Production and evaluation of transgenic sugarcane containing a Fiji disease virus (FDV) genome segment S9-derived synthetic resistance gene

2004 ◽  
Vol 55 (2) ◽  
pp. 139 ◽  
Author(s):  
R. B. McQualter ◽  
J. L. Dale ◽  
R. M. Harding ◽  
J. A. McMahon ◽  
G. R. Smith
Keyword(s):  
Resistance Gene ◽  
Disease Virus ◽  
Transgenic Line ◽  
Resistance Mechanism ◽  
Transcriptional Gene Silencing ◽  
Sugarcane Cultivar ◽  
Dna And Rna ◽  
Transgenic Sugarcane ◽  
Enhanced Resistance ◽  
Fiji Disease Virus

A transgenic line of the sugarcane cultivar Q124 with significantly enhanced resistance to Fiji disease was produced by microprojectile-mediated transformation with a transgene encoding a translatable version of Fiji disease virus (FDV) segment 9 ORF 1 under the control of the maize polyubiquitin promoter. Sixty-four transgenic lines were tested in glasshouse trials by caging the plants with viruliferous Perkinsiella saccharicida planthoppers. After 2 weeks, the planthoppers were removed and the plants monitored for symptoms. One transgenic line showed significantly enhanced resistance to Fiji disease compared with the Q124 parent and other lines showed varying levels of resistance. The molecular phenotypes of the transgenic plants at both the DNA and RNA levels were not entirely consistent with a resistance mechanism based on post-transcriptional gene silencing but were consistent with reports from other sugarcane-virus resistance systems. This is the first report of transgenic sugarcane containing an FDV-derived synthetic resistance gene showing resistance to FDV, although the mechanism of resistance has not yet been elucidated.

Improved Agrobacterium-mediated genetic transformation of GNA transgenic sugarcane

Biologia ◽  
2007 ◽  
Vol 62 (4) ◽  
Author(s):  
Dongting Zhangsun ◽  
Sulan Luo ◽  
Rukai Chen ◽  
Kexuan Tang
Keyword(s):  
Resistance Gene ◽  
Medium Composition ◽  
Marker Genes ◽  
35S Promoter ◽  
Selective Marker ◽  
Sugarcane Cultivar ◽  
Transgenic Sugarcane ◽  
Woolly Aphid ◽  
Ceratovacuna Lanigera ◽  
Screenable Marker

AbstractSix plasmids carrying a snowdrop lectin (Galanthus nivalis agglutinin, GNA) and one of three selection markers were successfully transferred into two sugarcane cultivars (FN81–745 and Badila) via Agrobacterium-mediated transformation. Agrobacterium strains LBA4404, EHA105 and A281 that harboured a super-binary vector were used for sugarcane transformation. The use of the hygromycin (Hyg) resistance gene (hpt II), phosphinothrincin (PPT) resistance gene (bar) or G418 resistance gene (npt II) as a screenable marker facilitated the initial selection of GNA transgenic sugarcane callus with different efficiencies and helped the rapid segregation of individual transformation events. All the three selective marker genes were controlled by CaMV 35S promoter, while GNA gene was controlled by promoter of RSs-1 (rice sucrose synthase-1) or Ubi (maize ubiquitin). Factors important to successful transformation mediated by Agrobacterium tumefaciens were optimized, which included concentration of A. tumefaciens, medium composition, co-cultivated methods with plant tissue, strain virulence and different selective marker genes. An efficient protocol for sugarcane transformation mediated by A. tumefaciens was established. The GNA gene has been integrated into sugarcane genome as demonstrated by PCR and Southern dot blotting detections. The preliminary results from bioassay demonstrated a significant resistance of the transgenic sugarcane plants to woolly aphid (Ceratovacuna lanigera Zehnther) indicating thus the possibility for obtaining a transgenic sugarcane cultivar with resistance to woolly aphid.

