Plant Regeneration and In Vitro Growth Performance of Male-Sterile Somatic Plantlets of Sugi (Japanese Cedar, Cryptomeria japonica) Derived from Different Embryogenic Cell Lines

With the spread of pollinosis caused by sugi (Japanese cedar, Cryptomeria japonica) pollen, the use of pollen-free somatic seedlings of sugi is expected in Japan. However, there is a lack of knowledge on the relationship between the abilities during somatic embryogenesis, initial in vitro growth traits, and subsequent growth of somatic seedlings. In the present study, we provide the first basic information on somatic embryo maturation efficiency, somatic embryo germination, and plantlet conversion frequencies, as well as on in vitro growth performance of pollen-free somatic plantlets derived from different embryogenic cell lines (ECLs). Somatic embryo maturation efficiency varied from 34 to 514 cotyledonary embryos per plate and the average for the 19 ECLs tested was 244 embryos per plate. Subsequently, the overall average rates of somatic embryo germination and conversion among ECLs were 87.8% and 85.3%, respectively. The results of in vitro growth performance of pollen-free somatic plantlets showed significant differences in growth rate among ECLs.

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Factors Influencing Somatic Embryo Maturation in Sugi (Japanese Cedar, Cryptomeria japonica (Thunb. ex L.f.) D. Don)

Plants ◽

10.3390/plants10050874 ◽

2021 ◽

Vol 10 (5) ◽

pp. 874

Author(s):

Tsuyoshi E. Maruyama ◽

Saneyoshi Ueno ◽

Hideki Mori ◽

Takumi Kaneeda ◽

Yoshinari Moriguchi

Keyword(s):

Somatic Embryo ◽

Cryptomeria Japonica ◽

Japanese Cedar ◽

Embryo Maturation ◽

Embryogenic Cell ◽

Factors Affecting ◽

Somatic Embryo Maturation ◽

Maturation Medium ◽

Embryogenic Cell Lines ◽

Basal Salts

This paper presents the results of several experiments identifying basal salts (BS) contained in maturation medium, polyethylene glycol (PEG) concentration, abscisic acid (ABA) concentration, additional supplementation with potassium chloride (KCl), amino acid (AA) concentration, and proliferation culture medium (PCM) as the main culture factors affecting somatic embryo maturation in sugi (Japanese cedar, Cryptomeria japonica, Cupressaceae). Highly efficient embryo maturation was achieved when embryogenic cell lines (ECLs) were cultured on media supplemented with a combination of PEG, ABA, and AAs. More than 1000 embryos per gram of fresh weight (FW) can be produced on EM maturation medium supplemented with 175 g L−1 PEG, 100 µM ABA, 2 g L−1 glutamine, 1 g L−1 asparagine, and 0.5 g L−1 arginine.

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Post-maturation treatment improves and synchronizes somatic embryo germination of three species of Japanese pines

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-012-0128-7 ◽

2012 ◽

Vol 110 (1) ◽

pp. 45-52 ◽

Cited By ~ 25

Author(s):

Tsuyoshi E. Maruyama ◽

Yoshihisa Hosoi

Keyword(s):

Somatic Embryo ◽

Embryo Germination ◽

Somatic Embryo Germination

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Root growth of somatic plants of hybrid Pinus strobus (L.) and P. wallichiana (A. B. Jacks.) is affected by the nitrogen composition of the somatic embryo germination medium

Trees ◽

10.1007/s00468-017-1635-2 ◽

2017 ◽

Vol 32 (2) ◽

pp. 371-381 ◽

Cited By ~ 3

Author(s):

M. T. Llebrés ◽

C. Avila ◽

F. M. Cánovas ◽

K. Klimaszewska

Keyword(s):

Root Growth ◽

Somatic Embryo ◽

Pinus Strobus ◽

Embryo Germination ◽

Germination Medium ◽

Somatic Plants ◽

Somatic Embryo Germination

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Induced Polyploidy in Diploid Ornamental Ginger (Hedychium muluense R. M. Smith) Using Colchicine and Oryzalin

HortScience ◽

10.21273/hortsci.44.7.1809 ◽

2009 ◽

Vol 44 (7) ◽

pp. 1809-1814 ◽

Cited By ~ 22

Author(s):

Hamidou F. Sakhanokho ◽

Kanniah Rajasekaran ◽

Rowena Y. Kelley ◽

Nurul Islam-Faridi

Keyword(s):

