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Efficient production of virus-free apple plantlets using the temporary immersion bioreactor system

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Abstract

Viral pathogens reduce the quality and yield of apple (Malus domestica) fruits by 30–50%. A mass production system of virus-free apple plantlets is needed to meet the demand of the domestic fruit tree industry. In this study, we compared the production of virus-free plants in different in vitro culture systems, including a temporary immersion bioreactor (TIB), a continuous immersion bioreactor, and conventional solid and liquid culture systems (controls). Apple plantlets were immersed in the TIB once every 3 h (TIB-3) or 6 h (TIB-6). The fresh weight of apple plants was the highest in the TIB-3 system and lowest in the liquid culture. Shoots were the longest in the TIB-3 system, approximately twofold longer than those in solid culture and liquid culture. Roots of apple plants were the longest in the TIB-3 system compared with solid and liquid cultures. Root number in the TIB-3 system was also higher than that in solid and liquid cultures. Moreover, leaf area was the highest in plants grown in the TIB-3 treatment. The total stem area of TIB-3 plants was the largest at 1.46 mm2. This study suggests that the airlift bioreactor is capable of producing a large number of virus-free plants in a short time compared with conventional culture systems. Additionally, secondary xylem was well developed in the stems of plants grown in the TIB-3 system. Therefore, this system shows a high potential for producing healthy plants suitable for acclimatization.

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References

  • Akdemir H, Süzerer V, Onay A, Tilkat E, Ersali Y, Çiftçi YO (2014) Micropropagation of the pistachio and its rootstocks by temporary immersion system. Plant Cell Tissue Organ 117:65–76

    Article  CAS  Google Scholar 

  • Alvarez C, Sáez P, Sáez K, Sánchez-Olate M, Ríos D (2012) Effects of light and ventilation on physiological parameters during in vitro acclimatization of Gevuina avellana mol. Plant Cell Tissue Organ 110:93–101

    Article  CAS  Google Scholar 

  • Ascough GD, Fennel CW (2004) The regeneration of plant growth and development in liquid culture. S Afr J Bot 70:181–190

    Article  CAS  Google Scholar 

  • Benelli C, Carlo AD (2018) In vitro multiplication and growth improvement of Olea europaea L. cv Canino with temporary immersion system (Plantform™). 3 Biotech 8:317. https://doi.org/10.1007/s13205-018-1346-4

    Article  PubMed  PubMed Central  Google Scholar 

  • Chen J, Tang HH, Li L, Qin SJ, Wang GP, Hong N (2017) Effect of virus infection on plant growth, root development and phytohormone levels in in vitro-cultured pear plants. Plant Cell Tissue Organ 131:359–368

    Article  CAS  Google Scholar 

  • Chen L, Wang MR, Li JW, Feng CH, Cui ZH, Zhao L (2019) Exogenous application of melatonin improves eradication of apple stem grooving virus from the infected in vitro shoots by shoot tip culture. Plant Pathol 68:997–1006

    Article  CAS  Google Scholar 

  • Cieślińska M, Rutkowski KP (2008) Effect of Apple chlorotic leaf spot virus on yield and quality of fruits from ‘Golden Delicious’ and ‘Sampion’ apple trees. Acta Hortic 781:119–124

    Article  Google Scholar 

  • Cuenca B, Sanchez C, Aldrey A, Bogo B, Blanco B, Correa B, Vidal N (2017) Micropropagation of axillary shoots of hybrid chestnut (Castanea sativa × C. crenata) in liquid medium in a continuous immersion system. Plant Cell Tissue Organ 131:307–320

    Article  CAS  Google Scholar 

  • Frometa OM, Morgado MME, Silva JAT, Morgado DTP, Gradaille MAD (2017) In vitro propagation of Gerbera jamesonii Bolus ex Hooker f. in a temporary immersion bioreactor. Plant Cell Tissue Organ 129:543–551

    Article  Google Scholar 

  • Gao M, Jiang W, Wei S, Lin Z, Cai B, Yang L, Luo C, He X, Tan J, Chen L (2015) High-efficiency propagation of Chinese water chestnut [Eleocharis dulcis (Burm. f.) Trin. ex Hensch] using a temporary immersion bioreactor system. Plant Cell Tissue Organ 121:761–772

