Indirect somatic embryogenesis and regeneration of Fraxinus mandshurica plants via callus tissue

Liu, Yang; Wei, Cheng; Wang, Hao; Ma, Xiaoyan; Shen, Hailong; Yang, Ling

doi:10.1007/s11676-020-01199-3

Cited by 19 publications

(11 citation statements)

References 32 publications

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“…To date, only one type of somatic embryogenetic pathway has been reported in some species. For instance, the DSE pathway was established in Minthostachys verticillata ( Bertero et al, 2020 ), and the ISE pathway was established in Fraxinus mandshurica ( Liu et al, 2020 ). Meanwhile, the poor synchrony of somatic embryo development, coupled with the slow growth rate and long cycle of woody plants, has further hampered the commercial application of raising SE seedlings.…”

Section: Introductionmentioning

confidence: 99%

Direct and Indirect Somatic Embryogenesis Induction in Camellia oleifera Abel

et al. 2021

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Camellia oleifera Abel. is an important woody oil species; however, the shortage of rapid and industrialized seedling culture is a large constraint on the development of the tea oil industry. Somatic embryogenesis (SE) is one of the main powerful biotechnological tools for plant mass regeneration, but the largely unknown SE in C. oleifera limits the scale production of clonal plants. In this study, we described a high-efficiency SE system via direct and indirect pathways in C. oleifera and investigated the effect of genotype, explant age and phytohormones on SE. In the direct pathway, somatic embryos were highly induced from immature seeds 220 days after full blossom, and the development of embryoids was achieved with a combination of 0.19 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mg/L thidiazuron (TDZ). In the indirect pathway, embryogenic calli were induced from the same explants in medium containing 1.5 mg/L 2,4-D, while 0.75 mg/L 2,4-D treatment led to high proliferation rates for embryogenic calli. The addition of 0.19 mg/L 2,4-D alone stimulated the production of globular embryos while causing a 75% loss of the induction rate in the heart embryo stage. Upon transfer of the globular embryos to phytohormone-free medium, an optimal induction rate of 62.37% from globular embryos to cotyledonary embryos was obtained. These data suggest that the subsequent differentiation process after the globular embryo stage in ISE is more similar to an endogenous phytohormones-driven process. Mature embryos germinated to produce intact plantlets on half-strength MS basal medium with a regeneration rate of 63.67%. Histological analysis confirmed the vascular bundle isolation of embryoids from the mother tissue. We further studied the different varieties and found that there were no significant genotype differences for SE induction efficiency in C. oleifera. Thus, we established a high-efficiency induction system for direct and indirect somatic embryogenesis (ISE) in C. oleifera and regenerated intact plantlets via SE, not organogenesis. ISE has a more complicated induction and regulatory mechanism than direct somatic embryogenesis. The improved protocol of SE would benefit mass propagation and genetic manipulation in C. oleifera.

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Section: Introductionmentioning

confidence: 99%

Direct and Indirect Somatic Embryogenesis Induction in Camellia oleifera Abel

et al. 2021

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“…Among the protocols in which cytokinins were used as the sole PGR for the induction of somatic embryogenesis, Kin and ZEA were the most frequently used cytokinins (Phillips and Garda 2019 ). In addition, ABA can induce somatic embryogenesis and regulate somatic embryo development in certain cases (Lü et al 2013 ; Liu et al 2021 ; Nishiwaki et al 2000 ; Su and Zhang 2014 ). Therefore, we utilized ABA, ZEA and Kin to test their effects on the differentiation of leaf and hypocotyl calli of B. chinense .…”

Section: Discussionmentioning

confidence: 99%

Plant regeneration via callus-mediated organogenesis in commercial variety of Chuanbeichai No. 1 in Bupleurum chinense DC

Zhao

Chen

et al. 2022

Plant Biotechnol Rep

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Bupleurum chinense DC is an important medicinal plant with many active ingredients that are used for the treatment of different types of diseases and valued in pharmaceutical markets. In vitro shoot regeneration can efficiently contribute to the improvement of B. chinense. In the present study, we investigated the effects of the explant type and plant growth regulators (PGRs) on embryogenic callus induction and plant regeneration in B. chinense . Our investigation demonstrated that 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) combined with 1 mg/L thidiazuron (TDZ) played a major role in promoting callus induction from leaf, hypocotyl and stem 2 explants, whereas the most effective treatment for stem 1 callus formation was Murashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L 6-benzyladenine (BA) and 0.5 mg/L kinetin (Kin). The highest shoot regeneration rate (57.14%) was obtained from hypocotyl-induced calli in MS medium with 0.5 mg/L Kin after 12 weeks of cultivation. This regeneration protocol can be used in large-scale cultivation and may be useful for future genetic modifications of B. chinense .

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“…A number of important species and crops can be genetically transformed by agrobacterium-mediated methods [15][16][17]. At present, F. mandshurica seeds have been used to induce embryogenic callus and establish a complete somatic embryogenesis system [18]. Calli culture systems of F. mandshurica and the genetic transformation of adventitious buds have been established, and while the potential effects of ultrasonic and vacuum treatment on transformation have been investigated, the transformation efficiency was found to be relatively low at only 7.31% [1].…”

Section: Introductionmentioning

confidence: 99%

A Stable and Efficient Genetic Transformation Method for Embryogenic Callus of Fraxinus mandshurica

Dong

Zhao

et al. 2023

Forests

Self Cite

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Fraxinus mandshurica is a widely used greening and ornamental tree species. However, its genetic transformation system has been hampered by problems such as low transformation efficiency, among others, which can hinder research related to molecular breeding and the analysis of functional genes. Thus, in this study, a novel genetic transformation method for efficient transformation of the embryonic callus of Fraxinus mandshurica was investigated. The method was optimized in terms of factors such as antibiotics, infection solution concentrations, co-culture time, and somatic embryo maturation. The results indicated that the optimal antibiotic concentration was 10 mg·L−1 of hygromycin (Hyg). At this point, the callus proliferation multiple was only 0.12. The highest transformation efficiency was found to be 93.93% when the absorbance of the infection solution concentration at OD600 was 0.4. Interestingly, transformation efficiency was found to be highest (77.9%) at 48 h of co-culture, with a GUS staining rate of 88.23%. The medium for somatic embryo maturation of transformed callus was half-strength MS medium (MS 1/2) containing 60 g·L−1 polyethylene glycol, 1 mg·L−1 abscisic acid, 400 mg·L−1 casein enzymatic hydrolysate (CH), 20 g·L−1 sucrose, 1 g·L−1 activated charcoal, and 5 g·L−1 gellan gum. The medium for somatic embryo germination was MS ½, containing 0.2 mg·L−1 of N-(Phenylmethyl)-9H-purin-6-amine(6-BA) and 5.0 mg·L−1 of gibberellin (GA). These results are of significance for the verification of the gene function and future genetic improvement of Fraxinus mandshurica.

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Indirect somatic embryogenesis and regeneration of Fraxinus mandshurica plants via callus tissue

Cited by 19 publications

References 32 publications

Direct and Indirect Somatic Embryogenesis Induction in Camellia oleifera Abel

Direct and Indirect Somatic Embryogenesis Induction in Camellia oleifera Abel

Plant regeneration via callus-mediated organogenesis in commercial variety of Chuanbeichai No. 1 in Bupleurum chinense DC

A Stable and Efficient Genetic Transformation Method for Embryogenic Callus of Fraxinus mandshurica

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