Effects of Ventilation and Sucrose Concentrations on the Growth and Plantlet Anatomy of Micropropagated Persian Walnut Plants

Document Type: Research paper

Authors

1 Department of Horticulture, College of Aburaihan, University of Tehran, Pakdasht, Iran.

2 Department of Horticulture, College of Aburaihan, University of Tehran, Pakdasht, Iran

3 Department of Plant Sciences, College of Sciences, University of Tehran, Tehran, Iran

4 National Clonal Germplasm Repository, USDA ARS, One Shields Drive, University of California, Davis, CA 95616

5 Department of Natural Resources Biotechnology, Research Institute of Forests and Rangelands, Tehran, Iran.

Abstract

Plantlets grown in conventional tissue culture systems usually encounter physiological and anatomical abnormalities including inability to photosynthesize, low chlorophyll content, open stomata, lack of a cuticle layer in the leaf, abnormal xylem parenchyma etc. Photoautotrophic and photomixotrophic systems could diminish these problems. The purpose of this study was to increase the chlorophyll content and photosynthetic ability of shoots grown in vitro and to improve the adaptation of walnut plantlets. Walnut apical leaf buds were cultured in vessels containing DKW medium supplemented with 0, 15, 30 and 45 g L-1of sucrose. The vessels were closed with a clear polypropylene lid with two syringe filters on the lid (V1) or with a 50 mm microporous polypropylene membrane (V2). Natural ventilation had a significant effect on most of the growth indices. Furthermore, different levels of sucrose had significant effects on growth characteristics. Natural ventilation increased chlorophyll content significantly. By using ventilated vessels containing 15 g L-1 sucrose, the total chlorophyll was significantly increased. Stomata under non-ventilated conditions were spherical with wide openings, whereas those in ventilated vessels were elliptical with narrow openings. As a result, ventilated vessels with 15 g L-1of sucrose produced healthy plantlets.

Keywords


1. Afreen, F. 2007. Physiological and Anatomical Characteristics of in Vitro Photoautotrophic Plants, pp 62-87. In: T. Kozai, F. Afreen and S.A.M. Zobayed (eds.) Photoautotrophic (Sugar-Free Medium) Micropropagation as a New Micropropagation and Transplant Production System, Springer Publishers, Netherlands.
2. Afreen, F., S.A.M. Zobayed, and T. Kozai. 2002. Photoautotrophic Culture of Coffea arabusta Somatic Embryos: Photosynthetic Ability and Growth of Different Stage Embryos. Ann. Bot. 90:11-19.
3. Avilés, F., D. Ríos, R. González, and M. Sánchez-Olate. 2009. Effect of Culture Medium in Callogenesis from Adult Walnut Leaves (Juglans regia L.). Chilean J. Agr. Res. 69:460-467.
4. Cui, Y., E. Hahn, T. Kozai, and K. Paek. 2000. Number of Air Exchanges, Sucrose Concentration, Photosynthetic Photon Flux, and Differences in Photoperiod and Dark Period Temperatures Affect Growth of Rehmannia glutinosa Plantlets in Vitro. Plant Cell Tissue Organ Cult. 62:219-226.
5. Driver, J.A., and A.H. Kuniyuki. 1984. In Vitro Propagation of Paradox Walnut Rootstock. Hort. Sci. 19:507–509.
6. Grout, B.W.W., and M.E. Donkin. 1987. Photosynthetic Activity of Cauliflower Meristem Cultures in Vitro and at Transplanting into Soil. Acta Hort. 212:323–327.
7. Hazarika, B.N. 2006. Morpho-Physiological Disorders in Vitro Culture of Plants. Scientia Hort. 108:105–120.
8. Hazarika, B.N., V.A. Parthasarathy, and V. Nagaraju. 2004. Influence of in Vitro Preconditioning of Citrus Sp. Microshoots with Sucrose on their ex Vitro Establishment. Indian J. Hort. 61:29–31.
9. Hdider, C. and, Y. Desjardins. 1994. Effect of Sucrose on Photosynthesis and Phosphoenol Pyruvate Carboxylase Activity of in Vitro Culture Strawberry Plantlets. Plant Cell Tissue Organ Cult. 36:27–33.
10. Hdider, C. and Y. Desjardins. 1995. Reduction of Ribolose-1, 5-Bisphosphate Carboxylase/Oxygenase Efficiency by the Presence of Sucrose during the Tissue Culture of Strawberry Plantlets. In Vitro Cellular Dev. Biol. Plant. 31:165-170.
11. Ibaraki, Y., Y. Iida, and K. Kurata. 1992. Effects of Air Currents on Gas Exchange of Culture Vessels. Acta Hort.319: 221-224.
12. Jackson, M.B., A. J. Abbott, A.R. Belcher, and K.C. Hall. 1987. Gas Exchange in Plant Tissue Cultures, pp. 57-71. In: M.B. Jackson, S.H. Mantell and J. Blake (eds.) Advances in the Chemical Manipulation of Plant Tissue Cultures. British Plant Growth Regulator Group Publishers, University of London.

