Preferencje help
Widoczny [Schowaj] Abstrakt
Liczba wyników
2014 | 36 | 09 |

Tytuł artykułu

Cold-induced genetic instability in micropropagated Pistacia lentiscus L. plantlets

Warianty tytułu

Języki publikacji



Genetic stability of plants during in vitro propagation and conservation is one of the important aspects of plant biotechnology. In the present study, micropropagated P. lentiscus L. shoot cultures, which are cultivated for the mastic resin, have been cold stored up to 12 months at 4°C in the dark for different durations (2, 4, 6, 8, 10 and 12 months) and genetic alterations in cold storage conditions were evaluated. Growth parameters such as proliferation rate, shoot numbers per explant, shoot lengths and shoot forming capacity were also calculated. Since the highest proliferation rate (100 %) was obtained in 6 month-stored shoot cultures without any severe influence of cold stress on proliferation ability, amplified fragment length polymorphism (AFLP) and inter-retrotransposon amplified polymorphism (IRAP) marker systems were used to determine genetic stability in the plantlets after this storage period. Totally, 702 scorable bands were produced by 10 AFLP primer pairs. Genetic similarity value of the non-stored (control) plant and coldstored clones ranged from 0.66 to 0.84 with a mean of 0.74. In the case of IRAP, 159 bands were produced by 8 IRAP primers. Genetic similarity value of the non-stored plant and cold-stored clones varied from 0.65 to 0.83 and the average genetic similarity value was determined as 0.72. The genetic similarity indices revealed that genetic variability was similar in both techniques. Our results showed that tissue culture and especially cold storage of P. lentiscus L. may result transposons activation, thus could cause genetic instability.








Opis fizyczny



  • Department of Molecular Biology and Genetics, Faculty of Science, Gebze Institute of Technology, 41400 Gebze Kocaeli, Turkey
  • Department of Molecular Biology and Genetics, Faculty of Science, Gebze Institute of Technology, 41400 Gebze Kocaeli, Turkey
  • Department of Biology, Faculty of Science, Dicle University, Diyarbakir, Turkey
  • Department of Molecular Biology and Genetics, Faculty of Science, Gebze Institute of Technology, 41400 Gebze Kocaeli, Turkey


