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CLC number: S436.661.1+1

On-line Access: 2014-01-28

Received: 2013-08-12

Revision Accepted: 2013-12-06

Crosschecked: 2014-01-17

Cited: 3

Clicked: 5891

Citations:  Bibtex RefMan EndNote GB/T7714

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Journal of Zhejiang University SCIENCE B 2014 Vol.15 No.2 P.116-124

http://doi.org/10.1631/jzus.B1300213


Improvement of a gene targeting system for genetic manipulation in Penicillium digitatum * #


Author(s):  Qian Xu, Cong-yi Zhu, Ming-shang Wang, Xue-peng Sun, Hong-ye Li

Affiliation(s):  . Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   hyli@zju.edu.cn

Key Words:  Penicillium digitatum , Efficiency, Gene targeting, Non-homologous end-joining (NHEJ) pathway, Ku80


Qian Xu, Cong-yi Zhu, Ming-shang Wang, Xue-peng Sun, Hong-ye Li. Improvement of a gene targeting system for genetic manipulation in Penicillium digitatum[J]. Journal of Zhejiang University Science B, 2014, 15(2): 116-124.

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author="Qian Xu, Cong-yi Zhu, Ming-shang Wang, Xue-peng Sun, Hong-ye Li",
journal="Journal of Zhejiang University Science B",
volume="15",
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pages="116-124",
year="2014",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1300213"
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%0 Journal Article
%T Improvement of a gene targeting system for genetic manipulation in Penicillium digitatum
%A Qian Xu
%A Cong-yi Zhu
%A Ming-shang Wang
%A Xue-peng Sun
%A Hong-ye Li
%J Journal of Zhejiang University SCIENCE B
%V 15
%N 2
%P 116-124
%@ 1673-1581
%D 2014
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1300213

TY - JOUR
T1 - Improvement of a gene targeting system for genetic manipulation in Penicillium digitatum
A1 - Qian Xu
A1 - Cong-yi Zhu
A1 - Ming-shang Wang
A1 - Xue-peng Sun
A1 - Hong-ye Li
J0 - Journal of Zhejiang University Science B
VL - 15
IS - 2
SP - 116
EP - 124
%@ 1673-1581
Y1 - 2014
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1300213


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 (ΔPdKu80) of P. digitatum. However, using ΔPdKu80 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 ΔPdKu80 strain would be a good candidate for rapid gene function analysis in P. digitatum.

一种高效的柑橘绿霉菌基因敲除体系的构建

研究目的:提高柑橘绿霉菌基因敲除效率。
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PdbrlAPdmpkA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。

关键词:指状青霉;基因敲除;非同源末端链接(NHEJ);Ku80

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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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

Please provide your name, email address and a comment





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