GB/T 33119-2016 in English

In-depth interpretation of quarantine and identification methods for grapevine damping-off pathogen

This standard specifies the quarantine and identification methods for grapevine damping-off pathogen (Eutypa lata (Pers.) Tul. et C. Tul.), and is applicable to the quarantine and identification of this pathogen in host plants such as grapes.

Comparison of standard frameworks

Analysis of the background of standard formulation and technological evolution

Laboratory quarantine and identification process

1. PCR gel electrophoresis screening test: Specific primers (ED1 and ED2) were used for amplification, and the expected product size was 643bp.
2. Pathogen isolation and culture: Potato dextrose agar medium was used with the addition of selective antibiotics (streptomycin sulfate, chlortetracycline hydrochloride, chloramphenicol).
3. Morphological identification: Observe the characteristics of ascocarp and record its size and structure.

Result judgment and sample preservation

Judgment basis: Comprehensive analysis of PCR initial screening results, isolate culture characteristics and ascocarp characteristics.

Sample preservation: Positive samples need to be stored in a 4℃ refrigerator for at least 3 months. The strain should be inoculated on a potato dextrose agar slant and stored at 4℃ after 72 hours of culture at 24℃.

Implementation recommendations

• Quarantine stations: Set up rapid testing points at major ports and airports and equip them with PCR equipment.
• Training: Provide regular technical training for front-line quarantine personnel to ensure standardized operations.
• Data sharing: Establish a national disease database to achieve information exchange.