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First Report of Krymsk® 5 (cv. VSL 2) Cherry Rootstock In Vitro Propagation: Studying the Effect of Cytokinins, Auxins and Endogenous Sugars

Athanasios TSAFOUROS, Peter A. ROUSSOS

Abstract


Krymsk® 5 (VSL-2) is a semi-dwarf cherry rootstock adaptable to a range of climates. The present study aimed to establish the first efficient in vitro propagation protocol for this rootstock. Therefore, six cytokinines, four adenine type (6-benzyladenine, 2-isopentenyladenine, kinetin and meta-topolin) and two phenylureas (thidiazuron and forchlorfenuron) at three (2.4 μΜ, 4.8 μΜ and 9.6 μΜ) concentrations plus three (0.24 μΜ, 0.48 μΜ, 0.96 μΜ) for thidiazuron only were tested during the multiplication stage. 6-Benzyladenine was the most efficient cytokinin, based on the number of shoots produced (3.5 shoots at 9.6 μΜ) and the number of nodes per explant (10 nodes at 9.6 μΜ) whereas the other aromatic adenine tested, i.e. meta-topolin, presented the highest number of nodes per cm and node per shoot. During the rooting stage the synthetic auxins 1-naphthaleneacetic acid (1-NAA) and indolebutyric acid (IBA) were tested at concentrations of 0, 2.5, 5, 10 and 20 μΜ both separately and in all possible combinations. The percentage of successfully rooted explants reached 95% under the combination of 20 μΜ IBA plus 5 μΜ 1-NAA, whereas the highest number of roots recorded was 8.5 roots for the treatment 20 μΜ ΙΒΑ plus 2.5 μΜ 1-NAA. Furthermore, two different carbon sources were compared, the widely used sucrose and the endogenous sugar ratio of mother plant softwood shoot, sampled during late of May. Endogenous sugar ratio proved to be the preferable carbon source, since it increased the number of shoots produced and almost doubled the number of produced nodes per explant.

 

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In press - Online First. Article has been peer reviewed, accepted for publication and published online without pagination. It will receive pagination when the issue will be ready for publishing as a complete number (Volume 47, Issue 1, 2019). The article is searchable and citable by Digital Object Identifier (DOI). DOI number will become active after the article will be included in the complete issue.


Keywords


benzyladenine; carbohydrates; furfuryladenine; metaTopolin, micropropagation; Prunus

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References


Abu-Romman SM, Al-Hadid KA, Arabiyyat AR (2015). Kinetin is the most effective cytokinin on shoot multiplication from cucumber. The Journal of Agricultural Science 7:159-165.

Ahmad T, Akhtar Abbasi N, Hafiz IA, Ali A (2007). Comparison of sucrose and sorbitol as main carbon energy source in morphogenesis of peach rootstock GF-677. Pakistan Journal of Botany 39:1264-1275.

Akhtar G, Jaskani MJ, Sajjad Y, Akram A (2016). Effect of antioxidants, amino acids and plant growth regulators on in vitro propagation of Rosa centifolia. Iranian Journal of Biotechnology 14:e1152.

Alanagh EN, Garoosi GA, Haddad R (2010). The effect of PGRs on in vitro shoot multiplication of GF677 hybrid (Prunus persica × P. amygdalus) rootstock on GNH medium. Iranian Journal of Genetics and Plant Breeding 1:34-43.

Andreu P, Marin JA (2005). In vitro culture establishment and multiplication of the Prunus rootstock ‘Adesoto 101’ (P. insititia L.) as affected by the type of propagation of the donor plant and by the culture medium composition. Scientia Horticulturae 106:258-267.

Bianco RL, Rieger M (2002). Partitioning of sorbitol and sucrose catabolism within peach fruit. Journal of the American Society for Horticultural Science 127:115-121.

Borkowska B, Szczerba J (1991). Influence of different carbon sources on invertase activity and growth of sour cherry (Prunus cerasus L.) shoot cultures. Journal of Experimental Botany 42:911-915.

Brison M, de Boucauda MT, Dosbab F (1995). Cryopreservation of in vitro grown shoot tips of two interspecific Prunus rootstocks. Plant Science 105:235-242.

Caboni E, Biasi R, Delia G, Tonelli M (2009). Effect of CPPU on in vitro axillary shoot proliferation and adventitious shoot regeneration in kiwifruit. Plant Biosystems 143:456-461.

