Golden 2-like transcription factor contributes to the major QTL against rice black-streaked dwarf virus disease.

KEY MESSAGE: A major resistance QTL was identified on chromosome 6 in rice variety Wuke; both overexpression and knockdown experiments confirmed that OsGLK1 is the candidate gene for association with Rice black-streaked dwarf virus disease. Rice black-streaked dwarf virus disease is one of the most destructive rice viral diseases in China and East Asia. Progress has been limited in RBSDVD resistance breeding due to inadequate knowledge on the underlying functional genes. In this study, a major QTL for RBSDV (rice black-streaked dwarf virus) independent of SBPH (small brown planthopper) resistance was mapped in a 1.8 Mb interval on chromosome 6 by using an F2:3 population originated from resistant rice variety Wuke. Representative transcripts within this region were analysed and three genes showing amino acid sequence variation in functional domains were selected for transformation. Overexpression experiments showed that one gene exhibited significant enhanced resistance compared to control lines, encoding protein involving Myb domain and probable transcription factor Golden 2-like1 (GLK1). Furthermore, OsGLK1 knockdown rice lines were investigated and the resistance ability was significantly declined without this gene compared to the wild type. Taken together, both overexpression and knockdown experiments strongly suggested that OsGLK1 plays an important role for RBSDV resistance and contributes to the major QTL. The study paves the way for elucidating the molecular mechanism underlying RBSDVD resistance and the molecular markers associated with OsGLK1 may be used for marker-assisted selection.

Melatonin mitigates traumatic brain injury-induced depression-like behaviors through HO-1/CREB signal in rats.

In addition to significant antioxidant properties, melatonin exhibits neuroprotective effects against various neurological diseases including traumatic brain injury (TBI) and ischemic stroke. Several potential mechanisms have been reported in the neuroprotection of melatonin among patients with TBI. Notably, the heme oxygenase-1 (HO-1)/cAMP response element-binding protein (CREB) signaling pathway is implicated in the development of a depressive state. Moreover, the activity of CREB in the nucleus accumbens (NAc) participates in reward and motivation, further contributing to depression induced by TBI. This study aims to explore whether melatonin could mitigate TBI-induced depression by activating of HO-1/CREB signal in a rodent model of weight-drop. As a consequence, melatonin (10 mg/kg) attenuated TBI-induced elevated immobility time in the force swim test, decreased time spent sniffing the novel rat in 3-chambered social test, and downregulated phosphorylated CERB in the NAc. However, a special inhibitor of HO-1 (SnPP) via intracerebroventricular injection partially reversed the neuroprotective effects of melatonin. Furthermore, melatonin decreased the number of summarized intersects in the astrocyte, A1-type astrocytes, IL-6-positive astrocytes in the NAc after TBI exposure, nevertheless, these changes could partially be restored by SnPP. Therefore, our findings demonstrate a novel neuroprotective mechanism for melatonin against TBI which can be a potential neuroprotective agent for the treatment of TBI-induced depression.

Melatonin alleviates traumatic brain injury-induced anxiety-like behaviors in rats: Roles of the protein kinase A/cAMP-response element binding signaling pathway.

Melatonin is a hormone produced by the pineal gland. Given its capabilities of neuroprotection and low neurotoxicity, melatonin could be a therapeutic strategy for traumatic brain injury (TBI). The present study was conducted to determine the neuroprotective effects of melatonin on TBI-induced anxiety and the possible molecular mechanism. Rats were randomly divided into seven groups. The rodent model of TBI was established using the weight-drop method. Melatonin was administered by intraperitoneal injection at a dose of 10 mg/kg after TBI. H89 (0.02 mg/kg), a special protein kinase A (PKA) inhibitor, or dibutyryl-cyclic adenosine monophosphate (cAMP; 0.1 mg/kg), an activator of PKA, were administered by stereotactic injection of the brain to evaluate the roles of PKA and cAMP-response element-binding protein (CREB) in melatonin-related mood regulation, respectively. At 30 days post-TBI, the changes in anxiety-like behaviors in rats were measured using the open field and elevated plus maze tests. At 24 h post-TBI, the number of activated astrocytes and neuronal apoptosis were evaluated using immunofluorescence assay. The expression levels of inflammatory cytokines (TNF-α and IL-6) in the amygdala were measured using an enzyme-linked immunosorbent assay. The expression levels of PKA, phosphorylated (p)-PKA, CREB, p-CREB, NF-κB and p-NF-κB in the amygdala were detected using western blotting. It was revealed that melatonin partially reversed TBI-induced anxiety-like behavior in rats, and decreased the number of activated astrocytes and neuronal apoptosis in the amygdala induced by TBI. H89 partially blocked the neuroprotective effects of melatonin; while dibutyryl-cAMP not only reduced the H89-induced emotional disturbance but also enhanced the protective effects of melatonin against TBI. Overall, melatonin can alleviate TBI-induced anxiety-like behaviors in rats. Moreover, the underlying mechanism may be associated with the activation of the PKA/CREB signaling pathway.

