Species‐specific detection of Candida tropicalis using evolutionary conserved intein DNA sequences

Inteins (internal proteins) are self‐splicing transportable genetic elements present in conserved regions of housekeeping genes. The study highlights the importance of intein as a potential diagnostic marker for species‐specific identification of Candida tropicalis, a rapidly emerging opportunistic human pathogen. Initial steps of primer validation, sequence alignment, phylogenetic tree analysis, gel electrophoresis and real‐time polymerase chain reaction (PCR) assays were performed to confirm the specificity of the designed primers. The primers were selective for C. tropicalis with 100% inclusivity and showed no cross‐species or cross‐genera matches. The established technique is a prototype for developing multifaceted PCR assays and for point‐of‐care testing in near future.

Significance and Impact of the Study

Development of molecular markers for specific detection of microbial pathogens using real‐time polymerase chain reaction (PCR) is an appealing and challenging technique. A real‐time PCR is an emerging technology frequently used to detect the aetiologic agents. In recent times, designing species‐specific primers for pathogen detection is gaining momentum. The method offers rapid, accurate and cost‐effective strategy to identify the target, thus providing sufficient time to instigate appropriate chemotherapy. The study highlights the use of intein DNA sequence as molecular markers for species‐specific identification of Candida tropicalis. The study also offers a prototype model for developing multifaceted PCR assays using intein DNA sequences, and provides a developmental starting point for point‐of‐care testing in near future.     

In situ or ex situ seed conservation: which is the more effective way to maintain seed longevity of an endangered cactus?

With restricted populations and a small number of individuals, Discocactus bahiensis Britton & Rose (Cactaceae) is an endangered species in Brazil and its capacity for the formation of seed banks in the soil and the maintenance of seed viability remains unknown. Thus, the aim of the present study was to determine the most efficient way to maintain viability during storage of seeds of D. bahiensis . Seeds were stored in paper bags and either kept in a cold chamber (7 ± 2°C) in the dark (ex situ conservation) or buried in the soil to a depth of 5 cm in an area of natural occurrence of the species (in situ conservation). Germinability of the seed banks was evaluated monthly for 20 months. During the first 10 months of storage, germinability of the seeds conserved in situ and ex situ was similar to that of recently collected seeds. After this period, a 70% reduction in germinability was found for the seeds maintained in situ and there was nearly complete loss of viability after 12 months of storage in the field (germinability < 10% in the last 8 months of the experiment), indicating the ability to form persistent soil seed banks. In contrast, the seeds stored in the cold chamber maintained greater than 70% germinability throughout the entire analysis period, demonstrating that ex situ conservation is the most efficient way to maintain the viability of the seeds of this endangered species.