Detection and identification of phytoplasma associated with witches’ broom and little leaf disease in Arundo donax: first report from India

During the survey of two successive years 2012–2013, in nearby places of Gorakhpur districts, Uttar Pradesh, India, Arundo donax plants were found to be exhibiting witches’ broom, excessive branching accompanied with little leaf symptoms with considerable disease incidence. Nested PCR carried out with universal primers pair R16F2n/R16R2 employing the PCR (P1/P7) product as a template DNA (1:20) resulted in expected size positive amplification ~1.2 kb in all symptom-bearing plants suggested the association of phytoplasma with witches’ broom disease of Narkat plants. BLASTn analysis of the 16S rRNA gene sequence showed the highest (99%) sequence identity with Candidatus phytoplasma asteris (16SrI group). In phylogenetic analysis, the sequence data showed close relationships with the members of 16SrI phytoplasma and clustered within a single clade of 16SrI group and closed to B subgroup representatives. This is a first report of 16Sr I-B group phytoplasma associated with witches’ broom accompanied with little leaf disease of Narkat in India.     

Development of microsatellite markers in a clonal perennial herb,Convallaria keiskei

We developed eight polymorphic microsatellite simple sequence repeat (SSR) loci from genomic DNA of a clonal perennial herb, Convallaria keiskei, using a dual‐suppression‐polymerase chain reaction (PCR) technique and an improved technique. These markers with four to 10 alleles per locus identified 29 genotypes in 82 samples collected from a population in Hokkaido, Japan. The observed and expected heterozygosities ranged from 0.241 to 0.862 and from 0.639 to 0.825, respectively. These SSR markers will be available to identify genets and evaluate genetic diversity of C. keiskei.     

The occurrence of haemosporidian parasites in the Fennoscandian bluethroat ( Luscinia svecica) population

A total of 86 adult bluethroats (Luscinia svecica) from nine different localities, covering the full length of the Fennoscandian mountain range, were screened for blood parasites of the three genera Haemoproteus, Plasmodium and Leucocytozoon using a recently developed polymerase chain reaction method. The overall occurrence of infection was 59.3%. Prevalence of Leucocytozoon spp. (47.7%), Plasmodium spp. (23.3%) and Haemoproteus spp. (1.2%) was detected. Of the infected birds, 15.1% carried mixed infections. Five different mitochondrial DNA-lineages of Leucocytozoon spp., eight lineages of Plasmodium spp. and one lineage of Haemoproteus spp. were found. Due to large sequence divergence these corresponded to at least five different species, but with the possibility of all 14 being independent evolutionary units with the potential of evolving different effects on the host. Of the lineages of Leucocytozoon spp., the most common was found throughout the range. The occurrence of the second most common lineage of Leucocytozoon spp. showed significant variation in prevalence between sites. The data also showed molecular evidence of one lineage of Leucocytozoon sp. existing in more than one species of avian host, thus challenging the use of host taxon as a taxonomic character when distinguishing between different species leucocytozoids.Communicated by F. Bairlein     

Variability of phytoplasma associated with few ornamental crop species in Uttar Pradesh,India

Abstract

The economic importance of ornamentals has been advancing significantly in many countries due to continuous international demand. Like other crops, ornamental plants are also affected by a number of pathogens and the phytoplasmas diseases are equally important group of pathogens which is responsible for damaging the different ornamental plant species and thereby affecting their market value. In the present investigation, total 24 symptomatic leaf samples of 8 ornamental plant species (C. roseus, R. rubiginosa, C. variegatum, Z. elegans, J. gendurusa, T. erecta, H. rosa sinensis, J. sambrac) were analyzed which were found positive in nested PCR assays with R16F2n/R16R2 primers. The phytoplasma associated with ornamental plant species collected from different districts of UP in the present investigation are belongs to 16Sr I, II, VI and XIV group.     

Targeted and passive environmental DNA approaches outperform established methods for detection of quagga mussels,Dreissena rostriformis bugensis in flowing water

1. The early detection of invasive non‐native species (INNS) is important for informing management actions. Established monitoring methods require the collection or observation of specimens, which is unlikely at the beginning of an invasion when densities are likely to be low. Environmental DNA (eDNA) analysis is a highly promising technique for the detection of INNS—particularly during the early stages of an invasion.
2. Here, we compared the use of traditional kick‐net sampling with two eDNA approaches (targeted detection using both conventional and quantitative PCR and passive detection via metabarcoding with conserved primers) for detection of quagga mussel, Dreissena rostriformis bugensis, a high priority INNS, along a density gradient on the River Wraysbury, UK.
3. All three molecular tools outperformed traditional sampling in terms of detection. Conventional PCR and qPCR both had 100% detection rate in all samples and outperformed metabarcoding when the target species was at low densities. Additionally, quagga mussel DNA copy number (qPCR) and relative read count (metabarcoding) were significantly influenced by both mussel density and distance from source population, with distance being the most significant predictor.
4. Synthesis and application. All three molecular approaches were more sensitive than traditional kick‐net sampling for the detection of the quagga mussel in flowing water, and both qPCR and metabarcoding enabled estimates of relative abundance. Targeted approaches were more sensitive than metabarcoding, but metabarcoding has the advantage of providing information on the wider community and consequently the impacts of INNS.

     

Multiplex PCR system applied for analysing microsatellite loci of Schlegel's black rockfish,Sebastes schlegeli

Two multiplex PCR amplifications were performed to analyse six microsatellite loci of Schlegel's black rockfish, Sebastes schlegeli, an important commercial fish in the northern part of Japan and an important species for the stock enhancement program in this area. We analysed 67 wild samples from Yamada Bay, Iwate Prefecture, Japan. The observed genotype frequencies agreed with the Hardy–Weinberg expectations at all loci, and the observed heterozygosities ranged from 0.072 to 0.897.     

Surgical and Molecular Evaluation of Pediatric Hydatid Cyst Cases in Eastern Turkey

Cystic echinococcosis (CE) caused by Echinococcus granulosus is a major public health problem worldwide, including Turkey. The aim of the current study was to identify the strains and to estimate the potential risk factors of E. granulosus in operated pediatric cases in eastern Turkey. Ten pediatric patients (7 boys and 3 girls) living in rural areas, with ages ranging from 3 to 15 years old and various clinical histories, were included in this study. Eight patients had only liver hydatid cyst, while 1 patient had liver and lung hydatid cyst and the other liver, lung, and spleen, together. There were 2 ruptured liver cysts. After surgery, during follow-up, no increase was observed in hemagglutination levels, there were no mortalities, and there was no evidence of recurrence at 2 years post operation in all patients. Molecular analysis was performed on hydatid cyst samples obtained from the 10 pediatric cases. According to mt-12S rRNA PCR results, all cases were found to be G1/G3 cluster of E. granulosus sensu stricto.