Canas LA (1988). In vitro culture of the olive tree (Olea europaea L.): present aspects and prospects. Bulletin de la Société botanique de France 135:263-277.

Cheong EJ, An C (2015). Effect of carbohydrates on in vitro shoot growth of various Prunus species. Korean Journal of Plant Resources 28: 357-362.

D'Angeli S, Lauri P, Caboni E, Dewitte W, Van Onckelen H (2001). Factors affecting in vitro shoot formation from vegetative shoot apices of apple and relationship between organogenic response and cytokinin localisation. Plant Biosystems 135:95-100.

De Klerk G-J, Brugge JT, Marinova S (1997). Effectiveness of indoleacetic acid, indolebutyric acid and naphthaleneacetic acid during adventitious root formation in vitro in Malus ‘Jork 9’. Plant Cell Tissue and Organ Culture 49:39-44.

Dejampour J, Majidi I, Khosravi S, Farhadi S, Shadmehr A (2011). In vitro propagation of HS314 rootstock (Prunus amygdalus × P. persica). HortScience 46:928-931.

Denaxa N-K, Vemmos SN, Roussos PA (2012). The role of endogenous carbohydrates and seasonal variation in rooting ability of cuttings of an easy and a hard to root olive cultivars (Olea europaea L.). Scientia Horticulturae 143:19-28.

Dolcet-Sanjuan R, Claveria E, Gruselle R, Meier-Dinkel A, Jay-Allemand C, Gaspar T (2004). Practical factors controlling in vitro adventitious root formation from walnut shoot microcuttings. Journal of the American Society for Horticultural Science 129:198-203.

Doric D, Ognjanov V, Ljubojevic M, Barac G, Dulic J, Pranjic A, Dugalic K (2014). Rapid propagation of sweet and sour cherry rootstock. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 42:488-494.

Escalona M, Cejas I, Gonzalez-Olmedo J, Capote I, Roels S, Canal MJ, Rodr?guez R, Sandoval J, Debergh P (2003). The effect of meta-topolin on plantain propagation using a temporary immersion bioreactor. InfoMusa 12:28-30.

Escobar-Gutiérrez AJ, Gaudillére J-P (1994). Variability in Sorbitol: Sucrose ratios in mature leaves of different peach cultivars. Journal of the American Society for Horticultural Science 119:321-324.

Fallahpour M, Miri SM, Bouzari N (2015). In vitro propagation of ‘Gisela 5’ rootstock as affected by mineral composition of media and plant growth regulators. Journal of Horticultural Research 23:57-64.

Fotopoulos S, Sotiropoulos E (2004). In vitro propagation of the peach rootstock: the effect of different carbon sources and types of sealing material on rooting. Biologia Plantarum 48:629-631.

Fotopoulos S, Sotiropoulos TE (2005). In vitro rooting of PR 204/84 rootstock (Prunus persica × P. amygdalus) as influenced by mineral concentration of the culture medium and exposure to darkness for period. Agronomy Research 3:3-8.

Fuentes SRL, Calheiros MBP, Manetti-Filho J, Vieira LGE (2000). The effects of silver nitrate and different carbohydrate sources on somatic embryogenesis in Coffea canephora. Plant Cell Tissue and Organ Culture 60:5-13.

Gentile A, Jaquez Gutierrez M, Martinez J, Frattarelli A, Nota P, Caboni E (2014). Effect of meta-Topolin on micropropagation and adventitious shoot regeneration in Prunus rootstocks. Plant Cell Tissue and Organ Culture 118:373-381.

George EF, Hall MA, Klerk GJD (2008). Plant Propagation by Tissue Culture. Springer, Netherlands.

Godoy S, Tapia E, Seit P, Andrade D, Sánchez E, Andrade P, Almeida AM, Prieto H (2017). Temporary immersion systems for the mass propagation of sweet cherry cultivars and cherry rootstocks: development of a micropropagation procedure and effect of culture conditions on plant quality. In Vitro Cellular & Developmental Biology-Plant 53:494-504.

Gonbad RA, Sinniah UR, Aziz MA, Mohamad R (2014). Influence of cytokinins in combination with GA3 on shoot multiplication and elongation of tea clone Iran 100 (Camellia sinensis (L.) O. Kuntze). The Scientific World Journal 5:149-168.

