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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

© The Korean Society of Plant Biotechnology

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

윤여진・양용준

상명대학교 식물식품공학과

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

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

Copyright © The Korean Society of Plant Biotechnology.

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

윤여진・양용준

상명대학교 식물식품공학과

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

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, &lsquo,Ohblang&rsquo,, Shoot organogenesis, Plant regeneration, Silver nitrate

Fig 1.

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.

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.

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.

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 culturez.

PGRSurvival rate (%)Callus formation (%)Bud formation (%)No. of buds/explantsShoot formation (%)No. of shoots/explants
BA (mg/L)NAA (mg/L)
0.50.2100.0 ± 0.0y100.0 ± 0.015.0 ± 6.1dex0.2 ± 0.1de15.0 ± 6.1ef0.2 ± 0.1de
0.50.5100.0 ± 0.0100.0 ± 0.055.0 ± 18.4bc0.7 ± 0.2cd55.0 ± 18.4abc1.0 ± 0.3d
0.51.0100.0 ± 0.0100.0 ± 0.090.0 ± 6.1a1.2 ± 0.2b85.0 ± 6.1a1.8 ± 0.2b
0.52.0100.0 ± 0.0100.0 ± 0.020.0 ± 9.4de0.3 ± 0.2de20.0 ± 9.4ef0.2 ± 0.1de
1.00.2100.0 ± 0.0100.0 ± 0.035.0 ± 12.7cd0.4 ± 0.1de35.0 ± 12.7cde0.6 ± 0.2cde
1.00.5100.0 ± 0.0100.0 ± 0.040.0 ± 6.1cd0.5 ± 0.1d35.0 ± 6.1cde0.7 ± 0.cde
1.01.0100.0 ± 0.0100.0 ± 0.090.0 ± 6.1a1.8 ± 0.2a75.0 ± 7.9ab3.2 ± 0.4a
1.02.0100.0 ± 0.0100.0 ± 0.055.0 ± 12.2bc0.6 ± 0.1d45.0 ± 12.2bcde0.6 ± 0.2cde
2.00.2100.0 ± 0.0100.0 ± 0.00.0 ± 0.00.0 ± 0.00.0 ± 0.00.0 ± 0.0
2.00.5100.0 ± 0.0100.0 ± 0.025.0 ± 7.9de0.3 ± 0.1de25.0 ± 7.9def0.5 ± 0.2cde
2.01.0100.0 ± 0.0100.0 ± 0.095.0 ± 5.0a1.8 ± 0.2a60.0 ± 10.0abc1.0 ± 0.1c
2.02.0100.0 ± 0.0100.0 ± 0.080.0 ± 9.4ab1.1 ± 0.1bc45.0 ± 14.6bcde0.9 ± 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/explantsShoot formation (%)No. of shoots/explants
0.0100.0 ± 0.0y100.0 ± 0.080.0 ± 5.0ax2.7 ± 0.4a60.0 ± 12.7ab1.2 ± 0.4ab
1.0100.0 ± 0.0100.0 ± 0.085.0 ± 6.1a3.4 ± 0.7a80.0 ± 9.4a2.6 ± 0.6a
3.0100.0 ± 0.0100.0 ± 0.080.0 ± 9.4a4.1 ± 1.2a55.0 ± 16.6ab1.5 ± 0.4ab
5.0100.0 ± 0.0100.0 ± 0.065.0 ± 10.0a3.1 ± 1.0a30.0 ± 9.4b0.9 ± 0.4b
7.0100.0 ± 0.0100.0 ± 0.065.0 ± 18.7a2.1 ± 0.8a40.0 ± 15.0ab1.2 ± 0.5ab
10.0100.0 ± 0.0100.0 ± 0.070.0 ± 14.6a4.7 ± 1.1a50.0 ± 15.8ab1.7 ± 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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