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Abstract Grapevine (Vitis vinifera) is an economically important fruit crop worldwide. The widely cultivated grapevine is susceptible to powdery mildew caused by Erysiphe necator. In this study, we used CRISPR-Cas9 to simultaneously knock out VviWRKY10 and VviWRKY30 encoding two transcription factors reported to be implicated in defense regulation. We generated 53 wrky10 single mutant transgenic plants and 15 wrky10 wrky30 double mutant transgenic plants. In a 2-yr field evaluation of powdery mildew resistance, the wrky10 mutants showed strong resistance, while the wrky10 wrky30 double mutants showed moderate resistance. Further analyses revealed that salicylic acid (SA) and reactive oxygen species contents in the leaves of wrky10 and wrky10 wrky30 were substantially increased, as was the ethylene (ET) content in the leaves of wrky10. The results from dual luciferase reporter assays, electrophoretic mobility shift assays and chromatin immunoprecipitation (ChIP) assays demonstrated that VviWRKY10 could directly bind to the W-boxes in the promoter of SA-related defense genes and inhibit their transcription, supporting its role as a negative regulator of SA-dependent defense. By contrast, VviWRKY30 could directly bind to the W-boxes in the promoter of ET-related defense genes and promote their transcription, playing a positive role in ET production and ET-dependent defense. Moreover, VviWRKY10 and VviWRKY30 can bind to each other's promoters and mutually inhibit each other's transcription. Taken together, our results reveal a complex mechanism of regulation by VviWRKY10 and VviWRKY30 for activation of measured and balanced defense responses against powdery mildew in grapevine. 

We propose the use of interactive vignettes as an alternative to traditional text- and video-based vignettes for conducting large-scale Human-Robot Interaction (HRI) studies. Interactive vignettes maintain the advantages of traditional vignettes while offering additional affordances for participant interaction and data collection through interactive elements. We discuss the core affordances of interactive vignettes, including explorability, responsiveness, and non-linearity, and look into how these affordances can enable HRI research with more complex scenarios. To demonstrate the strength of the approach, we present a case study of our own research project with N=87 participants and show the data we collect through interactive vignettes. We suggest that the use of interactive vignettes can benefit HRI researchers in learning how participants interact with, respond to, and perceive a robot’s behavior in pre-defined scenarios. 

Hurricanes often induce catastrophic flooding due to both storm surge near the coast, and pluvial and fluvial flooding further inland. In an effort to contribute to uncertainty quantification of impending flood events, we propose a probabilistic scenario generation scheme for hurricane flooding using state-of-art hydrological models to forecast both inland and coastal flooding. The hurricane scenario generation scheme incorporates locational uncertainty in hurricane landfall locations. For an impending hurricane, we develop a method to generate multiple scenarios by the predicated landfall location and adjusting corresponding meteorological characteristics such as precipitation. By combining inland and coastal flooding models, we seek to provide a comprehensive understanding of potential flood scenarios for an impending hurricane. To demonstrate the modeling approach, we use real-world data from the Southeast Texas region in our case study. 

This paper proposes a two-stage stochastic mixed integer programming framework for patient evacuation. While minimizing the expected total cost of patient evacuation operations, the model determines the location of staging areas and the number of emergency medical service (EMS) vehicles to mobilize in the first stage, and the EMS vehicle routing assignments in the second stage. A real-world data from Southeast Texas region is used to demonstrate our modeling approach. To provide a more pragmatic solution to the patient evacuation problem, we attempt to create comprehensive hurricane instances by integrating the publicly available state-of-art hydrology models for surge, Sea, Lake Ocean and Overland Surge for Hurricanes (SLOSH), and for streamflow, National Water Model (NWM), prediction. The surge product captures potential flooding in coastal region while the streamflow product predicts inland flooding. The results exhibit the importance of the integrated approach in patient evacuation planning, provide guidance on flood mapping and prove the potential benefit of comprehensive approach in scenario generation. 

Erysiphe necator is an economically important biotrophic fungal pathogen responsible for powdery mildew disease on grapevine. Currently, genome sequences are available for only a few E. necator isolates from the United States. Based on the combination of Nanopore and Illumina sequencing technologies, we present here the complete genome assembly for an isolate of E. necator, NAFU1, identified in China. We acquired a total of 15.93 Gb of raw reads. These reads were processed into a 61.12-Mb genome assembly containing 73 contigs with an N 50 of 2.06 Mb and a maximum length of 6.05 Mb. Combining the results of three gene-prediction modules (i.e., an evidence-based gene modeler [EVidenceModeler], an ab initio gene modeler, and a homology-based gene modeler), we predicted 7,235 protein-coding genes in the assembled genome of E. necator NAFU1. This information will facilitate studies of genome evolution and pathogenicity mechanisms of E. necator and other powdery mildew species through comparative genome sequence analysis and other molecular genetic tools. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license . 

Summary Calcium‐dependent protein kinases (CDPKs) play vital roles in metabolic regulations and stimuli responses in plants. However, little is known about their function in grapevine.Here, we report thatVpCDPK9andVpCDPK13, two paralogousCDPKsfromVitis pseudoreticulataaccession Baihe‐35‐1, appear to positively regulate powdery mildew resistance. The transcription of them in leaves of ‘Baihe‐35‐1’ were differentially induced upon powdery mildew infection. Overexpression ofVpCDPK9‐YFPorVpCDPK13‐YFPin theV. viniferasusceptible cultivar Thompson Seedless resulted in enhanced resistance to powdery mildew (YFP, yellow fluorescent protein). This might be due to elevation of SA and ethylene production, and excess accumulation of H₂O₂and callose in penetrated epidermal cells and/or the mesophyll cells underneath.Ectopic expression ofVpCDPK9‐YFPin Arabidopsis resulted in varied degrees of reduced stature, pre‐mature senescence and enhanced powdery mildew resistance. However, these phenotypes were abolished inVpCDPK9‐YFPtransgenic lines impaired in SA signaling (pad4sid2) or ethylene signaling (ein2). Moreover, both of VpCDPK9 and VpCDPK13 were found to interact with and potentially phosphorylate VpMAPK3, VpMAPK6, VpACS1 and VpACS2in vivo(ACS, 1‐aminocyclopropane‐1‐carboxylic acid (ACC) synthase; MAPK, mitogen‐activated protein kinase).These results suggest thatVpCDPK9andVpCDPK13contribute to powdery mildew resistance via positively regulating SA and ethylene signaling in grapevine.