Molecular probes for red-tide forming dinoflagellates found in HK  

Project Title: Molecular probes for red-tide forming dinoflagellates found in Hong Kong

Applicant: Hong Kong University of Science and Technology

Total Approved Grant: $593,700 (Actual Expenditure: $591,448.67)

Duration: 1.1.1999 - 31.12.2001

Project Status/Remarks: completed

Scope: To develop fast and accurate detection and identification methods for the major redtide forming dinoflagellates in Hong Kong.

Summary of the Findings/Outcomes:

The aim of the proposed research project was to develop fast and accurate detection and identification methods, based on molecular biological techniques, for the major red-tide forming dinoflagellates in Hong Kong. We were able to generate a database for sequence data of the commonly used gene (LsurDNA) for the dinoflagellates. We isolated strains of dinoflagellate strains and compared with the existing data from overseas strains. The data for the Alexandrium strains demonstrated the possible use of sequence data in tracing the origin of blooms. The data from Gymnodinium suggested that we have a new species of Gymnodinium cf. breve. We tested several protocols for the use of molecular probes and were able to improve on the methodology for use of local strains. We further optimised the techniques for detecting one single cell, the single-cell PCR technology that we previously developed. Using the accumulated sequence data, we were also able to design molecular probes to differentiate at least one species of Alexandrium catenella and the data obtained was positive. We held a workshop to demonstrate the techniques in detecting single dinoflagellate cell (single-cell PCR) to personnel from regulatory agencies. The results of the present project will be useful for further development of probes and DNA gene chip technology.

The project has achieved all the four objectives (see next section) stated in the original proposal, including the running of workshop for the regulatory agencies. The results of the project were also presented in four different occasions at two international/regional conferences. The publication of the results of this project also increased the awareness of the application of molecular probes in environmental monitoring.

Objectives achieved.

  • Sequence data gathering, including sequences from newly isolated Hong Kong species:

  • Probe design and manufacture; Using the sequence alignment in (a), we successfully designed a molecular probe for use in single-cell PCR.

  • Development of protocols for local species: we have developed new protocols for both Fluorescence-probe based hybridization method and single-cell PCR methods

  • Workshop to demonstrate the use of molecular probes/single-cell PCR for monitoring agencies concerned (EPD & AFCD) on 14 Dec 2001.