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

2020 ◽  
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.

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 Marker-Assisted Selection of Trees with MALE STERILITY 1 in Cryptomeria japonica D. Don

Forests ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 734 ◽  
Author(s):  
Yoshinari Moriguchi ◽  
Saneyoshi Ueno ◽  
Yoichi Hasegawa ◽  
Takumi Tadama ◽  
Masahiro Watanabe ◽  
...  
Keyword(s):  
Male Sterility ◽  
Linkage Map ◽  
Marker Assisted Selection ◽  
Tree Breeding ◽  
Segregation Ratio ◽  
Linkage Maps ◽  
Male Sterile ◽  
Miyagi Prefecture ◽  
Time Required ◽  
Selection Of

The practical use of marker-assisted selection (MAS) is limited in conifers because of the difficulty with developing markers due to a rapid decrease in linkage disequilibrium, the limited genomic information available, and the diverse genetic backgrounds among the breeding material collections. First, in this study, two families were produced by artificial crossing between two male-sterile trees, ‘Shindai11’ and ‘Shindai12’, and a plus tree, ‘Suzu-2’ (Ms1/ms1) (S11-S and S12-S families, respectively). The segregation ratio between the male-sterile and male-fertile trees did not deviate significantly from the expected 1:1 ratio in either family. These results clearly suggested that the male-sterile gene of ‘Shindai11’ and ‘Shindai12’ is MALE STERILITY 1 (MS1). Since it is difficult to understand the relative positions of each marker, due to the lack of a linkage map which all the closely linked markers previously reported are mapped on, we constructed a partial linkage map of the region encompassing MS1 using the S11-S and S12-S families. For the S11-S and S12-S families, 19 and 18 markers were mapped onto the partial linkage maps of the MS1 region, respectively. There was collinearity (conserved gene order) between the two partial linkage maps. Two markers (CJt020762_ms1-1 and reCj19250_2335) were mapped to the same position as the MS1 locus on both maps. Of these markers, we used CJt020762 for the MAS in this study. According to the MAS results for 650 trees from six prefectures of Japan (603 trees from breeding materials and 47 trees from the Ishinomaki natural population), five trees in Niigata Prefecture and one tree in Yamagata Prefecture had heterozygous ms1-1, and three trees in Miyagi Prefecture had heterozygous ms1-2. The results obtained in this study suggested that ms1-1 and ms1-2 have different geographical distributions. Since MAS can be used effectively to reduce the labor and time required for selection of trees with a male-sterile gene, the research should help ensure that the quantity of breeding materials will increase to assist future tree-breeding efforts.

scholarly journals Genetic evaluation of Cryptomeria japonica breeding materials for male-sterile trees

2019 ◽  
Vol 68 (1) ◽  
pp. 67-72
Author(s):  
Takumi Tadama ◽  
Satoko Totsuka ◽  
Junji Iwai ◽  
Kentaro Uchiyama ◽  
Yoichi Hasegawa ◽  
...  
Keyword(s):  
Male Sterility ◽  
Inbreeding Depression ◽  
Cryptomeria Japonica ◽  
Genetic Relatedness ◽  
Snp Markers ◽  
Male Sterile ◽  
Negative Effects ◽  
Niigata Prefecture ◽  
Genome Wide ◽  
Selection Of

Abstract Pyramiding of male-sterile genes in Cryptomeria japonica is currently being carried out in Niigata prefecture, Japan. This is the first attempt to apply pyramid breeding to forest trees. As the breeding materials for male sterility are limited, special attention must be given to increased genetic relatedness in the process of pyramid breeding to avoid the effects of inbreeding depression as much as possible. In this study, we estimated genetic relatedness based on 246 genome-wide SNP markers for male-sterile individuals in Niigata Prefecture (n = 6) and individuals doubly heterozygous for two male-sterile genes (hereafter referred to as “double-hetero”) produced by marker- assisted selection (n = 124). The pairwise relatedness estimates between male-sterile individuals selected from the same area in Niigata Prefecture were low (−0.01 ± 0.08, mean ± standard deviation), suggesting that there will be almost no negative effects even if the F1 of these male-sterile individuals is used for artificial crossing. On the other hand, the pairwise relatedness between double-hetero individuals in this study was higher than the theoretical relatedness values, as individuals with the slightly higher relatedness were used as parents in artificial crossings. However, there was a large variance in pairwise relatedness for double-hetero individuals. This result suggested that it may be possible to avoid the adverse effects of inbreeding depression by using a pair of double-heteros with lower relatedness for artificial crossing, when we produce a double-homo using the limited breeding materials of male- sterile individuals. It will also be important to continue additional selection of new breeding material for male sterility.

