Somatic embryogenesis of hybrid Pinus elliottii var. elliottii × P. caribaea var. hondurensis and ploidy assessment of somatic plants

The somatic embryogenesis process was evaluated as a potential tool for operational vegetative propagation using individuals from families currently used in the J.D. Irving, Ltd. black spruce tree improvement program. Most families were responsive although the number of individuals within families capable of producing embryogenic tissue (ET) varied greatly (1–70%). Seventy-four percent of the ET clones produced mature embryos and most of these germinated. Greenhouse survival was initially low (11%) but improved in subsequent experiments to 45% as growing regimes were refined. Demonstration plantings of the resulting somatic plants were established at two sites in New Brunswick. A total of 206 clones were cryopreserved. The potential for integrating somatic embryogenesis techniques into tree improvement and stock production programs is discussed. Key words: tree improvement, somatic embryogenesis, clonal propagation, black spruce, biotechnology

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Somatic Embryogenesis in Slash Pine (Pinus Elliottii Engelm.)

Somatic Embryogenesis in Woody Plants - Forestry Sciences ◽

10.1007/978-94-011-0960-4_11 ◽

1995 ◽

pp. 183-195 ◽

Cited By ~ 17

Author(s):

R. J. Newton ◽

K. A. Marek-Swize ◽

M. E. Magallanes-Cedeno ◽

N. Dong ◽

S. Sen ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Pinus Elliottii ◽

Slash Pine

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Somatic embryogenesis in slash pine (Pinus elliottii) from immature embryos cultured in vitro

Plant Science ◽

10.1016/0168-9452(89)90070-8 ◽

1989 ◽

Vol 65 (2) ◽

pp. 233-241 ◽

Cited By ~ 56

Author(s):

S. Mohan Jain ◽

Niu Dong ◽

R.J. Newton

Keyword(s):

Somatic Embryogenesis ◽

Pinus Elliottii ◽

Slash Pine ◽

Immature Embryos

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Somatic Embryogenesis and Plant Regeneration from Sugi (Japanese Cedar, Cryptomeria japonica D. Don, Cupressaceae) Seed Families by Marker Assisted Selection for the Male Sterility Allele ms1

Plants ◽

10.3390/plants9081029 ◽

2020 ◽

Vol 9 (8) ◽

pp. 1029 ◽

Cited By ~ 1

Author(s):

Tsuyoshi E. Maruyama ◽

Saneyoshi Ueno ◽

Satoko Hirayama ◽

Takumi Kaneeda ◽

Yoshinari Moriguchi

Keyword(s):

Somatic Embryogenesis ◽

Male Sterility ◽

Marker Assisted Selection ◽

Cryptomeria Japonica ◽

Public Health Problem ◽

Efficient Production ◽

Initial Growth ◽

Male Sterile ◽

Embryogenic Cell ◽

Somatic Plants

One of the possible countermeasures for pollinosis caused by sugi (Cryptomeria japonica), a serious public health problem in Japan, is the use of male sterile plants (MSPs; pollen-free plants). However, the production efficiencies of MSPs raised by conventional methods are extremely poor, time consuming, and resulting in a high seedling cost. Here, we report the development of a novel technique for efficient production of MSPs, which combines marker-assisted selection (MAS) and somatic embryogenesis (SE). SE from four full sib seed families of sugi, carrying the male sterility gene MS1, was initiated using megagametophyte explants that originated from four seed collections taken at one-week intervals during the month of July 2017. Embryogenic cell lines (ECLs) were achieved in all families, with initiation rates varying from 0.6% to 59%. Somatic embryos were produced from genetic marker-selected male sterile ECLs on medium containing maltose, abscisic acid (ABA), polyethylene glycol (PEG), and activated charcoal (AC). Subsequently, high frequencies of germination and plant conversion (≥76%) were obtained on plant growth regulator-free medium. Regenerated plantlets were acclimatized successfully, and the initial growth of male sterile somatic plants was monitored in the field.

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Induction of Radiata Pine Somatic Embryogenesis at High Temperatures Provokes a Long-Term Decrease in DNA Methylation/Hydroxymethylation and Differential Expression of Stress-Related Genes

Plants ◽

10.3390/plants9121762 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1762

Author(s):

Ander Castander-Olarieta ◽

Cátia Pereira ◽

Ester Sales ◽

Mónica Meijón ◽

Isabel Arrillaga ◽

...

Keyword(s):

Somatic Embryogenesis ◽

High Temperatures ◽

Stress Responses ◽

Radiata Pine ◽

Global Methylation ◽

Protein Coding ◽

Somatic Plants ◽

Stress Related Genes ◽

As Stress ◽

Stress Mediators

Based on the hypothesis that embryo development is a crucial stage for the formation of stable epigenetic marks that could modulate the behaviour of the resulting plants, in this study, radiata pine somatic embryogenesis was induced at high temperatures (23 °C, eight weeks, control; 40 °C, 4 h; 60 °C, 5 min) and the global methylation and hydroxymethylation levels of emerging embryonal masses and somatic plants were analysed using LC-ESI-MS/ MS-MRM. In this context, the expression pattern of six genes previously described as stress-mediators was studied throughout the embryogenic process until plant level to assess whether the observed epigenetic changes could have provoked a sustained alteration of the transcriptome. Results indicated that the highest temperatures led to hypomethylation of both embryonal masses and somatic plants. Moreover, we detected for the first time in a pine species the presence of 5-hydroxymethylcytosine, and revealed its tissue specificity and potential involvement in heat-stress responses. Additionally, a heat shock protein-coding gene showed a down-regulation tendency along the process, with a special emphasis given to embryonal masses at first subculture and ex vitro somatic plants. Likewise, the transcripts of several proteins related with translation, oxidative stress response, and drought resilience were differentially expressed.