Molecular Analysis of Fiji Disease Virus Segments 2, 4 and 7 Completes the Genome Sequence

Virus Genes ◽  
2006 ◽  
Vol 32 (1) ◽  
pp. 43-47 ◽  
Author(s):  
Robert M. Harding ◽  
Parichart Burns ◽  
Robert J. Geijskes ◽  
Richard M. McQualter ◽  
James L. Dale ◽  
...  
Keyword(s):  
Disease Virus ◽  
Molecular Analysis ◽  
Genome Sequence ◽  
Fiji Disease Virus

Characterization and Expression Analysis of Resistance Gene Analogues in Elite Sugarcane Genotypes

2021 ◽  
Vol 28 ◽  
Author(s):  
Aqsa Parvaiz ◽  
Ghulam Mustafa ◽  
Muhammad Sarwar Khan ◽  
Muhammad Amjad Ali
Keyword(s):  
Phylogenetic Analysis ◽  
Disease Resistance ◽  
Resistance Gene ◽  
Critical Role ◽  
Colletotrichum Falcatum ◽  
Resistance Response ◽  
Sequence Characterization ◽  
Dna And Rna ◽  
Disease Resistant ◽  
Cellular Dna

Background: Resistance Gene Analogues (RGAs) are an important source of disease resistance in crop plants and have been extensively studies for their identification, tagging and mapping of Quantitative Trait Loci (QTLs). Tracking these RGAs in sugarcane can be of great help for the selection and screening of disease resistant clones. Objective: In the present study expression of different Resistance Gene Analogues (RGAs) was assessed in indigenous elite sugarcane genotypes which include resistant, highly resistant, susceptible and highly susceptible to disease infestation. Methods: Total cellular DNA and RNA were isolated from fourteen indigenous elite sugarcane genotypes. PCR, semi-quantitative RT PCR and real time qPCR analyses were performed. The resultant amplicons were sequence characterized, chromosomal localization and phylogenetic analysis were performed. Result: All of the 15 RGA primers resulted in amplification of single or multiple fragments from genomic DNA whereas only five RGA primers resulted in amplification from cDNA. Sequence characterization of amplified fragments revealed 86-99% similarity with disease resistance proteins indicating their potential role in disease resistance response. Phylogenetic analysis also validated these findings. Further, expression of RGA-012, RGA-087, RGA-118, RGA-533 and RGA-542 appeared to be upregulated and down regulated in disease resistant and susceptible genotypes, respectively, after inoculation with Colletotrichum falcatum. Conclusion: RGAs are present in most of our indigenous genotypes. Anyhow, differential expression of five RGAs indicated that they have some critical role in disease resistance. So, the retrieved results can not only be employed to devise molecular markers for the screening of disease resistant genotypes but can also be used to develop disease resistant plants through transgenic technology.

scholarly journals High-resolution mapping of Rym14Hb, a wild relative resistance gene to barley yellow mosaic disease

2020 ◽  
Author(s):  
Hélène Pidon ◽  
Neele Wendler ◽  
Antje Habekuβ ◽  
Anja Maasberg ◽  
Brigitte Ruge-Wehling ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Resistance Gene ◽  
Resistance Mechanism ◽  
Mosaic Disease ◽  
Yellow Mosaic Virus ◽  
Linkage Drag ◽  
Resistance Locus ◽  
Wild Relative ◽  
Yellow Mosaic Disease ◽  
Barley Breeding

Abstract Key message We mapped the Rym14Hb resistance locus to barley yellow mosaic disease in a 2Mbp interval. The co-segregating markers will be instrumental for marker-assisted selection in barley breeding. Abstract Barley yellow mosaic disease is caused by Barley yellow mosaic virus and Barley mild mosaic virus and leads to severe yield losses in barley (Hordeum vulgare) in Central Europe and East-Asia. Several resistance loci are used in barley breeding. However, cases of resistance-breaking viral strains are known, raising concerns about the durability of those genes. Rym14Hb is a dominant major resistance gene on chromosome 6HS, originating from barley’s secondary genepool wild relative Hordeum bulbosum. As such, the resistance mechanism may represent a case of non-host resistance, which could enhance its durability. A susceptible barley variety and a resistant H. bulbosum introgression line were crossed to produce a large F2 mapping population (n = 7500), to compensate for a ten-fold reduction in recombination rate compared to intraspecific barley crosses. After high-throughput genotyping, the Rym14Hb locus was assigned to a 2Mbp telomeric interval on chromosome 6HS. The co-segregating markers developed in this study can be used for marker-assisted introgression of this locus into barley elite germplasm with a minimum of linkage drag.

