scholarly journals Distinguishing resistances of transgenic sugarcane generated from RNA interference and pathogen‐derived resistance approaches to combating sugarcane mosaic virus

2021 ◽  
Vol 26 (2) ◽  
pp. 107
Author(s):  
Weny Nailul Hidayati ◽  
Retnosari Apriasti ◽  
Hardian Susilo Addy ◽  
Bambang Sugiharto
Keyword(s):  
Rna Interference ◽  
Mosaic Virus ◽  
Plant Viruses ◽  
Sugarcane Mosaic Virus ◽  
Pcr Analysis ◽  
Post Inoculation ◽  
Gene Encoding ◽  
Transgenic Sugarcane ◽  
Transgenic Lines ◽  
Pathogen Derived Resistance

Sugarcane mosaic virus (SCMV) is a causative agent that reduces growth and productivity in sugarcane. Pathogen‐derived resistance (PDR) and RNA interference (RNAi) are the most common approaches to generating resis‐ tance against plant viruses. Two types of transgenic sugarcane have been obtained by PDR and RNAi methods using a gene‐encoding coat protein (CP) of SCMV (SCMVCp). This research aimed to distinguish resistance of the two transgenic sugarcanes in combating SCMV through artificial viral inoculation. The experiment was conducted using transgenic sugar‐ cane lines validated by PCR analysis. Insertion of gene‐encoding CP in the transgenic lines was confirmed by amplification of 702 bp of DNA fragment of SCMVCp. After viral inoculation, mosaic symptoms appeared earlier, at 21 days post inoculation (dpi) in PDR transgenic lines, but was at 26 dpi in RNAi transgenic lines. Symptom observation showed that 77.8% and 50% of the inoculated plants developed mosaic symptoms in PDR and RNAi transgenic lines, respectively. RT‐PCR analysis revealed that the nuclear inclusion protein b (Nib) gene of SCMV was amplified in the symptomatic leaves in plants classified as susceptible lines. Immunoblot analysis confirmed presence of viral CP with a molecular size of 37 kDa in the susceptible lines. Collectively, these results indicated that the RNAi approach targeting the gene for CP effectively produces more resistance against the SCMV infection in transgenic sugarcane compared to the PDR approach.

Full sequence of the coat protein gene is required for the induction of pathogen-derived resistance against sugarcane mosaic virus in transgenic sugarcane

2018 ◽  
Vol 45 (6) ◽  
pp. 2749-2758 ◽  
Author(s):  
Retnosari Apriasti ◽  
Suvia Widyaningrum ◽  
Weny N. Hidayati ◽  
Widhi D. Sawitri ◽  
Nurmalasari Darsono ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Sugarcane Mosaic Virus ◽  
Full Sequence ◽  
Transgenic Sugarcane ◽  
Pathogen Derived Resistance

scholarly journals Transgenic Sugarcane Resistant toSorghum mosaic virusBased on Coat Protein Gene Silencing by RNA Interference

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Jinlong Guo ◽  
Shiwu Gao ◽  
Qinliang Lin ◽  
Hengbo Wang ◽  
Youxiong Que ◽  
...  
Keyword(s):  
Rna Interference ◽  
Coat Protein ◽  
Mosaic Virus ◽  
Resistance Rate ◽  
Mosaic Disease ◽  
Artificial Inoculation ◽  
Multiple Sequence ◽  
Sugarcane Cultivar ◽  
Resistant Lines ◽  
Transgenic Lines

As one of the critical diseases of sugarcane, sugarcane mosaic disease can lead to serious decline in stalk yield and sucrose content. It is mainly caused byPotyvirus sugarcane mosaic virus(SCMV) and/orSorghum mosaic virus(SrMV), with additional differences in viral strains. RNA interference (RNAi) is a novel strategy for producing viral resistant plants. In this study, based on multiple sequence alignment conducted on genomic sequences of different strains and isolates of SrMV, the conserved region of coat protein (CP) genes was selected as the target gene and the interference sequence with size of 423 bp in length was obtained through PCR amplification. The RNAi vector pGII00-HACP with an expression cassette containing both hairpin interference sequence andcp4-epspsherbicide-tolerant gene was transferred to sugarcane cultivar ROC22 viaAgrobacterium-mediated transformation. After herbicide screening, PCR molecular identification, and artificial inoculation challenge, anti-SrMV positive transgenic lines were successfully obtained. SrMV resistance rate of the transgenic lines with the interference sequence was 87.5% based on SrMV challenge by artificial inoculation. The genetically modified SrMV-resistant lines of cultivar ROC22 provide resistant germplasm for breeding lines and can also serve as resistant lines having the same genetic background for study of resistance mechanisms.

