Abstract
The low survival rates of in vitro-propagated plantlets under ex vitro conditions greatly inhibits the production of virus-free apple rootstock plantlets and necessitates tight control of ex vitro environments during plantlet acclimatization. Accordingly, this study investigated the effects of light intensity on the ex vitro acclimation of apple plantlets. In vitro-propagated ‘M9’ apple plantlets were acclimatized for 6 weeks under different light treatments: 60 μmol m−2 s−1 (L), 100 μmol m−2 s−1 (M), 140 μmol m−2 s−1 (H), 180 μmol m−2 s−1 (VH), 60 → 100 μmol m−2 s−1 at 2 weeks (L2M4) or 4 weeks (L4M2), 60 → 100 → 140 μmol m−2 s−1 (L2M2H2), and 60 → 140 μmol m−2 s−1 at 4 weeks (L4H2). Survival rate, maximum quantum yield of photosystem II (Fv/Fm), growth-related parameters, and photosynthetic rate were measured. The H and VH treatments yielded the lowest survival rates (78 and 71%, respectively), whereas the M treatment yielded the highest (95%). Meanwhile, the Fv/Fm ratio at 6 weeks after transplanting decreased with increasing light intensity at 4 and 5 weeks, whereas photosynthetic rate at 5 weeks after transplanting and stem diameter at 6 weeks after transplanting increased with increasing light intensity. Furthermore, the M treatment yielded greater relative growth rates than the other treatments at 2–4 weeks, and both the M and L2M2H2 treatments yielded significantly greater relative growth rates at 4–6 weeks. These results suggest that the M and L2M2H2 treatments are appropriate for the acclimatization of in vitro-propagated ‘M9’ apple plantlets.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ali MB, Hahn EJ, Paek KY (2005) Effects of light intensities on antioxidant enzymes and malondialdehyde content during short-term acclimatization on micropropagated Phalaenopsis plantlet. Environ Exp Bot 54:109–120. https://doi.org/10.1016/j.envexpbot.2004.06.005
Baťková P, Pospíšilová J, Synkova H (2008) Production of reactive oxygen species and development of antioxidative systems during in vitro growth and ex vitro transfer. Biol Plant 52:413–422. https://doi.org/10.1007/s10535-008-0085-5
Campbell AI (1962) Apple virus inactivation by heat therapy and tip propagation. Nature 195:520. https://doi.org/10.1038/195520a0
Carvalho LC, Osório ML, Chaves MM, Amâncio S (2001) Chlorophyll fluorescence as an indicator of photosynthetic functioning of in vitro grapevine and chestnut plantlets under ex vitro acclimatization. Plant Cell Tissue Organ Cult 67:271–280
Carvalho LC, Amâncio S (2002) Antioxidant defense system in plantlets transferred from in vitro to ex vitro: effects of increasing light intensity and CO2 concentration. Plant Sci 162:33–40. https://doi.org/10.1016/s0168-9452(01)00524-6
Cembali T, Folwell RJ, Wandschneider P, Eastwell KC, Howell WE (2003) Economic implications of a virus prevention program in deciduous tree fruits in the US. Crop Prot 22:1149–1156. https://doi.org/10.1016/s0261-2194(03)00156-x
Chatenet M, Delage C, Ripolles M, Irey M, Lockhart BEL, Rott P (2001) Detection of Sugarcane yellow leaf virus in quarantine and production of virus-free sugarcane by apical meristem culture. Plant Dis 85:1177–1180. https://doi.org/10.1094/pdis.2001.85.11.1177
Dami I, Hughes HG (1997) Effects of PEG-induced water stress on in vitro hardening of ‘Valiant’grape. Plant Cell Tissue Organ Cult 47:97–101. https://doi.org/10.1007/bf02318944
Escobar-Gutiérrez AJ, Gaudillère JP (1997) Carbon partitioning in source leaves of peach, a sorbitol-synthesizing species, is modified by photosynthetic rate. Physiol Planta 100:353–360. https://doi.org/10.1034/j.1399-3054.1997.1000218.x
Estrada-Luna AA, Davies FT, Egilla JN (2001) Physiological changes and growth of micropropagated chile ancho pepper plantlets during acclimatization and post-acclimatization. Plant Cell Tissue Organ Cult 66:17–24
