AJPS  Vol.7 No.14 , October 2016
Effect of Photon Flux Density and Exogenous Sucrose on the Photosynthetic Performance during In Vitro Culture of Castanea sativa
Abstract: The low photon flux density (PFD) under in vitro conditions and sucrose added to the culture medium negatively limits the photochemical activity and photoprotective mechanisms of microshoots. In this work we hypothesize that decreasing sucrose in the culture medium in combination with increasing irradiance, could improve the photosynthesis and consequently the in vitro growth. We evaluated the effect of exogenous sucrose (30 and 5 g·L-1, HS and LS, respectively), under different PFD (50 and 150 μmol photons m-2·s-1, LL and HL, respectively) on the photosynthetic performance and growth of Castanea sativa microshoots. Decreasing sucrose negatively affected the physiological attributes evaluated. Only chloroplast ultrastructure was improved by LS; however this did not lead to an improved in photosynthesis or growth. HL HS produced an increase in photosynthetic activity and chlorophyll contents, reaching under these conditions a higher proliferation rate and biomass production. Additionally, the photochemical activity (electron transport rate and non-photochemical quenching) was improved by HL. Thus, our results suggest that, at least for C. sativa HL is beneficial during the in vitro culture, improving photosynthetic performance as well as growth, but this is only possible in the presence of moderate concentrations of sucrose added to the culture medium.
Cite this paper: Sáez, P. , Bravo, L. , Sánchez-Olate, M. , Bravo, P. and Ríos, D. (2016) Effect of Photon Flux Density and Exogenous Sucrose on the Photosynthetic Performance during In Vitro Culture of Castanea sativa. American Journal of Plant Sciences, 7, 2087-2105. doi: 10.4236/ajps.2016.714187.

[1]   Lal, M., Tiwari, A.K. and Gupta, G.N. (2015) Commercial Scale Micropropagation of Sugarcane: Constraints and Remedies. Sugar Tech, 4, 339-347.

[2]   Kozai, T. and Kubota, C. (2001) Development a Photoautotrophic Micropropagation System for Woody Plants. Journal of Plant Research, 114, 525-537.

[3]   Xiao, Y., Niu, G. and Kozai, T. (2011) Development and Application of Photoautotrophic Micropropagation Plant System. Plant Cell, Tissue and Organ Culture, 105, 149-158.

[4]   Kozai, T. and Kubota, C. (2005) Concepts, Definitions, Ventilation Methods, Advantages and Disadvantages. In: Kozai, T., Afreen, F. and Zobayed, S.M.A., Eds., Photoautotrophic (Sugar-Free Medium) Micropropagation as a New Propagation and Transplant Production System, Springer, Dordrecht, 19-30.

[5]   Waman, A., Bohra. P., Sathyanarayana, B., Umesha, K., Mukunda, G.K., Ashok, T.H. and Gowda, B. (2015) Optimization of Factors Affecting In Vitro Establishment, Ex Vitro Rooting and Hardening for Commercial Scale Multiplication of Silk Banana (Musa AAB). Erwerbs-Obstbau, 57, 153-164.

[6]   Sáez, P., Bravo, L., Sáez, K., Sánchez-Olate, M., Latsague, M. and Ríos, D. (2012) Photosynthetic and Leaf Anatomical Characteristics of Castanea sativa: A Comparison between In Vitro and Nursery Plants. Biologia Plantarum, 56, 15-24.

[7]   Sáez, P., Bravo, L., Latsague, M., Toneatti, M., Sánchez-Olate, M. and Ríos, D. (2013) Light Energy Management in Micropropagated Plants of Castanea sativa, Effects of Photoinhibition. Plant Science, 201-202, 12-24.

[8]   Horton, P. and Ruban, A. (2005) Molecular Design of the Photosystem II Light-Harvesting Antenna: Photosynthesis and Photoprotection. Journal of Experimental Botany, 56, 365-373.

[9]   Walters, R.G. (2005) Towards an Understanding of Photosynthetic Acclimation. Journal of Experimental Botany, 56, 435-441.

[10]   Taiz, L. and Zeiger, E. (2002) Plant Physiology. 3rd Edition, Sinauer Associates, Inc. Publishers, Sunderland, MA, USA.

