Genome-Wide Analysis of von Willebrand Factor A Gene Family in Rice for Its Role in Imparting Biotic Stress Resistance with Emphasis on Rice Blast Disease

von Willebrand factor A(vWA)genes are well characterized in humans except for few BONZAI genes,but the vWA genes are least explored in plants.Considering the novelty and vital role of vWA genes,this study aimed at characterization of vWA superfamily in rice.Rice genome was found to have 40 vWA genes distributed across all the 12 chromosomes,and 20 of the 40 vWA genes were unique while the remaining shared large fragment similarities with each other,indicating gene duplication.In addition to vWA domain,vWA proteins possess other different motifs or domains,such as ubiquitin interacting motif in protein degradation pathway,and RING finger in protein-protein interaction.Expression analysis of vWA genes in available expression data suggested that they probably function in biotic and abiotic stress responses including hormonal response and signaling.The frequency of transposon elements in the entire 3K rice germplasm was negligible except for 9 vWA genes,indicating the importance of these genes in rice.Structural and functional diversities showed that the vWA genes in a blast-resistant rice variety Tetep had huge variations compared to blast-susceptible rice varieties HP2216 and Nipponbare.qRT-PCR analysis of vWA genes in Magnaporthe oryzae infected rice tissues indicated OsvWA9,OsvWA36,OsvWA37 and OsvWA18 as the optimal candidate genes for disease resistance.This is the first attempt to characterize vWA gene family in plant species.

Application Technology of Tebuconazole WG against Sheath Blight 75% Trifloxystrobin · Rice Blast and Rice

[ Objective] The paper was to confirm the best application period and frequency of 75% trifloxystrobin ·tebuconazole WG against rice blast and rice sheath blight. [ Method] Influences of different dosages and different application periods of 75% trifloxystrobin · tebuconazole WG on control effects against rice blast and rice sheath blight, as well as their effects on rice yields were studied in the paper. [Result] The control effects of three different fungicides application treatments against rice sheath blight were 80.24%, 83.0% and 67.99%, and the control effects against rice blast were 56.4%, 49.11% and 61.1%, respective- ly. Advanced application of fungicide for two times had good prevention effect against rice sheath blight ; properly delayed application of fungicides for two times was conducive to improving the control effect against rice blast, and one time application of sufficient fungicide during middle booting stage had higher control effect than application for two times. Effective panicle number per unit area, total grain number, 1 000-grain weight and moisture content of various fungicide application treat- ments did not have significant difference with those of the treatments without fungicide application, but grain number per panicle in treatments applied with fungicide was higher than the treatment without application. [ Conclusion] 75% Trifloxystrobin ·tebuconazole WG has better control effect on rice blast and sheath blight, which helps to promote the formation of rice grain and increases yield significantly. The fungicide application against rice sheath blight should be appropriately ad- vanced, and application for one time against panicle blast after middle booting stage is helpful to improve the control effect.

In-vitro vs in-vivo Inoculation: Screening for Resistance of Australian Rice Genotypes Against Blast Fungus

To assist with rapid screening for rice blast resistance as a precursor in a breeding program, the susceptibility to rice blast of 13 rice genotypes from Australia was evaluated in May to June 2013 using three distinct inoculation methods（spot, filter paper and standard methods） at seedling, vegetative and reproductive stages. The results revealed that the spot and filter paper inoculation methods were successful in discerning susceptibility to the rice blast disease（P ≤ 0.05）. Disease susceptibility declined significantly from the vegetative to reproductive stages. The standard method was conducted at three different stages for pot plants grown inside the mist house. However, low temperatures did not produce disease symptoms except in a few genotypes. Among the 13 rice genotypes screened, AAT9 expressed a highly resistant response, and AAT4, AAT6, AAT10, AAT11, AAT13, AAT17 and AAT18 expressed resistance at various stages. The results will be useful for selecting elite genotypes for disease tolerance where rice blast is prevalent. In addition, the resistant genotypes can serve as a gene pool used in breeding programmes to develop new resistant genotypes.

The ANIP1-OsWRKY62 module regulates both basal defense and Pi9-mediated immunity against Magnaporthe oryzae in rice

During effector-triggered immunity(ETI)against the devastating rice blast pathogen Magnaporthe oryzae,Pi9 functions as an intracellular resistance protein sensing the pathogen-secreted effector AvrPi9 in rice.Importantly,the underlying recognition mechanism(s)between Pi9 and AvrPi9 remains elusive.In this study,We identified a rice ubiquitin-like domain-containing protein(UDP),AVRPI9-INTERACTING PROTEIN 1(ANP1),which is directly targeted by AvrPi9 and also binds to Pi9 in plants.Phenotypic analysis of anip1 mu-tants and plants overexpressing ANIP1 revealed that ANIP1 negatively modulates rice basal defense against M.oryzae.ANiP1 undergoes 26S proteasome-mediated degradation,which can be blocked by both AvrPi9 and Pi9.Moreover,ANIP1 physically associates with the rice WRKY transcription factor OsWRKY62,which also interacts with AvrPi9 and Pi9 in plants.In the absence of Pi9,ANIP1 negatively regulates OsWRKY62 abundance,which can be promoted by AvrPi9.Accordingly,knocking out of OsWRKY62 in a non-Pi9 back-ground decreased immunity against M.oryzae.However,we also observed that OsWRKY62 plays negative roles in defense against a compatible M.oryzae strain in Pi9-harboring rice.Pi9 binds to ANiP1 and OsWRKY62 to form a complex,which may help to keep Pi9 in an inactive state and weaken rice immunity.Furthermore,using competitive binding assays,we showed that AvrPi9 promotes Pi9 dissociation from ANiP1,which could be an important step toward ETI activation.Taken together,our results reveal an immune strategy whereby a UDP-WRKY module,targeted by a fungal effector,modulates rice immunity in distinct ways in the presence or absence of the corresponding resistance protein.