  • Acar İ (1988) Physico-chemical investigation of gum of Pistacia lentiscus L. var. Chia as a base for development of gum production. Forestry Research Institute Technical, Report, Turkey, No: 35
  • Akdemir H, Suzerer V, Tilkat E, Yildirim H, Onay A, Ozden Tokatlı Y (2013) In vitro conservation and cryopreservation of mature pistachio (Pistacia vera L.) germplasm. J Plant Biochem Biotech 21:43–51
  • Archak S, Gaikwad AB, Gautam D, Rao EVVB, Swamy KRM, Karihaloo JL (2003) Comparative assessment of DNA fingerprinting techniques (RAPD, ISSR and AFLP) for genetic analysis of cashew (Anacardium occidentale L.) accessions of India. Genome 46:362–369
  • Barakat MN, Abdel-Latif TH (1996) In vitro selection of wheat callus tolerant to high levels of salt and plant regeneration. Euphytica 91:127–140
  • Barra A, Coroneo V, Dessi S, Cabras P, Angioni A (2007) Characterization of the volatile constituents in the essential oil of Pistacia lentiscus L. from different origins and its antifungal and antioxidant activity. J Agric Food Chem 55(17):7093–7098
  • Bayram E, Yilmaz S, Hamat-Mecbur H, Kartal-Alacam G, Gozukirmizi N (2012) Nikita retrotransposon movements in callus cultures of barley (Hordeum vulgare L.). Plant. OMICS 5(3):211–215
  • Bednarek PT, Orłowska R, Koebner RM, Zimny J (2007) Quantification of the tissue-culture induced variation in barley (Hordeum vulgare L.). BMC Plant Biol 7(1):10
  • Botstein D, White RL, Skolnick M, Davis RW (1980) Construction of a genetic linkage map using restriction fragment length polymorphisms. Am J Hum Genet 32:314–331
  • Bowler C, Fluhr R (2000) The role of calcium and activated oxygens as signals for controlling cross-tolerance. Trends Plant Sci 5:241–246
  • Cassells AC, Curry RF (2001) Oxidative stress and physiological, epigenetic and genetic variability in plant tissue culture: implications for micropropagators and genetic engineers. Plant Cell Tissue Organ Cult 64:145–167
  • Castola V, Bighelli A, Casanova J (2000) Intraspecific chemical variability of the essential oil of Pistacia lentiscus L. from Corsica. Biochem Syst Ecol 28:79–88
  • Correia OA, Catarino FM (1994) Seasonal changes in soil-to-leaf resistance in Cistus sp. and Pistacia lentiscus. Acta Oecol 15:289–300
  • Cuesta C, Ordás RJ, Rodríguez A, Fernández B (2010) PCR-based molecular markers for assessment of somaclonal variation in Pinus pinea clones micropropagated in vitro. Biol Plant 54:435–442
  • Dabos KJ, Sfikaa E, Vlattaa LJ, Frantzia D, Amygdalosb GI, Giannikopoulos G (2010) Is Chios mastic gum effective in the treatment of functional dyspepsia? A prospective randomised double-blind placebo controlled trial. J Ethnopharmacol 127:205–209
  • Dogan Y, Baslar S, Aydın H, Mert HH (2003) A study of the soilplant interactions of Pistacia lentiscus L. distributed in the western Anatolian part of Turkey. Acta Bot Croat 62:73–88
  • Evrensel C, Yılmaz S, Temel A, Gozukirmizi N (2011) Variations in BARE-1 insertion patterns in barley callus cultures. Genet Mol Res 10(2):980–987
  • Fascella G, Airò M, Zizzo GV, Ruffoni B (2004) Prime osservazioni sulla coltivazione in vitro di Lentisco (Pistacia lentiscus L.). Italus Hortus 11(4):141–143
  • Gardeli C, Vassiliki P, Athanasios M, Kibouris T, Michael K (2008) Essential oil composition of Pistacia lentiscus L. and Myrtus communis L.: evaluation of antioxidant capacity of methanolic extracts. Food Chem 107:1120–1130
  • Gozukirmizi N, Arı S, Oraler G, Okatan Y, Unsal N (1990) Callus induction, plant regeneration and chromosomal variations in barley. Acta Bot Neerl 39:379–387
  • Grandbastien MA, Lucas H, Morel JB et al (1997) The expression of the tobacco Tnt1 retrotransposon is linked to plant defense responses. Genetica 100:241–252
  • Grandbastien MA, Audeon CE, Bonnivard JM, Casacuberta B et al (2005) Stress activation and genomic impact of Tnt1 retrotransposons in Solanaceae. Cytogenet Genome Res 110:229–241
  • Hao YJ, Deng XX (2003) Genetically stable regeneration of apple plants from slow growth. Plant Cell Tissue Organ Cult 72:253–260
  • Hao YJ, Wen XP, Deng XX (2004) Genetic and epigenetic evaluations of citrus calluses recovered from slow-growth culture. J Plant Physiol 161:479–484
  • Hirochika H (1995) Activation of plant retrotransposons by stress. In: Oono K, Takaiwa F (eds) Modification of gene expression and non-Mendelian inheritance. NIAR, Japan, pp 15–22
  • Hirochika H, Sugimoto K, Otsuki Y, Tsugawa H, Kanda M (1996) Retrotransposons of rice involved in mutations induced by tissue culture. Proc Natl Acad Sci USA 93:7783–7788
  • Jaccard P (1908) Nouvelles recherches sur la distribution florale. Bull Soc Vaud Sci Nat 44:223–270
