Virus-infected Plants Research Articles

Combinatorial interactions between viral proteins expand the potential functional landscape of the tomato yellow leaf curl virus proteome.

Viruses manipulate the cells they infect in order to replicate and spread. Due to strict size restrictions, viral genomes have reduced genetic space; how the action of the limited number of viral proteins results in the cell reprogramming observed during the infection is a long-standing question. Here, we explore the hypothesis that combinatorial interactions may expand the functional landscape of the viral proteome. We show that the proteins encoded by a plant-infecting DNA virus, the geminivirus tomato yellow leaf curl virus (TYLCV), physically associate with one another in an intricate network, as detected by a number of protein-protein interaction techniques. Importantly, our results indicate that intra-viral protein-protein interactions can modify the subcellular localization of the proteins involved. Using one particular pairwise interaction, that between the virus-encoded C2 and CP proteins, as proof-of-concept, we demonstrate that the combination of viral proteins leads to novel transcriptional effects on the host cell. Taken together, our results underscore the importance of studying viral protein function in the context of the infection. We propose a model in which viral proteins might have evolved to extensively interact with other elements within the viral proteome, enlarging the potential functional landscape available to the pathogen.

Ocimum basilicum-Mediated Synthesis of Silver Nanoparticles Induces Innate Immune Responses against Cucumber Mosaic Virus in Squash.

Cucumber mosaic virus (CMV) causes a significant threat to crop output sustainability and human nutrition worldwide, since it is one of the most prevalent plant viruses infecting most kinds of plants. Nowadays, different types of nanomaterials are applied as a control agent against different phytopathogens. However, their effects against viral infections are still limited. In the current study, the antiviral activities of the biosynthesized silver nanoparticles (Ag-NPs) mediated by aqueous extract of Ocimum basilicum against cucumber mosaic virus in squash (Cucurbita pepo L.) were investigated. The prepared Ag-NPs were characterized using scanning electron microscopy (SEM), dynamic light scattering (DLS), transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR) and zeta potential distribution techniques. DLS, SEM, and TEM analyses showed that the Ag-NPs were spherical, with sizes ranging from 26.3 to 83 nm with an average particle size of about 32.6 nm. FTIR identified different functional groups responsible for the capping and stability of Ag-NPs. The zeta potential was reported as being −11.1 mV. Under greenhouse conditions, foliar sprays of Ag-NPs (100 µg/mL) promoted growth, delayed disease symptom development, and significantly reduced CMV accumulation levels of treated plants compared to non-treated plants. Treatment with Ag-NPs 24 h before or after CMV infection reduced CMV accumulation levels by 92% and 86%, respectively. There was also a significant increase in total soluble carbohydrates, free radical scavenging activity, antioxidant enzymes (PPO, SOD, and POX), as well as total phenolic and flavonoid content. Furthermore, systemic resistance was induced by significantly increasing the expression levels of pathogenesis-related genes (PR-1 and PR-5) and polyphenolic pathway genes (HCT and CHI). These findings suggest that Ag-NPs produced by O. basilicum could be used as an elicitor agent and as a control agent in the induction and management of plant viral infections.

Dynamic changes in virus-induced volatiles in cotton modulate the orientation and oviposition behavior of the whitefly Bemisia tabaci.

Manipulation of insect vector behavior by virus-induced plant volatiles is well known. But how the viral disease progression alters the plant volatiles and its effect on vector behavior remains less explored. Our studies tracked changes in volatile profile in progressive infection stages of cotton leaf curl virus (CLCuV) infected plants and their effect on B. tabaci behavior. Significant differences in virus titers were noticed between progressive infection stages showing distinct symptoms. Whiteflies initially settled on CLCuV infected plants, but their preference was shifted to healthy plants over time. GC-MS analysis revealed subtle quantitative/qualitative changes in volatile organic compounds (VOCs) between the healthy and selected CLCuV infection stages. VOCs such as hexanal, (E)-2-hexen-1-ol, (+)-α-pinene, (−)-β-pinene, (Z)-3-hexen-1-ol, (+)-sylvestrene, and (1S,2E,6E, 10R)-3,7,11,11-tetramethylbicycloundeca-2,6-diene (Bicyclogermacrene) were associated with the infection stage showing upward curling of leaves; (E)-2-hexen-1-ol, β-myrcene, β-ocimene, and copaene were associated with the infection stage showing downward curling. Validation studies with eight synthetic VOCs indicated that γ-terpinene elicited attraction to B. tabaci (Olfactometric Preference Index (OPI) = 1.65), while β-ocimene exhibited strong repellence (OPI = 0.64) and oviposition reduction (66.01%–92.55%). Our studies have demonstrated that progression of CLCuV disease in cotton was associated with dynamic changes in volatile profile which influences the behavioural responses of whitefly, B.tabaci. Results have shown that VOCs such as (+)-α-pinene, (−)-β-pinene γ-Terpinene, α-guaiene; 4- hydroxy- 4 methyl-2- pentanone and β-ocimene emitted from Begomovirus infected plants could be the driving force for early attraction and later repellence/oviposition deterrence of B. tabaci on virus-infected plants. The findings of this study offer scope for the management of whitefly, B. tabaci through semiochemicals.