[11]   Yasodha, R., Kamala, S., Kumar, S.P.A., Kumar, D.P. and Kalaiarasi, K. (2008) Effect of Glucose on In Vitro Rooting of Mature Plants of Bambusa nutans. Scientia Horticulturae, 116, 113-116.

[12]   Mohamed, M. and Alsadon, A.A. (2010) Influence of Ventilation and Sucrose on Growth and Leaf Anatomy of Micropropaged Potato Plantlets. Scientia Horticulturae, 123, 295-300.

[13]   Santamaría, J.M., Lavergne, D., Trabelsi, S., Verdeil, J.L., Rival, A., Hamon, S. and Nato, A. (1999) Effect of Sucrose in the Medium, on the Photosynthetic Capacity of Coconut Vitroplants Derived from Zygotic Embryos. In: Recent Advances in Coconut Biotechnology, Kluwer Academic Publishers, Dordrecht, The Netherlands, 371-382.

[14]   Fuentes, G., Talavera, C., Oropeza, C., Desjardins, Y. and Santamaria, J. (2005) Exogenous Sucrose Can Decrease In Vitro Photosynthesis but Improve Field Survival and Growth of Coconut (Cocos nucifera L.) In Vitro Plantlets. In Vitro Cellular & Developmental Biology-Plant, 41, 69-76.

[15]   Koch, K.E. (1996) Carbohydrate-Modulated Gene Expression in Plants. Annual Review of Plant Physiology and Plant Molecular Biology, 47, 509-540.

[16]   Paul, M. and Stitt, M. (1993) Effect of Nitrogen and Phosphorous Deficiences on Levels of Carbohydrates, Respiratory Enzymes and Metabolites in Seedlings of Tobacoo and Their Response to Exogenoues Sucrose. Plant, Cell & Environment, 16, 1047-1057.

[17]   Tichá, I., Cáp, F., Pacosvská, D., Hofman, P., Haisel, D., Capková, V. and Schafer, C. (1998) Culture on Sugar Medium Enhances Photosynthesis Capacity and High Light Resistance of Plantlets Grown In Vitro. Physiologia Plantarum, 102, 155-162.

[18]   Carvalho, L. and Amancio, S. (2002) Effect of Ex Vitro Conditions on Growth and Adcquisition of Autotrophic Behaviour during the Acclimatisation of Chestnut Regenerated In Vitro. Scientia Horticulturae, 95, 151-164.

[19]   Sáez, P., Bravo, L., Latsague, M., Sánchez-Olate, M. and Ríos, D. (2012b) Increased Light Intensity during In Vitro Culture Improves Water Loss Control and Photosynthetic Performance of Castanea sativa Grown in Ventilated Vessels. Scientia Horticulturae, 138, 7-16.

[20]   Hasbun, R. (2006) Micropropagación de cultivares de Castanea sativa Mill., partir de material embrionario. Facultad de Ciencias Forestales, Universidad de Concepción, Concepción, Chile.

[21]   Lichtenthaler, H. and Wellburn, A. (1983) Determinations of Total Carotenoids and Chlorophylls a and b of Leaf Extracts in Different Solvents. Biochemical Society Transactions, 603, 591-592.

[22]   Rosenqvist, E. and Van Kooten, O. (2003) Chlorophyll Fluorescence: A General Description and Nomenclature. In: DeEll, J.R. and Toivonen, P.M.A., Eds., Practical Applications of Chlorophyll Fluorescence in Plant Biology, Kluwer Academic Publishers, 31-37.

[23]   Genty, B., Briantais, J.M. and Baker, N.R. (1989) The Relationship between the Quantum Yield of Photosynthetic Electron Transport and Quenching of Chlorophyll Fluorescence. Biochimica et Biophysica Acta (BBA)-General Subjects, 990, 87-92.

[24]   Kramer, D., Johnson, G., Kiirats, O. and Edwards, G. (2004) New Fluorescence Parameters for the Determination of QA Redox State and Excitation Energy Fluxes. Photosynthesis Research, 79, 209-218.

[25]   Maxwell, K. and Johnson, G.N. (2000) Chlorophyll Fluorescence: A Practical Guide. Journal of Experimental Botany, 51, 659-668.

