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Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus caryophyllus semperflorens flore pleno) Cultivars Cover

Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus caryophyllus semperflorens flore pleno) Cultivars

Open Access
|Dec 2021

Figures & Tables

Figure 1

Different stages of organogenesis from leaf explants in carnation cultivars: A, B – explant type and callus induction after 21 days; C – green spots on the surface of the callus 25 days after sub-culture on the regeneration medium; D – efficient shoot induction on the MS medium supplemented with 3 mg·dm−3 of BA, 0.6 mg·dm−3 of NAA, 5 mg·dm−3 of AgNO3, and 40 mg·dm−3 of As two months after incubation; E – hyperhydric shoots regenerated in most explants; F – recovery from hyperhydricity on the modified MS (0.275 mg·dm−3 of NH4NO3) containing 0.5 mg·dm−3 of GA3, 0.3 mg·dm−3 of BA, and 0.1 mg·dm−3 of NAA after 2–3 weeks; G – shoot elongation on the MS medium with 1 mg·dm−3 of KIN and 0.1 mg·dm−3 of NAA; H – rooting on the ½ MS macro- and full micronutrients and 1.5 mg·dm−3 of IBA; I – the acclimatization of plantlets in the greenhouse with controlled climate system and high relative humidity; J, K – acclimatized plantlets transferred to the soil in the greenhouse (scale bars = 1 cm)
Different stages of organogenesis from leaf explants in carnation cultivars: A, B – explant type and callus induction after 21 days; C – green spots on the surface of the callus 25 days after sub-culture on the regeneration medium; D – efficient shoot induction on the MS medium supplemented with 3 mg·dm−3 of BA, 0.6 mg·dm−3 of NAA, 5 mg·dm−3 of AgNO3, and 40 mg·dm−3 of As two months after incubation; E – hyperhydric shoots regenerated in most explants; F – recovery from hyperhydricity on the modified MS (0.275 mg·dm−3 of NH4NO3) containing 0.5 mg·dm−3 of GA3, 0.3 mg·dm−3 of BA, and 0.1 mg·dm−3 of NAA after 2–3 weeks; G – shoot elongation on the MS medium with 1 mg·dm−3 of KIN and 0.1 mg·dm−3 of NAA; H – rooting on the ½ MS macro- and full micronutrients and 1.5 mg·dm−3 of IBA; I – the acclimatization of plantlets in the greenhouse with controlled climate system and high relative humidity; J, K – acclimatized plantlets transferred to the soil in the greenhouse (scale bars = 1 cm)

Figure 2

Ploidy level of the regenerated (right) and mother plants (left) of carnation cultivars through flow cytometry analysis. DNA content histogram in ‘Tabor’, ‘Mariposa’, and ‘Cameron’ cultivars showed no significant differences. The nuclei were isolated from the leaves of the mother and regenerated plants
Ploidy level of the regenerated (right) and mother plants (left) of carnation cultivars through flow cytometry analysis. DNA content histogram in ‘Tabor’, ‘Mariposa’, and ‘Cameron’ cultivars showed no significant differences. The nuclei were isolated from the leaves of the mother and regenerated plants

Callus induction response to treatments among different carnation cultivars based on Duncan’s multiple range grouping

Callus induction in different carnation cultivars
Dependent variableVisual scoreSS‘Mariposa’‘Eskimo’‘Cameron’‘Tabasco’‘Tabor’‘Noblesse’‘Liberty’‘White Liberty’‘Grand Slam’
CF 0–100%excellent≥ 86 91.67% 90.33%
good51–8572.33 62.25%
medium25–50 38.92% 37.72%
weak≤ 25 18.67%22.73% 19.67%
DCI 2–30 dayexcellent≤ 9 6 5
good10–19 14 14
medium20–2925 21 21 21
weak≥ 30 33
CGR score 1–5excellent5 5 5
good4 4 4
medium3 3 33
weak≤ 22 1
CC score 1–5green555555 5
green - yellow444444
white3 33
brown≤ 2 1

Effects of different PGR treatments on regeneration frequency for plant regeneration in different carnation cultivars

