Türkiye Florasında Doğal Olarak Yetişen Üç Farklı Vaccinium Türünün Sürgün Çoğaltımı Üzerine Besi ortamı pH’sının Etkisi

Year 2016, Volume: 1 Issue: 2, 52 - 58, 29.12.2016

Mustafa Cüce Tuba Bekircan Atalay Sökmen

Abstract

Besi ortamlarının pH’sı bitki hücre kültürlerini etkileyen en önemli faktörlerden biridir ve genellikle bunun kontrolüne çok az önem verilmiştir. Bu çalışma V. arctostaphylos, V. myrtillus ve V. uliginosum’un sürgün çoğaltımında besi ortamı pH değişimlerinin etkilerini belirlemek amacıyla dizayn edilmiştir. Sürgün çoğaltımı, sürgün sayısı, sürgün boyu, nod sayısı ve kallus oluşum yüzdeleri açısından değerlendirilmiştir. Bunun için pH’sı otoklavdan önce 4.0, 4.5, 5.0, 5.5 ve 6.0’a ayarlanmış ve 1.0/0.1 mg/L zeatin/IBA ile desteklenmiş WPM, temel besi ortamı olarak kullanılmıştır. Nod segmentlerinin eksplant olarak kullanıldığı in vitro çalışmalarda sürgün çoğaltımı besi ortamı pH’sının yanı sıra türe bağlı olarak da büyük ölçüde etkilenmiştir. Besi ortamlarının son pH değişimleri 3.66 ve 5.42 olarak belirlenmiştir. En yüksek sürgün sayısı eksplant başına 5.03 ile 5.5 pH içeren besi ortamdaki V. arctostaphylos’tan, en yüksek sürgün boyu ise 40.78 mm ile 4.5 pH içeren besi ortamdaki V. myrtillus’tan elde edilmiştir. V. arctostaphylos ve V. uliginosum için en yüksek nod sayısını sürgün başına 10.83 ile pH’sı 6.0 olan ortam vermiştir. V. arctostaphylos’da en yüksek sürgün boyu ve en yüksek nod sayısı açısından 6.0 pH daha üstün bulunurken, en yüksek sürgün sayısı için 5.5 pH daha uygun olmuştur. V. myrtillus’da tüm parametrelerde en yüksek değerler 4.5 pH içeren ortamdan elde edilmiştir. pH değerleri bir artış eğiliminde olduğunda V. uliginosum’da sürgün çoğaltım oranları önemli bir şekilde azalmıştır.

Keywords

Mikroçoğaltım, pH, Vaccinium arctostaphylos, V. myrtillus, V. uliginosum, zeatin

Effect of Medium pH on Shoot Regeneration of Three Vaccinium Species Naturally Growing in Turkish Flora

Abstract

Medium pH is one of the crucial factors affecting plant cell culture, and generally very little attention has been given to the control of it. The present study was designed to monitor the effects of pH changes on the course of shoot regeneration of Vaccinium arctostaphylos L., Vaccinium myrtillus L. and Vaccinium uliginosum L. Shoot regeneration was assessed by evaluating shoot numbers, shoot length, nod numbers, and callus formation. For this, McCown Woody Plant Medium (WPM) supplemented with 1.0/0.1 mg/L zeatin/indole-3- butyric acid (IBA) was employed as basal media being adjusted with initial pH at 4.0, 4.5, 5.0, 5.5 and 6.0 before autoclaving. When in vitro established nodal segments were employed, shoot regeneration was dramatically influenced depending on the species as well as the medium pH. The final pH changes were determined 3.66 and 5.42. The highest shoot number was obtained 5.03 (shoots per explant) at pH 5.5 in V. arctostaphylos whereas the highest shoot length (in average) was obtained (40.78 mm) at pH 4.5 pH in V. myrtillus. Both media having a pH of 6.0 gave the highest nodes number with 10.83 (nodes/explant) for V. arctostaphylos and V. uliginosum. In V. arctostaphylos, pH 5.5 was the most favorable for shoot number while pH 6.0 was found to be superior to the shoot length and nod number. In V. myrtillus, the highest values for all parameters were obtained at pH 4.5. When the pH is fluctuated tending an increase, the shoot multiplication rates significantly decreased in V. uliginosum.

Keywords

Micropropagation, pH, Vaccinium arctostaphylos, V. myrtillus, V. uliginosum, zeatin

