Mitochondrial DNA and RAPD polymorphisms in the haploid mite Brevipalpus phoenicis (Acari: Tenuipalpidae)

Abstract

Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) is recognized as the vector of citrus leprosis virus that is a significant problem in several South American countries. Citrus leprosis has been reported from Florida in the past but no longer occurs on citrus in North America. The disease was recently reported in Central America, suggesting that B. phoenicis constitutes a potential threat to the citrus industries of North America and the Caribbean. Besides B. phoenicis, B. obovatus Donnadieu, and B. californicus (Banks) have been incriminated as vectors of citrus leprosis virus and each species has hundreds of host plants. In this study, Brevipalpus mite specimens were collected from different plants, especially citrus, in the States of Florida (USA) and São Paulo (Brazil), and reared on citrus fruit under standard laboratory conditions. Mites were taken from these colonies for DNA extraction and for morphological species identification. One hundred and two Random Amplified Polymorphic DNA (RAPD) markers were scored along with amplification and sequencing of a mitochondrial cytochrome oxidase subunit I gene fragment (374 bp). Variability among the colonies was detected with consistent congruence between both molecular data sets. The mites from the Florida and Brazilian colonies were morphologically identified as belonging to B. phoenicis, and comprise a monophyletic group. These colonies could be further diagnosed and subdivided geographically by mitochondrial DNA analysis.

Testing prey DNA fingerprinting on Amblyseius largoensis (Acari: Phytoseiidae) predation of Raoiella indica (Acari: Tenuipalpidae)

“…This limits the interpretation of the results, which have to be evaluated only as proportional detection of positive SUs relative to the total SUs collected. DNA extraction was conducted by macerating mite specimens in a cetyltrimethylammonium bromide (CTAB) buffer, following the extraction procedure in Rodrigues et al (2004). A positive (R. indica tissue) and a negative (without DNA as a contamination control) control were included for each extraction and amplification.…”

“…PCR reactions were done in a final volume of 25 ll, with 8 ll of DNA extraction product, 0.7 ll of each primer solution at 50 lM, and 12.5 ll of the Promega PCR master mix reaction buffer. The PCR reaction program started with an initial denaturation step of 4 min at 92°C, followed by 35 cycles of 1 min denaturation at 92°C, 1 min primer annealing at 50°C, and 1 min of elongation at 72°C, with a final extension step of 10 min at 72°C (following Rodrigues et al 2004). Fragment visualization was done by running 20 ll of the PCR product at 60 V for 40-50 min on a 1 % agarose gel stained with ethidium bromide.…”

DNA-based methods for eriophyoid mite studies: review, critical aspects, prospects and challenges

“…This contrasts with the common use of single mites which has been reported for other groups of plant mites, e.g. Tetranychidae and Tenuipalpidae (Navajas et al 2001;Rodrigues et al 2004). However, the bias to analyze several mites as a single sample seems to be less important in eriophyoids considering that many species reproduce by arrenotokous parthenogenesis (Helle and Wysoki 1996).…”

Reduced Diversity in the Bacteriome of the Phytophagous Mite Brevipalpus yothersi (Acari: Tenuipalpidae)

“…Those specimens were originally collected from three different hosts in Puerto Rico: Sweet orange ( Citrus sinensis , Rutaceae), Tahiti lime ( Citrus latifolia , Rutaceae), and glory-bower ( Clerodendrum thomsoniae , Lamiaceae). Mite colonies were established by a single egg and maintained in fruit-arenas of “Valencia” sweet orange for 10 generations in an environmental chamber (Precision Model 818; Thermo Scientific, Waltham, MA, USA) at 25 °C and 75% relative humidity [25]. As outgroups, all in the Tetranychoidea superfamily, we included samples of adult Tetranychus evansi obtained from tomato plants ( Solanum lycopersicum , Solanaceae) in the field, as well as colonies of Raoiella indica from coconut ( Cocos nucifera , Arecaceae), and an Oligonychus mite (from dry beans, Phaseolus vulgaris , Fabaceae) collected at the Rio Piedras Agricultural Station.…”