Gene-for-gene interactions are required for disease resistance mediated by virus transgene

2002 ◽  
Vol 38 (SI 1 - 6th Conf EFPP 2002) ◽  
pp. 177-179
Author(s):  
M. Ravelonandro
Keyword(s):  
Disease Resistance ◽  
Rna Interference ◽  
Molecular Mechanisms ◽  
Plant Viruses ◽  
Economic Losses ◽  
Gene Interactions ◽  
Severe Damage ◽  
Breeding Programs ◽  
Pathogen Derived Resistance ◽  
Potential Use

Plant viruses cause severe damage and significant economic losses to agriculture. Control of virus usually consist of<br />the elimination of virus vectors (insects, nematodes, fungi, etc), improvement of the sanitary status of the propagation<br />material, the use of resistance sources in breeding programs. The application of the pathogen-derived resistance strategy<br />has opened new avenues to protect plants against viruses. Two molecular mechanisms seem to underlie the engineered<br />protection, the virus transgene-derived protein and the transgene-RNA interference. A few examples that support the<br />efficiencies of these two molecular mechanisms are reviewed here and discussed in light of the potential use of virusresistant<br />transgenic plants in agriculture.

scholarly journals Noncoding RNAs of Plant Viruses and Viroids: Sponges of Host Translation and RNA Interference Machinery

2016 ◽  
Vol 29 (3) ◽  
pp. 156-164 ◽  
Author(s):  
W. Allen Miller ◽  
Ruizhong Shen ◽  
William Staplin ◽  
Pulkit Kanodia
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Mosaic Virus ◽  
Barley Yellow Dwarf Virus ◽  
Plant Viruses ◽  
Cauliflower Mosaic Virus ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Noncoding Sequences ◽  
Yellow Dwarf Virus

Noncoding sequences in plant viral genomes are well-known to control viral replication and gene expression in cis. However, plant viral and viroid noncoding (nc)RNA sequences can also regulate gene expression acting in trans, often acting like ‘sponges’ that bind and sequester host cellular machinery to favor viral infection. Noncoding sequences of small subgenomic (sg)RNAs of Barley yellow dwarf virus (BYDV) and Red clover necrotic mosaic virus (RCNMV) contain a cap-independent translation element that binds translation initiation factor eIF4G. We provide new evidence that a sgRNA of BYDV can globally attenuate host translation, probably by sponging eIF4G. Subgenomic ncRNA of RCNMV is generated via 5′ to 3′ degradation by a host exonuclease. The similar noncoding subgenomic flavivirus (sf)RNA, inhibits the innate immune response, enhancing viral pathogenesis. Cauliflower mosaic virus transcribes massive amounts of a 600-nt ncRNA, which is processed into small RNAs that overwhelm the host’s RNA interference (RNAi) system. Viroids use the host RNAi machinery to generate viroid-derived ncRNAs that inhibit expression of host defense genes by mimicking a microRNA. More examples of plant viral and viroid ncRNAs are likely to be discovered, revealing fascinating new weaponry in the host-virus arms race.

scholarly journals Molecular cloning, expression and characterization of Bmserpin-2 gene from Bombyx mori.

2009 ◽  
Vol 56 (4) ◽  
Author(s):  
Ye Pan ◽  
Hengchuan Xia ◽  
Peng Lü ◽  
KePing Chen ◽  
Qin Yao ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Developmental Stages ◽  
Pcr Analysis ◽  
Post Inoculation ◽  
Amino Acid Residues ◽  
Gene Encoding ◽  
Reading Frame ◽  
E Coli ◽  
Real Time Quantitative Pcr ◽  
Localization Analysis

Serpins are a broadly distributed family of protease inhibitors. In this study, the gene encoding Bombyx mori serpin-2 (Bmserpin-2) was cloned and expressed in E. coli. The Bmserpin-2 cDNA contains a 1125 bp open reading frame (ORF). The deduced protein has 374 amino-acid residues, contains a conserved SERPIN domain and shares extensive homology with other invertebrate serpins. RT-PCR analysis showed that Bmserpin-2 was expressed in all developmental stages of B. mori larvae and various larval tissues. Subcellular localization analysis indicated that Bmserpin-2 protein was located in the cytoplasm. Interestingly, real-time quantitative PCR revealed that the expression of Bmserpin-2 in the midgut of susceptible B. mori strain 306 significantly increased at 72 hours post inoculation (hpi) when infected with BmNPV. However, there was no significant increase of the Bmserpin-2 expression in resistant strain NB infected with BmNPV. Thus, our data indicates that Bmserpin-2 may be involved in B. mori antiviral response.

scholarly journals Field Performance of Transgenic Sugarcane Lines Resistant to Sugarcane Mosaic Virus

2017 ◽  
Vol 8 ◽  
Author(s):  
Wei Yao ◽  
Miaohong Ruan ◽  
Lifang Qin ◽  
Chuanyu Yang ◽  
Rukai Chen ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Field Performance ◽  
Sugarcane Mosaic Virus ◽  
Transgenic Sugarcane

A virus-derived short hairpin RNA confers resistance against sugarcane mosaic virus in transgenic sugarcane

