Complementary functions of two recessive R-genes determine resistance durability of tobacco ‘Virgin A Mutant’ (VAM) to Potato virus Y

AbstractTobaccos VAM and NC745 carry the recessive va gene that confers resistance to PVYNN. However, they exhibit different levels of resistance durability. Upon virus inoculation, only NC745 developed sporadic systemic symptoms caused by emerging resistance-breaking variants that easily infected both NC745 and VAM genotypes. To identify the differential host conditions associated with this phenomenon, cellular accumulation, cell-to-cell movement, vascular translocation, and foliar content of PVYNN were comparatively evaluated. Virus cell-to-cell movement was restricted and its transit through the vasculature boundaries was completely blocked in both tobacco varieties. However, an additional defense mechanism operating only in tobacco VAM drastically reduced the in situ cellular virus accumulation. Genetic analyses of hybrid plant progenies indicate that VAM-type resistance was conditioned by at least two recessive genes: va and a newly reported va2 locus. Moreover, segregant plant progenies that restricted virus movement but permitted normal virus accumulation were prone to develop resistance-breaking infections

Independent modulation of individual genomic component transcription and a cis-acting element related to high transcriptional activity in a multipartite DNA virus

Background: The genome of Banana bunchy top virus (BBTV) consists of at least six circular, single-stranded DNA components of 1kb in length. Some BBTV isolates may also carry satellite DNA molecules that are not essential for BBTV infection. The relation between multipartite DNA virus replication and their transcriptional levels and the underlying mechanism remain unclear. Results: To understand the coordinated replication and transcription of the multiple genomic components, the absolute amounts of each BBTV DNA component were measured by real-time PCR (qPCR), and their transcriptional levels were determined by RNAseq and reverse transcription-qPCR (qRT-PCR). Significant differences were found in the absolute amounts of individual BBTV genomic components. Transcriptional levels of each BBTV genomic component obtained from the RNAseq data matched closely to those obtained from qRT-PCR, but did not correspond to the absolute amount of each DNA component. The ratio of transcript over DNA copies ranged from 46.21 to 1059.44%, which was possibly regulated by the promoter region in the intergenic region of each component. To further determine this speculation, the promoter region of the DNA-S, -M or -N was constructed to the upstream of green fluorescent protein (GFP) gene for transient expression by agrobacterium-mediated transformation method. The qRT-PCR showed the highest transcriptional activity was promoted by DNA-N promoter, about 386.58% activity comparing with CaMV 35S promoter. Confocal microscopy observation showed that the intensity of green fluorescence was corresponding to that of qRT-PCR. Conclusions: Our data clearly showed that BBTV was able to control the transcriptional level of each DNA component independently by through the promoter sequences in the intergenic region. Moreover, a cis-acting element from DNA-N component had a high transcriptional activity.National Natural Science Foundation of China [31401709]; Hainan Provincial Natural Science Foundation [20153130]; Central Public-interest Scientific Institution Basal Research Fund for Chinese Academy of Tropical Agricultural Sciences [19CXTD-33]; Young Elite Scientists Sponsorship Program, CSTC [CSTC-QN201704]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Persistent Infection and Promiscuous Recombination of Multiple Genotypes of an RNA Virus within a Single Host Generate Extensive Diversity

Recombination and reassortment of viral genomes are major processes contributing to the creation of new, emerging viruses. These processes are especially significant in long-term persistent infections where multiple viral genotypes co-replicate in a single host, generating abundant genotypic variants, some of which may possess novel host-colonizing and pathogenicity traits. In some plants, successive vegetative propagation of infected tissues and introduction of new genotypes of a virus by vector transmission allows for viral populations to increase in complexity for hundreds of years allowing co-replication and subsequent recombination of the multiple viral genotypes. Using a resequencing microarray, we examined a persistent infection by a Citrus tristeza virus (CTV) complex in citrus, a vegetatively propagated, globally important fruit crop, and found that the complex comprised three major and a number of minor genotypes. Subsequent deep sequencing analysis of the viral population confirmed the presence of the three major CTV genotypes and, in addition, revealed that the minor genotypes consisted of an extraordinarily large number of genetic variants generated by promiscuous recombination between the major genotypes. Further analysis provided evidence that some of the recombinants underwent subsequent divergence, further increasing the genotypic complexity. These data demonstrate that persistent infection of multiple viral genotypes within a host organism is sufficient to drive the large-scale production of viral genetic variants that may evolve into new and emerging viruses

Three discontinuous loop nucleotides in the 3′ terminal stem-loop are required for Red clover necrotic mosaic virus RNA-2 replication

AbstractThe genome of Red clover necrotic mosaic virus (RCNMV) consists of positive-sense, single-stranded RNA-1 and RNA-2. The 29 nucleotides at the 3′ termini of both RNAs are nearly identical and are predicted to form a stable stem-loop (SL) structure, which is required for RCNMV RNA replication. Here we performed a systematic mutagenesis of the RNA-2 3′ SL to identify the nucleotides critical for replication. Infectivity and RNA replication assays indicated that the secondary structure of the 3′ SL and its loop sequence UAUAA were required for RNA replication. Single-nucleotide substitution analyses of the loop further pinpointed three discontinuous nucleotides (L1U, L2A, and L4A) that were vital for RNA replication. A 3-D model of the 3′ SL predicted the existence of a pocket formed by these three nucleotides that could be involved in RNA–protein interaction. The functional groups of the bases participating in this interaction at these positions are discussed

Relationship of Viroids to Macrophylla Decline

A physiological characterization has established that vascular changes in Macrophylla decline affected trees are not similar in character to xyloporosis affected trees. In addition, a survey of Macrophylla decline affected citrus did not establish any genetic similarity between Macrophylla decline and xyloporosis. We report diagnosis of either CCV or CEV by reverse transcription-polymerase chain reaction (RT-PCR), as well as diagnosis of Macrophylla decline or xyloporosis by Zn-distribution, water conductivity, accumulation of decline- specific proteins and examination of phloem morphology in lemon trees on the Macrophylla rootstock.Citrus Research Counci

Cotton Leaf Curl Virus, A Threat to Arizona Cotton?

A serious virus disease of cotton in Pakistan is distantly related to cotton leaf crumple in Arizona. It is much more destructive on cotton than leaf crumple, and has never been found in the western hemisphere. Cotton leaf crumple in Arizona causes only modestly damaging midseason infections, while leaf curl, has had a major impact on the crop in Pakistan. Modern transportation and the increasing movement of living plants in global trade has resulted in them recent introduction of a similar disease of another crop to the western hemisphere

Diseases of Citrus in Arizona

Revised10 pp

Chaos caused by a topologically mixing map

SIGLEITItal