Judith K Brown
Work Summary
Unravel the phylodynamics and transmission-specific determinants of emerging plant virus/fastidious bacteria-insect vector complexes, and translate new knowledge to abate pathogen spread in food systems.
Unravel the phylodynamics and transmission-specific determinants of emerging plant virus/fastidious bacteria-insect vector complexes, and translate new knowledge to abate pathogen spread in food systems.
Abstract:
A biochemical approach was used to characterize acetylcholinesterase (ACHE) insecticide insensitivity in several sweetpotato whitefly (Bemisia tabaci; SPW) populations. Discriminating doses of insecticide were established to differentiate between sensitive and insensitive SPW strains and to genotype individual whitefly. This technique was then used to examine the frequency of insensitive AChE alleles in several SPW populations and to isolate a line homozygous for insensitive AChE from a heterogenous B-type population. Inheritance of putative altered AChE genotypes was consistent with the proposed haplo-diploid status of B. tabaci. This biochemical diagnostic was also employed to determine the role of insensitive AChE in the observed resistance profiles of several laboratory populations subjected to different selection regimes. In keeping with previous studies on insecticide resistance in SPW, resistance does not appear to be uniquely associated with the B-type but rather with SPW populations found in crop systems.
PMID: 15826909;Abstract:
Cotton leaf curl Gezira virus (CLCuGV), a species of the genus Begomovirus (family Geminiviridae), was recently cloned from cotton, okra, and Sida alba plants exhibiting leaf-curling and vein-thickening symptoms in Sudan. Here, we describe a previously unknown lineage of single-stranded DNA satellite (satDNA) molecules, which are associated with CLCuGV, and are required for development of characteristic disease symptoms. Co-inoculation of cotton and Nicotiana benthamiana plants with satDNAs cloned from cotton, okra, and S. alba, together with CLCuGV as the 'helper virus' resulted in the development of characteristic leaf-curling and vein-thickening symptoms in both hosts. An anatomical study of symptomatic, virus-infected cotton leaves revealed that spongy parenchyma cells had developed instead of collenchyma cells at the sites of vein thickening. Phylogenetically, the CLCuGV-associated satDNAs from Sudan, together with their closest relatives from Egypt, form a new satDNA lineage comprising only satDNAs from the Upper and Lower Nile Basins. Analysis of satellites and their helper virus sequences identified a predicted REP-binding site consisting of the directly repeated sequence, 'CGGTACTCA', and an inverted repeated sequence, 'TGAGTACCG', which occur in the context of a 17-nucleotide motif. The conserved REP-binding motif identified herein, together with strict geographic isolation, and apparent host-restriction, may be the collective hallmark of these new satDNA-begomovirus lineages, extant in the Nile Basin. © 2004 Elsevier B.V. All rights reserved.
Abstract:
Despite a large number of investigations on the molecular genetics and population structure of the whitefly Bemisia tabaci (Gennadius) complex, no such study had been conducted in Iran. The genetic variation of B. tabaci was examined using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) for 18 field collections from cucumber, eggplant, and tomato in four provinces of Iran. PCR amplification and restriction digestion with two enzymes detected 388 RFLP fragments, of which 16 fragments showed polymorphisms. Cluster analysis of these data placed all B. tabaci individuals within a single group, and there was no evidence for between- or within-population genetic variation. Phylogenetic (Clustal W) analysis of 42 B. tabaci mtCOI sequences (n = 21 field collections) from Iran, and a comparison with well-studied haplotype or biotype reference sequences available in public sequence databases, revealed that the Iranian B. tabaci populations were most closely related to the B biotype at 0-1.2% nucleotide identity. The B biotype is a well-known member of a sister clade from the Middle East-North African region of the world, owing to its nearly worldwide distribution and invasive characteristics. This report indicates that a single major haplotype of B biotype is prevalent in Iran and that its closest relative is the B biotype. Also, given the extent of known variation in the Middle East and African continent, data indicate somewhat surprisingly that the B. tabaci collections sampled in Iran had limited genetic variation and population substructure. Knowledge that the B biotype of B. tabaci predominates in Iran is important for designing effective pest management strategies given that biotypes of B. tabaci are known to differ greatly with respect to insecticide resistance, host range, virus-vector interactions, and other key biological characteristics. © Springer-Verlag 2008.
Abstract:
The recent emergence of whitefiy species of the genus Bemisia as virus vectors and pests worldwide has stimulated investigations into their biology. The invasion of pantropical agroecosystems by an exotic whitefiy and increased pressures by indigenous whiteflies elsewhere have led to the examination of Bemisia tabaci biology with a new perspective. The concept of host races or biotypes was proposed in the 1950s to describe select B. tabaci populations with definitive host associations and specific virus-vector capabilities. However, little attention has been given to the mechanisms involved. Biochemical, molecular, and whole-system approaches are underway to examine the underlying diversity among reproductively isolated populations or biotypes of B. tabaci.
PMID: 19768650;Abstract:
Partial genome segments of a begomovirus were previously amplified from Wissadula amplissima exhibiting yellow-mosaic and leaf-curl symptoms in the parish of St. Thomas, Jamaica and this isolate assigned to a tentative begomovirus species, Wissadula golden mosaic St. Thomas virus. To clone the complete genome of this isolate of Wissadula golden mosaic St. Thomas virus, abutting primers were designed to PCR amplify its full-length DNA-A and DNA-B components. Sequence analysis of the complete begomovirus genome obtained, confirmed that it belongs to a distinct begomovirus species and this isolate was named Wissadula golden mosaic St. Thomas virus-[Jamaica:Albion:2005] (WGMSTV-[JM:Alb:05]). The genome of WGMSTV-[JM:Alb:05] is organized similar to that of other bipartite Western Hemisphere begomoviruses. Phylogenetic analyses placed the genome components of WGMSTV-[JM:Alb:05] in the Abutilon mosaic virus clade and showed that the DNA-A component is most closely related to four begomovirus species from Cuba, Tobacco leaf curl Cuba virus, Tobacco leaf rugose virus, Tobacco mottle leaf curl virus, and Tomato yellow distortion leaf virus. The putative Rep-binding-site motif in the common region of WGMSTV-[JM:Alb:05] was observed to be identical to that of Chino del tomate virus-Tomato [Mexico:Sinaloa:1983], Sida yellow mosaic Yucatan virus-[Mexico:Yucatan:2005], and Tomato leaf curl Sinaloa virus-[Nicaragua:Santa Lucia], suggesting that WGMSTV-[JM:Alb:05] is capable of forming viable pseudo-recombinants with these begomoviruses, but not with other members of the Abutilon mosaic virus clade. Biolistic inoculation of test plant species with partial dimers of the WGMSTV-[JM:Alb:05] DNA-A and DNA-B components showed that the virus was infectious to Nicotiana benthamiana and W. amplissima and the cultivated species Phaseolus vulgaris (kidney bean) and Lycopersicon esculentum (tomato). Infected W. amplissima plants developed symptoms similar to symptoms observed under field conditions, confirming that this virus is a causal agent of Wissadula yellow mosaic disease in W. amplissima. © 2009 Springer Science+Business Media, LLC.