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2016 Vol.34, Issue 2 Preview Page
Development of a Transient ihpRNA-induced Gene Silencing System for Functional Analysis in Persimmon (Diospyros kaki Thunb.)
Rongli Mo1
,
Na Zhang1
,
Sichao Yang1
,
Qinglin Zhang12
,
Zhengrong Luo1,2*
11Key Laboratory of Horticultural Plant Biology, Huazhong Agricultural University
2Hubei Collaborative Innovation Center for the Characteristic Resources Exploitation of Dabie Mountains
*교신저자. *Corresponding Author. 
30 April 2016. pp. 314-323
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Abstract

A transient ihpRNA-induced gene silencing system based on Agrobacterium-mediated injection infiltration has been established to evaluate candidate genes involved in proanthocyanidin (PAs) biosynthesis in persimmon (Diospyros kaki Thunb.). We chose DkPDS (phytoene desaturase) as a gene-silencing target to evaluate the newly developed transient gene silencing system. Our qRT-PCR analysis indicated that two ihpRNA constructs (pHG-PDS5’ and pHG-PDS3’) targeted DkPDS, which also led to significantly reduce expression of DkPDS in ‘Mopanshi’ persimmon leaves. To further confirm the reliability of the system, we successfully utilized it for DkLAR (leucoanthocyanidin reductase) gene silencing. The expression levels of DkLAR in ‘Mopanshi’ and ‘Eshi 1’ leaves were ca. 6-fold and ca. 5-fold lower than those in leaves harboring empty vector (pHG-GFP), respectively. DMACA (4-dimethylaminocinnamaldehyde) staining and the Folin-Ciocalteau assay showed that the accumulation of PAs was markedly inhibited in ‘Mopanshi’, ‘Eshi 1’ and ‘Youhou’ leaves. These results indicate that DkLAR plays an important role in the accumulation of PAs in persimmon. The transient ihpRNA-induced gene silencing method developed in this study is a highly efficient and useful tool for functional analysis of persimmon genes involved in PA biosynthesis.

Keywords
Agrobacterium tumefaciens
DkLAR
DkPDS
proanthocyanidin

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Information
  • Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
  • Publisher(Ko) :한국원예학회
  • Journal Title :Horticultural Science and Technology
  • Journal Title(Ko) :원예과학기술지
  • Volume : 34
  • No :2
  • Pages :314-323
  • Received Date : 2015-10-13
  • Revised Date : 2015-12-14
  • Accepted Date : 2016-03-23
  • DOI :https://doi.org/10.12972/kjhst.20160032
Journal Informaiton Horticultural Science and Technology Horticultural Science and Technology
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