An aspartic protease 47 causes quantitative recessive resistance to rice black-streaked dwarf virus disease and southern rice black-streaked dwarf virus disease.

Rice black-streaked dwarf virus disease (RBSDVD) and southern rice black-streaked dwarf virus disease (SRBSDVD) are the most destructive viral diseases in rice. Progress is limited in breeding due to lack of resistance resource and inadequate knowledge on the underlying functional gene. Using genome-wide association study (GWAS), linkage disequilibrium (LD) decay analyses, RNA-sequencing, and genome editing, we identified a highly RBSDVD-resistant variety and its first functional gene. A highly RBSDVD-resistant variety W44 was identified through extensive evaluation of a diverse international rice panel. Seventeen quantitative trait loci (QTLs) were identified among which qRBSDV6-1 had the largest phenotypic effect. It was finely mapped to a 0.8-1.2 Mb region on chromosome 6, with 62 annotated genes. Analysis of the candidate genes underlying qRBSDV6-1 showed high expression of aspartic proteinase 47 (OsAP47) in a susceptible variety, W122, and a low resistance variety, W44. OsAP47 overexpressing lines exhibited significantly reduced resistance, while the knockout mutants exhibited significantly reduced SRBSDVD and RBSDVD severity. Furthermore, the resistant allele Hap1 of OsAP47 is almost exclusive to Indica, but rare in Japonica. Results suggest that OsAP47 knockout by editing is effective for improving RBSDVD and SRBSDVD resistance. This study provides genetic information for breeding resistant cultivars.

ARGONAUTE 2 increases rice susceptibility to rice black-streaked dwarf virus infection by epigenetically regulating HEXOKINASE 1 expression.

ARGONAUTE (AGO) proteins play crucial roles in plant defence against virus invasion. To date, the role of OsAGO2 in rice antiviral defence remains largely unknown. In this study, we determined that the expression of OsAGO2 in rice was induced upon rice black-streaked dwarf virus (RBSDV) infection. Using transgenic rice plants overexpressing OsAGO2 and Osago2 mutants generated through transposon-insertion or CRISPR/Cas9 technology, we found that overexpression of OsAGO2 enhanced rice susceptibility to RBSDV infection. Osago2 mutant lines exhibited strong resistance to RBSDV infection through the elicitation of an early defence response, including reprogramming defence gene expression and production of reactive oxygen species (ROS). Compared to Nipponbare control, the expression level of OsHXK1 (HEXOKINASE 1) increased significantly, and the methylation levels of its promoter decreased in the Osago2 mutant on RBSDV infection. The expression profile of OsHXK1 was the opposite to that of OsAGO2 during RBSDV infection. Overexpression of OsHXK1 in rice also induced ROS production and enhanced rice resistance to RBSDV infection. These results indicate that OsHXK1 controls ROS accumulation and is regulated by OsAGO2 through epigenetic regulation. It is noteworthy that the Osago2 mutant plants are also resistant to southern rice black-streaked dwarf virus infection, another member of the genus Fijivirus. Based on the results presented in this paper, we conclude that OsAGO2 modulates rice susceptibility to fijivirus infection by suppressing OsHXK1 expression, leading to the onset of ROS-mediated resistance. This discovery may benefit future rice breeding programmes for virus resistance.

