of apple leaves cultured in vitro. Plant Cell, Tissue and Organ Culture, 19: 85- 89.
• Brainerd KE, Fuchigami LJ, 1981. Acclimatization of aseptically cultured apple plants to low relative humidity. Journal of the American Society for Horticultural Science, 106(4):515- 518.
• Bustamante-Garcia MA, 1984. Micropropagation and Rejuvenation of Pistacia species and the mechanism by which light influences root initiation. University of California, Davis, USA, Ph.D. Thesis (Printed).
• Chandra Dang J, Kumaria S, Kumar S and Tandon P, 2011. Micropropagation of Ilex khasiana, a critically endangered and endemic holly of Northeast India. AoB PLANTS, Plro 012. DOI:10. 1093/aobpla/plr012.
• Desjardins Y, Laforge C, Lussier C, Gosselin A, 1988. Effect of CO2 enrichment and high photosynthetic photon flux on the development of autotrophy and growth of tissue-cultured strawberry, raspberry and asparagus plants. Acta Horticulturae, 230: 45- 53.
• Gonzales A, Frutos D, 1990. In vitro culture of Pistacia vera L. embryos and aged tree explants. En Plant Aging: Basic and Applied Approaches. No:186, pp. 335- 338. Plenum Publishing Corporation, New York.
• Joshi P, Joshi N, Purohit SD, 2006. Stomatal characteristics during micropropagation of Wrightia tomentosa. Biologia Plantarum, 50(2): 275- 278.
• Kubota C, Fujiwara K, Kitaya Y, Kozai T, 1997. Recent advances in environmental control in micropropagation. In: Goto E, Kurata K, Hayashi M, Sasa S, eds. Plant production in closed ecosystems. Dordrecht: Kluwer Academic Publishers, 153–169.
• Marin JA, Gella R, Herrero M, 1988. Stomatal structure and functioning as a response to environmental changes in acclimatized micropropagated Prunus cerasifera L. Annual Review Botanic, 62: 662- 670.
• Martinelli A, 1988. Use of in vitro techniques for selection and cloning of different Pistacia species. Acta Horticulture, 227. 436- 437.
• Mišalová A, Jurkovič J, Mamoňová M, Priwitzer T, Lengyelová A, Hladká D, Lux A, 2009. Changes in leaf organisation, photosynthetic performance and wood formation during ex vitro acclimatisation of black mulberry (Morus nigra L.). Plant Biology, 11(5): 686–693.
• Mohamed MAH, Alsadon AA, 2010. Influence of ventilation and sucrose on growth and leaf anatomy of micropogated potato plantlets. Scientia Horticulturae, 123.295- 300.
• Moncalean P, Fal MA, Castanon S, Fernandez B, Rodriguez A, 2009. Relative water content, in vitro proliferation, and growth of Actinidia deliciosa plantlets are affected by benzyladenine. New Zealand Journal of Crop and Horticultural Science, 37: 351–359.
• Moyo M, Finnie JF, Staden JV, 2012. Microculture effects on leaf epidermis and root structure in Sclerocarya birrea subsp. caffra, South African Journal of Botany, 78: 170–177.
• Murashige T, Skoog F, 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473- 497.
• Namli S, Ayaz E, 2007. Influence of different cytokinins used in in vitro culture of the stoma morphology of pistachi. African Journal of Biotechnology, 6(5):561- 563.
• Onay A, 1996. In vitro organogenesis and embryogenesis of pistachio, Pistacia vera L. University of Edinburgh, UK. Ph.D. Thesis (Printed).
• Onay A, Fırat M Z, Namlı O, 1997. An improved method for embling production in Pistachio, Pistacia vera L. using liquid medium matured somatic embryos. Turkish Journal of Biology, 21: 159- 174.
• Onay A, 2000. Micropropagation of pistachio from mature trees. Plant Cell, Tissue and Organ Culture, 60, 159- 162.
• Ozden-Tokatlı Y, Ozudogru EA, Akcın A, 2005. In vitro response of pistachio nodal explants to silver nitrate. Scientia Horticulturae, 106: 415- 426.
• Salisbury EJ, 1927. On the causes and ecological significance of stomatal frequency with special reference to woodland flora. Philosophical Transactions of the Royal Society of London, series B. 216 (1928): 1- 65.
• Sutter E, Langhans RW, 1979. Epicuticular wax formation on carnation plantlets from shoot tip culture. Journal of the American Society for Horticultural Science, 104: 493- 496.
• Te chato S, Petsut P, Nuchum P, 2005. Effect of gelling agents on shoot growth and multiple shoot formation of mangosteen. Songklanakarin Journal of Science and Technology, 27: 637- 643.
• Tilkat E, Onay A, Yıldırım H, Ozen HC, 2008. Micropropagation of mature male pistachio Pistacia vera L., The Journal of Horticultural Science & Biotechnology, 83(3):328-333.
• Tilkat E, Onay A, 2009. Direct shoot organogenesis from in vitro derived mature leaf explants of pistachio, In Vitro Cellular & Developmental Biology – Plant, 45: 92- 98.
• 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.
• Tilkat E. 2006, Micropropagation of Male Pistacia vera L. via Apical Shoot Tip Culture, Institute of Science, University of Dicle, Ph.D. Thesis (Printed).
• Yang Z, Ludders P, 1993. In vitro propagation of Pistachio (Pistacia vera L.). Gartenbauwissenschaft 59: 30- 34.
• Zacchini M, Morini S, Vitagliano C, 1997. Effect of photoperiod on some stomatal characteristics of in vitro cultured fruit tree shoots. Plant Cell, Tissue and Organ Culture, 49: 195- 200.
• Ziv M, Schwartz A, Fleminger D, 1987. Malfunctioning stomata in vitreous leaves of carnation (Dianthus caryophyllus) plants propagated in vitro; implications for hardening. Plant Science Letters, 52: 127- 134.
• Zobayed SMA, Armstrong J, Armstrong W, 2001. Leaf anatomy of in vitro tobacco and cauliflower plantlets as affected by different types of ventilation. Plant Science, 161: 537–548.