2018 ◽  
Vol 27 (2) ◽  
pp. 203-210 ◽  
Author(s):  
Usman Aslam ◽  
Bushra Tabassum ◽  
Idrees Ahmad Nasir ◽  
Anwar Khan ◽  
Tayyab Husnain
Keyword(s):  
Mosaic Virus ◽  
Short Hairpin Rna ◽  
Sugarcane Mosaic Virus ◽  
Hairpin Rna ◽  
Transgenic Sugarcane ◽  
Short Hairpin

scholarly journals Expression and purification of recombinant coat protein of sugarcane mosaic virus from Indonesian isolate as an antigen for antibody production

2019 ◽  
Vol 24 (1) ◽  
pp. 57
Author(s):  
Natalia Tri Astuti ◽  
Nurmalasari Darsono ◽  
Suvia Widyaningrum ◽  
Widhi Dyah Sawitri ◽  
Sri Puji Astuti ◽  
...  
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Polyclonal Antibodies ◽  
Molecular Study ◽  
Insoluble Fraction ◽  
Sugarcane Mosaic Virus ◽  
Gene Encoding ◽  
Iptg Induction ◽  
Sds Page ◽  
Saccharum Sp

Sugarcane mosaic virus (SCMV, genus Potyvirus, family Potyviridae) is a prominent pathogen of sugarcane (Saccharum sp. hybrids). It can cause losses in susceptible varieties, in crop as well as sugar production, economically. Although it has been studied in major sugar-producing countries, research on the definement of SCMV from Indonesian isolates based on molecular study has been very limited. This study aimed to obtain a proper recombinant antigens emanating from coat protein of SCMV from Indonesian isolate in order to produce polyclonal antibodies that cann be used for immunodiagnosis assays in a subsequent study. A gene-encoding coat protein of SCMV (CP-SCMV) was amplified using RT-PCR and cloned into vector pJET1.2. The cDNA was inserted into 6X His-tag expression plasmid of pET28a(+) and over-expressed in Escherichia coli BL21(DE3) to produce a recombinant protein. The highest expression was found in 0.1M IPTG induction media for 5 h at 37oC. SDS-PAGE analysis clarified that the recombinant CP-SCMV remained as an insoluble fraction. Purifications was carried out by the affinity Ni-NTA resin, followed by electroelution to obtain a highly purified protein. To meet the quality requirements of a proper antigen, the highly purified protein was concentrated. A molecular weight of the rCP-SCMV (approximately 40 kDa) was clearly observed by 10% SDS-PAGE at the concentration of 16.184 mg/mL. 

scholarly journals First Report of Sugarcane Mosaic Virus in Zea mays L. in Ukraine

2021 ◽  
Vol 83 (5) ◽  
pp. 58-66
Author(s):  
H. Snihur ◽  
◽  
A. Kharina ◽  
M. Kaliuzhna ◽  
V. Chumak ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mosaic Virus ◽  
Rhopalosiphum Padi ◽  
Plant Viruses ◽  
Causal Agent ◽  
Sugarcane Mosaic Virus ◽  
Maize Dwarf Mosaic Virus ◽  
Maize Crop ◽  
First Report ◽  
Maize Plants

Maize viral diseases especially maize dwarf mosaic disease (MDMD), which is caused by potyviruses, lead to significant crop losses worldwide. Aim. The aim of this work was to identify the causal agent of mosaic symptoms, observed on maize plants during 2018—2020 in Kyiv region. Methods. Enzyme-linked immunosorbent assay in the DAS-ELISA modification using commercial Loewe Biochemica test systems for Maize dwarf mosaic virus (MDMV), Sugarcane mosaic virus (SCMV), Wheat streak mosaic virus (WSMV) were applied to identify the causal agent of maize disease in collected samples. Transmission electron microscopy was used in order to direct viral particle visualisation. Aphids, which are natural vectors of plant viruses, were found on diseased plants. Results. Plants with typical mosaic symptoms were observed in corn crops of the Kyiv region in early June 2018. The pathogen was transmitted by mechanical inoculation to maize and sweet maize plants with the manifestation of mosaic symptoms. Electron microscopy of the sap from diseased plants revealed the presence of flexible filamentous virions 750 nm long and 13 nm in diameter, typical for the genus Potyvirus. In August, mosaic symptoms and aphids Rhopalosiphum padi were found on previously healthy plants in the same maize crop. In 2020, in the same sown area, maize plants were free of viral infection during inspection in June, but a re-inspection in September revealed mosaic symptoms on maize crop and the presence of aphids in the leaf axils. The presence of SCMV in maize samples collected in June and August/September 2018 and 2020, as well as in inoculated maize and sweet maize plants, was confirmed by ELISA using a commercial test system. The obtained data allow suggesting that Rhopalosiphum padi is a natural vector of SCMV in agrocenoses of Ukraine. It should be noted that co-infection with MDMV and WSMV in the affected plants was not detected. Conclusions. This study presents the first report of SCMV in maize in Ukraine.

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