The Streptomyces chromofuscus Strain RFS-23 Induces Systemic Resistance and Activates Plant Defense Responses against Tomato Yellow Leaf Curl Virus Infection

Insect-vectored plant viruses pose a serious threat to sustainable production of economically important crops worldwide. This demands a continuous search for environmentally-friendly, sustainable and efficient approaches based on biological agents to address the mounting challenges of viral disease management. To date, the efficacy of actinomycetes bacteria against DNA plant viruses remains unknown. Here, through comparative analyses, we demonstrate that the RFS-23 strain of Streptomyces cellulase possesses protective activity as it positively regulated the plant growth and development. and diminished the severity, of disease symptoms, together with reduced accumulation of Tomato yellow leaf curl virus (TYLCV) DNA. The RFS-23 strain maintained relative chlorophyll contents by promoting the expression of genes (CLH1, HEMA1 and PORA) associated with chlorophyll biogenesis. As compared to another strain, CTF-20, the RSF-23 induced a significantly higher expression of plant defense-related genes (NbCIS and NbNCED) associated with biogenesis and accumulation of salicylic acid and abscisic acid. Additionally, the activity of antioxidant enzymes (SOD, CAT, POD and MDA) was significantly enhanced by RSF-23 treatment, despite the presence of viral infection. These findings suggest that RSF-23 is a novel biocontrol agent with protective activity, and it could be a potential candidate for the management of plant viral infections.

Ecotype-specific blockage of tasiARF production by two different RNA viruses in Arabidopsis.

Arabidopsis thaliana is one of the most studied model organisms of plant biology with hundreds of geographical variants called ecotypes. One might expect that this enormous genetic variety could result in differential response to pathogens. Indeed, we observed previously that the Bur ecotype develops much more severe symptoms (upward curling leaves and wavy leaf margins) upon infection with two positive-strand RNA viruses of different families (turnip vein-clearing virus, TVCV, and turnip mosaic virus, TuMV). To find the genes potentially responsible for the ecotype-specific response, we performed a differential expression analysis of the mRNA and sRNA pools of TVCV and TuMV-infected Bur and Col plants along with the corresponding mock controls. We focused on the genes and sRNAs that showed an induced or reduced expression selectively in the Bur virus samples in both virus series. We found that the two ecotypes respond to the viral infection differently, yet both viruses selectively block the production of the TAS3-derived small RNA specimen called tasiARF only in the virus-infected Bur plants. The tasiARF normally forms a gradient through the adaxial and abaxial parts of the leaf (being more abundant in the adaxial part) and post-transcriptionally regulates ARF4, a major leaf polarity determinant in plants. The lack of tasiARF-mediated silencing could lead to an ectopically expressed ARF4 in the adaxial part of the leaf where the misregulation of auxin-dependent signaling would result in an irregular growth of the leaf blade manifesting as upward curling leaf and wavy leaf margin. QTL mapping using Recombinant Inbred Lines (RILs) suggests that the observed symptoms are the result of a multigenic interaction that allows the symptoms to develop only in the Bur ecotype. The particular nature of genetic differences leading to the ecotype-specific symptoms remains obscure and needs further study.