Androgen is responsible for enhanced susceptibility of melatonin against traumatic brain injury in females.

BACKGROUND: Numerous publications have demonstrated that melatonin administration is associated with mortality reduction and improvement in neurological outcomes after traumatic brain injury (TBI). However, there are significant sex differences in several diseases associated with melatonin. We aimed to determine whether androgen was responsible for enhanced susceptibility of melatonin against TBI in females, as well as potential molecular mechanisms. METHODS: Weight-drop was used to establish a rodent model of TBI. Melatonin (10 mg/kg) and testosterone (1 mg/kg) were administered three times every day for three days after TBI using subcutaneous injection, respectively. Seven days after TBI, an open field assay was used to evaluate locomotor and exploratory activities. Neuronal amount, neuronal apoptosis, and expression of phosphorylated extracellularly regulated protein kinases 1/2 (ERK1/2), c-jun N-terminal kinase 1/2 (JNK1/2), and p38 mitogen-activated protein kinase (p38MAPK) in neurons were assessed using immunofluorescence assay seven days after TBI. The expression of caspase-3, Bax, and Bcl-2 in the frontal cortex was detected using western blot. RESULTS: Compared with female rats, melatonin administration exhibited more neuroprotective effects (including improved locomotor and exploratory activities, elevated neuronal amount, and reduced neuronal apoptosis) in male rats exposed to TBI. Moreover, testosterone significantly improved locomotor and exploratory activities, elevated neuronal amount, decreased neuronal apoptosis, downregulated phosphorylation of JNK1/2- and p38MAPK-positive neurons, but upregulated phosphorylation of ERK1/2-positive neurons in the frontal cortex, and reduced the expressions of cleaved caspase-3, Bax, but increased Bcl-2 expressions in female rats exposed to TBI. CONCLUSIONS: Androgen was responsible for the enhanced susceptibility to TBI under melatonin supplementation in females through a mechanism that may be associated with MAPK pathway regulation.

Tomato Yellow Leaf Curl Virus V2 Protein Plays a Critical Role in the Nuclear Export of V1 Protein and Viral Systemic Infection.

Geminiviruses are an important group of circular, single-stranded DNA viruses that cause devastating diseases in crops. Geminiviruses replicate their genomic DNA in the nucleus and the newly synthesized viral DNA is subsequently transported to the cytoplasm for further cell-to-cell and long-distance movement to establish systemic infection. Thus, nucleocytoplasmic transportation is crucial for successful infection by geminiviruses. For Tomato yellow leaf curl virus (TYLCV), the V1 protein is known to bind and shuttle viral genomic DNA, however, the role of the V2 protein in this process is still unclear. Here, we report that the V1 protein is primarily localized in the nucleus when expressed but the nucleus-localized V1 protein dramatically decreases when co-expressed with V2 protein. Moreover, the V2-facilitated nuclear export of V1 protein depends on host exportin-α and a specific V1-V2 interaction. Chemical inhibition of exportin-α or a substitution at cysteine 85 of the V2 protein, which abolishes the V1-V2 interaction, blocks redistribution of the V1 protein to the perinuclear region and the cytoplasm. When the V2C85S mutation is incorporated into a TYLCV infectious clone, the TYLCV-C85S causes delayed onset of very mild symptoms compared to wild-type TYLCV, suggesting that the V1-V2 interaction and, thus, the V2-mediated nuclear export of the V1 protein is crucial for viral spread and systemic infection. Our data point to a critical role of the V2 protein in promoting the nuclear export of the V1 protein and viral systemic infection, likely by promoting V1 protein-mediated nucleocytoplasmic transportation of TYLCV genomic DNA.

R-DEHM: CSI-Based Robust Duration Estimation of Human Motion with WiFi.

