Arman PAZUKI, Fatemeh AFLAKI, Songül GÜREL, Ali ERGÜL, Ekrem GÜREL

The effects of proline on in vitro proliferation and propagation of doubled haploid sugar beet (Beta vulgaris)

Doubled haploid induction is one of the available methods normally used for sugar beet breeding. Gynogenic haploid explants of sugar beet induced with 1 or 2 mg L-1 6-benzylaminopurine were treated with 5 g L-1 colchicine, then subcultured on a solidified MS medium plus 0.2 mg L-1 kinetin. Colchicine doubled the chromosome number of 27.7% of the treated haploid explants. With the aim of increasing the number of doubled haploid explants, the effects of five levels of proline (0.0, 0.1, 0.2, 0.3, or 0.4 mM) on the explants' proliferation, propagation, and shoot length were compared. With a large effect size (ES), proline at 0.3 mM induced the highest amount of proliferation, while proline-free medium resulted in the lowest amount of it. The highest propagation rates were observed for the explants treated on media with 0.2 and 0.3 mM proline (very large ES). Proline at 0.3 mM induced the shortest shoots (medium ES). A very strong positive correlation between proliferation and propagation, a moderate negative correlation between proliferation and length, and a strong negative correlation between propagation and length were observed. For the first time our results show beneficial effects of proline on in vitro proliferation and propagation of sugar beet.

Keywords:

