Chinese Bulletin of Botany >

2023 , Vol. 58 >Issue 5: 750 - 759

DOI: https://doi.org/10.11983/CBB22106

TECHNIQUES AND METHODS

Efficient Plant Regeneration via Somatic Embryogenesis in Alocasia reginula cv. ‘Black Velvet’

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  • Guangdong Key Lab of Ornamental Plant Germplasm Innovation and Utilization, Key Laboratory of Urban Agriculture in South China, Ministry of Agriculture and Rural Affairs, Environmental Horticulture Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Received date: 2022-05-24

  Accepted date: 2022-11-09

  Online published: 2023-01-12

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Abstract

In this study, a plant regeneration system via somatic embryogenesis was established in Alocasia genus. We obtained embryogenic cell suspension cultures of A. reginula through embryogenic calli induced from petioles, and achieved a high frequency of plant regeneration using embryogenic cell aggregates. Efficiency of embryogenic calli induced from petiole explants was highest (81.3%) on a Murashige and Skoog (MS) medium supplemented with 2.0 mg·L-1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg·L-1 thidiazuron (TDZ). The embryogenic calli were crushed into cell aggregates and then transferred to liquid MS media supplemented with 2.0 mg·L-1 2,4-D and 1.0 mg·L-1 TDZ for suspension cultivation. By subculturing biweekly, lots of cell aggregates were gained from embryogenic suspension cultures after 12 weeks. The cell aggregates within 28 weeks of suspension culture were transferred to solid 1/2MS media without plant growth regulator for differentiation culture, with an average of 2.3-2.5 plantlets regenerated from each cell aggregate. The formation and germination of somatic embryos were observed by optical microscopy and scanning electron microscopy (SEM). After the regenerated plantlets were transplanted to a greenhouse for 4 months, the achieved survival ratio was 95.3%. Flow cytometry (FCM) demonstrated that there was no chromosome ploidy variation in randomly selected 50 surviving plants. In addition, the nuclear DNA content was estimated at 10.94 pg·(2C)-1, and the genome size was 5 290.12 Mb·C-1. There was no significant variation in their phenotypes from the time the plants were transplanted to the greenhouse until they bloomed spontaneously. These results provide good technical support for the commercial production of seedlings and biotechnological breeding of A. reginula.

Key words： Alocasia reginula; embryogenic calli; embryogenic cell suspension; plant regeneration

Cite this article

Liu Xiaofei, Sun Yingbo, Huang Lili, Yang Yuchai, Zhu Genfa, Yu Bo . Efficient Plant Regeneration via Somatic Embryogenesis in Alocasia reginula cv. ‘Black Velvet’[J]. Chinese Bulletin of Botany, 2023 , 58(5) : 750 -759 . DOI: 10.11983/CBB22106

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