As wireless sensing has developed, wireless behavior recognition has become a promising research area, in which human motion duration is one of the basic and significant parameters to measure human behavior. At present, however, there is no consideration of the duration estimation of human motion leveraging wireless signals. In this paper, we propose a novel system for robust duration estimation of human motion (R-DEHM) with WiFi in the area of interest. To achieve this, we first collect channel statement information (CSI) measurements on commodity WiFi devices and extract robust features from the CSI amplitude. Then, the back propagation neural network (BPNN) algorithm is introduced for detection by seeking a cutting line of the features for different states, i.e., moving human presence and absence. Instead of directly estimating the duration of human motion, we transform the complex and continuous duration estimation problem into a simple and discrete human motion detection by segmenting the CSI sequences. Furthermore, R-DEHM is implemented and evaluated in detail. The results of our experiments show that R-DEHM achieves the human motion detection and duration estimation with the average detection rate for human motion more than 94% and the average error rate for duration estimation less than 8%, respectively.

Single amino acid in V2 encoded by TYLCV is responsible for its self-interaction, aggregates and pathogenicity.

The V2 protein encoded by Begomovirus is essential for virus infection and is involved in multiple functions, such as virus movement and suppression of the host defence response. In this study, we reported that V2 encoded by the Tomato yellow leaf curl virus (TYLCV), which is one of the most devastating tomato-infecting begomoviruses, could interact with itself and a S71A mutation of V2 (V2S71A) abolished its self-interaction. Fluorescence results showed that V2 localized primarily in the cytoplasm and around the nucleus. Site-directed mutagenesis V2S71A had the similar subcellular localization, but V2S71A formed fewer large aggregates in the cytoplasm compared to wild-type V2, whereas the level of aggregates came to a similar after treatment with MG132, which indicates that the S71A mutation might affect 26S proteasome-mediated degradation of V2 aggregates. Meanwhile, heterologous expression of V2S71A from a Potato virus X vector induced mild symptoms compared to wild-type V2, delay of virus infection associated with mild symptoms was observed in plants inoculated with TYLCV-S71A, which indicates that the amino acid on position 71 is also involved in the pathogenicity of V2. To the best of our knowledge, this report is the first to state that the S71A mutation of V2 encoded by TYLCV affects the self-interaction, aggregate formation and pathogenicity of V2.

A simple method for obtaining rice black-streaked dwarf virus-infected small brown planthopper nymphs.

Rice black-streaked dwarf virus (RBSDV), an important rice virus, is transmitted by vector small brown planthopper (SBPH) in a persistent manner, but not transovarial transmission. In order to obtain viruliferous SBPH nymphs for relevant research, a simple and reliable method was developed, through allowing SBPH adults laying eggs on RBSDV-infected rice plants. The results showed the hatching nymphs on diseased plants could early acquire virus, and the virus was detected in 2nd-instar nymphs from the spawning method, which was earlier than insect feed on diseased plant. The average viruliferous rate of SBPH from the spawning method was 32.9%, which was not lower than the feeding diseased plant method. The novel method was very easy to operate and time-saving, facilitating the study on the interaction between RBSDV and SBPH nymphs (especially young 2nd-4th instar nymphs), such as, the effect of RBSDV on nymph development, host plant orientation preference of viruliferous nymph, identification of viral interacting protein in nymph, etc.

Identification of virus-derived siRNAs and their targets in RBSDV-infected rice by deep sequencing.

RNA interference (RNAi) is a conserved mechanism against viruses in plants and animals. It is thought to inactivate the viral genome by producing virus-derived small interfering RNAs (vsiRNAs). Rice black-streaked dwarf virus (RBSDV) is transmitted to plants by the small brown planthopper (Laodelphax striatellus), and seriously threatens production of rice in East Asia, particularly Oryza sativa japonica subspecies. Through deep sequencing, genome-wide comparisons of RBSDV-derived vsiRNAs were made between the japonica variety Nipponbare, and the indica variety 9311. Four small RNA libraries were constructed from the leaves and shoots of each variety. We found 659,756 unique vsiRNAs in the four samples, and only 43,485 reads were commonly shared. The size distributions of vsiRNAs were mostly 21- and 22-nt long, and A/U bias (66-68%) existed at the first nucleotide of vsiRNAs. Additionally, vsiRNAs were continuously but heterogeneously distributed along S1-S10 segments of the RBSDV genome. Distribution profiles of vsiRNA hotspots were similar in different hosts and tissues, and the 5'- and 3'-terminal regions of S4, S5, and S8 had more hotspots. Distribution and abundance of RBSDV vsiRNAs could be useful in designing efficient targets for exploiting RNA interference for virus resistance. Degradome analysis found 25 and 11 host genes appeared to be targeted by vsiRNAs in 9311 and Nipponbare. We report for the first time vsiRNAs derived from RBSDV-infected rice.

