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SN/T 4643-2016: Standards for sampling, detecting and preserving specimen of nematodes
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SN/T 4643-2016: Standards for sampling, detecting and preserving specimen of nematodes

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Standards for sampling, detecting and preserving specimen of nematodes People's Republic of China Exit Inspection and Quarantine standards Collection and preservation of nematode specimens 2016-08-23 released 2017-03-01 Implementation People's Republic of China General Administration of Quality Supervision, Inspection and Quarantine released

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009. This standard is proposed and managed by the National Certification and Accreditation Administration Committee. This standard was drafted by Ningbo Entry-Exit Inspection and Quarantine Bureau of People's Republic of China. The main drafters of this standard. Gu Jian Feng, Chen Xianfeng, Zhang Huili, He Jie. Collection and preservation of nematode specimens

1 Scope

This standard specifies the exit phytosanitary nematode specimens collection, production and preservation methods. This standard applies to entry and exit inspection and quarantine departments of nematode specimens collection, production and preservation.

2 method principle

According to different parasitic modes of plant pathogenic nematodes, samples showing symptoms such as tumor, leaf spot, necrosis, discoloration and deformity are collected or collected The main plant roots and rhizosphere soil or media, using improved funnel separation method such as separation of nematodes, and then soaked to save or make a permanent slide to reach The purpose of preservation of specimens of nematodes. 3 instruments, reagents and appliances 3.1 instruments Stereo microscope (10 × above, with a translucent light source), incubator, pipette (20μL ~.200μL), centrifuge (2000r/min Above), freezers, scales, Fenwick floaters and more. 3.2 Reagents Formalin (4% formaldehyde), glycerol, ethanol, sucrose, encapsulated tablets (neutral gum, gum acacia, nail polish), paraffin, solution I Ethanol/glycerol/distilled water volume ratio of 20. 1. 79), solution II (glycerol/96% ethanol volume ratio 1.19), ethylenediaminetetraacetic acid disodium, Sulfoxide, sodium chloride, sodium hydroxide. DESS solution preparation method see Appendix A. 3.3 appliances Clock dish or dish, cell culture dish, cover glass, glass slide, scissors, nematode pick needle, alcohol lamp, double tissue, nematode filter paper, gauze, 500 μL microcentrifuge tube, closed plastic box, funnel, funnel rack, latex tube, water stop clip, heating plate, sieve mesh (10 mesh, 60 mesh, 300 mesh, 500 heads), sampling shovels, plastic bags, buckets, markers, etc.

4 sample collection

4.1 lesions or soil samples collected The phytopathogenic nematodes can be divided into endoparasites and ectoparasite nematodes according to different ways of parasitism. Most parasitic nematodes endanger the host plant's roots And underground stems, such as root knot nematode Meloidogyne parasitic in the root, cyst nematode Heterodera part of the exposed roots. But there is also Some nematodes (eg, Anguina, Aphelenchoides, Bursaphelenchus, etc.) Plants on the ground buds, leaves, stems, seeds, ears and so on. Part of the plant by the nematode hazards, sometimes can be expressed as gall tumor, leaf spot, necrosis, discoloration, Deformity and other symptoms, but the symptoms are not obvious on the ground. After the plant is endangered by the nematode, the site where the obvious lesion occurs is often also the site where the pathogenic nematode exists. Such as granulosus nematodes in the grain insects ...