A balance between vector survival and virus transmission is achieved through JAK/STAT signaling inhibition by a plant virus

Viruses pose a great threat to animal and plant health worldwide, with many being dependent on insect vectors for transmission between hosts. While the virus-host arms race has been well established, how viruses and insect vectors adapt to each other remains poorly understood. Begomoviruses comprise the largest genus of plant-infecting DNA viruses and are exclusively transmitted by the whitefly Bemisia tabaci. Here, we show that the vector Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway plays an important role in mediating the adaptation between the begomovirus tomato yellow leaf curl virus (TYLCV) and whiteflies. We found that the JAK/STAT pathway in B. tabaci functions as an antiviral mechanism against TYLCV infection in whiteflies as evidenced by the increase in viral DNA and coat protein (CP) levels after inhibiting JAK/STAT signaling. Two STAT-activated effector genes, BtCD109-2 and BtCD109-3, mediate this anti-TYLCV activity. To counteract this vector immunity, TYLCV has evolved strategies that impair the whitefly JAK/STAT pathway. Infection of TYLCV is associated with a reduction of JAK/STAT pathway activity in whiteflies. Moreover, TYLCV CP binds to STAT and blocks its nuclear translocation, thus, abrogating the STAT-dependent transactivation of target genes. We further show that inhibition of the whitefly JAK/STAT pathway facilitates TYLCV transmission but reduces whitefly survival and fecundity, indicating that this JAK/STAT-dependent TYLCV-whitefly interaction plays an important role in keeping a balance between whitefly fitness and TYLCV transmission. This study reveals a mechanism of plant virus-insect vector coadaptation in relation to vector survival and virus transmission.

Characteristics of Virus Symptoms in Chili Plants (Capsicum frutescens) Based on RGB Image Analysis

Virus infection in chili plants may cause various symptoms. The complexity of the symptoms and human vision ability often become limiting factors during disease investigations. Digital image analysis is expected to become a method to assist in comprehensively describing the symptoms of plant viruses. A disease survey was conducted on cayenne pepper fields in Southeast Sulawesi Province to observe symptoms of virus infection virtually and to record the symptomatic plant using an RGB camera. The split-channel method is used to process images, followed by multidimensional scaling statistical analysis. Later on, viruses associated with plants were detected serologically. Single or mixed infection of Tobacco mosaic virus, Cucumber mosaic virus, Chili veinal mottle virus, and Pepper mottle virus was confirmed by plant leaves showing yellow-mosaic and mottle symptoms. The digital image analysis method could show variations in the characteristics of symptoms based on digital numbers in that cannot be recognized based on the observation of visual symptoms. A new approach to study the interactions between plant infecting viruses and their effects based on image analysis has also been developed during this research. This method needs to be further validated through testing under controlled conditions, such as inoculating plants with a predetermined type of virus.

Complete genome sequence of Edgeworthia chrysantha mosaic-associated virus, a tentative new member of the genus Coguvirus (family Phenuiviridae).

A new negative-strand RNA (nsRNA) virus genome was discovered in Edgeworthia chrysantha Lindl. This virus, tentatively named "Edgeworthia chrysantha mosaic-associated virus" (ECMaV), has a bipartite genome that comprises (i) a nsRNA1, encoding the viral RNA-dependent RNA polymerase (RdRp), and (ii) an ambisense RNA2, coding for the putative movement protein (MP) and nucleocapsid protein (NP), with the open reading frames separated by a long AU-rich intergenic region (IR). Sequence comparisons and phylogenetic analysis showed that the RdRp is closely related to those of other recently discovered plant-infecting nsRNA viruses in the new genus Coguvirus and that ECMaV can be classified as a member of a novel species.

Foliar Application of Nanoclay Promotes Potato (Solanum tuberosum L.) Growth and Induces Systemic Resistance against Potato Virus Y.

Potato virus Y (PVY) is one of the most harmful phytopathogens. It causes big problems for potatoes and other important crops around the world. Nanoclays have been extensively studied for various biomedical applications. However, reports on their interactions with phytopathogens, particularly viral infections, are still limited. In this study, the protective activity of Egyptian nanoclay (CE) and standard nanoclay (CS) against PVY was evaluated on potato (Solanum tuberosum L.) plants. Their physicochemical and morphological properties were examined with scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), and energy dispersive spectrometer (EDS). SEM and TEM analyses revealed that CE has a spherical and hexagonal structure ranging from 20 to 80 nm in size, while CS has boulder-like and tubular structures of about 320 nm in size. FTIR and EDS showed that both nanoclay types have different functional groups and contain many vital plant nutrients that are necessary for every stage and process of the plant, including development, productivity, and metabolism. Under greenhouse conditions, a 1% nanoclay foliar application enhanced potato growth, reduced disease symptoms, and reduced PVY accumulation levels compared with non-treated plants. Significant increases in levels of antioxidant enzymes (PPO and POX) and considerable decreases in oxidative stress markers (MDA and H2O2) were also reported. Moreover, a significant increase in the transcriptional levels of defense-related genes (PAL-1, PR-5, and CHI-2) was observed. All experiment and analysis results indicate that the CE type is more effective than the CS type against PVY infection. Based on these results, the foliar applications of nanoclay could be used to manage plant viral infections in a way that is both effective and environmentally friendly. To our knowledge, this is the first report of the antiviral activity of the foliar application of nanoclay against PVY infection.