Faisal M, Anis M (2009) Changes in photosynthetic activity, pigment composition, electrolyte leakage, lipid peroxidation, and antioxidant enzymes during ex vitro establishment of micropropagated Rauvolfia tetraphylla plantlets. Plant Cell Tissue Organ Cult 99:125–132. https://doi.org/10.1007/s11240-009-9584-0
Faisal M, Anis M (2010) Effect of light irradiations on photosynthetic machinery and antioxidative enzymes during ex vitro acclimatization of Tylophora indica plantlets. J. Plant Interact 5:21–27. https://doi.org/10.1080/17429140903137652
Fila G, Ghashghaie J, Hoarau J, Cornic G (1998) Photosynthesis, leaf conductance and water relations of in vitro cultured grapevine rootstock in relation to acclimatisation. Physiol Plant 102:411–418. https://doi.org/10.1034/j.1399-3054.1998.1020309.x
Fu W, Li P, Wu Y (2012) Effects of different light intensities on chlorophyll fluorescence characteristics and yield in lettuce. Sci Hortic 135:45–51. https://doi.org/10.1016/j.scienta.2011.12.004
Grout BWW, Aston MJ (1977) Transplanting of cauliflower plants regenerated from meristem culture. I. Water loss and water transfer related to changes in leaf wax and to xylem regeneration. Hortic Res 17:1–7
Guan QZ, Guo YH, Sui XL, Li W, Zhang ZX (2008) Changes in photosynthetic capacity and antioxidant enzymatic systems in micropropagated Zingiber officinale plantlets during their acclimation. Photosynthetica 46:193. https://doi.org/10.1007/s11099-008-0031-y
Hansen AJ, Lane WD (1985) Elimination of apple chlorotic leafspot virus from apple shoot cultures by ribavirin. Plant Dis 69:134–135
Hayashi M, Nakayama M, Kozai T (1988) An application of the acclimatization unit for growth of carnation explants, and for rooting and acclimatization of the plantlets. Acta Hortic 230:189–194. https://doi.org/10.17660/actahortic.1988.230.22
Hazarika BN (2003) Acclimatization of tissue-cultured plants. Curr Sci 85:1704–1712
Hu G, Dong Y, Zhang Z, Fan X, Ren F, Zhou J (2015) Virus elimination from in vitro apple by thermotherapy combined with chemotherapy. Plant Cell Tissue Organ Cult 121:435–443. https://doi.org/10.1007/s11240-015-0714-6
James DJ, Thurbon IJ (1979) Rapid in vitro rooting of the apple rootstock M.9. J Hortic Sci 54:309–311. https://doi.org/10.1080/00221589.1979.11514887
Jeon MW, Ali MB, Hahn EJ, Paek KY (2006) Photosynthetic pigments, morphology and leaf gas exchange during ex vitro acclimatization of micropropagated CAM Doritaenopsis plantlets under relative humidity and air temperature. Environ Exp Bot 55:183–194. https://doi.org/10.1016/j.envexpbot.2004.10.014
Kim JH, Kim CC, Ko KC, Kim KR, Lee JC (1998). The particular of pomology, Ed 4. Hyangmunsa, Korea, pp 41–45
Ko SM, Lee JH, Oh MM (2018a) Control of relative humidity and root-zone water content for acclimation of in vitro-propagated ‘M9’ apple rootstock plantlets. Hortic Environ Biotechnol 59:303–313. https://doi.org/10.1007/s13580-018-0038-7
Ko SM, Lee JH, Oh MM (2018b) Development of nutrient solution for in vitro propagation of ‘M9’apple rootstock plantlets. Hortic Sci Technol 36:202–214. https://doi.org/10.12972/kjhst.20180021
Kumar R, Kumar N (2003) High irradiance-induces changes in carotenoids compositions and increase in non-photochemical quenching of Chl fluorescence in primary wheat leaves. J Plant Physiol 160:1141–1146. https://doi.org/10.1078/0176-1617-01069
Noctor G, Gomez L, Vanacker H, Foyer CH (2002) Interactions between biosynthesis, compartmentation and transport in the control of glutathione homeostasis and signaling. J Exp Bot 53:1283–1304. https://doi.org/10.1093/jexbot/53.372.1283
Miller GL (1959) Modified DNS method for reducing sugars. Anal Chem 31:426–428
Mengel K, Friedrich B, Judel GK (1985) Effect of light intensity on the concentrations of phytohormones in developing wheat grains. J Plant Physiol 120:255–266. https://doi.org/10.1016/s0176-1617(85)80112-7
Mengin V, Pyl ET, Alexandre Moraes T, Sulpice R, Krohn N, Encke B, Stitt M (2017) Photosynthate partitioning to starch in Arabidopsis thaliana is insensitive to light intensity but sensitive to photoperiod due to a restriction on growth in the light in short photoperiods. Plant Cell Environ 40:2608–2627. https://doi.org/10.1111/pce.13000
