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

PGRs (mg·dm⁻³)	Treatment number
PGRs (mg·dm⁻³)	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

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)

Table 2

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

[i] CF – callus frequency, DCI – days to callus induction (showing the number of days required for visible callus induction), CGR – callus growth rate scored from 1 for lowest to 5 for highest, CC – callus color scored from 1 for brown to 5 for green, SS – statistical score

Table 3

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	‘Mariposa’	‘Eskimo’	‘Cameron’	‘Tabasco’	‘Tabor’	‘Noblesse’	‘Liberty’	‘White Liberty’	‘Grand Slam’	Mean for all cultivars
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

[i] Different letters in a column indicate significant differences according to Duncan’s multiple range test (p < 0.05); values represent mean ± SE

Table 4

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	Mariposa’	‘Eskimo’	‘Cameron’	‘Tabasco’	‘Tabor’	‘Noblesse’	‘Liberty’	‘White Liberty’	‘Grand Slam’	Mean for all cultivars
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

[i] Note: See Table 1

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

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DOI: https://doi.org/10.2478/johr-2021-0020 | Journal eISSN: 2353-3978 (formerly 2300-5009) | Journal ISSN: 2300-5009

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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

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© 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.

Volume 29 (2021): Issue 2 (December 2021)

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

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