The Ubiquitin-Proteasome System (UPS) and Viral Infection in Plants.

The ubiquitin–proteasome system (UPS) is crucial in maintaining cellular physiological balance. The UPS performs quality control and degrades proteins that have already fulfilled their regulatory purpose. The UPS is essential for cellular and organic homeostasis, and its functions regulate DNA repair, gene transcription, protein activation, and receptor trafficking. Besides that, the UPS protects cellular immunity and acts on the host’s defense system. In order to produce successful infections, viruses frequently need to manipulate the UPS to maintain the proper level of viral proteins and hijack defense mechanisms. This review highlights and updates the mechanisms and strategies used by plant viruses to subvert the defenses of their hosts. Proteins involved in these mechanisms are important clues for biotechnological approaches in viral resistance.

Incidence and Genetic Diversity of Grapevine Virus G in Croatian Vineyards.

Grapevine virus G (GVG) is a recently discovered vitivirus infecting grapevines. Historically, viruses in the genus Vitivirus have been associated with the grapevine rugose wood disease. Based on new and previously reported GVG isolates, primers and probes were developed for real-time RT-PCR. The developed assay successfully detected the virus in infected plants during dormancy and the growing season. A field study of 4327 grapevines from Croatian continental and coastal wine-growing regions confirmed the presence of GVG in 456 (~10.5%) grapevines from three collection plantations and 77 commercial vineyards, with infection rates ranging from 2% to 100%. Interestingly, the virus was confirmed only in vines considered to be Croatian autochthonous cultivars, but not in introduced cultivars. A 564-nucleotide long portion of the coat protein gene from previously known and newly characterized GVG isolates had nucleotide and amino acid identities ranging from 89% to 100% and from 96.8% to 100%, respectively. Phylogenetic analysis revealed five distinct groups, with isolates originating from the same site being close to each other, indicating possible local infection. The information presented in this manuscript sets the stage for future studies to better understand the ecology and epidemiology of GVG and the possible need for inclusion in certification schemes.

Chlorophyll Fluorescence Imaging (CFI) and Laser-Induced Breakdown Spectroscopy (LIBS) Applied to Investigate Tomato Plants Infected by the Root Knot Nematode (RKN) Meloidogyne incognita and Tobacco Plants Infected by Cymbidium Ringspot Virus

Recently, studies on climate change have highlighted the central role of photosynthetic mechanisms in the defense response of plants to abiotic and biotic stresses. Photo-sensing and photo-activation are innovative technologies applied for the early detection of plant pathogens in order to prevent the dramatic impact they may have on plants. Chlorophyll Fluorescence Imaging (CFI) and Laser-Induced Breakdown Spectroscopy (LIBS) analytical techniques can be used to evaluate the amount of chlorophyll in plants, which can be altered in the case of biotic and abiotic stresses. In this work, both techniques were applied to two pathogenic model systems, i.e., roots of susceptible tomato plants infected by Meloidogyne incognita and Nicotiana benthamiana plants infected by cymbidium ringspot virus. Experimental evidence is provided and discussed showing that specific application protocols of both methods can be used successfully for the early detection of symptoms of the pathogen attacks of Meloidogyne incognita on tomato roots and of cymbidium ringspot virus infected plants. In particular, a decrease in chlorophyll content was measured by fluorescence imaging, and an increase in Mg++ content was determined by LIBS in both the leaves and stems of infected tomato plants and the leaves of infected plants, with respect to control (non-infected) plants. Thus, the two techniques used have been shown to be able to discriminate satisfactorily between control and infected plants and to provide some insight on the underlying mechanisms of plant defenses again nematodes and viruses.

Turnip Mosaic Virus Infection Differentially Modifies Cabbage Aphid Probing Behavior in Spring and Winter Oilseed Rape (Brassica napus).