Embryogenic Callus ◽

Stomatal Frequency ◽

Antimitotic Agent ◽

Embryogenic Cell ◽

Induction Frequency ◽

Embryogenic Cell Lines ◽

Agent Flow ◽

Stomatal Length ◽

First Time

The ploidy level of H. muluense, a diploid (2n = 2x = 34) and dwarf ornamental ginger species, has been determined and is reported for the first time. Oryzalin and colchicine were successfully used to induce polyploidy in Hedychium muluense in vitro. Embryogenic cell lines were treated with oryzalin (30, 60, or 120 μM) and colchicine (2.5, 5, or 10 mm) for 24, 48, or 72 h. The control contained no antimitotic agent. Flow cytometry, chloroplast count, and stomatal frequency were more effective and reliable than stomatal length as methods for assessing ploidy. Overall, oryzalin was more effective than colchicine in inducing polyploidy. The highest induction frequency (15%) of tetraploidy was achieved when embryogenic callus was exposed to 60 μM oryzalin for 72 h. For colchicine, exposure of embryogenic callus to the 2.5 mm colchicine for 24 h was the most effective in creating tetraploid (13%) plants.

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Effects of Polyethylene Glycol on Development of Grape (Vitis vinifera L. `Thompson Seedless') Somatic Embryos

HortScience ◽

10.21273/hortsci.33.3.557c ◽

1998 ◽

Vol 33 (3) ◽

pp. 557c-557

Author(s):

Fred K. Westphal ◽

Michael E. Compton

Keyword(s):

Polyethylene Glycol ◽

Somatic Embryo ◽

Somatic Embryos ◽

Vitis Vinifera L ◽

Zygotic Embryos ◽

Embryo Germination ◽

Thompson Seedless ◽

Germination Medium ◽

Peg Treatment ◽

Somatic Embryo Germination

Torpedo-stage somatic embryos were selected from actively growing cultures and trasferred to embryo maintenance medium [MS with (per liter) 412.5 mg NH4NO3, 475 mg KNO3, 1 g myo-inositol, 90 g sucrose, 2 g activated charcoal, and 7 g TC agar] supplemented with either 0%, 2.5%, 5%, 7.5%, or 10% polyethylene glycol (PEG) for 4, 8, or 12 weeks. Embryos placed on treatment media were transferred directly to grape somatic embryo germination medium [MS with (per liter) 1 g myo-inositol, 30 g sucrose, 1 M benzyladenine, and 7 g TC agar] once their PEG treatment was terminated. The number of embryos that germinated was recorded 4 weeks after transfer to somatic embryo germination medium. The number of germinated embryos that differentiated into plants was recorded at 8 weeks. There was no difference in germination rates and embryo differentiation among embryos incubated on medium with or without PEG for 4 weeks. A difference in embryo growth rate was observed after 8 weeks on medium with PEG. Embryo grew fastest on media containing 5% or 7.5% PEG. In addition, embryos grown on medium with 5% or 7.5% PEG were morphologically similar to zygotic embryos.

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Somatic embryo maturation, germination, and soil establishment of plants of black and white spruce (Picea mariana and Picea glauca)

Canadian Journal of Botany ◽

10.1139/b90-326 ◽

1990 ◽

Vol 68 (12) ◽

pp. 2583-2589 ◽

Cited By ~ 33

Author(s):

S. M. Attree ◽

T. E. Tautorus ◽

D. I. Dunstan ◽

L. C. Fowke

Keyword(s):

Abscisic Acid ◽

Somatic Embryo ◽

Picea Mariana ◽

Picea Glauca ◽

Black Spruce ◽

White Spruce ◽

Embryo Maturation ◽

Black And White ◽

Somatic Embryo Maturation

Somatic embryo maturation, germination, and soil establishment frequencies were compared for two conifer species, white and black spruce (Picea glauca and Picea mariana). The comparison of the two species regenerated and established in soil under the same conditions showed black spruce to be the most responsive. Shorter exposure times to 32 μM abscisic acid were not as effective as maturation on a medium containing 16 μM abscisic acid for 28 days. This gave similar maturation frequencies for the two species (6–8%), and germination frequencies of 64% for white spruce and over 73% for black spruce. Over 1800 black and white spruce plantlets were recovered, and more than 400 were transferred from in vitro to nonsterile conditions. Sixty percent (160) of the black spruce plantlets survived transfer and continued to grow vigorously. By comparison only 18% (29) of the white spruce plantlets survived, and half of these rapidly produced dormant buds and underwent no further shoot growth. White spruce plants that did not produce dormant buds grew vigorously. These results indicate that there are large differences in the ability of these closely related species to respond to plantlet establishment following regeneration from somatic embryos, and that black spruce is highly responsive to micropropagation by this method. Key words: Picea glauca, Picea mariana, somatic embryogenesis, maturation, germination, soil establishment.

