Special Issue "Molecular and Biotechnological Approaches for Characterization and Control of Plant Virus Disease"

A special issue of Agronomy (ISSN 2073-4395).

Deadline for manuscript submissions: closed (10 November 2017)

Special Issue Editor

Guest Editor
Prof. Dr. Jozef J. Bujarski

Plant Molecular Biology Center and the Department of Biological Sciences, Northern Illinois University, DeKalb, IL 60115, USA
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Phone: +1-815-753-0601

Special Issue Information

Dear Colleagues,

Viral diseases have a deep impact on the productivity of agriculture plants, affecting the crop yield and quality. Such human activities as the extensive crop management, monocultures in large fields, extensive breeding programs, the use of GMOs, but also climate changes—all enhance the effect of viral diseases on global agriculture. It is then essential to foster new ways of control of plant virus impacts.

The advances of recent years have progressed with the understanding of mechanisms of viral diseases. Methods of introducing anti-viral resistance have been developed, either through plant breeding or via genetic engineering in corn, soybean, cotton, cassava and other harvests. However, many crop plants are still susceptible to viral diseases while high mutation rates create new variants that enable plant viruses to surpass the resistance barriers.

The topic of this Special Issue will be “Molecular and Biotechnological Approaches for Characterization and Control of Plant Virus Disease”. We accept research articles, reviews and opinions, concerning novel resistance genes, plant–virus interactions, interference with vector transmission, management and sustainable control, transgenic resistance/biotechnological strategies, model systems, detection and diagnosis, or new casual agents/emerging viruses.

Prof. Dr. Jozef J. Bujarski
Guest Editor

Manuscript Submission Information

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Keywords

  • crop-virus interactions
  • antiviral control
  • crop biotechnology
  • virus diagnosis
  • new plant viruses
  • plant molecular virology

Published Papers (4 papers)