References

• Andersone U, Ievinsh G. 2008. Medium pH affects regeneration capacity and oxidative enzyme activity of Pinus sylvestris in tissue culture. Acta Univ Lat, 745: 25-35.
• Cain JC. 1952. A comparison of ammonium and nitrate nitrogen for blueberries. Proc. Amer Soc Hort Sci, 59: 161-166.
• Clapa D, Fira A, Vescan LA. 2012. Aspects regarding the in vitro culture and ex vitro rooting in Vaccinium macrocarpon Cultivar ‘Pilgrim’. Bulletin UASVM Animal Science and Biotechnologies 69: 226-234.
• Coville FV. 1910. Experiments in blueberry culture. US Dept Agr Bul, 193.
• Cüce M, Sökmen A. 2015. Micropropagation of Vaccinium myrtillus L. (Bilberry) naturally growing in the Turkish Flora, Turk J Biol, 39: 233-240.
• Cüce M, Bektas E, Sökmen A. 2013. Micropropagation of Vaccinium arctostaphylos L. via lateral-bud culture. Turk J Agric For, 37(1): 40-44.
• Debnath SC, McRae KB. 2001b. In vitro culture of lingonberry (Vaccinium vitis-idaea L.): The influence of cytokinins and media types on propagation. Small Fruits Rev, 1(3): 3-19.
• Debnath SC. 2005. Micropropagation of lingonberry: influence of genotype, explant orientation, and overcoming TDZ-induce inhibition of shoot elongation using zeatin. Hortscience, 40(1): 185-188.
• Druart P, De Wulf O. 1993. Activated charcoal catalysis sucrose hydrolysis during autoclaving. Plant Cell Tiss Org, 32: 97-99.
• Eccher T, Noe N. 1989. Comparison between 2iP and zeatin in the micropropagation of highbush blueberry (Vaccinium corymbosum). Acta Hortic, 441: 185-190.
• Galletta GJ. 1975. Blueberries and cranberries; p. 154-196. In: J. Janick and J.N. Moore (eds.). Advances in fruit breeding. Purdue Univ. Press. West Lafayette, Ind.
• Gomez MP, Segura J. 1995. Axillary shoot proliferation in cultures of explants from mature Juniperus oxycedrus trees. Tree Physiol, 15(9): 625-628.
• Gürel S, Gülsen Y. 1998. The effects of different sucrose, agar and pH levels on in vitro shoot production of almond Amygdalus communis L.). Turk J Bot, 22: 363-373.
• Harmer P. 1944. The effect of varying the reaction of organic soil on the growth and production of the domesticated blueberry. Soil Sci Soc, Proc 9: 133-141.
• Henry PH, Blazich FA, Hinesley LE. 1992. Vegetative propagation of eastern redcedar by stem cuttings. HortScience, 27(12): 1272-1274
• Huda KMK, Bhuiyan MSR, Zeba N, Banu SA, Mahmud F, Khatun A. 2009. Effect of FeSO4 and pH on shoot regeneration from the cotyledonary explants of Tossa Jute. Plant Omics Journal 2(5): 190-196.
• Hung CD, Hong CH, Kim SK, Lee KH, Park JY, Nam MW, Choi DH, Lee HI. 2016. LED light for in vitro and ex vitro efficient growth of economically important highbush blueberry (Vaccinium corymbosum L.). Acta Physiol Plant, 38(6): 1-9.
• Jaakola L, Tolvanen A, Laine K, Hohtola A. 2001. Effect of N6-isopentenyladenine concentration on growth initiation in vitro and rooting of bilberry and lingonberry microshoots. Plant Cell Tiss Org, 66: 73-77.
• Kaldmäe H, Starast M, Karp K, Paal T. 2006. Effect of donor plant physiological condition on in vitro establishment of Vaccinium angustifolium shoot explants. Acta Hortic, 715: 433-438.
• Korcak RF, Galletta GJ, Draper AD. 1982. Response of blueberry seedlings to a range of soil types. J Amer Soc Hort Sci, 107: 1153-1160.
• Laukkanen H, Soini H, Kontunen-Soppela S, Hohtola A, Viljanen M. 2000. A mycobacterium isolated from tissue cultures of mature Pinus sylvestris interferes with growth of Scots pine seedlings. Tree Physiol, 20: 915-920.
• Liu C, Callow P, Rowland LJ, Hancock JF, Song GQ. 2010. Adventitious shoot regeneration from leaf explants of southern highbush blueberry cultivars. Plant Cell Tiss Org, 103(1): 137-144.
• Lloyd G, McCown B. 1980. Commercially-feasible micropropagation of mountain laurel (Kalmia latifolia) by use of shoot tip culture. Com Proc Int Plant Prop Soc, 30: 421–427.
• Meiners J, Schwab M, Szankowski I. 2007. Efficient in vitro regeneration systems for Vaccinium species, Plant Cell Tiss Org, 89: 169-176.
• Ostrolucká MG, Gajdošová A, Ondrušková E, Lateèková M, Libiaková G. 2010. Effect of medium pH on axillary shoot proliferation of selected Vaccinium vitis-idaea L. cultivars. Acta Biol Cracov Bot, 52(2): 92-96.
• Ostrolucká MG, Libiaková G, Ondrušková E, Gajdošová A. 2004. In vitro propagation of Vaccinium species. Acta Univ Lat, Biology 676: 207-212.
• Owen HR, Wengerd D, Miller AR. 1991. Culture medium pH is influenced by basal medium, carbohydrate source, gelling agent, activated charcoal, and medium storage method. Plant Cell Rep, 10: 583-586.
• Selby C, Lee R, Harvey BMR. 1989. The effects of culture medium rigidity on adventitious bud production and tissue vitrification in needle cultures of Sitka spruce (Picea sitchensis (Boog.) Carr.). New Phytol, 113: 203-210.
• Stark R, Hall IV, Hendrickson PA. 1978. The partridgeberry of Newfoundland. Canadex (Hort Crops) 230.
• Zhao X, Zhan L, Zou X. 2011. In vitro high-frequency regeneration of half-highbush “Northland” blueberry, New Zeal J Crop Hort, 39: 51-59.
• Zheng W, Wang SY. 2003. Oxygen radical absorbing capacity of phenolics in blueberries, cranberries, chokeberries and lingonberries. J Agric Food Chem, 51: 502-509.