Comparative analysis, diversification, and functional validation of plant nucleotide-binding site domain genes.

Nucleotide-binding site (NBS) domain genes are one of the superfamily of resistance genes involved in plant responses to pathogens. The current study identified 12,820 NBS-domain-containing genes across 34 species covering from mosses to monocots and dicots. These identified genes are classified into 168 classes with several novel domain architecture patterns encompassing significant diversity among plant species. Several classical (NBS, NBS-LRR, TIR-NBS, TIR-NBS-LRR, etc.) and species-specific structural patterns (TIR-NBS-TIR-Cupin_1-Cupin_1, TIR-NBS-Prenyltransf, Sugar_tr-NBS etc.) were discovered. We observed 603 orthogroups (OGs) with some core (most common orthogroups; OG0, OG1, OG2, etc.) and unique (highly specific to species; OG80, OG82, etc.) OGs with tandem duplications. The expression profiling presented the putative upregulation of OG2, OG6, and OG15 in different tissues under various biotic and abiotic stresses in susceptible and tolerant plants to cotton leaf curl disease (CLCuD). The genetic variation between susceptible (Coker 312) and tolerant (Mac7) Gossypium hirsutum accessions identified several unique variants in NBS genes of Mac7 (6583 variants) and Coker312 (5173 variants). The protein-ligand and proteins-protein interaction showed a strong interaction of some putative NBS proteins with ADP/ATP and different core proteins of the cotton leaf curl disease virus. The silencing of GaNBS (OG2) in resistant cotton through virus-induced gene silencing (VIGS) demonstrated its putative role in virus tittering. The presented study will be further helpful in understanding the plant adaptation mechanism.

Comparative phylogenetic analysis of aquaporins provides insight into the gene family expansion and evolution in plants and their role in drought tolerant and susceptible chickpea cultivars.

Aquaporins (AQPs) are water channel proteins that play a significant role in drought stress. Although the AQPs identified in multiple plant species, there is no detailed evolutionary and comparative study of AQPs regarding chickpea plant. The current study involved evolutionary analyses coupled with promoter and expression analyses of chickpea AQPs (CaAQPs). A total of 924 non-redundant AQPs were studied in 24 plant species including algae, mosses, lycophytes, monocots and dicots. Phylogenetic analysis demonstrated a clear divergence of eight AQP subfamilies (LIPs, SIPs, GIPs, NIPs, XIPs, PIPs, HIPs and TIPs). The comparative phylogenetic trees of AQP subfamilies among Arabidopsis, soybean, common bean, maize and chickpea demonstrated that the AQPs were highly species-specific. Interestingly, the dual NPA motif was conserved in all species. However, the ar/R selectivity filter signatures [W/T/S/N/G/A]-[V/S/L/I/A]-[S/G/A]-R (in NIPs), F-H-T-R (in PIPs), [H/N/Q/S]-[A/I/L/S/V]-[A/G]-[A/C/L/M/R/V] (in TIPs) and [V/I/L/M]-[V/I/A/F/M]-[A/S/F/C]-[N/F/L/I/A/S (in SIPs) were found in five species. Moreover, the Froger's positions (P1-P5) were found as [F/L/Y]-[S/T]-A-Y-[L/I/M/V/F] (in NIPs), [Q/E/M]-S-A-F-W (in PIPs), [A/L/S/T/V]-[A/C/N/S/T/V]-[P/R/S]-[Y/N/F]-[W/Q] (in TIPs) and [I/M/F]-[A/V]-[A/V]-Y-W (in SIPs). The MEME motif analyses showed that most of the motifs were specific to subfamily and subgroups. Tissue-specific expression profiling of CaAQPs revealed that CaTIPs and CaPIPs are highly expressed in most of the tissues, while CaNIPs and CaSIPs have low expression. In promoter analysis of CaAQPs, multiple stress-related cis-acting elements e.g. MYB, MYC, ABRE, etc. were found. Semi-quantitative RT-PCR analysis showed that CaPIP2;3 and CaNIP3;1 are positive regulator, while CaSIP1;1 and CaPIP2;1 have a negative role in drought tolerance. The findings and implications of this study are discussed in detail.

Controlling Aedes aegypti in Cryptic Environments with Manually Carried Ultra-Low Volume and Mist Blower Pesticide Applications.