[26]   Le Van, Q., Samson, G. and Desjardins, Y. (2001) Opposite Effects of Exogenous Sucrose on Growth, Photosynthesis and Carbon Metabolism of in Vitro Plantlets of Tomato (L. esculentum Mill.) Grown under Two Levels of Irradiances and CO2 Concentrations. Journal of Plant Physiology, 158, 599-605.

[27]   Mosaleeyanon, K., Cha-um, S. and Kirdmanee, C. (2004) Enhanced Growth and Photosynthesis of Rain Tree (Samanea saman Merr.) Plantlets in Vitro under a CO2-Enriched Condition with Decreased Sucrose Concentration in the Medium. Scientia Horticulturae, 103, 51-63.

[28]   Kovtun, Y. and Daie, J. (1995) End-Product Control of Carbon Metabolism in Culture Sugar Beet Plants. Plant Physiology, 108, 1647-1656.

[29]   Von Caemmerer, S. and Farquhar, G.D. (1981) Some Relationship between the Biochemistry of Photosynthesis and the Gas Exchange of Leaves. Planta, 153, 376-387.

[30]   Hdider, C. and Desjardins, Y. (1994) Reduction of Ribulose-1,5-Biphosphater Carboxylase/ Oxigenase Efficiency by the Presence of Sucrose during the Tissue Culture of Strawberry Plants. In Vitro Cellular & Developmental Biology-Plant, 31, 165-170.

[31]   Desjardins, Y. (1995) Photosynthesis in Vitro: On the Factor Regulating CO2 Assimilation in Micropropagation Systems. ISHS Acta Horticulturae, 393, 45-61.

[32]   Portis, A.R. (1992) Regulation of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase actIvity. Annual Review of Plant Physiology and Plant Molecular Biology, 43, 415-437.

[33]   Galmés, J., Aranjuelo, I., Medrano, H. and Flexas, J. (2013) Variation in Rubisco Content and Activity under Variable Climatic Factors. Photosynthesis Research, 117, 73-90.

[34]   Serret, M.D., Trillas, M.I., Mata, J. and Araus, J.L. (1996) Development of Photoautotrophy and Photoinhibition of Gardenia jasminoides Plantlets during Micropropagation. Plant Cell, Tissue and Organ Culture, 45, 1-16.

[35]   Posada, L., Padrón, Y., González, J., Rodríguez, R., Norman, O., Barbón, R., Hurtado, O., Rodriguez, R., Daniels, C. and Gómez-Kosky, R. (2015) Effects of Different Culture Conditions (Photoautotrophic, Photomixotrophic) and the Auxin Indole-Butyric Acid on the in Vitro Acclimatization of Papaya (Carica papaya L. var. Red Maradol) Plants Using Zeolite as Support. African Journal of Biotechnology, 14, 2622-2635.

[36]   Shao, Q.S., Wang, H.Z., Guo, H.P., Zhou, A., Huang, Y.Q., Sun, Y.L. and Li, M.Y. (2014) Effects of Shade Treatments on Photosynthetic Characteristics, Chloroplast Ultrastructure, and Physiology of Anoectochilus roxburghii. PLoS ONE, 2, e85996.

[37]   Mao, L.Z., Lu, H.F., Wang, Q. and Cai, M.M. (2007) Comparative Photosynthesis Characteristics of Calycanthus chinensis and Chimonanthus praecox. Photosynthetica, 45, 601-605.

[38]   De Riek, J., Piqueras, A. and Debergh, P. (1997) Sucrose Uptake and Metabolism in a Double Layer System for Micropropagation of Rosa multiflora. Plant Cell, Tissue and Organ Culture, 47, 269-278.

[39]   Eckstein, A., Zieba, P. and Gabrys, H. (2012) Sugar and Light Effects on the Condition of the Photosynthetic Apparatus of Arabidopsis thaliana Cultured in Vitro. Journal of Plant Growth Regulation, 31, 90-101.

[40]   Badr, A. and Desjardins, Y. (2007) Sugar Uptake and Metabolism in Tissue Cultured Potato Plantlets Cultured in Liquid Medium. ISHS Acta Horticulturae, 748, 265-273.

[41]   Pessoni, R.A., Tersarotto, C.C., Mateus, C.A., Zerlin, J.K., Simooes, K., de Cássia, L., Figueiredo-Ribeiro, R. and Braga, M.R. (2015) Fructose Affecting Morphology and Inducing β-Fructofuranosidases in Penicillium janczewskii. SpringerPlus, 4, 487.