13. Jackson, M.B., A.R. Belcher, and P. Brain. 1994. Measuring Shortcomings in Tissue Culture Aeration and their Consequences for Explants Development, pp. 191-203. In: P.J. Lumsden, J.R. Nicholas and W.J. Davies (eds.) Physiology, Growth and Development of Plants in Culture. Kluwer Academic, Dordrecht, Netherlands.
14. Jeong, B.R., T. Kozai, and K. Watanabe. 1996. Stem Elongation and Growth of Mentha rotundifolla in Vitro as Influenced by Photoperiod, Photosynthetic Photon Flux and Difference between Day and Night Temperatures. Acta Hort. 440:539-544.
15. Jo, M.H., I. K. Ham, A.M. Lee, M.E. Lee, H.N. Song, H.G. Han, and S.I. Woo. 2002. Effects of Sealing Materials and Photosynthetic Photon Flux of Culture Vessel on Growth and Vitrification in Carnation Plantlets in Vitro. J. Korean Soc. Hort. Sci. 43:133-136.
16. Kaul, K., and S. Sabharwal. 1971. Effects of Sucrose and Kinetin on Growth and Chlorophyll Synthesis in Tobacco Tissue Cultures. Plant Physiol. 47:691-695.
17. Koch, K.E. 1996. Carbohydrate Modulated Gene Expression in Plants. Ann. Rev. Plant Physiol. Plant Mol. Biol. 47:509–540.
18. Kozai, T. and S.M.A. Zobayed. 2001. Acclimatization, pp 1-12. In: R. Spier (Ed.) Encyclopedia of Cell Technology. Wiley Publishers, New York.
19. Kozai, T., and C. Kubota. 2001. Developing a Photoautotrophic Micropropagation System for Woody Plants. J. Plant Res. 114:525-53
20. Kozai, T., K. Tanaka, B R. Jeong, and K. Fujiwara. 1993. Effect of Relative Humidity in the Culture Vessel on the Growth and Shoot Elongation of Potato (Solanum tuberosum L.) Plantlets in Vitro. J. Jpn. Soc. Hort. Sci. 62:413–417.
21. Kozai, T., Y. Koyama, and I. Watanabe. 2002. Multiplication of Potato Plantlets in Vitro with Sugar-Free Medium Under High Photosynthetic Photon Flux. Acta. Hort. 230:121-128.
22. Kubota, C. 2002. Photoautotrophic Micropropagation: Importance of Controlled Environment in Plant Tissue Culture. Proc. Intl. Plant Prop. Soc. 52:906– 913.
23. Larcher, W. 1995. Physiological Plant Ecology, pp. 424-426. Springer Verlag, Berlin, Heidelberg.
24. Mohamed, M.H. and A.A. Alsadon. 2010. Influence of Ventilation and Sucrose on Growth
and Leaf Anatomy of Micropropagated Potato Plantlets. Sci. Hort. 123:295–300.
25. Murashige, T., and F. Skoog. 1962. A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures. Physiol. Plant. 15:473–479.
26. Park, S.W., J.H. Jeon, H.S. Kim, Y.M. Park, C. Aswath, and H. Joung. 2004. Effect of Sealed and Vented Gaseous Microenvironments on the Hyperhydricity of Potato Shoots in Vitro. Sci. Hort. 99:199-205.
27. Payghamzadeh, K. and S.K. Kazemitabar. 2011. In vitro propagation of walnut - A review. African J. Biotechnol. 10:290-311.