  • Kaity A, Ashmore SE, Drew RA, Dullo ME (2008) Assessment of genetic and epigenetic changes following cryopreservation in papaya. Plant Cell Rep 27:1529–1539
  • Kalendar R (2011) The use of retrotransposon-based molecular markers to analyze genetic diversity. Ratar Povrt Field Veg Crop Res 48:261–274
  • Kalendar R, Schulman AH (2006) IRAP and REMAP for retrotransposon-based genotyping and fingerprinting. Nat Protoc 1:2478–2484
  • Karakır MN, İsfendiyaroğlu M (1999) Sakız Ağacı (Pistacia lentiscus L.)’nın vegetatif yöntemlerle çoğaltılması ve kök oluşumunun anatomik-fizyolojik incelenmesi üzerine araştırmalar. İzmir, TUBITAK, TOGTAG-1511 Nolu proje, s: 97
  • Koç İ, Özden Çiftçi Y (2012) Development of efficient in vitro propagation and conservation techniques for Lentisk. In Vitro Cell Dev Biol Animal, Bellevue, Washington, USA, S83-P2077, doi:10.1007/s11626-012-9504-1
  • Kour GL, Kour B, Kaul S, Dhar MK (2009) Genetic and epigenetic instability of amplification-prone sequences of a novel B chromosome induced by tissue culture in Plantago lagopus. Plant Cell Rep 28:1857–1867
  • Kovach WL (1999) MVSP-A Multi Variante Statistical Package for Windows, ver. 3.1. Kovach computing Services, Pentraeth
  • Kripkyy O, Kerkeb L, Molina A, Belver A, Rosales Rodrigues, Donaire PJ (2001) Effects of salt-adaptation and salt-stress on extracellular acidification and microsome phosphohydrolase activities in tomato cell suspensions. Plant Cell Tissue Organ Cult 66:41–47
  • Kumar A, Bennetzen JL (1999) Plant retrotransposons. Annu Rev Genet 33:479–532
  • Lambardi M, Sharma KK, Thorpe TA (1993) Optimization of in vitro bud induction and plantlet formation from mature embryos of Aleppo pine (Pinus halepensis Mill.). In Vitro Cell Dev Biol 29:189–199
  • Lamiri A, Lhaloui S, Benjilali B, Berrada M (2001) Insecticidal effects of essential oils against Hessian fly, Mayetiola destructor (Say). Field Crops Res 71:9–15
  • Larkin PJ, Scowcroft WR (1981) Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214
  • Lemonakis N, Magiatis P, Kostomitsopoulos N, Skaltsounis A, Tamvakopoulos C (2011) Oral administration of Chios mastic gum or extracts in mice: quantification of triterpenic acids by liquid chromatography–tandem mass spectrometry. Planta Med 77:1916–1923
  • Liu ZL, Han FP, Tan M, Shan XH, Dong YZ, Wang XZ, Fedak G, Hao S, Liu B (2004) Activation of a rice endogenous retrotransposon Tos17 in tissue culture is accompanied by cytosine demethylation and causes heritable alteration in methylation pattern of flanking genomic regions. Theor Appl Genet 109:200–209
  • Lodhi MA, Guang-Ning Y, Weeden NF, Reisch BI (1994) A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species. Plant Mol Biol Rep 26:6–13
  • Machczynska J, Orłowska R, Zimny J, Bednarek PT (2014) Extended metAFLP approach in studies of the tissue culture induced variation (TCIV) in case of triticale. Mol Breed. doi:10.1007/s11032-014-0079-2
  • Marascuilo LA, Mc Sweeney M (1977) Post-hoc multiple comparisons in sample preparations for test of homogenesity. In: McSweeney M, Marascuilo LA (eds) Non-parametric and distribution free methods the social sciences. Books/Cole Publication, Belmont, pp 141–147
  • Mascarello C, Fascella G, Zizzo GV, Mantovani E, Ruffoni B (2007) In vivo and in vitro propagation of Pistacia lentiscus L. Acta Hortic 764:299–305
  • McGregor CE, Lambert CA, Greyliing MM, Louw JH, Warnich L (2000) A comparative assessment of DNA fingerprinting techniques (RAPD, ISSR, AFLP, SSR) in tetraploid potato (Solanum tuberosum L.) germplasm. Euphytica 113:135–144
  • Müller J, Day JG, Harding K, Hepperle D, Lorenz M, Friedl T (2007) Assessing genetic stability of a range of terrestrial microalgae after cryopreservation using amplified fragment length polymorphism (AFLP). Am J Bot 94:799–808
  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco cultures. Physiol Plant 15:473–497
  • Neelakandan AK, Wang K (2012) Recent progress in the understanding of tissue culture-induced genome level changes in plants and potential applications. Plant Cell Rep 31:597–620
  • Ochatt SJ, Marconi PL, Radice S, Arnozis PA, Caso OH (1999) In vitro recurrent selection of potato: production and characterization of salt tolerant cell lines and plants. Plant Cell Tissue Organ Cult 55:1–8
  • Ozden-Tokatli Y, Ozudogru EA, Akcin A (2005) In vitro response of pistachio nodal explants to silver nitrate. Sci Hortic 106:415–426
  • Ozden-Tokatli Y, Akdemir H, Tilkat E, Onay A (2010) Current status and conservation of Pistacia germplasm. Biotechnol Adv 28:130–141
  • Ozeki Y, Davies E, Takeda J (1997) Somatic variation during long term subculturing of plant cells caused by insertion of a transposable element in a phenylalanine ammonia-lyase (PAL) gene. Mol Gen Genet 254:407–416
