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GB/T 40627-2021: Viability test of Leptosphaeria maculans (Desm) Ces. et De Not.
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| GB/T 40627-2021 | English | 189 |
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Viability test of Leptosphaeria maculans (Desm) Ces. et De Not.
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GB/T 40627-2021
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Basic data | Standard ID | GB/T 40627-2021 (GB/T40627-2021) | | Description (Translated English) | Viability test of Leptosphaeria maculans (Desm) Ces. et De Not. | | Sector / Industry | National Standard (Recommended) | | Classification of Chinese Standard | B16 | | Word Count Estimation | 10,131 | | Issuing agency(ies) | State Administration for Market Regulation, China National Standardization Administration | GB/T 40627-2021: Viability test of Leptosphaeria maculans (Desm) Ces. et De Not.---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Viability test of Leptosphaeria maculans (Desm) Ces.et De Not.
ICS 65.020.01
CCSB16
National Standards of People's Republic of China
Method for detecting the activity of canola stalk canker in rapeseed
Released on 2021-10-11
2022-05-01 implementation
State Administration for Market Regulation
Issued by the National Standardization Management Committee
Foreword
This document is in accordance with the provisions of GB/T 1.1-2020 "Guidelines for Standardization Work Part 1.Structure and Drafting Rules of Standardization Documents"
Drafting.
Please note that some of the contents of this document may involve patents. The issuing agency of this document is not responsible for identifying patents.
This document was proposed and managed by the National Plant Quarantine Standardization Technical Committee (SAC/TC271).
Drafting organizations of this document. Shenzhen Customs of the People's Republic of China, Shenzhen Institute of Inspection and Quarantine.
The main drafters of this document. Gao Ruifang, Zhang Guiming, Wang Ying, Huang Heqing, Xiang Caiyu, Li Na.
Method for detecting the activity of canola stalk canker in rapeseed
1 Scope
This document describes the application of ordinary fluorescence microscopy or laser scanning confocal microscopy to rape stem canker bacteria [Leptosphaeria]
maculans (Desm.) Ces.etDeNot.] Conidia is a method for viability detection.
This document is applicable to the activity detection and identification of the conidia of the canola stem canker carried in the related host of the canola stem canker.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
There are no terms and definitions that need to be defined in this document.
4 Principle
The use of fluorescent dyes to dye the conidia of canola stem canker, according to propidium iodide (PI)
It can penetrate the dying or dead cell membrane, and cannot penetrate the viable cell membrane, causing the inactive spores to emit red fluorescence.
Light, active spores do not emit fluorescence, use ordinary fluorescence microscope or laser scanning confocal microscope to treat fluorescent dyes
The conidia were tested and analyzed for their activity. See Appendix A for the basic information of the stem canker of rapeseed.
5 Apparatus and reagents
5.1 Reagents
Propidium iodide (PI) solution preparation method. Weigh 0.01g PI powder, add 10mL sterile water, and prepare a concentration of 1mg/mL
The mother liquor should be stored in a brown bottle at 4℃ and protected from light. Prepare the required concentration before use.
Potato Dextrose Agar (PDA)..200g peeled potatoes cut into small cubes, add 1000mL tap water, and boil
15min, filter with four layers of gauze to obtain the filtrate, add 18g glucose and 18g agar powder to the filtrate, heat to dissolve and dilute the solution to
1000mL, aliquot into Erlenmeyer flasks, autoclave at 121°C for 20min.
5.2 Equipment
Laser scanning confocal microscope or ordinary fluorescence microscope, autoclave, biological safety cabinet, biochemical incubator, refrigerator, centrifuge.
6 Activity detection method
6.1 Test sample preparation
In the ultra-clean workbench, use a dissecting needle to pick the conidia of the canola stem canker into sterilized water, and shake it for 3 minutes.
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