Cassava mosaic disease (CMD) represents a serious threat to cassava, a major root crop for more than 300 million Africans. CMD is caused by single-stranded DNA begomoviruses that evolve rapidly, making it challenging to develop durable disease resistance. In addition to the evolutionary forces of mutation, recombination and reassortment, factors such as climate, agriculture practices and the presence of DNA satellites may impact viral diversity. To gain insight into the factors that alter and shape viral diversity in planta, we used high-throughput sequencing to characterize the accumulation of nucleotide diversity after inoculation of infectious clones corresponding to African cassava mosaic virus (ACMV) and East African cassava mosaic Cameroon virus (EACMCV) in the susceptible cassava landrace Kibandameno. We found that vegetative propagation had a significant effect on viral nucleotide diversity, while temperature and a satellite DNA did not have measurable impacts in our study. EACMCV diversity increased linearly with the number of vegetative propagation passages, while ACMV diversity increased for a time and then decreased in later passages. We observed a substitution bias toward C→T and G→A for mutations in the viral genomes consistent with field isolates. Non-coding regions excluding the promoter regions of genes showed the highest levels of nucleotide diversity for each genome component. Changes in the 5′ intergenic region of DNA-A resembled the sequence of the cognate DNA-B sequence. The majority of nucleotide changes in coding regions were non-synonymous, most with predicted deleterious effects on protein structure, indicative of relaxed selection pressure over six vegetative passages. Overall, these results underscore the importance of knowing how cropping practices affect viral evolution and disease progression.

Cotton leafroll dwarf virus (CLRDV) is a yield-limiting, aphid-transmitted virus that was identified in cotton, Gossypium hirsutum L., in the United States of America in 2017. CLRDV is currently classified in the genus Polerovirus, family Solemoviridae. Although 8 species of aphids (Hemiptera: Aphididae) are reported to infest cotton, Aphis gossypii Glover is the only known vector of CLRDV to this crop. Aphis gossypii transmits CLRDV in a persistent and nonpropagative manner, but acquisition and retention times have only been partially characterized in Brazil. The main objectives of this study were to characterize the acquisition access period, the inoculation access period, and retention times for a U.S. strain of CLRDV and A. gossypii population. A sub-objective was to test the vector competence of Myzus persicae Sulzer and Aphis craccivora Koch. In our study, A. gossypii apterous and alate morphs were able to acquire CLRDV in 30 min and 24 h, inoculate CLRDV in 45 and 15 min, and retain CLRDV for 15 and 23 days, respectively. Neither M. persicae nor A. craccivora acquired or transmitted CLRDV to cotton.

A 2-yr study in cotton (Gossypium hirsutum L.) was conducted to determine the abundance and species composition of thrips (Thysanoptera: Thripidae) on different plant parts throughout the season in Alabama, Georgia, North Carolina, South Carolina, and Virginia. Plant parts sampled included seedlings, terminals with two expanded leaves, leaves from the upper, middle, and lower sections of the canopy, white flowers, and medium-sized bolls. Adult thrips were significantly more abundant on seedlings and flowers in 2014, and on flowers followed by seedlings and leaves from the middle canopy in 2015. Immature thrips were significantly more abundant on seedlings, followed by flowers in 2014, and on seedlings followed by leaves from the lower canopy and flowers in 2015. Across locations and plant parts, thrips consisted of Frankliniella tritici (Fitch) (46.8%), Frankliniella fusca Hinds (23.5%), Frankliniella occidentalis (Pergande) (17.1%), Neohydatothrips variabilis (Beach) (7.4%), Thrips tabaci (Lindeman) (1.8%), and other species (3.4%). Frankliniella fusca represented 86.7% of all thrips on seedlings, while F. tritici was more abundant on terminals (51.6%), squares (57.5%), and flowers (75.1%). Across all leaf positions, F. fusca was the most abundant species (28.8%), followed by F. tritici (19.2%), N. variabilis (18.8%), F. occidentalis (12.9%), and T. tabaci (5.2%), as well as other species (15.0%). As neonicotinoid insecticides remain a primary tool to manage seedling infestations of F. fusca, our data indicate that mid- to late-season applications of neonicotinoid insecticides targeting other insect pests will intensify selection pressure for resistance on F. fusca, the primary pest of seedling cotton.

This study investigated the role of vector acquisition and transmission on the propagation of single and co-infections of tomato yellow leaf curl virus (TYLCV,) and tomato mottle virus (ToMoV) (Family: Geminiviridae, Genus: Begomovirus) by the whitefly vector Bemisia tabaci MEAM1 (Gennadius) in tomato. The aim of this research was to determine if the manner in which viruses are co-acquired and co-transmitted changes the probability of acquisition, transmission and new host infections. Whiteflies acquired virus by feeding on singly infected plants, co-infected plants, or by sequential feeding on singly infected plants. Viral titers were also quantified by qPCR in vector cohorts, in artificial diet, and plants after exposure to viruliferous vectors. Differences in transmission, infection status of plants, and titers of TYLCV and ToMoV were observed among treatments. All vector cohorts acquired both viruses, but co-acquisition/co-inoculation generally reduced transmission of both viruses as single and mixed infections. Co-inoculation of viruses by the vector also altered virus accumulation in plants regardless of whether one or both viruses were propagated in new hosts. These findings highlight the complex nature of vector-virus-plant interactions that influence the spread and replication of viruses as single and co-infections.