RNA-seq-based digital gene expression analysis reveals modification of host defense responses by rice stripe virus during disease symptom development in Arabidopsis.

BACKGROUND: Virus infection induces and suppresses host gene expression on a global level. Rice stripe virus (RSV) is the type species of the genus Tenuivirus and infects rice and Arabidopsis plants. Microarray-based and next generation sequencing-based transcriptomic approaches have been used to study rice-RSV interactions. However, our knowledge of the response of Arabidopsis plants to RSV infection is limited, and it requires further investigation to determine the similarities (or differences) in virus-host interactions between monocot and dicot hosts infected with RSV. METHODS: We characterized transcriptome changes in Arabidopsis thaliana infected with rice stripe virus (RSV) with RNA-seq based digital gene expression (DGE) analysis. The transcriptomes of RSV-infected samples were compared to those of mock-treated samples at 14 and 21 days post-infection (dpi) during different stages of symptom development. RESULTS: We identified 624 differentially expressed genes (DEGs) in Arabidopsis influenced by RSV at 14 dpi and 21 dpi, among which at 14 dpi, 255 transcripts were induced, and 38 were repressed; at 21 dpi, 146 were induced, and 237 were repressed. Functional annotation indicated that these DEGs were related to multiple biological functions, including defense response, secondary metabolism, protein amino acid phosphorylation and response to abiotic stress. CONCLUSIONS: Importantly, the transcription of genes related to host defense systems was activated by RSV infection at an early stage of symptom development (14 dpi), whereas over the infection period (21 dpi), the host defense response systems were suppressed. A total of 52 genes were continuously differentially expressed between the two time points, indicating that the majority of DEGs were transient and unique to a particular time point during symptom development. The DEGs, particularly the defense response genes, identified in this study are candidates suitable for further functional analysis during the RSV-Arabidopsis interaction.

Genetic analysis and molecular mapping of QTLs for resistance to rice black-streaked dwarf disease in rice.

Rice black-streaked dwarf disease, caused by rice black-streaked dwarf virus (RBSDV), is transmitted by small brown planthoppers (Laodelphax striatellus Fallén, SBPH) and causes severe yield loss in epidemic years in China and other East Asian countries. Breeding for resistance to RBSDV is a promising strategy to control the disease. We identified Tetep that showed resistance to RBSDV using a field test and artificial inoculation test. An evaluation of the resistance mechanism revealed that Tetep was resistant to RBSDV but not to SBPH. Genetic analysis showed that the resistance of Tetep to RBSDV was controlled by quantitative trait loci (QTLs). Three new QTLs for RBSDV resistance were identified in this study, i.e., qRBSDV-3, qRBSDV-10 and qRBSDV-11. The LOD scores of qRBSDV-3, qRBSDV-10 and qRBSDV-11 were 4.07, 2.24 and 2.21, accounting for 17.5%, 0.3% and 12.4% of the total phenotypic variation, respectively. All the resistance loci identified in this study were associated with virus resistance genes. The alleles for enhancing resistance on chromosomes 3 and 11 originated from Tetep, whereas the other allele on chromosome 10 originated from a susceptible parent. The identified new resistance QTLs in this study are useful resources for efficiently breeding resistant rice cultivars to RBSDV.

Genetic analysis and QTL detection for resistance to white tip disease in rice.

