Optimization of maturation process for somatic embryo production and cryopreservation of embryogenic tissue in Pinus koraiensis

Peng, Chunxue; Gao, Fang; Wang, Hao; Shen, Hailong; Yang, Ling

doi:10.1007/s11240-020-01918-y

Cited by 39 publications

(29 citation statements)

References 40 publications

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“…The effect of sucrose, glucose, and fructose as the most important carbohydrates have been widely studied in in vitro morphogenic responses of different plants [ 106 ]. While sucrose has resulted in the maximum in vitro organogenesis and embryogenesis in some plants (e.g., Agave angustifolia [ 107 ], Sapindus trifoliatus [ 108 ], and Pinus koraiensis [ 109 ]), other plants (e.g., Vitis Vinifera [ 110 ], Brassica napus [ 111 ], and Chrysanthemum ×grandiflorum [ 112 ]) had better in vitro morphogenic responses to glucose and fructose [ 106 ]. Therefore, it is essential to study the effect of different carbohydrate sources on cannabis micropropagation.…”

Section: In Vitro Culture In Cannabismentioning

confidence: 99%

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Hesami

Baiton

Alizadeh

et al. 2021

IJMS

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For a long time, Cannabis sativa has been used for therapeutic and industrial purposes. Due to its increasing demand in medicine, recreation, and industry, there is a dire need to apply new biotechnological tools to introduce new genotypes with desirable traits and enhanced secondary metabolite production. Micropropagation, conservation, cell suspension culture, hairy root culture, polyploidy manipulation, and Agrobacterium-mediated gene transformation have been studied and used in cannabis. However, some obstacles such as the low rate of transgenic plant regeneration and low efficiency of secondary metabolite production in hairy root culture and cell suspension culture have restricted the application of these approaches in cannabis. In the current review, in vitro culture and genetic engineering methods in cannabis along with other promising techniques such as morphogenic genes, new computational approaches, clustered regularly interspaced short palindromic repeats (CRISPR), CRISPR/Cas9-equipped Agrobacterium-mediated genome editing, and hairy root culture, that can help improve gene transformation and plant regeneration, as well as enhance secondary metabolite production, have been highlighted and discussed.

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Section: In Vitro Culture In Cannabismentioning

confidence: 99%

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Hesami

Baiton

Alizadeh

et al. 2021

IJMS

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“…However, in conifer somatic embryogenesis, problems, such as low embryonic tissue induction efficiency, low somatic embryo (SE) yield, low germination/conversion percentage, and slow growth of seedlings, limited the commercial development of this technology [ 3 , 4 , 5 ]. In an earlier study of Pinus koraiensis somatic embryogenesis, we discovered that numerous cell lines in the embryonic tissue were unable to generate SEs even if they met the ideal SE maturation culture conditions [ 6 ]. Such a phenomenon is also commonly seen in somatic embryogenesis of Araucaria angustifolia , P. taeda , and Picea abies [ 7 , 8 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

Morphological and Physiological Indicators for Screening Cell Lines with High Potential for Somatic Embryo Maturation at an Early Stage of Somatic Embryogenesis in Pinus Koraiensis

