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

Maruyama, Tsuyoshi; Ueno, S.; Mori, Hideki; Kaneeda, Takumi; Moriguchi, Yoshinari

doi:10.3390/plants10050874

Cited by 14 publications

(8 citation statements)

References 72 publications

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“…SE and plant regeneration of C. japonica were described in our previous reports (Maruyama et al, 2020(Maruyama et al, , 2021a(Maruyama et al, , 2021b. The extrusion of ECs from the explants could begin about 2 weeks after the start of culture (Figure 1A) and the establishment of stable lines with evident EC proliferation was most frequently observed 4 weeks after it (Figure 1B).…”

Section: Se and Plant Regenerationmentioning

confidence: 52%

“…However, for many coniferous species, in vitro clonal propagation is still difficult or inefficient (Bonga et al, 2010). Currently, the most well-established source of SE propagation in sugi is immature zygotic embryos, for which protocols have been established up to the stage of plant regeneration (Maruyama et al, 2020(Maruyama et al, , 2021a(Maruyama et al, , 2021b. Selecting a male-sterile strain as early as possible in this process will improve the efficiency of MSP production.…”

Section: Introductionmentioning

confidence: 99%

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Marker-Assisted Selection for Pollen-Free Somatic Plants of Sugi (Japanese Cedar, Cryptomeria japonica): A Simple and Effective Methodology for Selecting Male-Sterile Mutants With ms1-1 and ms1-2

et al. 2021

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Pollen allergy caused by sugi (Japanese cedar, Cryptomeria japonica) is a serious problem in Japan. One of the measures against pollinosis is the use of male-sterile plants (MSPs; pollen-free plants). In this context, the development of a novel technique for the efficient production of sugi MSPs, which combines marker-assisted selection (MAS) with somatic embryogenesis (SE), was recently reported by our research group. To improve the efficiency of MSP production, in this paper we report improved MAS for male-sterile individuals from embryogenic cells, cotyledonary embryos, and somatic plants of sugi using a newly developed marker in the form of the causative mutation of MS1 itself, selecting individuals with ms1-1 and ms1-2 male-sterile mutations. We also describe simplified methods for extracting DNA from different plant materials and for MAS using LAMP diagnostics. Finally, we show that MAS can be efficiently performed using the one-step indel genotyping (ING) marker developed in this study and using InstaGene for DNA extraction. The combination of SE and 100% accurate marker selection during the embryogenic cell stage enables the mass production of MS1 male-sterile sugi seedlings.

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Section: Se and Plant Regenerationmentioning

confidence: 52%

Section: Introductionmentioning

confidence: 99%

Marker-Assisted Selection for Pollen-Free Somatic Plants of Sugi (Japanese Cedar, Cryptomeria japonica): A Simple and Effective Methodology for Selecting Male-Sterile Mutants With ms1-1 and ms1-2

et al. 2021

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“…Somatic embryo maturation was carried out in accordance with the methodology described by Maruyama et al [20]. ECs from 19 ECLs that had proliferated for 2 weeks were cultured in clumps (5 masses per 90 × 20 mm plate, 100 mg each) on maturation medium containing the basal salt concentration of standard EM medium [10] and supplemented with 30 g L −1 maltose, 2 g L −1 activated charcoal (AC), 100 µM ABA, amino acids (in g L −1 : glutamine 2, asparagine 1, arginine 0.5, citrulline 0.079, ornithine 0.076, lysine 0.055, alanine 0.04, and proline 0.035), 175 g L −1 PEG (Av.…”

Section: Maturation Of Somatic Embryosmentioning

confidence: 99%

“…In recent years, our group reported an improved methodology for SE initiation from the analysis of results on the influence of plant material, explant collection time, explant genotype, and culture conditions [19]. Subsequently, an optimized protocol for somatic embryo maturation from studies on the effect of basal salts contained in maturation medium, polyethylene glycol (PEG) concentration, abscisic acid (ABA) concentration, additional supplementation of potassium chloride, amino acid concentration, and proliferation culture medium, as the main culture-related factors affecting somatic embryo maturation in sugi, was reported [20].…”

Section: Introductionmentioning

confidence: 99%

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

Tsuruta

Maruyama

Ueno

et al. 2021

Forests

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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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“…The development of this novel technique for the efficient production of sugi MSPs was first reported by our research group ( Maruyama et al, 2020 ). Then, we released improved methodologies for SE initiation ( Maruyama et al, 2021a ), somatic embryo production, and somatic plant regeneration ( Maruyama et al, 2021b ; Tsuruta et al, 2021b ), as well as for the early effective selection of male-sterile ECLs ( Hasegawa et al, 2021 ; Tsuruta et al, 2021a ).…”

Section: Introductionmentioning

confidence: 99%

An Improved and Simplified Propagation System for Pollen-Free Sugi (Cryptomeria japonica) via Somatic Embryogenesis

et al. 2022

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Sugi (Japanese cedar, Cryptomeria japonica) is the most important forestry tree species in Japan, covering 44% of the total artificial forest area. Large amounts of pollen released from these forests each spring cause allergic reactions in approximately 40% of the population, which are a serious social and public health problem in Japan. As a countermeasure, there is an urgent need to reforest using male-sterile plants (MSPs; pollen-free plants); however, the production of MSPs via conventional methods is inefficient, time consuming, and requires considerable resources in terms of labor and space. In the present paper, we described an improved and simplified methodology for the efficient propagation of pollen-free Japanese cedar, combining the use of genetic markers (marker-assisted selection or marker-aided selection) for the early selection of male-sterile genotypes and the use of somatic embryogenesis (SE) for the clonal mass propagation of seedlings. We describe all the stages involved in the production process of somatic seedlings. Our results demonstrated that this methodology easily and efficiently produces MSPs with a discrimination rate of 100% in a short period of time. Production of 243.6 ± 163.6 cotyledonary embryos per plate, somatic embryo germination, and plantlet conversion frequencies of 87.1 ± 11.9% and 84.8 ± 12.6%, respectively, and a 77.6 ± 12.1% survival rate after ex vitro acclimatization was achieved. Moreover, we also describe an easy method for the collection of somatic embryos prior to germination, as well as an efficient and practical method for their storage at 5°C. Finally, a representative schedule for the propagation of pollen-free sugi somatic seedlings is presented as a reference for practical uses. This methodology will definitively help to accelerate the production of C. japonica MSPs across Japan.

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

Cited by 14 publications

References 72 publications

Marker-Assisted Selection for Pollen-Free Somatic Plants of Sugi (Japanese Cedar, Cryptomeria japonica): A Simple and Effective Methodology for Selecting Male-Sterile Mutants With ms1-1 and ms1-2

Marker-Assisted Selection for Pollen-Free Somatic Plants of Sugi (Japanese Cedar, Cryptomeria japonica): A Simple and Effective Methodology for Selecting Male-Sterile Mutants With ms1-1 and ms1-2

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

An Improved and Simplified Propagation System for Pollen-Free Sugi (Cryptomeria japonica) via Somatic Embryogenesis

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