Improving Fiji disease resistance screening trials in sugarcane by considering virus transmission class and possible origin of Fiji disease virus

2004 ◽  
Vol 55 (6) ◽  
pp. 665 ◽  
Author(s):  
Grant R. Smith ◽  
Judith M. Candy
Keyword(s):  
Disease Virus ◽  
Disease Incidence ◽  
Virus Transmission ◽  
Rating System ◽  
Plant Host ◽  
Plant Infection ◽  
Disease Resistance Screening ◽  
Disease Rating ◽  
Fiji Disease Virus ◽  
Persistently Transmitted Viruses

Fiji disease virus is a propagative, persistently transmitted virus that multiplies in species of the delphacid planthopper genus Perkinsiella, and in sugarcane, the feeding host of the insect. Efforts to improve and modify the disease rating system for Fiji disease have largely focussed on the planthopper as individual vectors of the virus, rather than as a population of the principal, or at least an alternative, host of the virus. This perspective has resulted in key parameters of disease incidence resulting from plant infection by propagative, persistently transmitted viruses being largely overlooked or misunderstood during efforts to improve the rating system. These parameters include the relatively long acquisition, latency, and transmission times, the percentage of the population containing virus, or viruliferous, in the above periods, and the effects of population density and number of plants visited on disease incidence. Suggestions to modify trial design to improve virus transmission to the plant, based on the disease incidence parameters of the propagative, persistent transmission class, are presented and the practical difficulties of implementing these proposals are discussed. In the context of fully understanding the underlying biology of this virus–insect–plant system, the hypothesis that Fiji disease virus, as a plant-infecting member of the Reoviridae, is primarily an insect virus with a secondary plant host, and may have diverged from an insect-infecting virus relatively recently is proposed and compared with other members of the family Reoviridae.

scholarly journals RamA, a transcriptional regulator conferring florfenicol resistance in Leclercia adecarboxylata R25

2020 ◽  
Vol 65 (6) ◽  
pp. 1051-1060
Author(s):  
Cong Cheng ◽  
Yuanyuan Ying ◽  
Danying Zhou ◽  
Licheng Zhu ◽  
Junwan Lu ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Efflux Pump ◽  
Gene Knockout ◽  
Regulatory Gene ◽  
Resistance Mechanism ◽  
Agricultural Practice ◽  
The Novel ◽  
Inappropriate Use ◽  
Rnd Efflux Pump ◽  
High Level

AbstractDue to the inappropriate use of florfenicol in agricultural practice, florfenicol resistance has become increasingly serious. In this work, we studied the novel florfenicol resistance mechanism of an animal-derived Leclercia adecarboxylata strain R25 with high-level florfenicol resistance. A random genomic DNA library was constructed to screen the novel florfenicol resistance gene. Gene cloning, gene knockout, and complementation combined with the minimum inhibitory concentration (MIC) detection were conducted to determine the function of the resistance-related gene. Sequencing and bioinformatics methods were applied to analyze the structure of the resistance gene-related sequences. Finally, we obtained a regulatory gene of an RND (resistance-nodulation-cell division) system, ramA, that confers resistance to florfenicol and other antibiotics. The ramA-deleted variant (LA-R25ΔramA) decreased the level of resistance against florfenicol and several other antibiotics, while a ramA-complemented strain (pUCP24-prom-ramA/LA-R25ΔramA) restored the drug resistance. The whole-genome sequencing revealed that there were five RND efflux pump genes (mdtABC, acrAB, acrD, acrEF, and acrAB-like) encoded over the chromosome, and ramA located upstream of the acrAB-like genes. The results of this work suggest that ramA confers resistance to florfenicol and other structurally unrelated antibiotics, presumably by regulating the RND efflux pump genes in L. adecarboxylata R25.

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