scholarly journals 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

2021 ◽  
Vol 12 ◽  
Author(s):  
Momi Tsuruta ◽  
Tsuyoshi E. Maruyama ◽  
Saneyoshi Ueno ◽  
Yoichi Hasegawa ◽  
Yoshinari Moriguchi
Keyword(s):  
Marker Assisted Selection ◽  
Cryptomeria Japonica ◽  
Japanese Cedar ◽  
Causative Mutation ◽  
Efficient Production ◽  
Male Sterile ◽  
Plant Materials ◽  
Embryogenic Cells ◽  
Embryogenic Cell ◽  
Somatic Plants

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.

scholarly journals 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 ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 1029 ◽  
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.

scholarly journals Somatic Embryogenesis Initiation in Sugi (Japanese Cedar, Cryptomeria japonica D. Don): Responses from Male-Fertile, Male-Sterile, and Polycross-Pollinated-Derived Seed Explants

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 398
Author(s):  
Tsuyoshi E. Maruyama ◽  
Saneyoshi Ueno ◽  
Yoshihisa Hosoi ◽  
Shin-Ichi Miyazawa ◽  
Hideki Mori ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Cryptomeria Japonica ◽  
Japanese Cedar ◽  
Influential Factor ◽  
Male Sterile ◽  
Embryogenic Cell ◽  
Factors Affecting ◽  
Seed Collection ◽  
Fertile Male ◽  
Collection Time

This study aimed to obtain information from several embryogenic cell (EC) genotypes analyzing the factors that affect somatic embryogenesis (SE) initiation in sugi (Cryptomeria japonica, Cupressaceae) to apply them in the improvement of protocols for efficient induction of embryogenic cell lines (ECLs). The results of several years of experiments including studies on the influence of initial explant, seed collection time, and explant genotype as the main factors affecting SE initiation from male-fertile, male-sterile, and polycross-pollinated-derived seeds are described. Initiation frequencies depending on the plant genotype varied from 1.35 to 57.06%. The best induction efficiency was achieved when seeds were collected on mid-July using the entire megagametophyte as initial explants. The extrusion of ECs started approximately after 2 weeks of culture, and the establishment of ECLs was observed mostly 4 weeks after extrusion on media with or without plant growth regulators (PGRs). Subsequently, induced ECLs were maintained and proliferated on media with PGRs by 2–3-week-interval subculture routines. Although, the initial explant, collection time, and culture condition played important roles in ECL induction, the genotype of the plant material of sugi was the most influential factor in SE initiation.

scholarly journals Fine mapping of the male-sterile genes (MS1, MS2, MS3, and MS4) and development of SNP markers for marker-assisted selection in Japanese cedar (Cryptomeria japonica D. Don)

PLoS ONE ◽  
2018 ◽  
Vol 13 (11) ◽  
pp. e0206695 ◽  
Author(s):  
Yoichi Hasegawa ◽  
Saneyoshi Ueno ◽  
Asako Matsumoto ◽  
Tokuko Ujino-Ihara ◽  
Kentaro Uchiyama ◽  
...  
Keyword(s):  
Fine Mapping ◽  
Marker Assisted Selection ◽  
Cryptomeria Japonica ◽  
Japanese Cedar ◽  
Snp Markers ◽  
Male Sterile

scholarly journals In vitro culture of various genotypes of male sterile Japanese cedar (Cryptomeria japonica D. Don)

2011 ◽  
Vol 28 (1) ◽  
pp. 103-106 ◽  
Author(s):  
Katsuaki Ishii ◽  
Yoshihisa Hosoi ◽  
Toru Taniguchi ◽  
Miyoko Tsubomura ◽  
Teiji Kondo ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Cryptomeria Japonica ◽  
Japanese Cedar ◽  
Male Sterile

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

Plants ◽  
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.

scholarly journals Identification and genetic diversity analysis of a male-sterile gene (MS1) in Japanese cedar (Cryptomeria japonica D. Don)

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yoichi Hasegawa ◽  
Saneyoshi Ueno ◽  
Fu-Jin Wei ◽  
Asako Matsumoto ◽  
Kentaro Uchiyama ◽  
...  
Keyword(s):  
Cryptomeria Japonica ◽  
Lipid Transfer Protein ◽  
Pollen Dispersal ◽  
Japanese Cedar ◽  
Lipid Transfer ◽  
Male Sterile ◽  
Causative Gene ◽  
Transfer Protein ◽  
The Japanese Archipelago ◽  
And Function

AbstractIdentifying causative genes for a target trait in conifer reproduction is challenging for species lacking whole-genome sequences. In this study, we searched for the male-sterility gene (MS1) in Cryptomeria japonica, aiming to promote marker-assisted selection (MAS) of male-sterile C. japonica to reduce the pollinosis caused by pollen dispersal from artificial C. japonica forests in Japan. We searched for mRNA sequences expressed in male strobili and found the gene CJt020762, coding for a lipid transfer protein containing a 4-bp deletion specific to male-sterile individuals. We also found a 30-bp deletion by sequencing the entire gene of another individual with the ms1. All nine breeding materials with the allele ms1 had either a 4-bp or 30-bp deletion in gene CJt020762, both of which are expected to result in faulty gene transcription and function. Furthermore, the 30-bp deletion was detected from three of five individuals in the Ishinomaki natural forest. From our findings, CJt020762 was considered to be the causative gene of MS1. Thus, by performing MAS using two deletion mutations as a DNA marker, it will be possible to find novel breeding materials of C. japonica with the allele ms1 adapted to the unique environment of each region of the Japanese archipelago.

scholarly journals Identification and genetic diversity analysis of a male-sterile gene (MS1) in Japanese cedar (Cryptomeria japonica D. Don)

2020 ◽  
Author(s):  
Yoichi Hasegawa ◽  
Saneyoshi Ueno ◽  
Fu-Jin Wei ◽  
Asako Matsumoto ◽  
Kentaro Uchiyama ◽  
...  
Keyword(s):  
Cryptomeria Japonica ◽  
Lipid Transfer Protein ◽  
Pollen Dispersal ◽  
Japanese Cedar ◽  
Lipid Transfer ◽  
Male Sterile ◽  
Causative Gene ◽  
Transfer Protein ◽  
The Japanese Archipelago ◽  
And Function

AbstractIdentifying causative genes for a target trait in conifer reproduction is challenging for species lacking whole-genome sequences. In this study, we searched for the male-sterility gene (MS1) in Cryptomeria japonica, aiming to promote marker-assisted selection (MAS) of male-sterile C. japonica to reduce the pollinosis caused by pollen dispersal from artificial C. japonica forests in Japan. We searched for mRNA sequences expressed in male strobili and found the gene CJt020762, coding for a lipid transfer protein containing a 4-bp deletion specific to male-sterile individuals. We also found a 30-bp deletion by sequencing the entire gene of another individual with the ms1. All nine breeding materials with the allele ms1 had either a 4-bp or 30-bp deletion in gene CJt020762, both of which are expected to result in faulty gene transcription and function. Furthermore, the 30-bp deletion was detected from three of five individuals in the Ishinomaki natural forest. From our findings, CJt020762 was considered to be the causative gene of MS1. Thus, by performing MAS using two deletion mutations as a DNA marker, it will be possible to find novel breeding materials of C. japonica with the allele ms1 adapted to the unique environment of each region of the Japanese archipelago.

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