Optimal concentrations of plant growth regulators and AgNO3 for the improvement of regeneration efficiency in Chrysanthemum morifolium ‘Ohblang’

Yeo Jin Youn ･Yong Joon Yang

doi:10.5010/JPB.2023.50.021.169

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J Plant Biotechnol (2023) 50:169-175

Published online October 11, 2023

https://doi.org/10.5010/JPB.2023.50.021.169

국화 ‘오블랑’의 재생 효율 증진을 위한 식물생장조절제와 AgNO₃ 적정 농도 선별

윤여진･양용준

상명대학교 식물식품공학과

Received: September 14, 2023; Revised: September 18, 2023; Accepted: September 18, 2023

Optimal concentrations of plant growth regulators and AgNO₃ for the improvement of regeneration efficiency in Chrysanthemum morifolium ‘Ohblang’

Yeo Jin Youn ･Yong Joon Yang

Author Info. +

(Department of Plant and Food Sciences, Sangmyung University, Cheonan 31066, Korea)

Correspondence to : e-mail: yjyang@smu.ac.kr

Received: September 14, 2023; Revised: September 18, 2023; Accepted: September 18, 2023

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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Abstract

A plant regeneration system was developed through shoot organogenesis from in vitro leaf explants of Chrysanthemum morifolium ‘Ohblang’. The effects of different concentrations of plant growth regulators and AgNO₃ on efficient shoot regeneration and inhibition of browning were evaluated in chrysanthemum. The explants were cultured on MS shoot induction medium supplemented with 12 combination treatments of 6-benzyladenine (BA) 0.5, 1.0 and 2.0 mg/L, and α-naphthaleneacetic acid (NAA) 0.2, 0.5, 1.0 and 2.0 mg/L in darkness for 6 weeks and cultured under a 16/8 h photoperiod for 6 weeks. The highest shoot regeneration was obtained from the explants cultured on the medium with 1.0 mg/L BA and 1.0 mg/L NAA. Based on this result, AgNO₃ was added to a shoot induction medium containing MS salts, vitamins, 1.0 mg/L BA, 1.0 mg/L NAA, 30 g/L sucrose, and 6 g/L agar to reduce browning of chrysanthemum leaf explants. In the control treatment without AgNO₃, leaf explants turned brown at the cut edge; however, browning was not observed in AgNO₃ treatments. Shoot organogenesis was higher at low concentrations of AgNO₃ and decreased with an increase in AgNO₃ concentration. The explants cultured on shoot induction medium (MS salts, vitamins, 1.0 mg/L BA, 1.0 mg/L NAA) with 1 mg/L of AgNO₃ produced the highest shoot regeneration with 2.6 shoots per explants and a browning index of 0.7. When the regenerated shoots were detached from the explants and cultured on MS medium, the shoots were elongated and rooted successfully.

Keywords Chrysanthemum, ‘Ohblang’, Shoot organogenesis, Plant regeneration, Silver nitrate

Article

Research Article

J Plant Biotechnol 2023; 50(1): 169-175

Published online October 11, 2023 https://doi.org/10.5010/JPB.2023.50.021.169

국화 ‘오블랑’의 재생 효율 증진을 위한 식물생장조절제와 AgNO₃ 적정 농도 선별

윤여진･양용준

상명대학교 식물식품공학과

Received: September 14, 2023; Revised: September 18, 2023; Accepted: September 18, 2023

Optimal concentrations of plant growth regulators and AgNO₃ for the improvement of regeneration efficiency in Chrysanthemum morifolium ‘Ohblang’

Yeo Jin Youn ･Yong Joon Yang

(Department of Plant and Food Sciences, Sangmyung University, Cheonan 31066, Korea)

Correspondence to:e-mail: yjyang@smu.ac.kr

Received: September 14, 2023; Revised: September 18, 2023; Accepted: September 18, 2023

Abstract

Keywords: Chrysanthemum, &lsquo,Ohblang&rsquo,, Shoot organogenesis, Plant regeneration, Silver nitrate

Fig 1.

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Figure 1.Plant regeneration via adventitious shoot organogenesis from leaf explants of chrysanthemum ‘Ohblang’. (A, D) Callus and bud formation from leaf explants after 6 weeks in MS medium supplemented with 1.0 mg/L BA and 1.0 mg/L IAA. (B, E) Shoot formation from leaf explants cultured on MS medium with 1.0 mg/L BA and 1.0 mg/L NAA after 6 weeks (12 weeks in total). (C, F) Shoot multiplication and elongation after 8 weeks (16 weeks in total) in culture. (G) Rooting of regenerated shoot in MS medium. Scale bar: 1 cm

Journal of Plant Biotechnology 2023; 50: 169-175https://doi.org/10.5010/JPB.2023.50.021.169

Fig 2.

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Figure 2.Effect of the BA and NAA growth regulators on the in vitro shoot regeneration of chrysanthemum ‘Ohblang’ at 12 weeks of culture

Journal of Plant Biotechnology 2023; 50: 169-175https://doi.org/10.5010/JPB.2023.50.021.169

Fig 3.

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Figure 3.Comparisons of AgNO₃ concentrations on adventitious shoot formation from leaf explants in chrysanthemum ‘Ohblang’. The explants were cultured on shoot induction medium, composed of MS salts and vitamins with 1.0 mg/L BA and 1.0 mg/L NAA, for 6 weeks and then transferred to the same medium and cultured for 6 weeks

Journal of Plant Biotechnology 2023; 50: 169-175https://doi.org/10.5010/JPB.2023.50.021.169

Fig 4.

