GB/T 18086-2000 in English

《GB/T 18086-2000植物检疫 烟霜霉病菌检疫鉴定方法》由TC271（全国植物检疫标准化技术委员会）归口，主管部门为国家标准化管理委员会。
本标准规定了进境植物检疫中对烟霜霉病菌的检疫鉴定方法。本标准适用于进境干烟叶和烟属植物种子、种苗的烟霜霉病菌检疫和鉴定。

1 Scope

This standard specifies the method for inspection and identification of Peronospora hyoscyami de Bary f. sp. tabacina (Adam) Skalichy in import plant quarantine.

This standard is applicable to inspection and identification of Peronospora hyoscyami de Bary f. sp. tabacina (Adam) Skalichy in import dry tobacco leaves and seeds and seedlings of nicotiana plants.

2 Principle

Peronospora hyoscyami de Bary f. sp. tabacina (Adam) Skalichy is an obligate parasite. After tobacco leaves are infected, nearly round spots with blurred edges and chlorosis will appear, and later pale brown necrotic spots that can fall off will form. Under suitable temperature and humidity conditions, light blue mold [namely, the sporangiophore and sporangium of Peronospora hyoscyami de Bary f. sp. tabacina (Adam) Skalichy] will be formed on the necrotic spot on the back of leaves; under certain conditions, oospore may be formed in diseased tissues. Parasitic specialization and morphological characteristics of pathogenic bacteria are the basis for the identification of Peronospora hyoscyami de Bary f. sp. tabacina (Adam) Skalichy.

3 Instruments and reagents

3.1 Handhold magnifier.

3.2 Magnifier with light source.

3.3 Binocular dissecting microscope (10×~50×).

3.4 Microscope (100×~1 000×).

3.5 Centrifuge (1 000r/min ~ 4 000r/min).

3.6 Stainless steel metal screen (aperture size of 60μm).

3.7 Oscillator (200r/min).

3.8 Phosphate buffer solution, 0.02mol/L (pH=7).

3.9 Aniline blue.

3.10 Phenol glycerin solution.

Preparation of phenol glycerin solution: mixture of 20mL of phenol crystal (dissolved by heating in water bath), 20mL of lactic acid, 40mL of glycerinum and 20mL of distilled water.

3.11 10% potassium hydroxide solution

Preparation of potassium hydroxide solution: taking 10g of potassium hydroxide and dissolving with 90mL of water to form 10% potassium hydroxide solution.

Foreword i
1 Scope
2 Principle
3 Instruments and reagents
4 Sampling
5 Inspection methods of tobacco leaves
6 Inspection methods for seeds and seedlings of nicotiana plant
7 Identification characteristics
8 Result judgment
Annex A (Informative) Oospore inspection by frozen homogenization method