SN/T 4732-2016 in English

VALID

Detection and identification of Xanthomonas campestris pv, musacearum

Issued on:2016-12-12
Implemented on:2017-07-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $250.00
$243.00

Standard No:	SN/T 4732-2016
Document status:	VALID
Title in English:	Detection and identification of Xanthomonas campestris pv, musacearum
Title in Chinese:	香蕉细菌性萎蔫病菌检疫鉴定方法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2016-12-12
Implemented on:	2017-07-01
Professional Classification:	SN-Import&Export Inspection
Related Keywords:	xanthomonas campestris pv detection method molecular detection species-specific detection triple detection system
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Introduction

Standard Background and Technological Evolution

SN/T 4732-2016 is my country's industry standard for quarantine and detection of banana bacterial wilt (Xanthomonas campestris pv. musacearum). This pathogen is listed as a quarantine pest for imported plants in my country and can cause devastating losses to the banana industry. The standard integrates a triple detection system using traditional culture, molecular biology, and mass spectrometry techniques, achieving improved specificity and sensitivity compared to the detection method first reported internationally in 2007.

Pathogen characteristic analysis

Characteristic dimensions	Biological characteristics	Identification points
Morphological characteristics	0.4-0.7×0.7-1.8μm short rods	Gram negative, monopolar flagellum
Cultural characteristics	Growing at 25-30℃, CC medium produces 3-4mm yellow colonies	YTSA-CC medium forms 2-3mm translucent colonies after 48h
Biochemical reactions	Oxidase negative, starch hydrolysis positive	Gelatin liquefaction test that distinguishes it from the closely related species X. vasicola

Detection Technology System

1. Selective Culture

The standard recommends two culture medium formulas: CCA (containing cefoperazone) and YTSA-CC:

CCA medium: Cultured at 25°C for 6 days, inhibiting bacteria by 90%
YTSA-CC: Adding triphenyl blue for specific color development improves accuracy by 30%

2. Molecular Detection

Using a duplex PCR system:

GspD gene target (265 bp): species-specific detection
16S rDNA (641 bp): internal control

Cycling parameters: 95°C for 3 min initial denaturation; 35 cycles (95°C for 20 s, 64°C for 15 s, 72°C for 13 s)

Implementation Recommendations

Sample Handling Guidelines

Disease tissue should be surface disinfected with 75% ethanol for 1-2 min, rinsed with sterile water, and then ground into vascular tissue. DNA was extracted using the CTAB method for PCR template preparation and stored at -20°C for no more than 30 days.

Result interpretation

Comprehensive judgment must meet the following requirements:

Typical colony morphology in selective culture medium
Simultaneous appearance of 265bp and 641bp bands in PCR
MALDI-TOF MS score ≥2.3

More samples: NB/T 47013.2-2015 DL/T 5154-2012 QC/T 942-2021

Sample only — not a preview of SN/T 4732-2016

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