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Development of an Efficient Protocol to Obtain Transgenic Coffee, Coffea arabica L., Expressing the Cry10Aa Toxin of Bacillus thuringiensis

International Journal of Molecular Sciences ◽

10.3390/ijms20215334 ◽

2019 ◽

Vol 20 (21) ◽

pp. 5334 ◽

Cited By ~ 2

Author(s):

Eliana Valencia-Lozano ◽

José L. Cabrera-Ponce ◽

Miguel A. Gómez-Lim ◽

Jorge E. Ibarra

Keyword(s):

Mass Spectrometry ◽

Bacillus Thuringiensis ◽

Somatic Embryo ◽

Somatic Embryos ◽

Leaf Tissue ◽

Stable Transformation ◽

Embryo Maturation ◽

Embryogenic Cell ◽

Somatic Embryo Maturation ◽

Coffee Plants

This report presents an efficient protocol of the stable genetic transformation of coffee plants expressing the Cry10Aa protein of Bacillus thuringiensis. Embryogenic cell lines with a high potential of propagation, somatic embryo maturation, and germination were used. Gene expression analysis of cytokinin signaling, homedomains, auxin responsive factor, and the master regulators of somatic embryogenesis genes involved in somatic embryo maturation were evaluated. Plasmid pMDC85 containing the cry10Aa gene was introduced into a Typica cultivar of C. arabica L. by biobalistic transformation. Transformation efficiency of 16.7% was achieved, according to the number of embryogenic aggregates and transgenic lines developed. Stable transformation was proven by hygromycin-resistant embryogenic lines, green fluorescent protein (GFP) expression, quantitative analyses of Cry10Aa by mass spectrometry, Western blot, ELISA, and Southern blot analyses. Cry10Aa showed variable expression levels in somatic embryos and the leaf tissue of transgenic plants, ranging from 76% to 90% of coverage of the protein by mass spectrometry and from 3.25 to 13.88 μg/g fresh tissue, with ELISA. qPCR-based 2−ΔΔCt trials revealed high transcription levels of cry10Aa in somatic embryos and leaf tissue. This is the first report about the stable transformation and expression of the Cry10Aa protein in coffee plants with the potential for controlling the coffee berry borer.

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Somatic embryogenesis of selected spruce species (Picea abies, P. omorika, P. pungens 'Glauca' and P. breweriana)

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.2008.023 ◽

2011 ◽

Vol 77 (3) ◽

pp. 189-199 ◽

Cited By ~ 5

Author(s):

Teresa Hazubska-Przybył ◽

Krystyna Bojarczyk

Keyword(s):

Somatic Embryogenesis ◽

Picea Abies ◽

Sucrose Concentration ◽

Medium Composition ◽

Embryogenic Tissue ◽

Zygotic Embryos ◽

Embryogenic Cell ◽

Nursery Production ◽

Embryogenic Cell Lines

Somatic embryogenesis was studied in four spruce species (Picea abies, P. omorika, P. pungens 'Glauca' and P. brewenana) to determine if this method can be used for in vitro propagation of coniferous trees. The highest frequency of initiation of embryogenic tissue was obtained when mature zygotic embryos were used as explants. It ranged then from 10.8% (P. brewenana) to 23.75% (P. omorika and P. pungens 'Glauca'). The frequency of embryogenic tissue initiation was strongly affected by medium composition, i.e. addition of appropriate auxins (2,4-D, NAA, Picloram) and sucrose concentration (10-20 g-1"1). A lower frequency was obtained in Picea omorika (10%) when megagametophytes (endosperms with immature zygotic embryos) were used as explants. No emryogenic tissue was produced from hypocotyls, cotyledons and needles. A satisfactory frequency was achieved with the use of somatic embryos of Picea abies (30%). The proliferation of embryogenic cell lines of spruces was affected by medium type. The experiments resulted in production of somatic plantlets of P. abies and P. omorika. This enables the application of this method of spruce micropropagation for genetic and breeding research or for nursery production.

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