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Figure 4.Effects of AgNO₃ on browning of leaf explants in chrysanthemum 'Ohblang’. The browning index was calculated as follows: 0 = none of the explants are brown, 1 = 1/4 of the explants are brown, 2 = 2/4, 3 = 3/4, 4 = 4/4). Different letters indicate a significant difference according to Duncan's multiple range test at p < 0.05

Journal of Plant Biotechnology 2023; 50: 169-175https://doi.org/10.5010/JPB.2023.50.021.169

Table 1 . Effects of plant growth regulators on shoot regeneration from leaf explants in chrysanthemum ‘Ohblang’ after 12 weeks of culture^z.

PGR		Survival rate (%)	Callus formation (%)	Bud formation (%)	No. of buds/explants	Shoot formation (%)	No. of shoots/explants
BA (mg/L)	NAA (mg/L)	Survival rate (%)	Callus formation (%)	Bud formation (%)	No. of buds/explants	Shoot formation (%)	No. of shoots/explants
0.5	0.2	100.0 ± 0.0^y	100.0 ± 0.0	15.0 ± 6.1de^x	0.2 ± 0.1de	15.0 ± 6.1ef	0.2 ± 0.1de
0.5	0.5	100.0 ± 0.0	100.0 ± 0.0	55.0 ± 18.4bc	0.7 ± 0.2cd	55.0 ± 18.4abc	1.0 ± 0.3d
0.5	1.0	100.0 ± 0.0	100.0 ± 0.0	90.0 ± 6.1a	1.2 ± 0.2b	85.0 ± 6.1a	1.8 ± 0.2b
0.5	2.0	100.0 ± 0.0	100.0 ± 0.0	20.0 ± 9.4de	0.3 ± 0.2de	20.0 ± 9.4ef	0.2 ± 0.1de
1.0	0.2	100.0 ± 0.0	100.0 ± 0.0	35.0 ± 12.7cd	0.4 ± 0.1de	35.0 ± 12.7cde	0.6 ± 0.2cde
1.0	0.5	100.0 ± 0.0	100.0 ± 0.0	40.0 ± 6.1cd	0.5 ± 0.1d	35.0 ± 6.1cde	0.7 ± 0.cde
1.0	1.0	100.0 ± 0.0	100.0 ± 0.0	90.0 ± 6.1a	1.8 ± 0.2a	75.0 ± 7.9ab	3.2 ± 0.4a
1.0	2.0	100.0 ± 0.0	100.0 ± 0.0	55.0 ± 12.2bc	0.6 ± 0.1d	45.0 ± 12.2bcde	0.6 ± 0.2cde
2.0	0.2	100.0 ± 0.0	100.0 ± 0.0	0.0 ± 0.0	0.0 ± 0.0	0.0 ± 0.0	0.0 ± 0.0
2.0	0.5	100.0 ± 0.0	100.0 ± 0.0	25.0 ± 7.9de	0.3 ± 0.1de	25.0 ± 7.9def	0.5 ± 0.2cde
2.0	1.0	100.0 ± 0.0	100.0 ± 0.0	95.0 ± 5.0a	1.8 ± 0.2a	60.0 ± 10.0abc	1.0 ± 0.1c
2.0	2.0	100.0 ± 0.0	100.0 ± 0.0	80.0 ± 9.4ab	1.1 ± 0.1bc	45.0 ± 14.6bcde	0.9 ± 0.3cd

^ZExplants were cultured in the dark for 6 weeks and then cultured under light for 6 weeks..

^yMean ± SE..

^xMeans within columns followed by different letters are significantly different according to Duncan's multiple range test at p < 0.05..

Table 2 . Effects of AgNO₃ on shoot regeneration from leaf explants after 12 weeks of culture in chrysanthemum ‘Ohblang’^z.

AgNO₃ (mg/L)	Survival rate (%)	Callus formation (%)	Bud formation (%)	No. of buds/explants	Shoot formation (%)	No. of shoots/explants
0.0	100.0 ± 0.0^y	100.0 ± 0.0	80.0 ± 5.0a^x	2.7 ± 0.4a	60.0 ± 12.7ab	1.2 ± 0.4ab
1.0	100.0 ± 0.0	100.0 ± 0.0	85.0 ± 6.1a	3.4 ± 0.7a	80.0 ± 9.4a	2.6 ± 0.6a
3.0	100.0 ± 0.0	100.0 ± 0.0	80.0 ± 9.4a	4.1 ± 1.2a	55.0 ± 16.6ab	1.5 ± 0.4ab
5.0	100.0 ± 0.0	100.0 ± 0.0	65.0 ± 10.0a	3.1 ± 1.0a	30.0 ± 9.4b	0.9 ± 0.4b
7.0	100.0 ± 0.0	100.0 ± 0.0	65.0 ± 18.7a	2.1 ± 0.8a	40.0 ± 15.0ab	1.2 ± 0.5ab
10.0	100.0 ± 0.0	100.0 ± 0.0	70.0 ± 14.6a	4.7 ± 1.1a	50.0 ± 15.8ab	1.7 ± 0.6ab

^ZExplants were cultured in the dark for 6 weeks and then cultured under light for 6 weeks..

^yMean ± SE..

^xMeans within columns followed by different letters are significantly different according to Duncan's multiple range test at p < 0.05..

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국화 ‘오블랑’의 재생 효율 증진을 위한 식물생장조절제와 AgNO₃ 적정 농도 선별

Optimal concentrations of plant growth regulators and AgNO₃ for the improvement of regeneration efficiency in Chrysanthemum morifolium ‘Ohblang’

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국화 ‘오블랑’의 재생 효율 증진을 위한 식물생장조절제와 AgNO₃ 적정 농도 선별

Optimal concentrations of plant growth regulators and AgNO₃ for the improvement of regeneration efficiency in Chrysanthemum morifolium ‘Ohblang’

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