Heteroplasmy is the existence of multiple mitochondrial DNA haplotypes within the cell. Although the number of reports of heteroplasmy is increasing for arthropods, the occurrence, number of variants, and origins are not well studied. In this research, the occurrence of heteroplasmy was investigated in Thrips tabaci, a putative species complex whose lineages can be distinguished by their mitochondrial DNA haplotypes. The results from this study showed that heteroplasmy was due to the occurrence of mitochondrial cytochrome oxydase I (mtCOI) haplotypes from two different T. tabaci lineages. An assay using flow cytometry and quantitative real-time PCR was then used to quantify the per cell copy number of the two mtCOI haplotypes present in individuals exhibiting heteroplasmy from nine geographically distant populations in India. All of the T. tabaci individuals in this study were found to exhibit heteroplasmy, and in every individual the per cell copy number of mtCOI from lineage 3 comprised 75-98% of the haplotypes detected and was variable among individuals tested. There was no evidence to suggest that the presense of lineage-specific haplotypes was due to nuclear introgression; however, further studies are needed to investigate nuclear introgression and paternal leakage during rare interbreeding between individuals from lineages 2 and 3.
The gut microbial community structure of adult Thrips tabaci collected from 10 different agro-climatically diverse locations of India was characterized by using the Illumina MiSeq platform to amplify the V3 region of the 16S rRNA gene of bacteria present in the sampled insects. Analyses were performed to study the bacterial communities associated with Thrips tabaci in India. The complete bacterial metagenome of T. tabaci was comprised of 1662 OTUs of which 62.25% belong to known and 37.7% of unidentified/unknown bacteria. These OTUs constituted 21 bacterial phyla of 276 identified genera. Phylum Proteobacteria was predominant, followed by Actinobacteria, Firmicutes, Bacteroidetes and Cyanobacteria. Additionally, the occurrence of the reproductive endosymbiont, Wolbachia was detected at two locations (0.56%) of the total known OTUs. There is high variation in diversity and species richness among the different locations. Alpha-diversity metrics indicated the higher gut bacterial diversity at Bangalore and lowest at Rahuri whereas higher bacterial species richness at T. tabaci samples from Imphal and lowest at Jhalawar. Beta diversity analyses comparing bacterial communities between the samples showed distinct differences in bacterial community composition of T. tabaci samples from different locations. This paper also constitutes the first record of detailed bacterial communities associated with T. tabaci. The location-wise variation in microbial metagenome profile of T. tabaci suggests that bacterial diversity might be governed by its population genetic structure, environment and habitat.
The nucleotide sequence of M- and S-RNA segments of an Indian iris yellow spot virus (IYSV) were determined. Sequence comparisons showed that both of these sequences shared less than 95 % identity with those other known IYSV isolates. Phylogenetic analysis revealed that the S- and M-RNA sequences of known IYSV isolates clustered with those of the tospoviruses, tomato yellow ring virus, polygonum ringspot virus and hippeastrum chlorotic ringspot virus. Further, multiple recombination detection methods detected inter- and intra-species recombination events that clustered primarily within the intergenic regions of S- and M-RNA, suggesting that these are possibly recombination hotspots in IYSV and closely related tospoviruses.
Allium tuberosum L., commonly known as garlic chives, is an important spice in northeastern India as well as in many other parts of the world. Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) is an important pathogen of onion (4) and other related Alliums such as garlic (3) and leek (2). During April 2013, symptoms potentially induced by IYSV such as chlorotic and straw-colored spindle-like lesions were observed on leaves of A. tuberosum accession Hanzong Winter (CGN 20779) plants in the wild species garden at the Directorate of Onion and Garlic Research (DOGR), Rajgurunagar, Pune, Maharashtra, India. Ten plant samples of A. tuberosum were randomly collected from the wild species garden and the upper, middle, and lower portions of the leaves were pooled and tested by double-antibody sandwich (DAS)-ELISA using a commercially available kit (Agdia Inc., Elkhart, IN) for IYSV. All of them showed positive results for IYSV incidence. Total RNA from the ELISA positive leaf samples of A. tuberosum was extracted using the RNeasy Plant Mini kit (Qiagen GmbH, Hilden, Germany). The primer pair IYSV-F (5′-TCAGAAATCGAGAAACTT-3′) and IYSV-R (5′-TAATTATATCTATCTTTCTTGG-3′) (1) was used for RT-PCR. The primer pair was specific to amplify 797 bp of the nucleocapsid (N) gene of IYSV. The amplified product derived from A. tuberosum isolate was purified by QIAquick PCR Purification Kit (Qiagen) and cloned using the vector pDrive (Qiagen). The recombinant clone was sequenced (Accession No. KF624624). Sequence analysis performed on CLC Main Workbench Version 6.8.4 confirmed that the fragment was of IYSV. Nucleotide sequence comparison of our virus with other IYSV isolates revealed that the highest nucleotide identity (99%) was with the IYSV garlic isolate (HM173691) from India. Further, maximum 96% protein identity was with IYSV onion isolate (ACA09432) and garlic isolate (ADK56108) from India. To our knowledge, this is the first report of IYSV naturally occurring on A. tuberosum in India. It is evident from previous studies that IYSV causes significant losses in onions (1) and from this study, that its symptoms have direct impact on quality of garlic chives. Further detailed studies are required to assess the magnitude of the impact of IYSV infection on yield and quality of A. tuberosum. References: (1) A. Bulajic et al. Plant Dis. 93:976, 2009. (2) M. C. Córdoba-Sellés et al. Plant Dis. 91:1365, 2007. (3) S. J. Gawande et al. Plant Dis. 94:1066, 2010. (4) B. Mandal et al. Plant Dis. 96:468, 2012.
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