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Journal of Zhejiang University SCIENCE B
ISSN 1673-1581(Print), 1862-1783(Online), Monthly
2014 Vol.15 No.2 P.116-124
Improvement of a gene targeting system for genetic manipulation in Penicillium digitatum
Abstract: Penicillium digitatum is the most important pathogen of postharvest citrus. Gene targeting can be done in P. digitatum using homologous recombination via Agrobacterium tumefaciens mediated transformation (ATMT), but the frequencies are often very low. In the present study, we replaced the Ku80 homolog (a gene of the non-homologous end-joining (NHEJ) pathway) with the hygromycin resistance cassette (hph) by ATMT. No significant change in vegetative growth, conidiation, or pathogenicity was observed in Ku80-deficient strain (?PdKu80) of P. digitatum. However, using ?PdKu80 as a targeting strain, the gene-targeting frequencies for both genes PdbrlA and PdmpkA were significantly increased. These results suggest that Ku80 plays an important role in homologous integration and the created ?PdKu80 strain would be a good candidate for rapid gene function analysis in P. digitatum.
Key words: Penicillium digitatum, Efficiency, Gene targeting, Non-homologous end-joining (NHEJ) pathway, Ku80
The online version of this article contains supplementary materials Fig. S1, Fig. S2, Fig. S3, and Table S1
创新要点:低效的基因敲除与丝状真菌非同源末端链接(NHEJ)的DNA双链断裂修复途径有关。为提高柑橘绿霉病菌基因敲除效率,本研究利用农杆菌介导的转化体系,获得NHEJ途径中关键因子Ku80的缺失突变体(ΔPdKu80)。
研究方法:与野生型菌株相比,以ΔPdKu80作为出发菌株,提高柑橘绿霉病菌PdbrlA和PdmpkA的基因敲除效率(见表1)。
重要结论:ΔPdKu80的营养生长、产孢和致病性与野生型菌株基本一致。ΔPdKu80作为出发菌株,能显著提高柑橘绿霉菌的敲除效率。
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Open peer comments: Debate/Discuss/Question/Opinion
<1>
Qing-qing Huang<819075736@qq.com>
2015-06-12 10:12:23
Dear author,
Where is the Table S1,I couldn't find it. I want to make it as a reference.I am looking forward to your help
DOI:
10.1631/jzus.B1300213
CLC number:
S436.661.1+1
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2024-08-27
Received:
2023-10-17
Revision Accepted:
2024-05-08
Crosschecked:
2014-01-17