Somatic Embryogenesis in Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae)

Guerra, Miguel Pedro; Steiner, Neusa; Farias-Soares, Francine L.; Vieira, Leila do Nascimento; Fraga, Hugo Pacheco de Freitas; Rogge-Renner, Gladys D.; Maldonado, Sara

doi:10.1007/978-1-4939-3061-6_24

Cited by 7 publications

(6 citation statements)

References 14 publications

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“…Several genome-wide methylation studies confirm DNA methylation changes in culture [19,40,66,75]. Accordingly, the present study introduces a stage-dependent paradigm, wherein we have identified genome-wide changes in the methylome and transcriptome of IR64 and TNG67, reflecting the stage of culture, cultivar-type and their interaction.…”

Section: Discussionsupporting

confidence: 52%

“…Abiotic stress-induced demethylation may result in chromatin remodeling, thereby activating transcription [65]. Demethylation may lead to the up-regulation of stress-related genes in culture, providing an adaptive advantage to the callus [66]. Thus, the success of regeneration may be associated with a hypomethylated state at loci related to stress tolerance.…”

Section: Differential Adaptive Response Of Cultivars In Culturementioning

confidence: 99%

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Dynamics of the Methylome and Transcriptome during the Regeneration of Rice

et al. 2018

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Oryza sativa indica (cv. IR64) and Oryza sativa japonica (cv. TNG67) vary in their regeneration efficiency. Such variation may occur in response to cultural environments that induce somaclonal variation. Somaclonal variations may arise from epigenetic factors, such as DNA methylation. We hypothesized that somaclonal variation may be associated with the differential regeneration efficiency between IR64 and TNG67 through changes in DNA methylation. We generated the stage-associated methylome and transcriptome profiles of the embryo, induced calli, sub-cultured calli, and regenerated calli (including both successful and failed regeneration) of IR64 and TNG67. We found that stage-associated changes are evident by the increase in the cytosine methylation of all contexts upon induction and decline upon regeneration. These changes in the methylome are largely random, but a few regions are consistently targeted at the later stages of culture. The expression profiles showed a dominant tissue-specific difference between the embryo and the calli. A prominent cultivar-associated divide in the global methylation pattern was observed, and a subset of cultivar-associated differentially methylated regions also showed stage-associated changes, implying a close association between differential methylation and the regeneration programs of these two rice cultivars. Based on these findings, we speculate that the differential epigenetic regulation of stress response and developmental pathways may be coupled with genetic differences, ultimately leading to differential regeneration efficiency. The present study elucidates the impact of tissue culture on callus formation and delineates the impact of stage and cultivar to determine the dynamics of the methylome and transcriptome in culture.

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Section: Discussionsupporting

confidence: 52%

Section: Differential Adaptive Response Of Cultivars In Culturementioning

confidence: 99%

Dynamics of the Methylome and Transcriptome during the Regeneration of Rice

et al. 2018

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“…Findings from the integrated cell biology studies may contribute to overcome the bottlenecks encountered with the SE. For example, morphological changes observed during EC proliferation resulted from the subsequent differential competence of the somatic embryos (Guerra et al 2016;Pereira-Dias et al 2020). Based on the observation of histological morphology, in conifers trees, the proliferations in cell cultures were generally driven by proembryogenic masses (PEMs), which divided cyclically into PEM I, II, and III, and development of PEMs signi cantly affected the ability of differentiation of the somatic embryo (Filonova et al 2000;Stasolla and Yeung 2003;Steiner et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Morphological and endogenous phytohormone changes during long-term embryogenic cultures in Korean pine

Liang

Shen

et al. 2022

Plant Cell Tiss Organ Cult

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Somatic embryogenesis (SE) is a biotechnological tool with high application potential for in vitro propagation and regeneration of conifers species. However, in SE of Korean pine, a dominant species of the natural mixed coniferous and broadleaf forests of the temperate forests in Northeast China, decreasing embryogenic potential was found in embryogenic callus (EC) with successive subcultures, especially when the cultures were maintained for more than 5-6 months, which is one of the most signi cant problems preventing commercial implementation of the SE technique. Aiming to investigate the morphological and the endogenous phytohormone changes associated with the decreasing embryogenic potential of Korean pine, here, we compared seven types of calli during long-term embryogenic culture. It was found that both the external morphological characteristics and internal cell structure were signi cantly different among the different types of calli. Speci cally, similar to their zygotic counterparts of conifer trees, the calli with high embryogenic potential were characterized by a higher number of designated proembryogenic masses (PEMs) structure. In addition, surface of this types of callus was covered with a large amount of mucilaginous extracellular matrix (ECM), which could further differentiate into globular embryos containing the embryonal masses (EM) and embryonic tube (ET) structure.In contrast, in those low embryogenic potential calli, we observed disintegrated PEMs structure, a large amount of irregularly shaped cells, and degraded cells, which failed to form somatic embryos. The endogenous hormone results showed that high levels of ABA were bene cial for promoting early somatic embryo differentiation in Korean pine, but maintaining high embryogenic potential required moderate levels of IAA and low levels of ZR. Additionally, a high ratio of IAA / ABA and MeJA / ABA was conducive to maintaining high embryogenic potential, whereas a low ratio of IAA / ABA and MeJA / ABA were effective for early somatic embryos morphological differentiation. This study demonstrates morphological and physiological characteristics of the embryogenesis potential of Korean pine calli.

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“…According to Steiner et al (2015), PEM III formation and polarization is a crucial step to form early somatic embryos. Thus, integrated cell biology studies are necessary to help overcome the bottlenecks observed with the somatic embryogenesis protocol (Guerra et al 2016).…”

Section: Introductionmentioning

confidence: 99%

Integrated proteomics and histochemical analysis of Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae) in embryogenic suspension culture

Dias

Steiner

Cangahaula-Inocente

et al. 2020

AFR

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t Cell suspension culture is a useful in vitro model-system for both scaling up and conserving the Brazilian conifer Araucaria angustifolia. In the present work, cell suspension of Araucaria was subjected to proteomics, biochemical and histochemical analyses. The results revealed new insights underlying the molecular mechanism of proembryogenic masses transition in cell suspension. Embryogenic cell cultures were cultivated in a basal liquid medium modified in a Steward apparatus (orbital agitator). Cell growth dynamics was evaluated using cell volume after sedimentation, fresh weight, mitotic index, conductivity, pH, and the number of proembryogenic masses (PEMs: I, II, III). Histochemical parameters, cell viability, and cell death analyses were performed to pinpoint growth rates. Proteomics analysis was performed using two-dimensional electrophoresis, and protein identification was carried out by MALDI-TOF-TOF tandem mass spectrometry. Cell growth dynamics showed a predominance of PEM III. Maximum slope of the exponential phase growth in fresh weight occurred at exponential phase after 15 days (optimal cultivation time), after which cell viability and pH decreased, thereby allowing the identification of stressrelated proteins. Several metabolism and growth proteins were abundant, such as: cytoskeletal, WOX1, cytokinin-related, and auxin-related proteins acting on cell wall modification, suspensor cell formation, and PEM I to PEM III transition.

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Somatic Embryogenesis in Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae)

Cited by 7 publications

References 14 publications

Dynamics of the Methylome and Transcriptome during the Regeneration of Rice

Dynamics of the Methylome and Transcriptome during the Regeneration of Rice

Morphological and endogenous phytohormone changes during long-term embryogenic cultures in Korean pine

Integrated proteomics and histochemical analysis of Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae) in embryogenic suspension culture

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