Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant Population
To further develop barley breeding and genetics, more information on gene functions based on the analysis of the mutants of each gene is needed. However, the mutant resources are not as well developed as the model plants, such as Arabidopsis and rice. Although genome editing techniques have been abl...
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| Format: | Article |
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MDPI AG
2020-09-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/9/9/1153 |
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| author | Yudai Kawamoto Hirotaka Toda Hiroshi Inoue Kappei Kobayashi Naoto Yamaoka Takuya Araki Takashi Yaeno |
| author_facet | Yudai Kawamoto Hirotaka Toda Hiroshi Inoue Kappei Kobayashi Naoto Yamaoka Takuya Araki Takashi Yaeno |
| author_sort | Yudai Kawamoto |
| collection | DOAJ |
| description | To further develop barley breeding and genetics, more information on gene functions based on the analysis of the mutants of each gene is needed. However, the mutant resources are not as well developed as the model plants, such as Arabidopsis and rice. Although genome editing techniques have been able to generate mutants, it is not yet an effective method as it can only be used to transform a limited number of cultivars. Here, we developed a mutant population using ‘Mannenboshi’, which produces good quality grains with high yields but is susceptible to disease, to establish a Targeting Induced Local Lesions IN Genomes (TILLING) system that can isolate mutants in a high-throughput manner. To evaluate the availability of the prepared 8043 M<sub>3</sub> lines, we investigated the frequency of mutant occurrence using a rapid, visually detectable waxy phenotype as an indicator. Four mutants were isolated and single nucleotide polymorphisms (SNPs) were identified in the <i>Waxy</i> gene as novel alleles. It was confirmed that the mutations could be easily detected using the mismatch endonuclease CELI, revealing that a sufficient number of mutants could be rapidly isolated from our TILLING population. |
| first_indexed | 2024-03-10T16:32:24Z |
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| id | doaj.art-f618182d04054601868aa0131a686356 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T16:32:24Z |
| publishDate | 2020-09-01 |
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| series | Plants |
| spelling | doaj.art-f618182d04054601868aa0131a6863562023-11-20T12:46:45ZengMDPI AGPlants2223-77472020-09-0199115310.3390/plants9091153Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant PopulationYudai Kawamoto0Hirotaka Toda1Hiroshi Inoue2Kappei Kobayashi3Naoto Yamaoka4Takuya Araki5Takashi Yaeno6Department of Agriculture, Ehime University, Tarumi, Matsuyama, Ehime 790-8566, JapanDepartment of Agriculture, Ehime University, Tarumi, Matsuyama, Ehime 790-8566, JapanDepartment of Agriculture, Ehime University, Tarumi, Matsuyama, Ehime 790-8566, JapanDepartment of Agriculture, Ehime University, Tarumi, Matsuyama, Ehime 790-8566, JapanDepartment of Agriculture, Ehime University, Tarumi, Matsuyama, Ehime 790-8566, JapanDepartment of Agriculture, Ehime University, Tarumi, Matsuyama, Ehime 790-8566, JapanDepartment of Agriculture, Ehime University, Tarumi, Matsuyama, Ehime 790-8566, JapanTo further develop barley breeding and genetics, more information on gene functions based on the analysis of the mutants of each gene is needed. However, the mutant resources are not as well developed as the model plants, such as Arabidopsis and rice. Although genome editing techniques have been able to generate mutants, it is not yet an effective method as it can only be used to transform a limited number of cultivars. Here, we developed a mutant population using ‘Mannenboshi’, which produces good quality grains with high yields but is susceptible to disease, to establish a Targeting Induced Local Lesions IN Genomes (TILLING) system that can isolate mutants in a high-throughput manner. To evaluate the availability of the prepared 8043 M<sub>3</sub> lines, we investigated the frequency of mutant occurrence using a rapid, visually detectable waxy phenotype as an indicator. Four mutants were isolated and single nucleotide polymorphisms (SNPs) were identified in the <i>Waxy</i> gene as novel alleles. It was confirmed that the mutations could be easily detected using the mismatch endonuclease CELI, revealing that a sufficient number of mutants could be rapidly isolated from our TILLING population.https://www.mdpi.com/2223-7747/9/9/1153barleyTILLINGwaxyCELI |
| spellingShingle | Yudai Kawamoto Hirotaka Toda Hiroshi Inoue Kappei Kobayashi Naoto Yamaoka Takuya Araki Takashi Yaeno Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant Population Plants barley TILLING waxy CELI |
| title | Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant Population |
| title_full | Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant Population |
| title_fullStr | Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant Population |
| title_full_unstemmed | Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant Population |
| title_short | Fast and Inexpensive Phenotyping and Genotyping Methods for Evaluation of Barley Mutant Population |
| title_sort | fast and inexpensive phenotyping and genotyping methods for evaluation of barley mutant population |
| topic | barley TILLING waxy CELI |
| url | https://www.mdpi.com/2223-7747/9/9/1153 |
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