Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.).
Mutagenesis is an important tool in crop improvement. However, the hexaploid genome of wheat (Triticum aestivum L.) presents problems in identifying desirable genetic changes based on phenotypic screening due to gene redundancy. TILLING (Targeting Induced Local Lesions IN Genomes), a powerful revers...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0041570&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850137342297243648 |
|---|---|
| author | Liang Chen Linzhou Huang Donghong Min Andy Phillips Shiqiang Wang Pippa J Madgwick Martin A J Parry Yin-Gang Hu |
| author_facet | Liang Chen Linzhou Huang Donghong Min Andy Phillips Shiqiang Wang Pippa J Madgwick Martin A J Parry Yin-Gang Hu |
| author_sort | Liang Chen |
| collection | DOAJ |
| description | Mutagenesis is an important tool in crop improvement. However, the hexaploid genome of wheat (Triticum aestivum L.) presents problems in identifying desirable genetic changes based on phenotypic screening due to gene redundancy. TILLING (Targeting Induced Local Lesions IN Genomes), a powerful reverse genetic strategy that allows the detection of induced point mutations in individuals of the mutagenized populations, can address the major challenge of linking sequence information to the biological function of genes and can also identify novel variation for crop breeding. Wheat is especially well-suited for TILLING due to the high mutation densities tolerated by polyploids. However, only a few wheat TILLING populations are currently available in the world, which is far from satisfying the requirement of researchers and breeders in different growing environments. In addition, current TILLING screening protocols require costly fluorescence detection systems, limiting their use, especially in developing countries. We developed a new TILLING resource comprising 2610 M(2) mutants in a common wheat cultivar 'Jinmai 47'. Numerous phenotypes with altered morphological and agronomic traits were observed from the M(2) and M(3) lines in the field. To simplify the procedure and decrease costs, we use unlabeled primers and either non-denaturing polyacrylamide gels or agarose gels for mutation detection. The value of this new resource was tested using PCR with RAPD and Intron-spliced junction (ISJ) primers, and also TILLING in three selected candidate genes, in 300 and 512 mutant lines, revealing high mutation densities of 1/34 kb by RAPD/ISJ analysis and 1/47 kb by TILLING. In total, 31 novel alleles were identified in the 3 targeted genes and confirmed by sequencing. The results indicate that this mutant population represents a useful resource for the wheat research community. We hope that the use of this reverse genetics resource will provide novel allelic diversity for wheat improvement and functional genomics. |
| format | Article |
| id | doaj-art-ac8a049d5dd34c0892f58989dfc343a4 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-ac8a049d5dd34c0892f58989dfc343a42025-08-20T02:30:51ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0177e4157010.1371/journal.pone.0041570Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.).Liang ChenLinzhou HuangDonghong MinAndy PhillipsShiqiang WangPippa J MadgwickMartin A J ParryYin-Gang HuMutagenesis is an important tool in crop improvement. However, the hexaploid genome of wheat (Triticum aestivum L.) presents problems in identifying desirable genetic changes based on phenotypic screening due to gene redundancy. TILLING (Targeting Induced Local Lesions IN Genomes), a powerful reverse genetic strategy that allows the detection of induced point mutations in individuals of the mutagenized populations, can address the major challenge of linking sequence information to the biological function of genes and can also identify novel variation for crop breeding. Wheat is especially well-suited for TILLING due to the high mutation densities tolerated by polyploids. However, only a few wheat TILLING populations are currently available in the world, which is far from satisfying the requirement of researchers and breeders in different growing environments. In addition, current TILLING screening protocols require costly fluorescence detection systems, limiting their use, especially in developing countries. We developed a new TILLING resource comprising 2610 M(2) mutants in a common wheat cultivar 'Jinmai 47'. Numerous phenotypes with altered morphological and agronomic traits were observed from the M(2) and M(3) lines in the field. To simplify the procedure and decrease costs, we use unlabeled primers and either non-denaturing polyacrylamide gels or agarose gels for mutation detection. The value of this new resource was tested using PCR with RAPD and Intron-spliced junction (ISJ) primers, and also TILLING in three selected candidate genes, in 300 and 512 mutant lines, revealing high mutation densities of 1/34 kb by RAPD/ISJ analysis and 1/47 kb by TILLING. In total, 31 novel alleles were identified in the 3 targeted genes and confirmed by sequencing. The results indicate that this mutant population represents a useful resource for the wheat research community. We hope that the use of this reverse genetics resource will provide novel allelic diversity for wheat improvement and functional genomics.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0041570&type=printable |
| spellingShingle | Liang Chen Linzhou Huang Donghong Min Andy Phillips Shiqiang Wang Pippa J Madgwick Martin A J Parry Yin-Gang Hu Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.). PLoS ONE |
| title | Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.). |
| title_full | Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.). |
| title_fullStr | Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.). |
| title_full_unstemmed | Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.). |
| title_short | Development and characterization of a new TILLING population of common bread wheat (Triticum aestivum L.). |
| title_sort | development and characterization of a new tilling population of common bread wheat triticum aestivum l |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0041570&type=printable |
| work_keys_str_mv | AT liangchen developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml AT linzhouhuang developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml AT donghongmin developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml AT andyphillips developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml AT shiqiangwang developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml AT pippajmadgwick developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml AT martinajparry developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml AT yinganghu developmentandcharacterizationofanewtillingpopulationofcommonbreadwheattriticumaestivuml |