The inheritance of resistance to white tip disease (WTDR) in rice (Oryza sativa L.) was analyzed with an artificial inoculation test in a segregating population derived from the cross between Tetep, a highly resistant variety that was identified in a previous study, and a susceptible cultivar. Three resistance-associated traits, including the number of Aphelenchoides besseyi (A. besseyi) individuals in 100 grains (NA), the loss rate of panicle weight (LRPW) and the loss rate of the total grains per panicle (LRGPP) were analyzed for the detection of the quantitative trait locus (QTL) in the population after construction of a genetic map. Six QTLs distributed on chromosomes 3, 5 and 9 were mapped. qNA3 and qNA9, conferring reproduction number of A. besseyi in the panicle, accounted for 16.91% and 12.54% of the total phenotypic variance, respectively. qDRPW5a and qDRPW5b, associated with yield loss, were located at two adjacent marker intervals on chromosome 5 and explained 14.15% and 14.59% of the total phenotypic variation and possessed LOD values of 3.40 and 3.39, respectively. qDRPW9 was considered as a minor QTL and only explained 1.02% of the phenotypic variation. qLRGPP5 contributed to the loss in the number of grains and explained 10.91% of the phenotypic variation. This study provides useful information for the breeding of resistant cultivars against white tip disease in rice.

A novel, in vivo, indoor method to preserve rice black-streaked dwarf virus in small brown planthopper using wheat seedling as a bridge host.

Rice black-streaked dwarf virus (RBSDV) naturally infects Gramineae plants through small brown planthopper (SBPH) as a vector. However, RBSDV cannot be transmitted to the SBPH offspring through transovarian transmission. Wheat plant, an important intermediate host in winter, is essential for the completion of the annual cycle of RBSDV in farm ecosystem. We developed a novel, in vivo, indoor method to preserve RBSDV in SBPH using wheat seedling as a bridge host. The temperature range of 23-27°C was initially selected to rear the insects and plants. Before initiating the scheme cycle, viruliferous SBPH was obtained by feeding the virus-free 1st to 2nd instar nymphs with RBSDV-infected rice plants. Four to six RBSDV-infected SBPH were placed per plant to inoculate wheat seedlings at two-to-four leaf stages. After 48 h of inoculation, the viruliferous SBPH were removed. Five mated, newly emerged virus-free SBPH females were then transferred onto each inoculated plant and allowed to lay eggs for 48 h. The newly hatched SBPH were raised on wheat seedlings until the 2nd instar nymph stage, and then transferred onto healthy rice seedlings for further development until 5th instar nymphs or adults. These newly obtained viruliferous SBPH can be used for inoculating new wheat seedlings in the succeeding maintenance cycles, or for further experiments. We discovered that the incubation period of RBSDV in wheat seedlings synchronized with the gestation period of SBPH eggs at four to six inoculated viruliferous SBPH per plant and lasted for approximately seven days. In addition, this period was optimal for enhancing the SBPH infection ratio because SBPH nymphs can only acquire the virus after they hatch. The RBSDV infection ratio of the SBPHs acquired through this method consistently exceeded 50%.

Eugenol enhances the resistance of tomato against tomato yellow leaf curl virus.

BACKGROUND: Tomato yellow leaf curl virus disease (TYLCVD) causes severe to economic losses in tomato crops in China. The control of TYLCVD is based primarily on the use of synthetic insecticide to control its vector whitefly (Bemisia tabaci). To look for an alternative method for disease control, we investigated the effect of eugenol on controlling TYLCVD. The potential of eugenol to trigger systemic acquired resistance (SAR) in tomato (Jiangsu 14) plants against TYLCV was also investigated. RESULTS: In greenhouse experiments, eugenol significantly reduced disease severity when applied as a foliar spray, thus demonstrating a systemic effect. The disease spread rapidly in control plants and by the end of the experiment almost all control plants showed severe symptoms. Eugenol also induced H2O2 accumulation in tomato plants. Activities of peroxidase (POD), polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) were significantly induced compared with those of control plants. As further consequences, increase of salicylic acid (SA) levels and expression of PR-1 proteins, a molecular marker of SAR in tomato, could also be observed. CONCLUSION: This is the first report of eugenol as an elicitor and its ability to suppress plant virus diseases under greenhouse conditions. It is suggested that eugenol has the potential to be an effective biocontrol agent against TYLCV in tomato plants.

Overexpression of rice black-streaked dwarf virus p7-1 in Arabidopsis results in male sterility due to non-dehiscent anthers.