Simple SummaryWinter oilseed rape accounts for about 90% of total oilseed rape planting area in China, with the majority of it concentrated in the Yangtze River basin. The remaining 10% is mostly found in the provinces of China’s northwest plateau. Winter oilseed rape areas in China have gradually expanded to the north in the last decade, resulting in cabbage aphids and turnip mosaic virus (TuMV). The aphid is a vector for TuMV and is gradually increasing on winter and spring oilseed rape. Quantifying the probing behaviors of the aphids on spring oilseed rape and winter oilseed rape helps us to understand TuMV regulation of the aphids. We found that compared to mock-inoculated plants, cabbage aphids on infected plants increased brief probing frequency, cell penetration frequency, intracellular probing time, decreased time to first probe and pathway duration, potentially promoting viral acquisition and minimizing viral loss and plant damage. Viruliferous aphids had reduced pathway duration, increased cell penetration frequency, increased intracellular probing time, increased salivation frequency, and ingested less sap compared with non-viruliferous aphids, primed for viral infection. TuMV infection also differentially modified aphid feeding behavior on winter and spring oilseed rape cultivars, primarily on uninfected plants.Direct and indirect effects of plant virus infection on vector behavior have been discovered to improve virus transmission efficiency, but the impact of plant cultivars in virus–vector–plant interactions has received little attention. Electropenetrography (EPG) allows real-time tracking and quantification of stylet penetration behaviors, pathogen transmission, and plant resistance mechanisms. Quantitative probing behaviors on a spring oilseed rape cultivar, ‘Xinyou17’, and a winter oilseed rape cultivar, ‘Zheping4’, were investigated using EPG to compare turnip mosaic virus (TuMV) regulation of cabbage aphid probing behavior. Results for indirect effects showed that compared to mock-inoculated plants, cabbage aphids on infected plants increased brief probing frequency, cell penetration frequency, intracellular probing time, and decreased time to first probe and pathway time, potentially promoting viral acquisition. TuMV also directly influences aphid probing behavior. Viruliferous aphids had reduced pathway time, increased cell penetration frequency, increased intracellular probing time, increased salivation frequency, and ingested less sap than non-viruliferous aphids, primed for viral infection. Although oilseed rape cultivars can also influence aphid behavior, the main effect of cultivars was not significant on TuMV-infected plants.

Induction of Systemic Resistance to Tobacco mosaic virus in Tomato through Foliar Application of Bacillus amyloliquefaciens Strain TBorg1 Culture Filtrate.

The application of microbe-derived products as natural biocontrol agents to boost systemic disease resistance to virus infections in plants is a prospective strategy to make agriculture more sustainable and environmentally friendly. In the current study, the rhizobacterium Bacillus amyloliquefaciens strain TBorg1 was identified based on 16S rRNA, rpoB, and gyrA gene sequences, and evaluated for its efficiency in conferring protection of tomato from infection by Tobacco mosaic virus (TMV). Under greenhouse circumstances, foliar sprays of TBorg1 culture filtrate (TBorg1-CF) promoted tomato growth, lowered disease severity, and significantly decreased TMV accumulation in systemically infected leaves of treated plants relative to untreated controls. TMV accumulation was reduced by 90% following the dual treatment, applied 24 h before and after TMV infection. Significant increases in levels of total soluble carbohydrates, proteins, and ascorbic acid were also found. In addition, a significant rise in activities of enzymes capable of scavenging reactive oxygen species (PPO and POX), as well as decreased levels of non-enzymatic oxidative stress markers (H2O2 and MDA) were observed, compared to untreated plants. Enhanced systemic resistance to TMV was indicated by significantly increased transcript accumulation of polyphenolic pathway (C4H, HCT, and CHI) and pathogenesis-related (PR-1 and PR-5) genes. Out of the 15 compounds identified in the GC-MS analysis, 1,2-benzenedicarboxylic acid mono(2-ethylhexyl) ester and phenol, 2,4-bis(1,1-dimethylethyl), as well as L-proline, N-valeryl-, and heptadecyl ester were present in the highest concentrations in the ethyl acetate extract of TBorg1-CF. In addition, significant amounts of n-hexadecanoic acid, pyrrolo [1,2-a] pyrazine-1,4-dione hexahydro-3-(2-methylpropyl)-, nonane, 5-butyl-, and eicosane were also detected. These compounds may act as inducers of systemic resistance to viral infection. Our findings indicate that the newly isolated B. amyloliquefaciens strain TBorg1 could be a potentially useful rhizobacterium for promoting plant growth and a possible source of biocontrol agents for combating plant virus infections.