Because Aedes aegypti , a vector of dengue, chikungunya, and Zika viruses, exhibits resting and reproductive behaviors that present challenges to control them, pesticide application equipment available for vector control must be evaluated for their ability to control this species in a variety of cryptic environments. Five types of pesticide sprayers, representing 3 spray technologies (1 mister, 2 ultra-low volume [ULV] cold foggers, and 2 ULV thermal foggers), were evaluated for their ability to control adult and immature stages of Ae. aegypti in indoor and outdoor environments. Cages holding adult mosquitoes and larvae were placed in cryptic sites for indoor sprays and placed among dense vegetation for outdoor sprays. Adult and pupal mortality data were recorded following applications of a mixture of synergized pyrethrins and pyriproxyfen. We found that no single sprayer was best suited for controlling Ae. aegypti in indoor and outdoor environments, nor was one best for controlling adult and immature mosquitoes. Sprayers producing larger droplets (misters and cold foggers) were more effective in controlling immature mosquitoes indoors and outdoors. Thermal fogging was more effective in controlling adults indoors, whereas cold fogs and misters were more effective for outdoor control.

Field evaluation of commercial off-the-shelf spatial repellents against the Asian tiger mosquito, Aedes Albopictus (Skuse), and the potential for use during deployment.

The Testing and Evaluation Department of the US Navy Entomology Center of Excellence (NECE), Naval Air Station, Jacksonville, Florida, is dedicated to the evaluation of novel equipment and vector control techniques to provide guidance on effective protection measures against human pathogens transmitted by blood-feeding arthropods. Personal protective measures (PPM), to include repellents, are part of a series of techniques that contribute toward reducing human-vector contact for globally and domestically deployed military forces. However, improper PPM use and limited availability has created vulnerabilities, causing troops to purchase spatial repellent products that are not approved by the Department of Defense. In order to ensure the most effective products are available, NECE has evaluated the spatial repellency response of Aedes albopictus (Skuse) to 4 commercial off-the-shelf (COTS) spatial repellents to provide technical guidance on proper use and effectiveness. The COTS products evaluated ThermaCELL, OFF! Clip On, Lentek Bite Shield, and Bug Button Mosquito Eliminator. A Biogents Sentinel (BGS) trap was placed in 5 locations with a spatial repellent device suspended at the level of the BGS trap opening over 4 of them (the fifth was control). Each trap catch was collected every 12 hours, at which time the spatial repellent device was rotated to the next position. Using this method, each spatial repellent device and control was rotated across each of the 5 locations a total of 6 times. Spatial repellent efficiency was evaluated by comparing the total number of mosquitoes collected in the BGS traps during a 12-hour period. The number of adult mosquitoes repelled by the ThermaCell spatial repellent was significantly more than other spatial repellents with the exception of OFF!. These data indicate that COTS products using repellent insecticide rather than botanicals are more effective at deterring Ae. albopictus from biting a host.

Homeobox genes d11-d13 and a13 control mouse autopod cortical bone and joint formation.

The molecular mechanisms that govern bone and joint formation are complex, involving an integrated network of signaling pathways and gene regulators. We investigated the role of Hox genes, which are known to specify individual segments of the skeleton, in the formation of autopod limb bones (i.e., the hands and feet) using the mouse mutant synpolydactyly homolog (spdh), which encodes a polyalanine expansion in Hoxd13. We found that no cortical bone was formed in the autopod in spdh/spdh mice; instead, these bones underwent trabecular ossification after birth. Spdh/spdh metacarpals acquired an ovoid shape and developed ectopic joints, indicating a loss of long bone characteristics and thus a transformation of metacarpals into carpal bones. The perichondrium of spdh/spdh mice showed abnormal morphology and decreased expression of Runt-related transcription factor 2 (Runx2), which was identified as a direct Hoxd13 transcriptional target. Hoxd11-/-Hoxd12-/-Hoxd13-/- triple-knockout mice and Hoxd13-/-Hoxa13+/- mice exhibited similar but less severe defects, suggesting that these Hox genes have similar and complementary functions and that the spdh allele acts as a dominant negative. This effect was shown to be due to sequestration of other polyalanine-containing transcription factors by the mutant Hoxd13 in the cytoplasm, leading to their degradation. These data indicate that Hox genes not only regulate patterning but also directly influence bone formation and the ossification pattern of bones, in part via Runx2.