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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2020.1
- 수록면
- 549 - 558 (10page)
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초록· 키워드
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The clustered regulatory interspaced short palindromic repeat/CRISPR-associated protein 9 (CRISPR/Cas9) system has been applied to edit the genomes of quite a few plant species, including perennial woody poplar trees. However, chimeras often exist in primary transgenic plants. For perennial woody trees such as poplar trees, it is difcult to obtain homozygous mutants by self-pollination for due to their long vegetative life and low-seed setting rates. In this study, we report an efec?tive approach to reduce the frequency of chimeric mutants of poplar trees by CRISPR/Cas9 with a second round of shoot regeneration using leaves as the explants. PdbPDS1 was used as the target gene, and only one homozygous PdbPDS1 mutant was obtained from 15 primary transgenic plantlets, which was verifed by both the phenotype and the DNA sequence of the PCR product. This indicates that the majority of primary transgenic plantlets in the T0 generation were chimeras. After the second round of shoot regeneration of the chimeric mutants generated by CRISPR/Cas9, approximately 27.0% or 19.1% of the regenerated shoots were homozygous mutants with or without kanamycin selection, respectively. The results showed that a second regeneration could produce homozygous mutant shoots at a high frequency and that kanamycin selection could increase the frequency of homozygous mutant shoots.
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참고문헌 (27)
참고문헌 신청
Brooks C / 2014 / Efficient gene editing in tomato in the first generation using the clustered regularly interspaced short palindromic repeats/CRISPR-associated9system / Plant Physiol 166:1292~1297
Cermak T / 2015 / High frequency, precise modification of the tomato genome / Genome Biol 16:232~245
Chen GQ / 2011 / Effective reduction of chimeric tissue in transgenics for the stable genetic transformation of Lesquerella fendleri / HortScience 46:86~90
Christou P / 1990 / Morphological description of transgenic soybean chimeras created by the delivery, integration and expression of foreign DNA using electric discharge particle acceleration / Ann Bot 66:379~386
Domínguez A / 2004 / Characterisation of regenerants obtained under selective conditions after Agrobacterium-mediated transformation of citrus explants reveals production of silenced and chimeric plants at unexpected high frequencies / Mol Breed 14:171~183
Cermak T / 2015 / High frequency, precise modification of the tomato genome / Genome Biol 16:232~245
Chen GQ / 2011 / Effective reduction of chimeric tissue in transgenics for the stable genetic transformation of Lesquerella fendleri / HortScience 46:86~90
Christou P / 1990 / Morphological description of transgenic soybean chimeras created by the delivery, integration and expression of foreign DNA using electric discharge particle acceleration / Ann Bot 66:379~386
Domínguez A / 2004 / Characterisation of regenerants obtained under selective conditions after Agrobacterium-mediated transformation of citrus explants reveals production of silenced and chimeric plants at unexpected high frequencies / Mol Breed 14:171~183
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