Proline, propagation, proliferation, doubled haploid, Beta vulgaris,

PDF

___

Aflaki F, Pazuki A, Gürel S, Stevanato S, Biancardi E, Gürel E (2017). Doubled haploid sugar beet: an integrated view of factors influencing the processes of gynogenesis and chromosome doubling. Int Sugar J 119: 884-895.
Aksakal O, Tabay D, Esringu A, Aksakal FI, Esim N (2017). Effect of proline on biochemical and molecular mechanisms in lettuce ( Lactuca sativa L.) exposed to UV-B radiation. Photochem Photobiol Sci 16: 246-254.
Barow M, Meister A (2003). Endopolyploidy in seed plants is differently correlated to systematics, organ, life strategy and genome size. Plant Cell Environ 26: 571-584.
Burritt DJ (2008). Efficient cryopreservation of adventitious shoots of Begonia x Erythrophylla using encapsulation–dehydration requires pretreatment with both ABA and proline. Plant Cell Tiss Organ Cult 95: 209-215.
Castro S, Romeiras MM, Castro M, Duarte MC, Loureiro J (2013). Hidden diversity in wild Beta taxa from Portugal: insights from genome size and ploidy level estimations using flow cytometry. Plant Sci 207: 72-78.
Cohen J (1988). Statistical Power Analysis for the Behavioral Sciences. 2nd ed. Hillsdale, NJ, USA: Lawrence Erlbaum Associates, Inc., pp. 24–27.
Couillerot JP, Windels D, Vazquez F, Michalski JC, Hilbert JL, Blervacq AS (2012). Pretreatments, conditioned medium and co-culture increase the incidence of somatic embryogenesis of different Cichorium species. Plant Signal Behav 7: 121-131.
Doležel J, Bartoš JAN (2005). Plant DNA flow cytometry and estimation of nuclear genome size. Ann Bot-London 95: 99- 110.
Dörffling K, Dörffling H, Luck E (2009). Improved frost tolerance and winter hardiness in proline overaccumulating winter wheat mutants obtained by in vitro-selection is associated with increased carbohydrate, soluble protein and abscisic acid (ABA) levels. Euphytica 165: 545-556.
Eujayl I, Strausbaugh C, Lu C (2016). Registration of sugarbeet doubled haploid line KDH13 with resistance to beet curly top. J Plant Regist 10: 93-96.
Field A (2013). Discovering Statistics Using IBM SPSS Statistics. 4th ed. London, UK: Sage.
Franck T, Kevers C, Gaspar T, Dommes J, Deby C, Greimers R, Serteyn D, Deby-Dupont G (2004). Hyperhydricity of Prunus avium shoots cultured on gelrite: a controlled stress response. Plant Physiol Bioch 42: 519-527.
Górecka K, Krzyżanowska D, Kowalska U, Kiszczak W, Podwyszyńska M (2017). Development of embryoids by microspore and anther cultures of red beet ( Beta vulgaris L. subsp. vulgaris). J Cent Eur Agr 18: 185-195.
Gürel E (1997). Callus and root development from leaf explants of sugar beet ( Beta vulgaris L.): variability at variety, plant and organ level. Turk J Bot 21: 131-136.
Gürel E, Gürel S, Lemaux PG (2008). Biotechnology applications for sugar beet. Crit Rev Plant Sci 27: 108-140.
Gürel S, Gürel E (2013). In vitro regeneration of sugar beet ( Beta vulgaris L.). In: Ramawatand KG, Merillon JM, editors. Bulbous Plants: Biotechnology. Boca Raton, FL, USA: CRC Press, pp. 113-151.
Gürel S, Gürel E, Kaya Z (2002). Protoplast fusion in sugar beet ( Beta vulgaris L.). Turk J Biol 26: 163-170.
Hagedorn F, Joseph J, Peter M, Luster J, Pritsch K, Geppert U, Kerner R, Molinier V, Egli S, Schaub M et al. (2016). Recovery of trees from drought depends on belowground sink control. Nature Plants 2: 16111.
Hansen AL, Gertz A, Joersbo M, Andersen SB (2000). Chromosome doubling in vitro with amiprophos-methyl in Beta vulgaris ovule culture. Acta Agr Scand B-S P 50: 89-95.
Howell DC (2012). Statistical Methods for Psychology. 8th ed. Belmont, CA, USA: Wadsworth.
Ivic-Haymes SD, Smigocki AC (2005). Identification of highly regenerative plants within sugar beet ( Beta vulgaris L.) breeding lines for molecular breeding. In Vitro Cell Dev Biol- Plant 41: 483-488.
Karimi-Ashtiyani R, Ishii T, Niessen M, Stein N, Heckmann S, Gurushidze M, Banaei-Moghaddam AM, Fuchs J, Schubert V, Koch K et al. (2015). Point mutation impairs centromeric CENH3 loading and induces haploid plants. P Natl Acad Sci USA 112: 11211-11216.
Klimek-Chodacka M, Baranski R (2013). Comparison of haploid and doubled haploid sugar beet clones in their ability to micropropagate and regenerate. Electron J Biotechnol 16: 1-10.
Maluszynski M, Kasha KJ, Forster BP, Szarejko I (2003). Doubled Haploid Production in Crop Plants: A Manual. Berlin, Germany: Springer.
Manjili FA, Sedghi M, Pessarakli M (2012). Effects of phytohormones on proline content and antioxidant enzymes of various wheat cultivars under salinity stress. J Plant Nutr 35: 1098-1111.
Mezei S, Kovacev L, Nagl N (2006). Sugar beet micropropagation. Biotechnol Biotec Eq 20: 9-14.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plantarum 15: 473-497.
Nanjo T, Kobayashi M, Yoshiba Y, Sanada Y, Wada K, Tsukaya H, Kakubari Y, Yamaguchi‐Shinozaki K, Shinozaki K (1999). Biological functions of proline in morphogenesis and osmotolerance revealed in antisense transgenic Arabidopsis thaliana . Plant J 18: 185-193.
Pazuki A, Aflaki F, Gürel E, Ergül A, Gürel S (2018a). Gynogenesis induction in sugar beet ( Beta vulgaris ) improved by 6-benzylaminopurine (BAP) and synergized with cold pretreatment. Sugar Tech 20: 69-77.
Pazuki A, Aflaki F, Gurel E, Ergul A, Gurel S (2017) A robust method for haploid sugar beet ( Beta vulgaris ) in vitro proliferation and hyperhydricity reduction. Folia Hort 29: 241-250.
Pazuki A, Aflaki F, Gürel S, Ergül A, Gürel E (2018b). Production of doubled haploids in sugar beet ( Beta vulgaris ): an efficient method by a multivariate experiment. Plant Cell Tiss Org Cult 132: 85-97.
Pazuki A, Asghari J, Sohani MM, Pessarakli M, Aflaki F (2015). Effects of some organic nitrogen sources and antibiotics on callus growth of indica rice cultivars. J Plant Nutr 38: 1231- 1240.
Pedersen HC, Keimer B (1996). Haploidy in sugar beet ( Beta vulgaris L). In: Jain SM, Sopory SK, Veilleux RE, editors. In Vitro Haploid Production in Higher Plants, Vol. 3. Important Selected Plants. Dordrecht, the Netherlands: Kluwer Academic Publishers, pp. 17-36.
Per TS, Khan NA, Reddy PS, Masood A, Hasanuzzaman M, Khan MI, Anjum NA (2017). Approaches in modulating proline metabolism in plants for salt and drought stress tolerance: phytohormones, mineral nutrients and transgenics. Plant Physiol Biochem 115: 126-140.
Perchlik M, Tegeder M (2017). Improving plant nitrogen use efficiency through alteration of amino acid transport processes. Plant Physiol 175: 235-247.
Pospíšilová J, Synková H, Rulcová J (2000). Cytokinins and water stress. Biol Plantarum 43: 321-328.
Putnik-Delic M, Maksimovic I, Venezia A, Nagl N (2013). Free proline accumulation in young sugar beet plants and in tissue culture explants under water deficiency as tools for assessment of drought tolerance. Rom Agric Res 30: 141-148.
Řezbová H, Maitah M, Škubna O, Smutka L (2016). The economic aspects of sugar beet production. Ver Sem Agr 25: 327–335.
Sliwinska E, Zielinska E, Jedrzejczyk I (2005). Are seeds suitable for flow cytometric estimation of plant genome size? Cytometry 64: 72-79.
Svirshchevskaya AM, Doleze FJ (2000). Production and performance of gynogenetic sugarbeet lines. J Sugar Beet Res 37: 117-133. Szabados L, Savoure A (2010). Proline: a multifunctional amino acid. Trends Plant Sci 15: 89-97.
Tian J, Cheng Y, Kong X, Liu M, Jiang Z, Wu F (2017). Induction of reactive oxygen species and the potential role of NADPH oxidase in hyperhydricity of garlic plantlets in vitro. Protoplasma 254: 379-388.
Tsai CJ, Saunders JW (1999). Evaluation of sole nitrogen sources for shoot and leaf disc cultures of sugarbeet. Plant Cell Tiss Organ Cult 59: 47-56.
van den Dries N, Giannì S, Czerednik A, Krens FA, de Klerk GJM (2013). Flooding of the apoplast is a key factor in the development of hyperhydricity. J Exp Bot 64: 5221-5230.
Verbruggen N, Hermans C (2008). Proline accumulation in plants: a review. Amino Acids 35: 753-759.
Weber J, Georgiev V, Haas C, Bley T, Pavlov A (2010). Ploidy levels in Beta vulgaris (red beet) plant organs and in vitro systems. Eng Life Sci 10: 139-147.