Regeneration frequency (%) for carnation cultivars
PGR (mg·dm−3)‘Mariposa’‘Eskimo’‘Cameron’‘Tabasco’‘Tabor’‘Noblesse’‘Liberty’‘White Liberty’‘Grand Slam’Mean for all cultivars
BANAAAsAgNO3
30.2004.76±0.74d23.81±0.74e23.81±0.74e28.57±1.3e19.05±0.74d23.81±0.74d19.05±0.74d23.81±0.74e4.76±0.74d19.05±0.95f
30.60019.05±0.74b66.67±0.74b71.43±1.3b66.67±0.74b66.67±0.74b42.86±2.24c66.67±0.74b66.67±0.74b19.05±0.74b53.96±2.38b
20.2004.76±0.74d14.29±1.3g4.76±0.74h14.29±0.14f9.53±0.74e14.29±1.3e9.53±0.74e4.76±0.74h9.53±0.74c9.53±0.46h
20.6000e38.1±0.74d9.53±1.5f38.1±0.74d0f23.81±1.98d0f9.53±1.5g0e13.23±1.79g
20.22059.53±1.4c38.1±0.74d33.34±0.74d38.1±0.74d9.53±1.44e33.34±0.74c9.53±1.5e23.81±1.98e23.81±1.98b24.34±1.36e
30.22059.53±0.74c33.34±1.5d76.19±0.74b38.1±1.98d33.34±1.5c80.95±0.74b76.19±0.74b38.1±1.98d23.81±1.5b45.51±2.86b
20.64054.76±0.74d57.14±1.27c61.91±1.5c61.91±1.5c42.86±0.79c61.91±1.5c33.34±1.5c42.86±1.3c9.53±0.74d41.80±2.46c
30.640538.1±0.74a90.47±0.74a95.24±0.74a95.24±0.74a80.95±0.74a95.24±0.74a90.47±0.74a90.47±0.74a33.34±0.74a78.83±2.76a
50.64054.76±0.74d23.81±0.74e66.67±0.74c61.91±0.74c28.57±2.24d14.29±1.3e28.57±2.24d19.05±1.98f9.53±0.74c28.57±2.42d
30.68050e19.05±0.74f19.05±0.74e19.05±0.74f33.34±1.5c0f28.57±1.3d19.05±0.74f4.76±0.74d15.87±1.32g

Effects of different treatments on shoot number per explant for plant regeneration in different carnation cultivars

Shoot number per explant for carnation cultivars
PGR (mg·dm−3)Mariposa’‘Eskimo’‘Cameron’‘Tabasco’‘Tabor’‘Noblesse’‘Liberty’‘White Liberty’‘Grand Slam’Mean for all cultivars
BANAAAsAgNO3
30.2000.43±0.07c1.81±0.08d3.85±0.2f1.81±0.78f2.81±0.12e1.81±0.08d2.24±0.7c1.81±0.077c0.43±0.07d1.89±0.12e
30.6001±0.02a1.15±0.022c8.53±0.12e8.5±0.11d7.76±0.12d1.91±0.13e1.91±0.13d5.86±0.18b1±0.02c4.18±0.38d
20.2000.29±0.05d0.67±0.055e0.29±0.044g1.05±0.1g1.05±.1f0.76±0.06e0.67±0.5e0.29±0.04d0.57±0.04d0.63±0.03f
20.6000e1.81±0.077d3.71±0.59g4.76±0.56g0g1.81±0.08d0f1.05±0.1c0 e1.46±0.20e
20.22050.76±0.12c2.29±0.0c20.67±0.91d3.14±0.066f0.76±0.12f5±0.33c0.76±0.12e1.53±0.12c1.95±0.17c4.22±0.85d
30.22050.76±0.06b7.09±017b28.3±0.29a21.24±1.29c21.24±1.29c6.47±0.21b7.81±0.21b7.09±0.18b2.28±0.22b11.36±1.08b
20.64052.71±0.34a5.76±0.37c24.19±0.38b27.81±0.66a24.19±0.38a5.76±0.38c11.57±0.31b21.81±0.38a2.71±0.34a14.06±1.15b
30.64051±0.033a22.33±0.11a23.76±0.17b22±0.1b24.14±0.12a19±0.36a20.19±0.11a22.33±0.38a0.67±0.22b17.27±1.05a
50.64050.19±0.033d6.4±0.1b7.81±0.21e16.19±0.51c4.76±0.41e2.9±0.36c5±0.55d4.76±0.41c0.62±0.06c5.4±0.53c
30.68050e0.76±0.033e1.05±0.01g5.09±0.01e1.57±0.04e0 f2.38±0.01c1.28±0.011c0.38±0.06c1.39±0.18e

Different PGR treatments used for callus induction from leaf segments of carnation

PGRs (mg·dm−3)Treatment number
12345678910111213141516
2,4-D0.50.50.50.50.20.2110.50.50.50.50.50.50.20.2
BA0.20.20.50.50.20.20.20.20.20.20.50.50000
NAA0.50.50.50.50.50.50.50.500000000
CH02000200020002000200020002000200
DOI: https://doi.org/10.2478/johr-2021-0020 | Journal eISSN: 2353-3978 | Journal ISSN: 2300-5009
Language: English
Page range: 65 - 74
Submitted on: Aug 1, 2021
Accepted on: Nov 1, 2021
Published on: Dec 31, 2021
Published by: National Institute of Horticultural Research
In partnership with: Paradigm Publishing Services
Publication frequency: 2 times per year

© 2021 Hamid Reza Sabaghi, Gholamreza Sharifi-Sirchi, Pejman Azadi, Mohammad Hossein Azimi, published by National Institute of Horticultural Research
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.