[42]   Herbers, K., Monke, G., Badur, R. and Sonnewald, U. (1995) A Simplified Procedure for the Subtractive cDNA Cloning of Photoassimilate-Responding Genes: Isolation of cDNAs Encoding a New Class of Pathogenesis-Related Proteins. Plant Molecular Biology, 29, 1027-1038.

[43]   Mita, S., Suzuki-Fujii, K. and Nakamura, K. (1995) Sugar-Inducible Expression of a Gene for β-Amylase in Arabidopsis thaliana. Plant Physiology, 107, 895-904.

[44]   Hazarika, B. (2006) Morpho-Physiological Disorders in in Vitro Culture of Plants. Scientia Horticulturae, 108, 105-120.

[45]   Ghosh, P.K., Ramesh, P., Bandyopadhay, K.K., Tripathi, A.K., Hati, K.M. and Misra, A.K. (2004) Comparative Effectiveness of Cattle Manure, Poultry Manure, Phosphocompost and Fertilizer-NPK on Three Cropping Systems in Vertisoils of Semi-Arid Tropic. I. Crop Yields and Systems in Performance. Bioresource Technology, 95, 77-83.

[46]   Sestak, Z. (1996) Limitations for Finding Linear Relationship between Chlorophyll Content and Photosynthetic Activity. Biologia Plantarum, 8, 336-346.

[47]   Rodríguez, R., Aragón, A., Escalona, M., González, J. and Desjardins, Y. (2008) Carbon Metabolism in Leaves of Micropropagated Sugarcane during Acclimatization Phase. In Vitro Cellular & Developmental Biology-Plant, 44, 533-539.

[48]   Demmig-Adams, B., Gilmore, A. and Adams, W. (1996) Carotenoids 3: In Vivo Functions of Carotenoids in Higher Plants. The FASEB Journal, 10, 403-412.

[49]   Osório, M., Osório, J. and Romano, A. (2010) Chlorophyll Fluorescence in Micropropagated Rhododendron ponticum subsp. baeticum Plants in Response to Different Irradiances. Biologia Plantarum, 54, 415-422.

[50]   álvarez, C., Sáez, P., Sánchez-Olate, M. and Ríos, D. (2012) Effects of Light and Ventilation on Physiological Parameters during in Vitro Acclimatization of Gevuina avellana mol. Plant Cell, Tissue and Organ Culture (PCTOC), 110, 93-101.

[51]   Dietzel, L., Brautigam, K. and Pfannschmidt, T. (2008) Photosynthetic Acclimation: State Transition and Adjustment of Photosystem Stoichiometry—Functional Relationship between Short-Term and Long-Term Light Quality Acclimation in Plants. The FEBS Journal, 275, 1080-1088.

[52]   Hüner, N.P.A., Oquist, G. and Sarhan, F. (1998) Energy Balance and Acclimation to Light and Cold. Trends in Plant Science, 3, 224-230.

[53]   Lee, N., Wetzstein, H.Y. and Sommer, H.E. (1985) Effects of Quantum Flux Density on Photosynthesis and Chloroplast Ultrastructure in Tissue-Cultured Plantlets and Seedlings of Liquidambar styraciflua L. towards Improved Acclimatization and Field Survival. Plant Physiology, 78, 637-641.

[54]   Serret, M.D. and Trillas, M.I. (2000) Effects of Light and Sucrose Levels on the Anatomy, Ultrastructure, and Photosynthesis of Gardenia jasminoides Ellis Leaflets Cultured in Vitro. International Journal of Plant Sciences, 161, 281-289.

[55]   Capellades, M., Fontarnau, R., Carulla, C. and Debergh, P. (1991) Environment Influences Anatomy of Stomata and Epidermal Cells in Tissue Cultured Rosa multiflora. Journal of the American Society for Horticultural Science, 115, 141-145.

[56]   Cournac, L., Dimon, B., Carrier, P., Lohou, A. and Chagvardieff, P. (1991) Growth and Photosynthetic Characteristic of Solanum tuberosum Plantlets Cultivated in Vitro under Different Conditions of Aeration, Sucrose Supply, and CO2 Enrichment. Plant Physiology, 97, 112-117.