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Assessment of penetrance and expressivity of RNAi‐mediated silencing of the Arabidopsis phytoene desaturase gene
Author(s) -
Wang Tao,
Iyer Lakshminarayan M.,
Pancholy Rakesh,
Shi Xiangyu,
Hall Timothy C.
Publication year - 2005
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/j.1469-8137.2005.01454.x
Subject(s) - rna interference , gene silencing , biology , penetrance , phytoene desaturase , genetics , gene knockdown , functional genomics , gene , arabidopsis , phenotype , expressivity , genome , rna , genomics , mutant
Summary• RNA interference (RNAi) is of great value in plant functional genomics. However, the absence of RNAi phenotypes and the lack of uniform level of RNAi silencing has complicated gene identification. Here, the penetrance and expressivity of RNAi‐mediated silencing of the phytoene desaturase ( PDS ) gene in Arabidopsis thaliana were examined quantitatively to provide a reference for the likely severity and distribution of silencing effects. • Arabidopsis plants were transformed with an RNAi construct targeting PDS . Transgenic plants were examined for frequency of RNAi‐mediated silencing and various silencing phenotypes. mRNA depletion level and RNAi expressivity were assayed by relative reverse transcription polymerase chain reaction (RT‐PCR). • High penetrance and variable expressivity of RNAi were demonstrated. An inverse correlation between PDS mRNA level and RNAi phenotype was seen. No direct relationship between copy number for the RNAi‐generating transgene and phenotype was evident. Decreased RNAi penetrance in T 2 plants was observed. • It is suggested that variability in RNAi expressivity and postmeiotic decrease in RNAi penetrance constitute barriers for high throughput plant gene characterization.