Isolation, culture of protoplasts of Angelica gigas Nakai and regeneration of plants via somatic embryogenesis

Lee, Han-Sol; Han, Jong-Eun; Jie, Eun Yee; Kim, Suk Weon; Kwon, Hyuk Joon; Lee, Gun-Myung; Lee, Hak Sung; Murthy, Hosakatte Niranjana; Park, So-Young

doi:10.1007/s11240-023-02666-5

Cited by 1 publication

(3 citation statements)

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“…Angelica gigas is an important medicinal plant that has many therapeutic values [ 16 ], earlier we have reported successful protoplast isolation using the embryogenic callus of A. gigas and reported the optimum duration of enzyme treatment, medium for culture of protoplasts and plant regeneration [ 19 ]. In the present study, we focused on the protoplast culture methods and plant growth regulators and the application of PSK for the promotion of cell division, callus induction, and plant regeneration.…”

Section: Discussionmentioning

confidence: 99%

“…Embryogenic callus of Angelica gigas Nakai which was induced on MS [ 61 ] medium supplemented with 3% (w/v) sucrose and 1.0 mg L − 1 2,4-D was used in the present studies [ 19 ].…”

Section: Methodsmentioning

confidence: 99%

“…Various factors such as source material used in protoplast isolation, culture medium, method of culture (liquid or semi-solid, or their combination), plant growth regulators (auxins, cytokinins, and their combination), plating density, supplements added to culture medium including polyamines, growth peptides are affecting a lot during protoplast division, micro-callus formation, and plant regeneration [ 1 ]. Recently, we were involved in protoplast isolation from embryogenic callus and plant regeneration via somatic embryogenesis in Angelica gigas [ 19 ] and evaluated the effect of the enzyme incubation period, culture method, and culture medium. However, the frequency of protoplast division was very low.…”

Section: Introductionmentioning

confidence: 99%

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Response surface methodology mediated optimization of phytosulfokine and plant growth regulators for enhanced protoplast division, callus induction, and somatic embryogenesis in Angelica Gigas Nakai

Lee,

Han,

Bae

et al. 2024

BMC Plant Biol

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Background Angelica Gigas (Purple parsnip) is an important medicinal plant that is cultivated and utilized in Korea, Japan, and China. It contains bioactive substances especially coumarins with anti-inflammatory, anti-platelet aggregation, anti-cancer, anti-diabetic, antimicrobial, anti-obesity, anti-oxidant, immunomodulatory, and neuroprotective properties. This medicinal crop can be genetically improved, and the metabolites can be obtained by embryonic stem cells. In this context, we established the protoplast-to-plant regeneration methodology in Angelica gigas. Results In the present investigation, we isolated the protoplast from the embryogenic callus by applying methods that we have developed earlier and established protoplast cultures using Murashige and Skoog (MS) liquid medium and by embedding the protoplast in thin alginate layer (TAL) methods. We supplemented the culture medium with growth regulators namely 2,4-dichlorophenoxyaceticacid (2,4-D, 0, 0.75, 1.5 mg L− 1), kinetin (KN, 0, 0.5, and 1.0 mg L− 1) and phytosulfokine (PSK, 0, 50, 100 nM) to induce protoplast division, microcolony formation, and embryogenic callus regeneration. We applied central composite design (CCD) and response surface methodology (RSM) for the optimization of 2,4-D, KN, and PSK levels during protoplast division, micro-callus formation, and induction of embryogenic callus stages. The results revealed that 0.04 mg L− 1 2,4-D + 0.5 mg L− 1 KN + 2 nM PSK, 0.5 mg L− 1 2,4-D + 0.9 mg L− 1 KN and 90 nM PSK, and 1.5 mg L− 1 2,4-D and 1 mg L− 1 KN were optimum for protoplast division, micro-callus formation and induction embryogenic callus. MS basal semi-solid medium without growth regulators was good for the development of embryos and plant regeneration. Conclusions This study demonstrated successful protoplast culture, protoplast division, micro-callus formation, induction embryogenic callus, somatic embryogenesis, and plant regeneration in A. gigas. The methodologies developed here are quite useful for the genetic improvement of this important medicinal plant.

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Section: Discussionmentioning

confidence: 99%

“…Embryogenic callus of Angelica gigas Nakai which was induced on MS [ 61 ] medium supplemented with 3% (w/v) sucrose and 1.0 mg L − 1 2,4-D was used in the present studies [ 19 ].…”

Section: Methodsmentioning

confidence: 99%