Müller P, Li XP, Nigogi KK (2001) Non-photochemical quenching. A response to excess light energy. Plant Physiol 125:1558–1566. https://doi.org/10.1104/pp.125.4.1558
Osório ML, Osório J, Romano A (2010) Chlorophyll fluorescence in micropropagated Rhododendron ponticum subsp. baeticum plants in response to different irradiances. Biol Plant 54:415–422. https://doi.org/10.1007/s10535-010-0076-1
Paprstein F, Sedlak J, Polak J, Svobodova L, Hassan M, Bryxiova M (2008) Results of in vitro thermotherapy of apple cultivars. Plant Cell Tissue Organ Cult 94:347–352. https://doi.org/10.1007/s11240-008-9342-8
Pinto G, Silva S, Loureiro J, Costa A, Dias MC, Araújo C, Neves L, Santos C (2011) Acclimatization of secondary somatic embryos derived plants of Eucalyptus globulus Labill.: an ultrastructural approach. Trees 25:383–392. https://doi.org/10.1007/s00468-010-0513-y
Pospíšilová J, Wilhelmová NA, Synková H, Čatský J, Krebs D, Tichá I, Hanáčková B, Snopek J (1998) Acclimation of tobacco plantlets to ex vitro conditions as affected by application of abscisic acid. J Exp Bot 49:863–869. https://doi.org/10.1093/jxb/49.322.863
Pospís̆ilová J, Synková H, Haisel D, Čatský J, Wilhelmová NA, Šrámek F (1999) Effect of elevated CO2 concentration on acclimation of tobacco plantlets to ex vitro conditions. J Exp Bot 50:119–126
Pospóšilová J, Tichá I, Kadleček P, Haisel D, Plzáková Š (1999) Acclimatization of micropropagated plants to ex vitro conditions. Biol Plant 42:481–497. https://doi.org/10.1093/jxb/50.330.119
Rodríguez-López NF, Martins SC, Cavatte PC, Silva PE, Morais LE, Pereira LF, Reis JV, Ávila RT, Godoy AG, Lavinski AO, DaMatta FM (2014) Morphological and physiological acclimations of coffee seedlings to growth over a range of fixed or changing light supplies. Environ Exp Bot 102:1–10. https://doi.org/10.1016/j.envexpbot.2014.01.008
Saez PL, Bravo LA, Sáez KL, Sánchez-Olate M, Latsague MI, Ríos DG (2012) Photosynthetic and leaf anatomical characteristics of Castanea sativa: a comparison between in vitro and nursery plants. Biol Plant 56:15–24. https://doi.org/10.1007/s10535-012-0010-9
Shi K, Gu X, Lu W, Lu D (2018) Effects of weak-light stress during grain filling on the physicochemical properties of normal maize starch. Carbohydr Polym 202:47–55. https://doi.org/10.1016/j.carbpol.2018.08.114
Stitt M, Zeeman SC (2012) Starch turnover: pathways, regulation and role in growth. Curr Opin Plant Biol 15:282–292. https://doi.org/10.1016/j.pbi.2012.03.016
Van Huylenbroeck JM, Piqueras A, Debergh PC (1998) Photosynthesis and carbon metabolism in leaves formed prior and during ex vitro acclimatization of micropropagated plants. Plant Sci 134:21–30. https://doi.org/10.1016/s0168-9452(98)00043-0
Walkey DGA (1968) The production of virus-free rhubarb by apical tip-culture. J Hortic Sci 43:283–287. https://doi.org/10.1080/00221589.1968.11514255
Acknowledgements
This work was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through the Agri-Bio Industry Technology Development Program, which is funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA; Grant No. 315003051SB020).
Author information
Authors and Affiliations
Contributions
GJ conducted the measurements and data analysis and drafted the manuscript. JH helped with some of the measurements. MM provided substantial guidance regarding experiment design and critically revised the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Communicated by Heakeun Yun, Ph.D.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Chung, GJ., Lee, JH. & Oh, MM. Growth and acclimation of in vitro-propagated ‘M9’ apple rootstock plantlets according to light intensity. Hortic. Environ. Biotechnol. 61, 501–510 (2020). https://doi.org/10.1007/s13580-020-00247-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13580-020-00247-5