Guo B, Zeb A, Abbasi BH, Zeb A, Xu LL, Wei YH (2011). Thidiazuron: A multi-dimensional plant growth regulator. African Journal of Biotechnology 10:8984-9000.

Hammatt N (1993). Micropropagation of fastigiate bird cherry (Prunus padus L.) and adventitious shoot formation from leaves. Journal of Horticultural Science 68:975-981.

Hart DS, Keightley A, Sappington D, Nguyen PTM, Chritton C, Seckinger GR, Torres KC (2016). Stability of adenine-based cytokinins in aqueous solution. In Vitro Cellular & Developmental Biology-Plant 52:1-9.

Hossini AD, Moghadam EG, Anahid S (2010). Effects of media cultures and plant growth regulators in micropropagation of Gisela 6 rootstock. Annals of Biological Research 1:135-141.

Huetteman CA, Preece JE (1993). Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue and Organ Culture 33:105-119.

Kadota M, Imizu K, Hirano T (2001). Double-phase in vitro culture using sorbitol increases shoot proliferation and reduces hyperhydricity in Japanese pear. Scientia Horticulturae 89:207-215.

Kadota M, Niim Y (2003). Effects of cytokinin types and their concentrations on shoot proliferation and hyperhydricity in in vitro pear cultivar shoots. Plant Cell Tissue and Organ Culture 72:261-265.

Kalinina A, Brown DCW (2007). Micropropagation of ornamental Prunus spp. and GF305 peach, a Prunus viral indicator. Plant Cell Reports 26:927-935.

Kanayama Y (2009). Physiological roles of polyols in horticultural crops. Journal of the Japanese Society for Horticultural Science 78:158-168.

Kieber JJ, Schaller GE (2004). Cytokinins. The Arabidopsis Book 12:e0168.

Long LE, Kaiser C (2010). Sweet Cherry Rootstocks for the Pacific Northwest. PNW 619.

Maas FM, Balkhoven-Baart J, van der Steeg PAH (2014) Evaluation of Krymsk® 5 (VSL-2) and Krymsk® 6 (LC-52) as rootstocks for sweet cherry ‘Kordia’. Acta Horticulture 1058:531-536.

Mangal M, Sharma M, Kumar S (2014). In vitro regeneration in olive (Olea europaea L.) cv. ‘Frontio’ from nodal segments. Indian Journal of Experimental Biology 52:912-916.

Marino G, Bertazza G, Magnanini E, Doro Altan A (1993). Comparative effects of sorbitol and sucrose as main carbon energy sources in micropropagation of apricot. Plant Cell Tissue and Organ Culture 34:235-244.

Moing A, Langlois N, Svanella L, Zanetto A, Gaudillère J-P (1997). Variability in Sorbitol:Sucrose ratio in mature leaves of different Prunus species. Journal of the American Society for Horticultural Science 122:83-90.

Moncousin C, Ribaux M, O'Rourke JO, Gavillet S (1992). Effects of type of carbohydrate during proliferation and rooting of microcuttings of Malus ‘Jork 9’. Agronomie 12:775-781.

Monticelli S, Gentile A, Frattarelli A, Caboni E (2017). Effects of the natural cytokinin meta-Topolin on in vitro shoot proliferation and acclimatization of Prunus spp. Acta Horticulture 1155:375-380.

Murai Y, Harada H, Yamashita H (1997). In vitro propagation of apricot (Prunus armeniaca L.) cv. ‘Bakuoh junkyou’. Journal of the Japanese Society for Horticultural Science 66:475-480.

Nacheva L, Gercheva P (2009). Micropropagation of Gisela 5 (cherry dwarf rootstock): The effect of the type and the concentration of the carbohydrates in the nutrient medium. Acta Horticulture 825:261-268.

Nowak B, Miczynski K, Hudy L (2004). Sugar uptake and utilisation during adventitious bud differentiation on in vitro leaf explants of ‘Wegierka Zwyk?a’ plum (Prunus domestica). Plant Cell Tissue and Organ Culture 76: 255-260.

Pruski K, Astatkie T, Nowak J (2005). Tissue culture propagation of Mongolian cherry (Prunus fruticosa) and Nanking cherry (Prunus tomentosa). Plant Cell Tissue and Organ Culture 82:207-211.