scholarly journals Selection of Trees with Male Sterile Genes Except for MALE STERILITY 1 in Cryptomeria japonica D. Don

2021 ◽  
Vol 103 (2) ◽  
pp. 161-167
Author(s):  
Satoko Hirayama ◽  
Junji Iwai ◽  
Yumi Higuchi ◽  
Takeo Kaneko ◽  
Yoshinari Moriguchi
Keyword(s):  
Male Sterility ◽  
Cryptomeria Japonica ◽  
Male Sterile ◽  
Selection Of

scholarly journals A single nucleotide polymorphism in an R2R3 MYB transcription factor gene triggers the male sterility in soybean ms6 (Ames1)

2021 ◽  
Author(s):  
Junping Yu ◽  
Guolong Zhao ◽  
Wei Li ◽  
Ying Zhang ◽  
Peng Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Glycine Max ◽  
Male Sterility ◽  
Bulked Segregant Analysis ◽  
Soybean Protein ◽  
Anther Development ◽  
Hybrid Breeding ◽  
Myb Transcription Factor ◽  
Male Sterile ◽  
R2r3 Myb

Abstract Key message Identification and functional analysis of the male sterile gene MS6 in Glycine max. Abstract Soybean (Glycine max (L.) Merr.) is an important crop providing vegetable oil and protein. The male sterility-based hybrid breeding is a promising method for improving soybean yield to meet the globally growing demand. In this research, we identified a soybean genic male sterile locus, MS6, by combining the bulked segregant analysis sequencing method and the map-based cloning technology. MS6, highly expressed in anther, encodes an R2R3 MYB transcription factor (GmTDF1-1) that is homologous to Tapetal Development and Function 1, a key factor for anther development in Arabidopsis and rice. In male sterile ms6 (Ames1), the mutant allele contains a missense mutation, leading to the 76th leucine substituted by histidine in the DNA binding domain of GmTDF1-1. The expression of soybean MS6 under the control of the AtTDF1 promoter could rescue the male sterility of attdf1 but ms6 could not. Additionally, ms6 overexpression in wild-type Arabidopsis did not affect anther development. These results evidence that GmTDF1-1 is a functional TDF1 homolog and L76H disrupts its function. Notably, GmTDF1-1 shows 92% sequence identity with another soybean protein termed as GmTDF1-2, whose active expression also restored the fertility of attdf1. However, GmTDF1-2 is constitutively expressed at a very low level in soybean, and therefore, not able to compensate for the MS6 deficiency. Analysis of the TDF1-involved anther development regulatory pathway showed that expressions of the genes downstream of TDF1 are significantly suppressed in ms6, unveiling that GmTDF1-1 is a core transcription factor regulating soybean anther development.

scholarly journals The Dynamics of Gynodioecy in Plantago lanceolatu L. II. Mode of Action and Frequencies of Restorer Alleles

Genetics ◽  
1997 ◽  
Vol 147 (3) ◽  
pp. 1317-1328
Author(s):  
Anita A de Haan ◽  
Hans P Koelewijn ◽  
Maria P J Hundscheid ◽  
Jos M M Van Damme
Keyword(s):  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Allele Frequency ◽  
Mode Of Action ◽  
Male Fertility ◽  
Fertility Restoration ◽  
Plantago Lanceolata ◽  
Allele Frequencies ◽  
Male Sterile ◽  
Male Fertility Restoration

Male fertility in Plantago lanceolata is controlled by the interaction of cytoplasmic and nuclear genes. Different cytoplasmic male sterility (CMS) types can be either male sterile or hermaphrodite, depending on the presence of nuclear restorer alleles. In three CMS types of P. lanceolata (CMSI, CMSIIa, and CMSIIb) the number of loci involved in male fertility restoration was determined. In each CMS type, male fertility was restored by multiple genes with either dominant or recessive action and capable either of restoring male fertility independently or in interaction with each other (epistasis). Restorer allele frequencies for CMSI, CMSIIa and CMSIIb were determined by crossing hermaphrodites with “standard” male steriles. Segregation of male steriles vs. non-male steriles was used to estimate overall restorer allele frequency. The frequency of restorer alleles was different for the CMS types: restorer alleles for CMSI were less frequent than for CMSIIa and CMSIIb. On the basis of the frequencies of male steriles and the CMS types an “expected” restorer allele frequency could be calculated. The correlation between estimated and expected restorer allele frequency was significant.

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