    Article  CAS  Google Scholar 

  • Gatti E, Sgarbi E, Ozudogru EA, Lambardi M (2017) The effect of Plantform™ bioreactor on micropropagation of Quercus robur in comparison to a conventional in vitro culture system on gelled medium, and assessment of the microenvironment influence on leaf structure. Plant Biosyst 151:1129–1136

    Article  Google Scholar 

  • George EF, Hall MA, De Klerk GJD (2008) Micropropagation: uses and methods. In: George EF, Hall MA, De Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, pp 29–64

    Google Scholar 

  • Georgiev V, Schumann A, Pavlov A, Bley T (2014) Temporary immersion systems in plant biotechnology. Eng Life Sci 14:607–621

    Article  CAS  Google Scholar 

  • Hadidi A, Barba M (2011) Economic impact of pome and stone fruit viruses and viroids. In: Hadidi A, Barba M, Candresse TH, Jelkmann W (eds) Virus and virus-like diseases of pome and stone fruits. APS Press, College Park, pp 1–7

    Chapter  Google Scholar 

  • Hu GJ, Dong YF, Zhang ZP, Fan XD, Ren F, Li ZN (2017) Efficacy of virus elimination from apple by thermotherapy coupled with in vivo shoot-tip grafting and in vitro meristem culture. J Phytopathol 165:701–706

    Article  CAS  Google Scholar 

  • Ko SM, Lee JH, Oh MM (2018) Development of nutrient solution for in vitro propagation of ‘M9’ apple rootstock plantlets. Hortic Sci Technol 36:202–214

    Google Scholar 

  • Kovalchuk I, Lyudvikova Y, Volgina M, Reed BM (2009) Medium, container and genotype all influence in vitro cold storage of apple germplasm. Plant Cell Tissue Organ 96:127–136

    Article  Google Scholar 

  • Kwon AR, Cui HY, Lee HS, Shin HN, Kang KS, Park SY (2015) Light quality affects shoot regeneration, cell division, and wood formation in elite clones of Populus euramericana. Acta Physiol Plant 37:65. https://doi.org/10.1007/s11738-015-1812-0

    Article  Google Scholar 

  • Larema L, da Cruz ACF, Saldanha CW, Dias LLC, Vieira RF, de Oliveira EJ, Otoni WC (2012) Photoautotrophic propagation of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen]. Plant Cell Tissue Organ 110:227–238

    Article  Google Scholar 

  • Le KC, Jeong CS, Lee H, Paek KY, Park SY (2019) Ginsenoside accumulation profiles in long- and short-term cell suspension and adventitious root cultures in Panax ginseng. Hortic Environ Biotechnol 60:125–134

    Article  CAS  Google Scholar 

  • Li BQ, Feng CH, Hu LY, Wang MR, Wang QC (2016) Shoot tip culture and cryopreservation for eradication of Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) from apple rootstocks ‘M9’ and ‘M26’. Ann Appl Biol 168:142–150

    Article  CAS  Google Scholar 

  • Lichtenthaler HK (1987) Chlorophyll fluorescence signatures of leaves during the autumnal chlorophyll breakdown. J Plant Physiol 131:101–110

    Article  CAS  Google Scholar 

  • Martínez-Estrada E, Islas-Luna B, Pérez-Sato JA, Bello-Bello JJ (2019) Temporary immersion improves in vitro multiplication and acclimatization of Anthurium andreanum Lind. Sci Hortic 249:185–191

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol 15:473–497

    Article  CAS  Google Scholar 

  • Park SY, Moon HK, Murthy HN, Kim YW (2011) Improved growth and acclimatization of somatic embryo-derived Oplopanax elatus plantlets by ventilated photoautotrophic culture. Biol Plant 55:559–562

    Article  Google Scholar 

  • Quiala E, Canal MJ, Meijon M, Rodriguez R, Chavez M, Valledor L, Feria M, Barbon R (2012) Morphological and physiological responses of proliferating shoots of teak to temporary immersion and BA treatments. Plant Cell Tissue Organ 109:223–234

    Article  CAS  Google Scholar 

  • Ramírez-Mosqueda MA, Iglesias-Andreu LG, Ramirez-Madero G, Hernandez-Rincon EU (2016) Micropropagation of Stevia rebaudiana Bert. in temporary immersion systems and evaluation of genetic fidelity. S Afr J Bot 106:238–243