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Research

Open AccessArticle
Production of a Polyclonal Antibody against the Recombinant Coat Protein of the Sugarcane Mosaic Virus and Its Application in the Immunodiagnostic of Sugarcane
Received: 3 May 2018 / Revised: 3 June 2018 / Accepted: 11 June 2018 / Published: 13 June 2018
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Abstract
Sugarcane mosaic virus (SCMV) is a mosaic disease that has spread over sugarcane plantations in Indonesia. The important step to overcome the disease is to detect the pathogen as early as possible. Detection of the pathogen can be achieved using the immunodiagnostic method [...] Read more.
Sugarcane mosaic virus (SCMV) is a mosaic disease that has spread over sugarcane plantations in Indonesia. The important step to overcome the disease is to detect the pathogen as early as possible. Detection of the pathogen can be achieved using the immunodiagnostic method by employing a specific antibody against the viral coat protein. The objective of this research was to produce a polyclonal antibody using the recombinant coat protein of SCMV, and to test its sensitivity for detection of SCMV in the symptomatic plant. The gene encoding of the coat protein was cloned using the RT-PCR Kit and total RNA isolated from symptomatic sugarcane leaves cultivar PS-881. Nucleotide sequences analysis of the cloned cDNA indicated that the cDNA contained 998 nucleotides and named SCMVCp-cDNA. The cDNA was then inserted into a His-tag expression plasmid of pET28a and overexpressed in Escherichia coli BL21 (DE3) to produce a recombinant protein. The recombinant fused protein SCMVCp was strongly expressed in an insoluble fraction, with a molecular size of around 44 kDa, without the addition of an IPTG inducer. Purification of the recombinant protein using an affinity Ni-NTA resin, followed by SDS-PAGE separation, resulted in a high purity of the protein and used as an antigen to raise the polyclonal antibody in a rabbit. The sensitivity of the antiserum determined by western blot analysis showed that the antiserum was able to detect the recombinant protein at a concentration of 10 ng. The western blot analysis also detected a clear single band of 36.7 kDa of the SCMV coat protein in symptomatic sugarcane leaves and not in healthy leaves. Interestingly, when the sample proteins were prepared using low-speed centrifugation, the corresponding coat protein was detected in a soluble fraction by western blot analysis. Thus, the antiserum was successfully used for indirect-ELISA analysis using the soluble protein fraction. The results provide an easy method to detect and diagnose SCMV infection using the immunodiagnostic. Full article
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Open AccessArticle
Molecular Cloning and Structure–Function Analysis of a Trypsin Inhibitor from Tartary Buckwheat and Its Application in Combating Phytopathogenic Fungi
Received: 10 January 2018 / Revised: 8 April 2018 / Accepted: 10 April 2018 / Published: 12 April 2018
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Abstract
Host plant protease inhibitors offer resistance to proteases from invading pathogens. Trypsin inhibitors (TIs), in particular, serve as protective agents against insect and pathogen attacks. In this study, we designed a pair of degenerate primers based on highly conserved motifs at the N- [...] Read more.
Host plant protease inhibitors offer resistance to proteases from invading pathogens. Trypsin inhibitors (TIs), in particular, serve as protective agents against insect and pathogen attacks. In this study, we designed a pair of degenerate primers based on highly conserved motifs at the N- and C-termini of the TI from tartary buckwheat (Fagopyrum tataricum; Ft) to clone the central portion. Genomic walking was performed to isolate the 5′ and 3′ flanking regions of FtTI. We demonstrated the successful PCR amplification of a 644 bp portion of FtTI. The full-length DNA of FtTI contains a complete open reading frame of 264 bp, encoding 87 amino acids with a mass of approximately 9.5 kDa. The FtTI protein sequence was 49% identical and 67% similar to potato protease inhibitors. Site-directed mutagenesis identified the residues, Asp67 and Arg68, as crucial for the inhibitory activity of the FtTI. Recombinant and mutant FtTI inhibited both the hyphal growth and spore germination of Alternaria solani. The calculated 50% inhibitory concentrations of FtTI ranged from 5–100 μg mL−1 for spore germination and 1–50 μg mL−1 for fungal growth. Thus, recombinant FtTI may function in host resistance against a variety of fungal plant pathogens. Full article
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Open AccessArticle
Different Approaches to Produce Transgenic Virus B Resistant Chrysanthemum
Received: 6 February 2018 / Revised: 25 February 2018 / Accepted: 2 March 2018 / Published: 8 March 2018
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Abstract
Chrysanthemum is a vegetative propagated culture in which viral transmission with planting material is important for its production. Chrysanthemum virus B (CVB) belongs to the viruses that strike this plant culture. Chrysanthemum virus B is found everywhere where chrysanthemum is cultivated. Damage to [...] Read more.
Chrysanthemum is a vegetative propagated culture in which viral transmission with planting material is important for its production. Chrysanthemum virus B (CVB) belongs to the viruses that strike this plant culture. Chrysanthemum virus B is found everywhere where chrysanthemum is cultivated. Damage to plants by CVB often leads to a complete loss of floral yield. Chrysanthemum (Chrysanthemum morifolium Ramat cv. White Snowdon) was transformed via Agrobacterium-mediated DNA delivery with the aim of improving resistance to CVB infection. Transformation vectors contain the nucleotide sequence of CVB coat proteins (CP) in sense, antisense, and double sense orientation. The transformative vectors also invert repeats of CVB coat protein gene fragments for the induction of RNA-interference. The transgenic chrysanthemum plants were successfully obtained. The integration of the target sequences in plant genomes was confirmed by polymerase chain reaction (PCR) and Southern blot analyses. Chrysanthemum lines were transformed with antisense, sense, and double sense CVB CP sequences, as well as with hairpin RNA-interference constructs that were assayed for resistance to CVB. Infection of transgenic plants by CVB through the grafting of infected scions shows resistance only among plants with carried double sense (16.7%) and hairpin (12.5%) constructs. The plants transformed by sense and double sense sequences were observed and classified as tolerant. Full article
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Open AccessArticle
Generation of Transgenic Rootstock Plum ((Prunus pumila L. × P. salicina Lindl.) × (P. cerasifera Ehrh.)) Using Hairpin-RNA Construct for Resistance to the Plum pox virus
Received: 9 November 2017 / Revised: 4 December 2017 / Accepted: 15 December 2017 / Published: 22 December 2017
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Abstract
The use of Prunus rootstocks that are resistant to plum pox virus (PPV) is an important agronomic strategy to combat the spread of the Sharka disease in nurseries and orchards. Despite remarkable progress in developing stone fruit rootstocks to adapt to various stresses, [...] Read more.
The use of Prunus rootstocks that are resistant to plum pox virus (PPV) is an important agronomic strategy to combat the spread of the Sharka disease in nurseries and orchards. Despite remarkable progress in developing stone fruit rootstocks to adapt to various stresses, breeding that ensures durable virus resistance has not yet been achieved. For this reason, the engineering of PPV resistant plants through genetic transformation is a very promising approach to control sharka disease. The aim of the present study is to produce transgenic plants of the clonal rootstock ‘Elita’, which is resistant to PPV using ribonucleic acid interference (RNAi) technology. The genetic construct containing the self-complementary fragments of the plum pox virus coat protein (PPV-CP) gene sequence were used to induce the mechanism of post-transcriptional gene silencing to ensure virus resistance. Transgenic plants have been produced after agrobacterium-mediated transformation of in vitro explanted leaves. The results of polymerase chain reaction (PCR) and Southern blotting analyses confirmed the stable genomic integration of the PPV-CP sense and antisense intron-hairpin-RNA sequence. The functionality of the introduced expression cassette was confirmed by the activity of including the uidA gene into the transferring T-DNA. To our knowledge, this is the first interspecific plum rootstock produced by genetic engineering to achieve PPV resistance. Full article
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