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

Hamid Reza Sabaghi
Hamid Reza Sabaghi
, 
Gholamreza Sharifi-Sirchi
Gholamreza Sharifi-Sirchi
, 
Pejman Azadi
Pejman Azadi
 oraz 
Mohammad Hossein Azimi
Mohammad Hossein Azimi
   | 31 gru 2021
Journal of Horticultural Research's Cover Image
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Data publikacji: 31 gru 2021
Zakres stron: 65 - 74
Otrzymano: 01 sie 2021
Przyjęty: 01 lis 2021
DOI: https://doi.org/10.2478/johr-2021-0020
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© 2021 Hamid Reza Sabaghi et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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 variable Visual score SS ‘Mariposa’ ‘Eskimo’ ‘Cameron’ ‘Tabasco’ ‘Tabor’ ‘Noblesse’ ‘Liberty’ ‘White Liberty’ ‘Grand Slam’
CF 0–100% excellent ≥ 86 91.67% 90.33%
good 51–85 72.33 62.25%
medium 25–50 38.92% 37.72%
weak ≤ 25 18.67% 22.73% 19.67%
DCI 2–30 day excellent ≤ 9 6 5
good 10–19 14 14
medium 20–29 25 21 21 21
weak ≥ 30 33
CGR score 1–5 excellent 5 5 5
good 4 4 4
medium 3 3 3 3
weak ≤ 2 2 1
CC score 1–5 green 5 5 5 5 5 5 5
green - yellow 4 4 4 4 4 4
white 3 3 3
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
BA NAA As AgNO3
3 0.2 0 0 4.76±0.74d 23.81±0.74e 23.81±0.74e 28.57±1.3e 19.05±0.74d 23.81±0.74d 19.05±0.74d 23.81±0.74e 4.76±0.74d 19.05±0.95f
3 0.6 0 0 19.05±0.74b 66.67±0.74b 71.43±1.3b 66.67±0.74b 66.67±0.74b 42.86±2.24c 66.67±0.74b 66.67±0.74b 19.05±0.74b 53.96±2.38b
2 0.2 0 0 4.76±0.74d 14.29±1.3g 4.76±0.74h 14.29±0.14f 9.53±0.74e 14.29±1.3e 9.53±0.74e 4.76±0.74h 9.53±0.74c 9.53±0.46h
2 0.6 0 0 0e 38.1±0.74d 9.53±1.5f 38.1±0.74d 0f 23.81±1.98d 0f 9.53±1.5g 0e 13.23±1.79g
2 0.2 20 5 9.53±1.4c 38.1±0.74d 33.34±0.74d 38.1±0.74d 9.53±1.44e 33.34±0.74c 9.53±1.5e 23.81±1.98e 23.81±1.98b 24.34±1.36e
3 0.2 20 5 9.53±0.74c 33.34±1.5d 76.19±0.74b 38.1±1.98d 33.34±1.5c 80.95±0.74b 76.19±0.74b 38.1±1.98d 23.81±1.5b 45.51±2.86b
2 0.6 40 5 4.76±0.74d 57.14±1.27c 61.91±1.5c 61.91±1.5c 42.86±0.79c 61.91±1.5c 33.34±1.5c 42.86±1.3c 9.53±0.74d 41.80±2.46c
3 0.6 40 5 38.1±0.74a 90.47±0.74a 95.24±0.74a 95.24±0.74a 80.95±0.74a 95.24±0.74a 90.47±0.74a 90.47±0.74a 33.34±0.74a 78.83±2.76a
5 0.6 40 5 4.76±0.74d 23.81±0.74e 66.67±0.74c 61.91±0.74c 28.57±2.24d 14.29±1.3e 28.57±2.24d 19.05±1.98f 9.53±0.74c 28.57±2.42d
3 0.6 80 5 0e 19.05±0.74f 19.05±0.74e 19.05±0.74f 33.34±1.5c 0f 28.57±1.3d 19.05±0.74f 4.76±0.74d 15.87±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
BA NAA As AgNO3
3 0.2 0 0 0.43±0.07c 1.81±0.08d 3.85±0.2f 1.81±0.78f 2.81±0.12e 1.81±0.08d 2.24±0.7c 1.81±0.077c 0.43±0.07d 1.89±0.12e
3 0.6 0 0 1±0.02a 1.15±0.022c 8.53±0.12e 8.5±0.11d 7.76±0.12d 1.91±0.13e 1.91±0.13d 5.86±0.18b 1±0.02c 4.18±0.38d
2 0.2 0 0 0.29±0.05d 0.67±0.055e 0.29±0.044g 1.05±0.1g 1.05±.1f 0.76±0.06e 0.67±0.5e 0.29±0.04d 0.57±0.04d 0.63±0.03f
2 0.6 0 0 0e 1.81±0.077d 3.71±0.59g 4.76±0.56g 0g 1.81±0.08d 0f 1.05±0.1c 0 e 1.46±0.20e
2 0.2 20 5 0.76±0.12c 2.29±0.0c 20.67±0.91d 3.14±0.066f 0.76±0.12f 5±0.33c 0.76±0.12e 1.53±0.12c 1.95±0.17c 4.22±0.85d
3 0.2 20 5 0.76±0.06b 7.09±017b 28.3±0.29a 21.24±1.29c 21.24±1.29c 6.47±0.21b 7.81±0.21b 7.09±0.18b 2.28±0.22b 11.36±1.08b
2 0.6 40 5 2.71±0.34a 5.76±0.37c 24.19±0.38b 27.81±0.66a 24.19±0.38a 5.76±0.38c 11.57±0.31b 21.81±0.38a 2.71±0.34a 14.06±1.15b
3 0.6 40 5 1±0.033a 22.33±0.11a 23.76±0.17b 22±0.1b 24.14±0.12a 19±0.36a 20.19±0.11a 22.33±0.38a 0.67±0.22b 17.27±1.05a
5 0.6 40 5 0.19±0.033d 6.4±0.1b 7.81±0.21e 16.19±0.51c 4.76±0.41e 2.9±0.36c 5±0.55d 4.76±0.41c 0.62±0.06c 5.4±0.53c
3 0.6 80 5 0e 0.76±0.033e 1.05±0.01g 5.09±0.01e 1.57±0.04e 0 f 2.38±0.01c 1.28±0.011c 0.38±0.06c 1.39±0.18e

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

PGRs (mg·dm−3) Treatment number
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
2,4-D 0.5 0.5 0.5 0.5 0.2 0.2 1 1 0.5 0.5 0.5 0.5 0.5 0.5 0.2 0.2
BA 0.2 0.2 0.5 0.5 0.2 0.2 0.2 0.2 0.2 0.2 0.5 0.5 0 0 0 0
NAA 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0 0 0 0 0 0 0 0
CH 0 200 0 200 0 200 0 200 0 200 0 200 0 200 0 200
eISSN:
2353-3978
Język:
Angielski
Częstotliwość wydawania:
2 razy w roku
Dziedziny czasopisma:
Life Sciences, Biotechnology, Plant Science, Ecology, other
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