Virus-induced gene silencing is an effective tool for assaying gene function in the basal eudicot species Papaver somniferum (opium poppy)

被引:157
作者
Hileman, LC
Drea, S
de Martino, G
Litt, A
Irish, VF
机构
[1] Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA
[2] Yale Univ, Dept Ecol & Evolutionary Biol, New Haven, CT 06520 USA
关键词
TRV-VIGS; virus-induced gene silencing; Papaver somniferum; poppy; phytoene desaturase;
D O I
10.1111/j.1365-313X.2005.02520.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Virus-induced gene silencing (VIGS) is an attractive method for assaying gene function in species that are resistant to conventional genetic approaches. However, VIGS has been shown to be effective in only a few, closely related plant species. Tobacco rattle virus (TRV), a bipartite RNA virus, has a wide host range and so in principle could serve as an efficient vector for VIGS in a diverse array of plant species. Here we show that a vector based on TRV sequences is effective at silencing the endogenous phytoene desaturase (PapsPDS) gene in Papaver somniferum (opium poppy). We show that this vector does not compromise the growth or reproduction of poppy and the plants did not display viral symptoms. The silencing of PapsPDS resulted in a significant reduction in PapsPDS mRNA and a concomitant photobleached phenotype. The ability to rapidly assay gene function in A somniferum will be valuable in manipulation of the opiate pathway in this pharmaceutically important species. We suggest that our vacuum infiltration method used to deliver TRV-based vectors into poppy is a promising approach for expanding VIGS to diverse angiosperm species in which traditional delivery methods fail to induce VIGS. Furthermore, these studies demonstrate the utility of TRV-VIGS for probing gene function in a basal eudicot species that is phylogenetically distant from model plant species.
引用
收藏
页码:334 / 341
页数:8
相关论文
共 47 条
[31]   Virus-induced gene silencing in plants [J].
Lu, R ;
Martin-Hernandez, AM ;
Peart, JR ;
Malcuit, I ;
Baulcombe, DC .
METHODS, 2003, 30 (04) :296-303
[32]   Molecular biology of the tobraviruses [J].
MacFarlane, SA .
JOURNAL OF GENERAL VIROLOGY, 1999, 80 :2799-2807
[33]   Cellular RNA-dependent RNA polymerase involved posttranscriptional gene silencing has two distinct activity modes [J].
Makeyev, EV ;
Bamford, DH .
MOLECULAR CELL, 2002, 10 (06) :1417-1427
[34]   Putting the opium in poppy to sleep [J].
Memelink, J .
NATURE BIOTECHNOLOGY, 2004, 22 (12) :1526-1527
[35]   Analgesia -: Morphine-pathway block in top1 poppies [J].
Millgate, AG ;
Pogson, BJ ;
Wilson, IW ;
Kutchan, TM ;
Zenk, MH ;
Gerlachll, WL ;
Fist, AJ ;
Larkin, PJ .
NATURE, 2004, 431 (7007) :413-414
[36]   An EDS1 orthologue is required for N-mediated resistance against tobacco mosaic virus [J].
Peart, JR ;
Cook, G ;
Feys, BJ ;
Parker, JE ;
Baulcombe, DC .
PLANT JOURNAL, 2002, 29 (05) :569-579
[37]   Ubiquitin ligase-associated protein SGT1 is required for host and nonhost disease resistance in plants [J].
Peart, JR ;
Lu, R ;
Sadanandom, A ;
Malcuit, I ;
Moffett, P ;
Brice, DC ;
Schauser, L ;
Jaggard, DAW ;
Xiao, SY ;
Coleman, MJ ;
Dow, M ;
Jones, JDG ;
Shirasu, K ;
Baulcombe, DC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (16) :10865-10869
[38]   Ectopic expression of rice OsMADS1 reveals a role in specifying the lemma and palea, grass floral organs analogous to sepals [J].
Prasad, K ;
Sriram, P ;
Kumar, CS ;
Kushalappa, K ;
Vijayraghavan, U .
DEVELOPMENT GENES AND EVOLUTION, 2001, 211 (06) :281-290
[39]   Tobacco rattle virus as a vector for analysis of gene function by silencing [J].
Ratcliff, F ;
Martin-Hernandez, AM ;
Baulcombe, DC .
PLANT JOURNAL, 2001, 25 (02) :237-245
[40]   RETRACTED: CITRX thioredoxin interacts with the tomato Cf-9 resistance protein and negatively regulates defence (Retracted article. See vol. 38, 2019) [J].
Rivas, S ;
Rougon-Cardoso, A ;
Smoker, M ;
Schauser, L ;
Yoshioka, H ;
Jones, JDG .
EMBO JOURNAL, 2004, 23 (10) :2156-2165