scholarly journals RNA Silencing of the Introduced Coat Protein Gene of Turnip mosaic virus Confers Broad-Spectrum Resistance in Transgenic Arabidopsis

2004 ◽  
Vol 94 (7) ◽  
pp. 730-736 ◽  
Author(s):  
Ken Nomura ◽  
Kazusato Ohshima ◽  
Toyoaki Anai ◽  
Hidetoshi Uekusa ◽  
Nobuhiro Kita

The coat protein (CP) gene derived from Turnip mosaic virus (TuMV) isolate JO was introduced into Arabidopsis thaliana and the resulting transgenic progenies were analyzed for resistance to TuMV. Transgenic Arabidopsis plants with no detectable transcripts of the introduced CP gene exhibited complete resistance to TuMV. There was no significant correlation between the resistance and the copy number of the transgene. Instead, small interfering RNAs (siRNAs) were detected in these resistant plants, indicating that the resistance is attributed to RNA silencing. The RNA-mediated resistance was not only inherited over successive generations but also effective against 17 worldwide TuMV isolates with different pathogenicity. Comparative analysis of the CP genes among the 17 TuMV isolates revealed that the 380-nt in the 3′ region is highly conserved, suggesting the importance of the 3′ conserved region for broad-spectrum resistance. These results indicate that introduction of the TuMV-CP gene into the target Brassicaceae plants followed by selecting transformants that show RNA silencing for the transgenes can be an effective and reliable strategy for developing crucifer crops with a broad spectrum of resistance to TuMV.

2003 ◽  
Vol 93 (1) ◽  
pp. 112-120 ◽  
Author(s):  
Huey-Jiunn Bau ◽  
Ying-Huey Cheng ◽  
Tsong-Ann Yu ◽  
Jiu-Sherng Yang ◽  
Shyi-Dong Yeh

Papaya ringspot virus (PRSV) is a major limiting factor for cultivation of papaya (Carica papaya) in tropical and subtropical areas throughout the world. Although the coat protein (CP) gene of PRSV has been transferred into papaya by particle bombardment and transgenic lines with high resistance to Hawaii strains have been obtained, they are susceptible to PRSV isolates outside of Hawaii. This strain-specific resistance limits the application of the transgenic lines in other areas of the world. In this investigation, the CP gene of a local strain isolated from Taiwan, designated PRSV YK, was transferred into papaya via Agrobacterium-mediated transformation. A total of 45 putative transgenic lines were obtained and the presence of the transgene in papaya was confirmed by polymerase chain reaction amplification. When the plants of transgenic lines were challenged with PRSV YK by mechanical inoculation, they showed different levels of resistance ranging from delay of symptom development to complete immunity. Molecular analysis of nine selected lines that exhibited different levels of resistance revealed that the expression level of the transgene is negatively correlated with the degree of resistance, suggesting that the resistance is manifested by a RNA-mediated mechanism. The segregation analysis showed that the transgene in the immune line 18-0-9 has an inheritance of two dominant loci and the other four highly resistant lines have a single dominant locus. Seven selected lines were tested further for resistance to three PRSV heterologous strains that originated in Hawaii, Thailand, and Mexico. Six of the seven lines showed varying degrees of resistance to the heterologous strains, and one line, 19-0-1, was immune not only to the homologous YK strain but also to the three heterologous strains. Thus, these CP-transgenic papaya lines with broad-spectrum resistance have great potential for use in Taiwan and other geographic areas to control PRSV.


2014 ◽  
Vol 27 (9) ◽  
pp. 944-955 ◽  
Author(s):  
Yi-Jung Kung ◽  
Pin-Chun Lin ◽  
Shyi-Dong Yeh ◽  
Syuan-Fei Hong ◽  
Nam-Hai Chua ◽  
...  

Cross-protection triggered by a mild strain of virus acts as a prophylaxis to prevent subsequent infections by related viruses in plants; however, the underling mechanisms are not fully understood. Through mutagenesis, we isolated a mutant strain of Turnip mosaic virus (TuMV), named Tu-GK, that contains an Arg182Lys substitution in helper component-proteinase (HC-ProK) that confers complete cross-protection against infection by a severe strain of TuMV in Nicotiana benthamiana, Arabidopsis thaliana Col-0, and the Arabidopsis dcl2-4/dcl4-1 double mutant defective in DICER-like ribonuclease (DCL)2/DCL4-mediated silencing. Our analyses showed that HC-ProK loses the ability to interfere with microRNA pathways, although it retains a partial capability for RNA silencing suppression triggered by DCL. We further showed that Tu-GK infection triggers strong salicylic acid (SA)-dependent and SA-independent innate immunity responses. Our data suggest that DCL2/4-dependent and –independent RNA silencing pathways are involved, and may crosstalk with basal innate immunity pathways, in host defense and in cross-protection.