Molecular basis of mutual benefits between Cucurbit chlorotic yellows virus (CCYV) transmission and imidacloprid resistance in Bemisia tabaci

Increased insecticide resistance among insect vectors and arboviruses results in significant economic losses to agricultural production and poses a continuous threat to food safety. To date, many studies have been conducted on how virus-infected host plants indirectly and plant virus directly change the physiological characteristics of insect vectors. However, the relationship between the insecticide resistance of the insect vector and the virulence of the viral pathogen has not received sufficient attention. Here, we investigated the effect of Cucurbit chlorotic yellows virus (CCYV), a plant virus transmitted by whitefly (Bemisia tabaci) in a semi-persistent manner, on the resistance of B. tabaci to the neonicotinoid imidacloprid. We found CCYV enhanced the resistance of B. tabaci to imidacloprid. The CYP6CM1 gene was significantly up-regulated in viruliferous susceptible B. tabaci compared with non-viruliferous B. tabaci after an acquisition access period of 10 d, including its upstream regulators. Silencing the CYP6CM1 gene by RNA interference reduced the ability of B. tabaci to acquire CCYV compared with the control. Additionally, imidacloprid-resistant B. tabaci outperformed sensitive adults in terms of virus transmission ability. These results suggest that CCYV enhances the ability of imidacloprid-sensitive B. tabaci to resist adverse external factors (insecticides). In turn, B. tabaci resistance affects virus transmission. Thus, to a certain extent, the plant virus and insect vector establish a mutually beneficial relationship that facilitates virus transmission.

Phylogenetic And Recombination Analysis Of Yellow Mosaic Disease In Soybean Plant

Numerous viruses have been discovered in plants, both domestic and wild, in recent years. The biodiversity of plant viruses has been studied using a variety of methods, including virus-specific DNA or RNA, followed by sequencing and bioinformatics analyses. All the analysis reveals more knowledge about plant viruses. Phylogenetic analyses are increasingly being used in epidemiology, to understand the pattern of DNA/RNA virus infection in soybean plants, while the Recombination detection program is also used to analyse nucleotide sequence data and to identify the evidence of genetic recombination between the viruses infecting soybean.This study have found evidence that recombination is an important impelling cause in the evolution and divergence ofmany plant viruses, including begomoviruses, potyviruses and other viruses.

Eukaryotic translation initiation factor 4E family member nCBP facilitates the accumulation of TGB-encoding viruses by recognizing the viral coat protein in potato and tobacco.

Due to their limited coding capacity, plant viruses have to depend on various host factors for successful infection of the host. Loss of function of these host factors will result in recessively inherited resistance, and therefore, these host factors are also described as susceptibility genes or recessive resistance genes. Most of the identified recessive resistance genes are members of the eukaryotic translation initiation factors 4E family (eIF4E) and its isoforms. Recently, an eIF4E-type gene, novel cap-binding protein (nCBP), was reported to be associated with the infection of several viruses encoding triple gene block proteins (TGBps) in Arabidopsis. Here, we, for the first time, report that the knockdown of nCBP in potato (StnCBP) compromises the accumulation of potato virus S (PVS) but not that of potato virus M (PVM) and potato virus X (PVX), which are three potato viruses encoding TGBps. Further assays demonstrated that StnCBP interacts with the coat proteins (CPs) of PVS and PVM but not with that of PVX, and substitution of PVS CP in the PVS infectious clone by PVM CP recovered the virus infection in StnCBP-silenced transgenic plants, suggesting that the recognition of PVS CP is crucial for StnCBP-mediated recessive resistance to PVS. Moreover, the knockdown of nCBP in Nicotiana benthamiana (NbnCBP) by virus-induced gene silencing suppressed PVX accumulation but not PVM, while NbnCBP interacted with the CPs of both PVX and PVM. Our results indicate that the nCBP orthologues in potato and tobacco have conserved function as in Arabidopsis in terms of recessive resistance against TGB-encoding viruses, and the interaction between nCBP and the CP of TGB-encoding virus is necessary but not sufficient to determine the function of nCBP as a susceptibility gene.