  • Palacio S, Milla R, Montserrat-Marti G (2005) A phenological hypothesis on the thermophilous distribution of Pistacia lentiscus L. Flora 200:527–553
  • Palevitch D, Yaniv Z (2000) The effects of aqueous extracts prepared from the leaves of Pistacia lentiscus in experimental liver disease. In: Ljubuncic et al (eds) Medicinal plants of the Holy Land. Modan Publishing House, Tel Aviv, pp 198–204 (J Ethnopharmacol)
  • Palombi MA, Damiano C (2002) Comparison between RAPD and SSR molecular markers in detecting variation in kiwifruit (Actinidia deliciosa A. Chev). Plant Cell Rep 20:1061–1066
  • Paraschos S, Magiatis P, Mitakou S, Petraki K, Kalliaropoulos A, Maragkoudakis A, Mentis A, Sgouras D, Skaltsounis AL (2007) In vitro and in vivo activities of Chios mastic gum extracts, and constituents against Helicobacter pylori. Antimicrob Agents Chemother 51:551–559
  • Paun O, Schönswetter P (2012) Amplified Fragment Length Polymorphism (AFLP)—an invaluable fingerprinting technique for genomic, transcriptomic and epigenetic studies. Methods Mol Biol 862:75–87
  • Peredo EL, Arroyo-Garcia R, Reed BM, Revilla MA (2008) Genetic and epigenetic stability of cryopreserved and cold-stored hops (Humulus lupulus L.). Cryobiology 57:234–241
  • Peschke VM, Phillips R, Gengenback BG (1987) Discovery of transposable element activity among progeny of tissue culture derived maize plants. Science 238:804–808
  • Prevost A, Wilkinson MJ (1999) A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor Appl Genet 98:107–112
  • Rahman MH, Krishnaraj S, Thorpe TA (1995) Selection for salt tolerance in vitro using microspore-derived embryos of Brassica napus cv. Topas, and the characterization of putative tolerant plants. In Vitro Cell Dev Biol Plant 31:116–121
  • Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan AK (2011) Developing stress tolerant plants through in vitro selection—an overview of the recent progress. Environ Exp Bot 71:89–98
  • Renau-Morata B, Arrillaga I, Segura J (2006) In vitro storage of cedar shoot cultures under minimal growth conditions. Plant Cell Rep 25(7):636–642
  • Rohlf FJ (1993) NTSYS-pc: numerical taxonomy and multivariate analysis system. Exeter Publishing, New York
  • Roldan-Ruiz I, Calsyn E, Gilliand TJ, Coll R, Vaneijk MJT, DeLoose M (2000) Estimating genetic conformity between related ryegrass (Lolium) varieties. 2. AFLP characterization. Mol Breed 6:593–600
  • Ruffoni B, Mascarello C, Fascella G, Airó M, Amoretti M (2004) La micro-propagazione del Lentisco (Pistacia lentiscus L.). Atti II° Convegno nazionale Piante mediterranee. Italus Hortus 11(4):141–143
  • Ryynänen L, Aronen T (2005) Genome fidelity during short and long term tissue culture and differentially cryostored meristems of silver birch (Betula pendula). Plant Cell Tissue Organ Cult 83:21–32
  • Saidi Y, Hasnaoui F, Hasnaoui B (2009) Production potentiality in fruits, biomass, oil and medicinal properties of the mastic tree (Pistacia lentiscus) in Kroumirie, N-W Tunusia. In: Palahi M, Birot Y, Bravo F, Gorriz E (ed) Modelling, valuing and managing Mediterranean Forest ecosystems for non-timber goods and services. Efi Proceedings No: 57, European Forest Institute, pp 185–195
  • Schulman AH, Flavell AJ, Ellis THN (2004) The application of LTR retrotransposons as molecular markers in plants. Methods Mol Biol 260:145–173
  • Slesak I, Libik M, Karpinska B, Karpinski S, Miszalski Z (2007) The role of hydrogen peroxide in regulation of plant metabolism and cellular signalling in response to environmental stresses. Acta Biochim Pol 54:39–50
  • Steward N, Ito M, Yamaguchi Y, Koizumi N, Sano H (2002) Periodic DNA methylation in maize nucleosomes and demethylation by environmental stress. J Biol Chem 277:37741–37746
  • Takeda S, Sugimoto K, Otsuki H, Hirochika H (1999) A 13-bp cisregulatory element in the LTR promoter of the tobacco retrotransposon Tto1 is involved in responsiveness to tissue culture, wounding, methyl jasmonate and fungal elicitors. Plant J 18:383–393
  • Taşkın T, İnal A (2005) Sakız ağacı (Pistacia lentiscus var. Chia Duhamel)’ nın in vitro mikroçoğaltımı üzerine araştırmalar. Ege Tarımsal Araştırma Enstitüsü, Turkey
  • Triantafyllou A, Bikineyeva A, Dikalova A, Nazarewicz R, Lerakis S, Dikalov S (2011) Anti-inflammatory activity of Chios mastic gum is associated with inhibition of TNF-alpha induced oxidative stres. Nutr J 64:1–9
  • Yıldırım H (2012) Micropropagation of Pistacia lentiscus L. from axenic seedlings-derived explants. Sci Hortic 137:29–35

Typ dokumentu



Identyfikator YADDA

JavaScript jest wyłączony w Twojej przeglądarce internetowej. Włącz go, a następnie odśwież stronę, aby móc w pełni z niej korzystać.