The failure to adequately identify plant pathogens from culture-based morphological techniques has led to the development of culture-independent molecular approaches. The timely and accurate detection of pathogens is a critical aid in the study of epidemiology and biology of plant diseases. In the case of regulated organisms, the availability of sensitive and reliable assay is essential when trying to achieve early detection of pathogens. We developed and tested the PCR assay for detection of Phytophthora colocasiae, an oomycetes pathogen of leaf blight of taro and of rotting of taro tubers. The method described here is specific for P. colocasiae when tested across fungal, bacterial, and other Phytophthora species. In conventional (single-round) PCR, the limit of detection was 20 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was 0.2 pg. In sampling studies, P. colocasiae-specific primers were used to detect leaf blight in infected leaves and tubers of taro cultivar. The causal pathogen P. colocasiae was detected by PCR from artificially infected tubers after 16 h of post inoculation, before any visible symptoms were present. The method was also tested to detect fungal DNA in infected leaves and infested soils. The PCR assay and direct tissue extraction methods provide tools which may be used to detect P. colocasiae pathogens in taro planting material and thus limit the transmission and spread of new, aggressive strains of P. colocasiae in taro-growing regions.
The Greater Yam (Dioscorea alata L.), a significant tropical tuber crop is highly affected by the anthracnose/dieback disease caused by Colletotrichum gloeosporioides, which greatly reduces the yield as well as market acceptability of the tubers. The different methods that could be used for proper identification of the pathogen by PCR were investigated. The studies indicate that species specific polymerase chain reaction assay based on highly conserved regions in ITS in the genome of the pathogen can be the best strategy for detection of this pathogen at species level. The use of genus specific primers was also successful in detection of Colletotrichum spp. The cloning and sequencing of evolutionarily conserved regions such as ITS, PelB and paralleling them with the available sequences in NCBI database is also costly but reliable approach. The various methods are elaborately tested and their use in diagnosis is discussed in detail.
Anthracnose caused by Colletotrichum gloeosporioides is an economically important disease which affects greater yam (Dioscorea alata L.) worldwide. Apart from airborne conidia, the pathogen propagules surviving in soil and planting material are the major sources of inoculum. A nested PCR assay has been developed for specific detection of C. gloeosporioides in soil and planting material. In conventional (single-round) PCR, the limit of detection was 20 pg, whereas in nested PCR the detection limit increased to 0.2 pg of DNA. The primers designed were found to be highly specific and could be used for accurate identification of the pathogen up to species level. The protocol was standardized for detection of the pathogen in artificially and naturally infected field samples.
VRK1 (vaccinia‐related kinase 1) was initially reported to be associated with spinal muscular atrophy–pontocerebellar hypoplasia (SMA‐PCH). Congenital or infantile‐onset progressive sensory‐motor neuropathy with microcephaly, adult‐onset distal SMA, and adult‐onset motor neuron disease are the other phenotypes described recently with VRK1. Since VRK1‐related complex motor disorders other than PCH is rare, we aim to depict the diverse clinical phenotypes and the genotypes of two patients with VRK1 variants from India. Proband‐1 is a 7‐year‐old girl who presented with distal muscle weakness and wasting of upper and lower limbs with brisk deep tendon reflexes (DTR), mild intellectual disability, and behavioral problems. She had a homozygous c.1159 + 1G > A pathogenic variant in VRK1, inherited from parents. Proband‐2 is a 25‐year‐old adopted male with sensory‐motor neuropathy and brisk DTR. He had a homozygous c.7C > T (p.R3C) missense variant of uncertain significance in VRK1 predicted to be damaging by multiple computational tools.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.