1991 ◽  
Vol 57 (4) ◽  
pp. 549-557 ◽  
Author(s):  
Hiroyuki NAKASHIMA ◽  
Nobumichi SAKO ◽  
Keiichiro JOH ◽  
Katsuji HORI ◽  
Fukuji NONAKA

2013 ◽  
Vol 19 (1) ◽  
pp. 22-27
Author(s):  
Marija Žižytė ◽  
Donatas Šneideris ◽  
Irena Zitikaitė ◽  
Laima Urbanavičienė ◽  
Juozas Staniulis

Abstract Two isolates of Pepino mosaic virus (PepMV) from tomato plants grown in different commercial greenhouses in Lithuania were characterized by coat protein (CP) gene sequence analysis. Comparison with other PepMV isolates from the GenBank database showed that both Lithuanian PepMV isolates share 78.3% nucleotide identity and belong to two distinct EU and CH2 genotypes of PepMV. This is the first report on characterization of two PepMV genotypes detected in Lithuania.


Plant Disease ◽  
2006 ◽  
Vol 90 (1) ◽  
pp. 109-109 ◽  
Author(s):  
J. Méndez-Lozano ◽  
E. Quintero-Zamora ◽  
M. P. Barbosa-Jasso ◽  
N. E. Leyva-López ◽  
J. A. Garzón-Tiznado ◽  
...  

Since June 2001, symptoms of yellowing, leaf curling, crumpling, and stunted growth were observed on soybean (Glycine max Merr.) plants in Sinaloa, Mexico. These symptoms and the presence of whiteflies (Bemisia tabaci Gennadius) in the affected fields suggested a viral etiology. Samples from symptomatic plants were collected from commercial fields and analyzed for the presence of begomoviruses using DNA hybridization, and as a probe, the DNA A of Pepper huasteco virus at low stringency (2). Thirty-five positive samples were subsequently used for polymerase chain reaction (PCR) amplification with the degenerate primers RepMot and CPMot (1). These primers direct the amplification of a DNA A segment comprising the entire intergenic region (IR) and the first 210 bp of the coat protein (CP) gene, which is highly variable in size and nucleotide sequence among begomoviruses. PCR products were obtained for 25 of 35 samples and five of these were cloned into the pGEM-T easy vector (Promega, Madison, WI) and sequenced. The 571-bp DNA sequence (GenBank Accession No. AY905553) was compared with sequences of other begomoviruses in GenBank using the Clustal alignment method (MegAlign, DNASTAR software, London). The sequence was 74 and 70% identical to the Pepper golden mosaic virus (PepGMV; GenBank Accession No. U57457) and Cabbage leaf curl virus (CaLCuV; GenBank Accession No. U65529) sequences, respectively. Interestingly, the partial coat protein gene sequence (210 nt) of this soybean-infecting virus was 98% identical to the CP gene of Tobacco apical stunt virus (TbASV; GenBank Accession No. AF076855). Nonetheless, the known sequence of TbASV intergenic region (GenBank Accession No. AF077744) is very different from the homologous region of the soybean virus (34% of nucleotide identity). Analysis of the soybean virus intergenic region revealed that it harbors almost identical iterons (i.e., Rep-binding sites) to PepGMV, suggesting a close relationship between these two viruses. Soybean-infecting geminiviruses have been previously reported only from Asia; however, the partial sequence of a begomovirus isolated from soybean in Brazil was recently deposited in Genbank (Accession No. AY436328). Sequence comparisons between the Brazilian and Mexican isolates showed these viruses are less related with a nucleotide identity of 46%. Taken together, our data indicate that the virus identified in this study might be either a different strain of PepGMV adapted to leguminous plants or a new begomovirus species. To our knowledge, this is the first report of a begomovirus infecting soybean in Mexico. References: (1) J. T. Ascencio-Ibañez et al. Plant Dis. 86:692, 2002. (2) J. Méndez-Lozano et al. Phytopathology 93:270, 2003.


Author(s):  
LISTIHANI ◽  
SRI HENDRASTUTI HIDAYAT ◽  
SURYO WIYONO ◽  
Tri Asmira Damayanti

Abstract. Listihani, Hidayat SH, Wiyono S, Damayanti TA. 2019. Characteristic of Tobacco mosaic virus isolated from cucumber and tobacco collected from East Java, Indonesia. Biodiversitas 20: 2937-2942. Tobacco mosaic virus (TMV) is a newly emerging virus infecting cucumbers in Java, Indonesia. The basic characters of the TMV isolated from cucumber need further study to investigate its differences with that from tobacco. Thus, the research aimed to study the character of both isolates based on their biological, symptomatology and nucleic acid of coat protein (CP) gene properties. The TMV isolates from both cucumber and tobacco were able to infect similar indicator plants with differing symptom expressions, especially on eggplant. Homology of nucleotide and amino acid of coat protein gene among isolates were about 90.3% and 91.0%, and homology to other isolates was about 87.6 to 93.8% and 89.3 to 96.8%, respectively. There were 17 amino acid differences in the CP gene which is presumed to differentiate those two isolates. Phylogenetic analysis CP gene sequences compared with corresponding isolates in GenBank showed the two isolates separated in different clades. The cucumber isolate from Kediri is closely related to tomato isolate from China in clade I, while tobacco isolate from Jember closely related to tobacco isolate from China in clade IIindicating the existence of two differ TMV variants. It will increase the difficulty to manage TMV in the fields.


2020 ◽  
Vol 21 (9) ◽  
pp. 1194-1211 ◽  
Author(s):  
Zhaoji Dai ◽  
Rongrong He ◽  
Mark A. Bernards ◽  
Aiming Wang

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