Rafique A, Anis M (2014). Rapid in vitro propagation system through shoot tip cultures of Vitex trifolia L. Physiology and Molecular Biology of Plants 20:385-392.

Ranney TG, Bassuk NL, Whitlow TH (1991). Osmotic Adjustment and Solute Constituents in Leaves and Roots of Water-stressed Cherry (Prunus) trees. Journal of the American Society for Horticultural Science 116:684-688.

Rossi F, Baraldi R, Facini O, Lercari B (1993). Photomorphogenic effects on in vitro rooting of Prunus roostock GF 655-2. Plant Cell Tissue and Organ Culture 32:145-151.

Roussos PA, Pontikis CA (2002). In vitro propagation of olive (Olea europaea L.) cv. Koroneiki. Plant Growth Regulators 37:295-304.

Roussos PA, Tolia-Marioli A, Pontikis CA, Kotsias D (1999). Rapid multiplication of jojoba seedlings by in vitro culture. Plant Cell Tissue and Organ Culture 57:133-137.

Rugini E, Tarini P, Rossodivita ME (1987). Control of shoot vitrification of almond and olive grown in vitro. Acta Horticulture 212:177-183.

Ružic DjV, Lazic T, Cerovic R (2008). Micropropagation of some Prunus and Pyrus genotypes in vitro as affected by different carbon sources. Acta Horticulture 795:413-418.

Ružic DjV, Vujovic TI (2008). The effects of cytokinin types and their concentration on in vitro multiplication of sweet cherry cv. ‘Lapins’ (Prunus avium L.). Horticultural Science - Prague 35:12-21.

Sadeghi F, Yadollahi A, Jafarkhani Kermani M, Eftekhari M (2015). Optimizing culture media for in vitro proliferation and rooting of Tetra (Prunus empyrean) rootstock. Journal of Genetic Engineering and Biotechnology 13:19-23.

Schmülling T, Werner T, Riefler M, Krupková E, Bartrina y Manns I (2003). Structure and function of cytokinin oxidase/dehydrogenase genes of maize, rice, Arabidopsis and other species. Journal of Plant Research 116:241-252.

Tang H, Ren Z, Reustle G, Krezal G (2001). Plant regeneration from leaves of sweet and sour cherry cultivars. Scientia Horticulturae 93:235-244.

Tiwari V, Tiwari KN, Singh BD (2001). Comparative studies of cytokinins on in vitro propagation of Bacopa monniera. Plant Cell Tissue and Organ Culture 66:9-16.

van Staden J, Crouch NR (1996). Benzyladenine and derivatives - their significance and interconversion in plants. Plant Growth Regulators 19:153-175.

Webster AD (1993). New dwarfing rootstocks for apple, pear, plum and sweet cherry - A brief review. Acta Horticulturae 349:145-153.

Wiszniewska A, Nowak B, Kolton A, Sitek E, Grabski K, Dziurka M, Dlugosz-Grochowska O, Dziurka K, Tukaj Z (2016). Rooting response of Prunus domestica L. microshoots in the presence of phytoactive medium supplements. Plant Cell Tissue and Organ Culture 125:163-176.

Wojtania A (2010). Effect of meta-topolin on in vitro propagation of Pelargonium × hortorum and Pelargonium × hederaefolium cultivars. Acta Societatis Botanicorum Poloniae 79:101-106.

Yancheva S, Kondakova V (2016). Plant Tissue Culture Technology: Present and Future Development In: Pavlov A, Bley T (Eds.) Bioprocessing of plant in vitro systems, Reference Series in Phytochemistry. Springer International Publishing AG.

Yaseen M, Ahmad T, Sablok G, Standardi A, Hafiz IA (2013). Review: role of carbon sources for in vitro plant growth and development. Molecular Biology Reports 40:2837-2849.

Zhang H, Horgan KJ, Reynolds PHS, Jameson PE (2010). 6-Benzyladenine metabolism during reinvigoration of mature Pinus radiata buds in vitro. Tree Physiology 30:514-526.




DOI: http://dx.doi.org/10.15835/nbha47111276

June 1, 2018: Notulae Botanicae Horti Agrobotanici Cluj-Napoca in Scopus – Elsevier CiteScore 2017=0.78, Horticulture; Agronomy and Crop Science; Plant Science


 
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