Selectable marker-free co-expression of Nicotiana rustica CN and Nicotiana tabacum HAK1 genes improves resistance to tobacco mosaic virus in tobacco

2015 ◽  
Vol 42 (8) ◽  
pp. 802 ◽  
Author(s):  
Li-Jun Qin ◽  
Dan Zhao ◽  
Yi Zhang ◽  
De-Gang Zhao
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Plant Cells ◽  
Viral Disease ◽  
Breeding Strategy ◽  
Pcr Analysis ◽  
Physiological Status ◽  
Plant Responses ◽  
Biotic Stresses ◽  
Marker Free

The viral disease caused by tobacco mosaic virus (TMV) is the most prevalent viral disease in many tobacco production areas. A breeding strategy based on resistance genes is an effective method for improving TMV resistance in tobacco. Also, the physiological status of plants is also critical to disease resistance improvement. Potassium ion is one of the most abundant inorganic nutrients in plant cells, and mediates plant responses to abiotic and biotic stresses. Improving K+ content in soil by fertilising can enhance diseases resistance of crops. However, the K+ absorption in plants depends mostly on K+ transporters located in cytoplasmic membrane. Therefore, the encoding genes for K+ transporters are putative candidates to target for improving tobacco mosaic virus resistance. In this work, the synergistic effect of a N-like resistance gene CN and a tobacco putative potassium transporter gene HAK1 was studied. The results showed that TMV-resistance in CN-HAK1-containing tobaccos was significantly enhanced though a of strengthening leaf thickness and reduction in the size of necrotic spots compared with only CN-containing plants, indicating the improvement of potassium nutrition in plant cells could increase the tobacco resistance to TMV by reducing the spread of the virus. Quantitative real-time polymerase chain reaction (qRT–PCR) analysis for TMV-CP expression in the inoculated leaf of the transgenic and wild-type plants also supported the conclusion. Further, the results of defence-related determination including antioxidative enzymes (AOEs) activity, salicylic acid (SA) content and the expression of resistance-related genes demonstrated CN with HAK1 synergistically enhanced TMV-resistance in transgenic tobaccos. Additionally, the HAK1- overexpression significantly improved the photosynthesis and K+-enriching ability in trans-CN-HAK1 tobaccos, compared with other counterparts. Finally, this work provides a method for screening new varieties of marker-free and safe transgenic antiviral tobacco.

scholarly journals Quantitative Real-Time PCR Analysis of Individual Flue-Cured Tobacco Seeds and Seedlings Reveals Seed Transmission of Tobacco Mosaic Virus

2020 ◽  
Vol 110 (1) ◽  
pp. 194-205 ◽  
Author(s):  
Madeleine D. Ellis ◽  
Jessica M. Hoak ◽  
Bradley W. Ellis ◽  
Jessica A. Brown ◽  
Tim L. Sit ◽  
...  
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Seed Coat ◽  
Rna Virus ◽  
Seed Transmission ◽  
Pcr Analysis ◽  
Infected Seed ◽  
Tobacco Seed ◽  
Individual Seed ◽  
Tobacco Seeds

Tobacco mosaic virus (TMV) is an extensively studied RNA virus known to infect tobacco (Nicotiana tabacum) and other solanaceous crops. TMV has been classified as a seedborne virus in tobacco, with infection of developing seedlings thought to occur from contact with the TMV-infected seed coat. The mechanism of TMV transmission through seed was studied in seed of the K 326 cultivar of flue-cured tobacco. Cross pollinations were performed to determine the effect of parental tissue on TMV infection in seed. Dissection of individual tobacco seeds into seed coat, endosperm, and embryo was performed to determine TMV location within a seed, while germination tests and separation of the developing seedling into seed coat, roots, and cotyledons were conducted to estimate the percent transmission of TMV. A reverse-transcriptase quantitative PCR (RT-qPCR) assay was developed and used to determine TMV concentrations in individual seed harvested from pods that formed on plants from TMV-infected and noninfected crosses. The results showed maternal transmission of TMV to tobacco seed and seedlings that developed from infected seed, not paternal transmission. RT-qPCR and endpoint PCR assays were also conducted on the separated seed coat, endosperm, and embryo of individual seed and separated cotyledons, roots, and seed coats of individual seedlings that developed from infected tobacco seed to identify the location of the virus in the seed and the subsequent path the virus takes to infect the developing seedling. RT-qPCR and endpoint PCR assay results showed evidence of TMV infection in the endosperm and embryo, as well as in the developing seedling roots and cotyledons within 10 days of initiating seed germination. To our knowledge, this is the first report of TMV being detected in embryos of tobacco seed, demonstrating that TMV is seedborne and seed-transmitted in flue-cured tobacco.

Resistance Mechanism of Cultured Plant Cells to Tobacco Mosaic Virus.

