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Use of coconut water and fertilizer for in vitro proliferation and plantlet production of Dendrobium ‘Gradita 31’

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Abstract

To improve the micropropagation of Dendrobium ‘Gradita 31’, coconut water (CW) and fertilizer media were used to enhance growth, proliferation, and germination of protocorm-like bodies (PLBs). PLBs formed when shoots approximately 0.4 cm long were cultured on semi-solid half-strength Murashige and Skoog (MS) basal medium containing 1 mg L−1 thidiazuron (TDZ) and 0.5 mg L−1 N6-benzyladenine (BA). After four rounds of 15-d subcultures, PLBs were initially proliferated in liquid half-strength MS medium supplemented with 0.3 mg L−1 TDZ and 0.1 mg L−1 α-naphthalene acetic acid (NAA) and subcultured monthly into fresh medium. When 15% (v/v) CW was added to half-strength MS medium, optimal growth and proliferation of PLBs resulted with the following principal findings: a maximum of 2.86 g total fresh weight (FW) of PLBs formed from 1 g of PLBs, an average of 0.46 g FW of PLBs added per subculture period, a total of 157.1 PLBs formed, an average of 25.3 PLBs added per subculture period, and a low percentage of PLB browning (20.7%). Half-strength MS medium containing 15% (v/v) CW with Rosasol® medium (liquid fertilizer; 1.5 g L−1 18 N:18P:18 K + 1.5 g L−1 25 N:10P:10 K + TE) resulted in a similar organogenic outcome with 25.5% PLB browning. PLBs germinated easily and rooted on Rosasol-supplemented plant growth regulator-free medium. In total, 180 Dendrobium ‘Gradita 31’ plantlets from all treatments were successfully acclimatized with 100% survival on a Cycas rumphii bulk substrate and grew well after repotting in a mixture of wood charcoal and C. rumphii bulk (1:1, v/v).

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Authors and Affiliations

  1. Indonesian Ornamental Crops Research Institute, Jln. Raya Ciherang, Pacet, Cianjur, 43253, West Java, Indonesia

    Budi Winarto

  2. P. O. Box 7, Miki-cho post office, Ikenobe 3011-2, Kagawa-ken, 761-0799, Japan

    Jaime A. Teixeira da Silva

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  1. Budi Winarto
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Correspondence to Budi Winarto or Jaime A. Teixeira da Silva.

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Supplementary Figure 1

Multiplication of D. ‘Gradita 31’ PLBs in the presence of different concentrations of coconut water (CW) (in mL L−1: 0, 75, 150, 300) over four subcultures (SC). Average FW of PLBs added in each SC. Means followed by the same letter in the same SC are not significantly different (Tukey’s multiple range test, P = 0.05). Coefficients of variation (%): 0% (SC-0), 4.32% (SC-1), 2.06% (SC-2), 1.74% (SC-3), 5.88% (SC-4). (DOCX 28 kb)

Supplementary Figure 2

Multiplication of D. ‘Gradita 31’ PLBs in the presence of different fertilizer media (FM) over four subcultures (SC). Average FW of PLBs added in each SC. Means followed by the same letter in the same SC are not significantly different (Tukey’s multiple range test, P = 0.05). ControL−1: half-strength MS medium; Control 2: VW medium; FM-1: Hyponex® medium (1.5 g L−1 25N:5P:20K + 1.5 g L−1 10N:40P:15K); FM-2: Hyponex® medium (3 g L−1 25N:5P:20K); FM-3: Growmore® medium (1.5 g L−1 32N:10P:10K + 1.5 g L−1 20N:20P:20K); FM-4: Growmore® medium (1.5 g L−1 20N:20P:20K + 1.5 g L−1 6N:30P:30K); FM-5: Rosasol® medium (1.5 g L−1 25N:10P:10K + TE + 1.5 g L−1 15N:10P:30K + TE); FM-6: Rosasol® medium (1.5 g L−1 18N:18P:18K + TE + 1.5 g L−1 25N:10P:10K + TE). Coefficients of variation (%): 0% (SC-0), 3.07% (SC-1), 1.16% (SC-2), 1.32% (SC-3), 2.74% (SC-4). (DOCX 32 kb)

Supplementary Figure 3

Multiplication of D. ‘Gradita 31’ PLBs in three selected media i.e. MS, VW and Rosasol medium in monthly periodical subcultures (SC). Average FW of PLBs added in each SC. Means followed by the same letter in the same SC are not significantly different (Tukey’s multiple range test, P = 0.05). Coefficients of variation (%): 0% (SC-0), 3.13% (SC-1), 1.38% (SC-2), 1.06% (SC-3), 1.56% (SC-4). (DOCX 29 kb)

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Winarto, B., da Silva, J.A.T. Use of coconut water and fertilizer for in vitro proliferation and plantlet production of Dendrobium ‘Gradita 31’. In Vitro Cell.Dev.Biol.-Plant 51, 303–314 (2015). https://doi.org/10.1007/s11627-015-9683-z

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