    Article  Google Scholar 

  • Ramos-Castellá A, Iglesias-Andreu LG, Bello-Bello J, Lee-Espinosa H (2014) Improved propagation of vanilla (Vanilla planifolia Jacks. ex Andrews) using a temporary immersion system. In Vitro Cell Dev Biol-Plant 50:576–581

    Article  Google Scholar 

  • Sáez PL, Bravo LA, Latsague MI, Sanchez-Olate ME, Ríos DG (2012) Increased light intensity during in vitro culture improves water loss control and photosynthetic performance of Castanea sativa grown in ventilated vessels. Sci Hortic 130:7–16

    Article  Google Scholar 

  • Schuetz M, Smith R, Ellis B (2012) Xylem tissue specification, patterning, and dirrerentiation mechanisms. J Exp Bot 64:11–31

    Article  Google Scholar 

  • Ševčíková H, Lhotáková Z, Hamet J, Lipavská H (2018) Mixotrophic in vitro cultivations: the way to go astray in plant physiology. Physiol Plant. https://doi.org/10.1111/ppl.12893

    Article  Google Scholar 

  • Sreedhar RV, Venkatachalam L, Neelwarne B (2009) Hyperhydricity-related morphologic and biochemical changes in vanilla (Vanilla planifolia). J Plant Growth Regul 28:46–57

    Article  CAS  Google Scholar 

  • Valdez-Tapia R, Capataz-Tafur J, López-Laredo AR, Trejo-Espino JL, Trejo-Tapia G (2014) Effect of immersion cycles on growth, phenolics content, and antioxidant properties of Castilleja tenuiflora shoots. Vitro Cell Dev Biol-Plant 50:471–477

    Article  CAS  Google Scholar 

  • Vives K, Andújar I, Lorenzo JC, Concepción O, Hernández M, Escalona M (2017) Comparison of different in vitro micropropagation methods of Stevia rebaudiana B. including temporary immersion bioreactor (BIT®). Plant Cell Tissue Organ 131:195–199

    Article  CAS  Google Scholar 

  • Yeung EC (1999) The use of histology in the study of plant tissue culture systems-some practical comments. In Vitro Cell Dev Biol Plant 35:137–143

    Article  Google Scholar 

  • Zhang B, Song L, Bekele LD, Shi J, Jia Q, Zhang B, Jin L, Duns GJ, Chen J (2018) Optimizing factors affecting development and propagation of Bletilla striata in a temporary immersion bioreactor system. Sci Hortic 232:121–126

    Article  CAS  Google Scholar 

  • Ziska LH, Panicker S, Wojno HL (2008) Recent and projected increases in atmospheric carbon dioxide and the potential impacts on growth and alkaloid production in wild poppy (Papaver setigerum DC.). Clim Change 91:395. https://doi.org/10.1007/s10584-008-9418-9

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through Agri-Bio industry Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (Grant Number 315003-5).

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Authors and Affiliations

  1. Division of Animal, Horticulture and Food Sciences, Department of Horticultural Science, Chungbuk National University, Cheongju, 28466, Republic of Korea

    Na-Young Kim, Ho-Dong Hwang, Jin-Ho Kim, Bo-Min Kwon, Daeil Kim & So-Young Park

  2. Useful Plant Resources Center, Korea National Arboretum, Pocheon, Gyunggi-do, 11186, Republic of Korea

    Jin-Ho Kim

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  1. Na-Young Kim
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  2. Ho-Dong Hwang
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  3. Jin-Ho Kim
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  4. Bo-Min Kwon
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  5. Daeil Kim
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Contributions

NYK contributed to the data acquisition and wrote the manuscript. HDH, JHK, and BMK participated in the experiment and sample analysis. DK participated in data interpretation and revising of the manuscript. S-YP made substantial contributions to data interpretation, revising of the manuscript, the conception, and design of this study.

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Correspondence to So-Young Park.

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Communicated by Heakeun Yun, Ph.D.

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Kim, NY., Hwang, HD., Kim, JH. et al. Efficient production of virus-free apple plantlets using the temporary immersion bioreactor system. Hortic. Environ. Biotechnol. 61, 779–785 (2020). https://doi.org/10.1007/s13580-020-00257-3

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  • DOI: https://doi.org/10.1007/s13580-020-00257-3

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