Peng

Gao

Wang

et al. 2022

Plants

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Many cell lines in the embryogenic callus cannot produce somatic embryos (SEs) even if they meet the optimal SE maturation culture conditions during conifer somatic embryogenesis. This phenomenon hinders the progress of the industrial-scale reproduction of conifers. Therefore, there is an urgent need to obtain morphological and physiological markers to screen embryogenic calli in response to SE maturation conditions. To detect cell lines with high somatic embryogenesis potential during the proliferation process, we counted the number of pro-embryos and early SEs (ESEs) in different cell lines and storage substances, endogenous hormones, and polyamine contents. The results showed that the yield of P. koraiensis SEs was heavily dependent on genotype (p = 0.001). There were high levels of PE III (pro-embryo III) number, ESE number, and soluble protein content, in the response cell lines (R cell lines), which were 1.6-, 3-, and 1.1-fold those of the obstructive cell lines (B cell lines), respectively. The B cell line had high levels of starch, auxin (IAA), Put, Spd, and putrescine: spermine (Put: Spm) compared to the R cell line. In addition, the numbers of PE III, ESEs, and soluble protein content were significantly positively correlated with SE yield. In contrast, the contents of starch, abscisic acid (ABA), Put, Spm, and Spd were significantly negatively correlated with SE yield. To ensure the accuracy of the results, we used nine cell lines to test the results. The PE III and ESE numbers and the Spm and Spd contents were positively correlated with SE yield, while the levels of starch, ABA, IAA, Put: Spd, and Put: Spm were negatively correlated with SE yield. Thus, we recommend using high PE III and ESEs as morphological indicators and low levels of starch, IAA, ABA, and Put: Spm as physiological markers to screen cell lines with a high somatic embryogenesis potential. In addition, we also found that the relationship between Spd, Spm, and SE yield was opposite in the two experimental results. Therefore, we speculate that the differences in Spd and Spm content are mainly affected by genotype. In conclusion, this study obtained the morphological and physiological markers of some high-somatic embryogenic cell lines by comparing the differences between nine somatic embryogenic cell lines. Our results can guide the improvement of conifer somatic embryogenesis technology and can provide a theoretical basis for accelerating the application of biotechnology in large-scale artificial breeding.

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“…Experiments have confirmed that the average number of L. olgensis somatic embryos was 202.06 g −1 when adding 20 mg L −1 ABA [24]. In Pinus koraiensis, increasing ABA (up to 80 µM L −1 ), gellan gum (up to 12 g L −1 ) and sucrose (0.2 m L −1 ) concentrations in maturation medium improved the yield of cotyledonary somatic embryo [25].…”

Section: Introductionmentioning

confidence: 75%

Plant Regeneration via Somatic Embryogenesis in Larix principis-rupprechtii Mayr

Jiang

Chen

Gao

et al. 2021

Forests

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Prince Rupprecht’s larch (Larix principis-rupprechtii Mayr) is a native conifer in North China with great economic and ecological values. Somatic embryogenesis (SE) is a powerful tool for the mass clonal propagation in plants. In this study, we described a high-efficiency SE system via indirect pathways and investigated the effect of genotype, culture conditions and phytohormones on SE. Immature zygotic embryos (IZEs) of L. principis-rupprechtii Mayr were used as explant materials. In the induction stage, embryogenic tissues (ETs) were induced on mLV medium supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 6-benzylaminopurine (6-BA). The initiation frequencies showed significant differences (p < 0.05) among 20 genotypes of open-pollinated mother trees with the highest induction frequency reaching 30%. For tissue proliferation, proliferation in liquid medium was more efficient compared with proliferation in semi-solid medium, providing a multiplication rate of 3.12 in an 8-day subculture period. As a necessary exogenous plant growth regulator (PGR) for somatic embryo maturation in conifers, abscisic acid (ABA) was optimized at 16 mg L−1 in this system. Next, an orthogonal test on osmotic pressure factors showed 50 g L−1 sucrose, 7 g L−1 phytagel and 75 g L−1 polyethylene glycol (PEG) was the optimal combination for somatic embryo maturation in L. principis-rupprechtii Mayr. Moreover, the dispersion culture method provided a more efficient somatic embryo maturation, up to 545 per gram of fresh weight (FW). Finally, 2 g L−1 of active charcoal (AC) was found to increase the somatic embryo germination rate to 63.46%. The improved protocol of SE will serve as a foundation for establishing mass propagation and genetic transformation of L. principis-rupprechtii Mayr.

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Optimization of maturation process for somatic embryo production and cryopreservation of embryogenic tissue in Pinus koraiensis

Cited by 39 publications

References 40 publications

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Morphological and Physiological Indicators for Screening Cell Lines with High Potential for Somatic Embryo Maturation at an Early Stage of Somatic Embryogenesis in Pinus Koraiensis

Plant Regeneration via Somatic Embryogenesis in Larix principis-rupprechtii Mayr

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