Rice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus in the family Reoviridae, is propagatively transmitted by the small brown planthopper (Laodelphax striatellus Fallén). RBSDV causes rice black-streaked dwarf and maize rough dwarf diseases, which lead to severe yield losses in crops in China. Although several RBSDV proteins have been studied in detail, the functions of the nonstructural protein P7-1 are still largely unknown. To investigate the role of the P7-1 protein in virus pathogenicity, transgenic Arabidopsis thaliana plants were generated in which the P7-1 gene was expressed under the control of the 35S promoter. The RBSDV P7-1-transgenic Arabidopsis plants (named P7-1-OE) were male sterility. Flowers and pollen from P7-1-transgenic plants were of normal size and shape, and anthers developed to the normal size but failed to dehisce. The non-dehiscent anthers observed in P7-1-OE were attributed to decreased lignin content in the anthers. Furthermore, the reactive oxygen species levels were quite low in the transgenic plants compared with the wild type. These results indicate that ectopic expression of the RBSDV P7-1 protein in A. thaliana causes male sterility, possibly through the disruption of the lignin biosynthesis and H2O2-dependent polymerization pathways.

Reverse transcription loop-mediated isothermal amplification of RNA for sensitive and rapid detection of southern rice black-streaked dwarf virus.

Southern rice black-streaked dwarf virus (SRBSDV) causes one of the most serious viral diseases of rice in Southeast Asia. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of SRBSDV using total RNA extracted from rice tissues and the insect pest, white-backed planthopper. The assay was based on a set of four primers matching a total of six sequences in the S9 region of SRBSDV genome. Presence of the virus could be detected in RT-LAMP reactions containing 1.2×10(-6)µg of a total RNA extract, which was ten times more sensitive than a classical RT-PCR assay. The SRBSDV could be distinguished from the closely related rice black-streaked dwarf virus (RBSDV) by this method, indicating a high degree of specificity. This simple and sensitive RT-LAMP assay shows potential for detection of SRBSDV in field samples of hosts or vectors at a relatively low cost.

Antifungal activity of the osthol derivative JS-B against Phytophthora capsici.

JS-B (C(12)H(10)O(3)) is a derivative compound of osthol. The antifungal properties of JS-B were tested against 10 economically important plant pathogens. JS-B was effective in inhibiting the mycelial growth of Phytophthora capsici, and its inhibition on different stages of the life cycle of P. capsici was observed. The 50% effective concentration (EC(50)) of JS-B on mycelial dry weight and zoospore germination of P. capsici was 43.74 and 86.03 microg/ml, respectively. The rupture of released zoospores induced by JS-B was reduced by the addition of 100 mM glucose. The ultrastructural study showed that JS-B caused destruction of most of the mitochondrions, the concentration of cell nuclear, and the existing vesicles. When compared with dimethomorph, the activity of JS-B on P. capsici was determined under pot conditions. The result showed that JS-B has a curative effect on pepper blight.

Efficacy of osthol, a potent coumarin compound, in controlling powdery mildew caused by Sphaerotheca fuliginea.

The efficacy of osthol, a natural coumarin compound, in controlling powdery mildew was evaluated in 2004-2005 in Anhui and Hebei Provinces of China. In both years, the treatments (osthol 15.0 and 18.0 g ai ha(- 1)) showed a stable control efficiency of 75.42, 81.24% and 76.36, 84.84%, respectively, at the Institutes of Plant Protection of Hebei Academy of Agricultural Sciences. In field experiments, osthol was as effective as difenoconazole in controlling powdery mildew and was more effective than triadimefon against Sphaerotheca fuliginea. Protection was expressed as a significant reduction (up to 87% compared with the control) in the mildewed leaf area in young pumpkin plants. Osthol strongly inhibited spore germination and mycelial growth of S. fuliginea in vitro, damaged the cell wall and the organelles of the pathogen. At 48 h after incubation, 50 microg ml(- 1) osthol could completely inhibit spore germination. These findings suggested that the effect of osthol on powdery mildew may be associated with the direct fungitoxic property against the pathogen. We conclude that osthol would be an attractive natural compound for practical agronomic use against powdery mildew.