28. Pruski, K., T. Astatkie, M. Mirza, and J. Nowak. 2002. Photoautotrophic Micropropagation of Russet Burbank Potato. Plant Cell Tissue Organ Cult. 69:197–200.
29. Rahman, M.H., and A.A. Alsadon. 2007. Photoautotrophic and Photomixotrophic Micropropagation of Three Potato Cultivars. J. Biol. Sci. 15:111-116.
30. Reverberi, M., G. Falasca, P. Lauri, E. Caboni, and M.M. Altamura. 2001. Indoleacetic Acid Induces Xylem Formation Instead Rooting in Walnut (Juglans regia L.) Microcuttings. Plant Biosyst. 135:71-77.
31. Richardson A.D., S.P. Duigan, and G.P. Berlyn. 2002. An Evaluation of Noninvasive Methods to Estimate Foliar Chlorophyll Content. New Phytol. 153:185-194.
32. Roberts, A.V., E.F. Smith, I. Horan, S. Walker, D. Matthews, and J. Mottley. 1994. Stage III Techniques for Improving Water Relations and Autotrophy in Micropropagated Plants, pp 314-322. In: P.J. Lumsden, J.R. Nicholas and W.J. Davies (eds.) Physiology, Growth and Development of Plants in Culture. Kluwer Academic Publishers, Netherlands.
33. Sampson, J. 1961. A Method of Replicating Dry or Moist Surfaces for Examination by Light Microscopy, Nature. 191:932–3.
34. Sha Valli, K.P.S., T. Kozai, Q.T. Nguyen, C. Kubota, and V. Dhawan. 2003. Growth and Water Relations of Paulownia fortunei under Photomixotrophic and Photoautotrophic Conditions. Biol. Plant 46:161–166.
35. Xiao, Y., and T. Kozai. 2004. Commercial Application of a Photoautotrophic Micropropagation System Using Large Vessels with Forced Ventilation: Plantlet Growth and Production Cost. Hort. Sci. 39:1387-1391.

36. Vahdati, K., C. Leslie, Z. Zamani, and G. McGranahan. 2004. Rooting and Acclimatization of in-Vitro Grown Shoots from Three Mature Persian Walnut Cultivars. HortScience 39: 324-327.
37. Zobayed S.M.A. 2000. In Vitro Propagation of Lagerstroemia Spp. From Nodal Explants and Gaseous Composition in the Culture Head Space. Environ. Control Biol. 38:1-11.
38. Zobayed, S.M.A. 2007. Ventilation in Micropropagation, pp 150-186. In: T. Kozai, F. Afreen and S.A.M. Zobayed (eds.) Photoautotrophic (Sugar-Free Medium)
Micropropagation as a New Micropropagation and Transplant Production System, Springer Publishers, Netherlands.
39. Zobayed, S.M.A., F. Afreen, and T. Kozai. 2001a. Physiology of Eucalyptus Plantlets Grown Photoautotrophically in a Scaled-Up Vessel. In Vitro Cellular Dev. Biol. Plant 37:807-813.
40. Zobayed, S.M.A., J. Armstrong, and W. Armstrong. 2001b. Leaf Anatomy of in Vitro Tobacco and Cauliflower Plantlets as Affected by Different Types of Ventilation. Plant Sci. 161:537-548.