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Heat Stress in Pinus halepensis Somatic Embryogenesis Induction: Effect in DNA Methylation and Differential Expression of Stress-Related Genes

Plants ◽

10.3390/plants10112333 ◽

2021 ◽

Vol 10 (11) ◽

pp. 2333

Author(s):

Cátia Pereira ◽

Ander Castander-Olarieta ◽

Ester Sales ◽

Itziar A. Montalbán ◽

Jorge Canhoto ◽

...

Keyword(s):

Dna Methylation ◽

Somatic Embryogenesis ◽

Heat Stress ◽

Differential Expression ◽

Pinus Halepensis ◽

Expression Patterns ◽

Induction Effect ◽

Somatic Plants ◽

Stress Related Genes

In the current context of climate change, plants need to develop different mechanisms of stress tolerance and adaptation to cope with changing environmental conditions. Temperature is one of the most important abiotic stresses that forest trees have to overcome. Recent research developed in our laboratory demonstrated that high temperatures during different stages of conifer somatic embryogenesis (SE) modify subsequent phases of the process and the behavior of the resulting ex vitro somatic plants. For this reason, Aleppo pine SE was induced under different heat stress treatments (40 °C for 4 h, 50 °C for 30 min, and 60 °C for 5 min) in order to analyze its effect on the global DNA methylation rates and the differential expression of four stress-related genes at different stages of the SE process. Results showed that a slight decrease of DNA methylation at proliferating embryonal masses (EMs) can correlate with the final efficiency of the process. Additionally, different expression patterns for stress-related genes were found in EMs and needles from the in vitro somatic plants obtained; the DEHYDRATION INDUCED PROTEIN 19 gene was up-regulated in response to heat at proliferating EMs, whereas HSP20 FAMILY PROTEIN and SUPEROXIDE DISMUTASE [Cu–Zn] were down-regulated in needles.

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Cytokinins are involved in drought tolerance of Pinus radiata plants originating from embryonal masses induced at high temperatures

Tree Physiology ◽

10.1093/treephys/tpaa055 ◽

2020 ◽

Cited By ~ 4

Author(s):

Ander Castander-Olarieta ◽

Paloma Moncaleán ◽

Catia Pereira ◽

Aleš Pěnčík ◽

Ivan Petřík ◽

...

Keyword(s):

Somatic Embryogenesis ◽

High Temperature ◽

High Temperatures ◽

Pinus Radiata ◽

Temperature Regime ◽

Zeatin Riboside ◽

Embryogenic Cell ◽

Temperature Regimes ◽

Extreme High Temperature ◽

Somatic Plants

Abstract Vegetative propagation through somatic embryogenesis is an effective method to produce elite varieties and can be applied as a tool to study the response of plants to different stresses. Several studies show that environmental changes during embryogenesis could determine future plant development. Moreover, we previously reported that physical and chemical conditions during somatic embryogenesis can determine the protein, hormone and metabolite profiles, as well as the micromorphological and ultrastructural organization of embryonal masses and somatic embryos. In this sense, phytohormones are key players throughout the somatic embryogenesis process as well as during numerous stress–adaptation responses. In this work, we first applied different high-temperature regimes (30 °C, 4 weeks; 40 °C, 4 days; 50 °C, 5 min) during induction of Pinus radiata D. Don somatic embryogenesis, together with control temperature (23 °C). Then, the somatic plants regenerated from initiated embryogenic cell lines and cultivated in greenhouse conditions were subjected to drought stress and control treatments to evaluate survival, growth and several physiological traits (relative water content, water potential, photosynthesis, stomatal conductance and transpiration). Based on those preliminary results, even more extreme high-temperature regimes were applied during induction (40 °C, 4 h; 50 °C, 30 min; 60 °C, 5 min) and the corresponding cytokinin profiles of initiated embryonal masses from different lines were analysed. The results showed that the temperature regime during induction had delayed negative effects on drought resilience of somatic plants as indicated by survival, photosynthetic activity and water- use efficiency. However, high temperatures for extended periods of time enhanced subsequent plant growth in well-watered conditions. High-temperature regime treatments induced significant differences in the profile of total cytokinin bases, N6-isopentenyladenine, cis-zeatin riboside and trans-zeatin riboside. We concluded that phytohormones could be potential regulators of stress-response processes during initial steps of somatic embryogenesis and that they may have delayed implications in further developmental processes, determining the performance of the generated plants.

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