Enrichment Book Design Based on Laboratory Results for Viruses on Plants Material

This study aims to determine the design of the development of an enrichment book based on laboratory results of infection in plants. The subtopics presented are infection in plants and detection of infected plants by using isozymes. The initial stage that is carried out is that researchers conduct research in the field and in the laboratory to obtain information in writing the manuscript or the content of the material to be presented in the enrichment book. The development of the enrichment book refers to the book development procedure by Sitepu (2012), are script writing, illustration, editing, cover design, and printing. The design for the development of the enrichment book is arranged in several parts, are the cover page, the initial page, the content section, the evaluation page, and the final page. The development of the enrichment book will be tested on high school students who have obtained infection material on plants. The development of this enrichment book is based on the lack of utilization of research results as teaching materials in schools. Therefore, this biological enrichment book was developed based on the results of research conducted, namely disease incidence (di), disease severity (ds), and isozyme characterization of virus-infected chili plants. The development of this enrichment book is expected to increase students' insight, knowledge, and curiosity and be used as a learning resource to understand the concept of plant infection material widely.

Impact of Watermelon bud necrosis virus (WBNV) infected plants on the volatile emission pattern in cowpea plants

Pathogens, including tospoviruses, are known to manipulate the behaviour of vectors after virus acquisition by plants to enhance virus transmission. Furthermore, as recently proven in the maize chlorotic mottle virus pathosystem, the vector's choice for virus-infected plants can change to a preference for noninfected plants after virus uptake by the vector. A similar trend was observed in the cowpea - Watermelon Bud Necrosis Virus (WBNV) - Thrips palmi (Karny) pathosystem. Similarly, in the no-choice bioassay, viruliferous T.palmi (carrying WBNV) settled preferentially more on healthy cowpea plants (56%) compared to virus-infected plants (47.3%), whereas non-viruliferous T.palmi settled preferentially more on WBNV infected (58.67%) cowpea plants compared to healthy plants (44%). The changes in preference of thrips towards host plants before and after virus acquisition may be due to the change of volatile cues. This study looked at the headspace volatile composition of healthy and WBNV-infected cowpea plants that attract thrips. Furthermore, the volatile analysis revealed that 1, 2-Propanediamine (0.62%) and Tuaminoheptane (0.55%) from healthy cowpea plants, as well as Tetradecane (0.35%) from WBNV-infected cowpea plants, both have a higher area percent than other volatiles. The amine (53%) and hydrocarbon (69%) groups of volatile organic compounds make up the majority of host volatiles found in healthy and virus-infected plants. The increased contact rates of viruliferous and non-viruliferous T.palmi towards healthy and WBNV-infected host plants could enhance virus transmission if thrips feed on them and acquire the pathogen prior to dispersal and the recorded host volatiles might be useful in vector management in future.

Homeobox 27, a Homeodomain Transcription Factor, Confers Tolerances to CMV by Associating with Cucumber Mosaic Virus 2b Protein.

Transcription factors (TFs) play an important role in plant development; however, their role during viral infection largely remains unknown. The present study was designed to uncover the role transcription factors play in Cucumber mosaic virus (CMV) infection. During the screening of an Arabidopsis thaliana (Col-0) transcription factor library, using the CMV 2b protein as bait in the yeast two-hybrid system, the 2b protein interacted with Homeobox protein 27 (HB27). HB27 belongs to the zinc finger homeodomain family and is known to have a regulatory role in flower development, and responses to biotic and abiotic stress. The interaction between CMV 2b and HB27 proteins was further validated using in planta (bimolecular fluorescence complementation assay) and in vitro far-Western blotting (FWB) methods. In the bimolecular fluorescence complementation assay, these proteins reconstituted YFP fluorescence in the nucleus and the cytoplasmic region as small fluorescent dots. In FWB, positive interaction was detected using bait anti-MYC antibody on the target HB27-HA protein. During CMV infection, upregulation (~3-fold) of the HB27 transcript was observed at 14 days post-infection (dpi) in A. thaliana plants, and expression declined to the same as healthy plants at 21 dpi. To understand the role of the HB27 protein during CMV infection, virus accumulation was determined in HB27-overexpressing (HB27 OE) and knockout mutants. In HB27-overexpressing lines, infected plants developed mild symptoms, accumulating a lower virus titer at 21 dpi compared to wild-type plants. Additionally, knockout HB27 mutants had more severe symptoms and a higher viral accumulation than wild-type plants. These results indicate that HB27 plays an important role in the regulation of plant defense against plant virus infection.