1985 ◽  
Vol 114 (2) ◽  
pp. 126-133 ◽  
Author(s):  
H. Toyoda ◽  
Y. Oishi ◽  
Y. Matsuda ◽  
K. Chatani ◽  
T. Hirai
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Resistance Mechanism ◽  
Plant Cells

scholarly journals Proteomic and Phosphoproteomic Analysis in Tobacco Mosaic Virus-Infected Tobacco (Nicotiana tabacum)

Biomolecules ◽  
2019 ◽  
Vol 9 (2) ◽  
pp. 39 ◽  
Author(s):  
Zi-Shu Lu ◽  
Qian-Si Chen ◽  
Qing-Xia Zheng ◽  
Juan-Juan Shen ◽  
Zhao-Peng Luo ◽  
...  
Keyword(s):  
Tobacco Mosaic Virus ◽  
Protein Phosphorylation ◽  
Mosaic Virus ◽  
Metal Ion ◽  
Common Source ◽  
Plant Responses ◽  
Phosphorylation Sites ◽  
Post Translational Modification ◽  
Biological Stress ◽  
Infected Tobacco

Tobacco mosaic virus (TMV) is a common source of biological stress that significantly affects plant growth and development. It is also useful as a model in studies designed to clarify the mechanisms involved in plant viral disease. Plant responses to abiotic stress were recently reported to be regulated by complex mechanisms at the post-translational modification (PTM) level. Protein phosphorylation is one of the most widespread and major PTMs in organisms. Using immobilized metal ion affinity chromatography (IMAC) enrichment, high-pH C18 chromatography fraction, and high-accuracy mass spectrometry (MS), a set of proteins and phosphopeptides in both TMV-infected tobacco and control tobacco were identified. A total of 4905 proteins and 3998 phosphopeptides with 3063 phosphorylation sites were identified. These 3998 phosphopeptides were assigned to 1311 phosphoproteins, as some proteins carried multiple phosphorylation sites. Among them, 530 proteins and 337 phosphopeptides corresponding to 277 phosphoproteins differed between the two groups. There were 43 upregulated phosphoproteins, including phosphoglycerate kinase, pyruvate phosphate dikinase, protein phosphatase 2C, and serine/threonine protein kinase. To the best of our knowledge, this is the first phosphoproteomic analysis of leaves from a tobacco cultivar, K326. The results of this study advance our understanding of tobacco development and TMV action at the protein phosphorylation level.

Quantitative Measurements on the Ion Etching of TMV

1973 ◽  
Vol 31 ◽  
pp. 336-337
Author(s):  
Irwin Bendet ◽  
Nabil Rizk
Keyword(s):  
Electron Microscope ◽  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Ion Current ◽  
Beam Diameter ◽  
Ion Etching ◽  
Preliminary Results ◽  
Quantitative Measurements ◽  
Monitoring Devices

Preliminary results reported last year on the ion etching of tobacco mosaic virus indicated that the diameter of the virus decreased more rapidly at 10KV than at 5KV, perhaps reaching a constant value before disappearing completely.In order to follow the effects of ion etching on TMV more quantitatively we have designed and built a second apparatus (Fig. 1), which incorporates monitoring devices for measuring ion current and vacuum as well as accelerating voltage. In addition, the beam diameter has been increased to approximately 1 cm., so that ten electron microscope grids can be exposed to the beam simultaneously.

Tobacco Mosaic Virus-Infected Tissue

1985 ◽  
Vol 43 ◽  
pp. 510-511
Author(s):  
Egbert W. Henry
Keyword(s):  
Nicotiana Tabacum ◽  
Tobacco Mosaic Virus ◽  
Chlorogenic Acid ◽  
Mosaic Virus ◽  
Host Resistance ◽  
Oxidative Metabolism ◽  
Leaf Tissue ◽  
Vein Clearing ◽  
Basal Septum ◽  
Concentration Levels

Tobacco mosaic virus (TMV) infection has been studied in several investigations of Nicotiana tabacum leaf tissue. Earlier studies have suggested that TMV infection does not have precise infective selectivity vs. specific types of tissues. Also, such tissue conditions as vein banding, vein clearing, liquification and suberization may result from causes other than direct TMV infection. At the present time, it is thought that the plasmodesmata, ectodesmata and perhaps the plasmodesmata of the basal septum may represent the actual or more precise sites of TMV infection.TMV infection has been implicated in elevated levels of oxidative metabolism; also, TMV infection may have a major role in host resistance vs. concentration levels of phenolic-type enzymes. Therefore, enzymes such as polyphenol oxidase, peroxidase and phenylalamine ammonia-lyase may show an increase in activity in response to TMV infection. It has been reported that TMV infection may cause a decrease in o-dihydric phenols (chlorogenic acid) in some tissues.

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