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SN/T 2398-2010 English PDF

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SN/T 2398-2010: Identification of candidatus phytoplasma mali
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Basic data

Standard ID SN/T 2398-2010 (SN/T2398-2010)
Description (Translated English) Identification of candidatus phytoplasma mali
Sector / Industry Commodity Inspection Standard (Recommended)
Classification of Chinese Standard B16
Classification of International Standard 07.080
Word Count Estimation 10,156
Date of Issue 2010-01-10
Date of Implementation 2010-07-16
Adopted Standard EPPO PM7/62 (1), NEQ
Regulation (derived from) National Quality Inspection [2010] No. 13
Issuing agency(ies) General Administration of Customs
Summary This standard specifies the Apple profusion phytoplasma quarantine and identification methods. This standard applies to inbound and outbound apple apple seedlings and scion in profusion phytoplasma quarantine and identification.

SN/T 2398-2010: Identification of candidatus phytoplasma mali

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Identification of candidatus phytoplasma mali Exit inspection and quarantine industry standard book People's Republic of China Apple tufted quarantine and identification of phytoplasma Issued on. 2010-01-10 2010-07-16 implementation People's Republic of China The State Administration of Quality Supervision, Inspection and Quarantine released

Foreword

This standard is part of EPPO phytosanitary standards (PM7/62 (1)), the main technical differences are as follows. --- Remove ELISA detection method; --- Remove indicator plant identification methods; --- Added electron microscopy methods. The Standard Appendix B, Appendix C, Appendix D is a normative appendix, Appendix A is an information appendix. This standard is proposed and managed by the National Certification and Accreditation Administration Committee. This standard is drafted by. China Inspection and Quarantine Science Research Institute. Participated in the drafting of this standard. People's Republic of China Beijing Entry-Exit-Entry Inspection and Xinjiang People's Republic of China Quarantine Bureau, People's Republic of China Liaoning Province Exit Inspection and Quarantine. The main drafters of this standard. Huang, Mu Haiqing, from Deng Liang, Yang Xuguang, Zhang Xianglin, Wangyou Fu, Zhu Shuifang. This standard is the first release of the entry-exit inspection and quarantine industry standards. Apple tufted quarantine and identification of phytoplasma

1 Scope

This standard specifies the overgrown apple quarantine and identification of phytoplasma. This standard applies to entry and exit in apple apple seedlings and scion overgrown plant quarantine and identification of the original body. Principle 2 2.1 taxonomic status Doors, flexible membrane mycetes without steroids Mycoplasma mesh, without steroids MLO Section, phytoplasma belong, 16SrX phytoplasma group. 2.2 Detection Methods Its molecular biology and gene sequence information and other biological characteristics is based on the quarantine and identification methods. DAPI staining method is simple, low cost, in this standard as a primary screening method phytoplasma; PCR technology Ling Sensitive and high specificity, RFLP technology is currently the main method of identification and classification of phytoplasma international, PCR technology in this standard and RFLP technique combined with an accurate method tufted apple phytoplasma as identification.

3 equipment and reagents

PCR instrument, clean bench, sterilizer, ice maker, nucleic acid protein analyzer, high-speed refrigerated centrifuge, small desktop centrifuge, ultra-low temperature Refrigerator, conventional refrigerator, whirlpool oscillator, micro-injector, electrophoresis, gel imaging system. DAPI, PCR buffer, magnesium chloride, dNTP, T Rao q DNA polymerase, primers and the like.

4 Detection Methods

4.1 Observation of Symptoms Careful observation apple seeds, fruits, leaves and other symptoms described in Appendix A. 4.2 DAPI staining Select young tissues (new leaf stems, shoots, stems and roots phloem tissue), slices, 1μg/mLDAPI solution (4 ', 6-diamidino Indole-2-phenylindole) staining, 460nm excitation conditions, fluorescence microscopy, in phloem sieve portion obvious blue fluorescent signal It indicates a phytoplasma presence. Within the plant due to the uneven distribution of phytoplasma, so each sample required to select different parts of the tender material for testing. 4.3 universal primer PCR and RFLP gel electrophoresis detection Universal primer PCR and RFLP gel electrophoresis detected in Appendix B. 4.4-specific primers PCR gel electrophoresis Specific primers PCR gel electrophoresis in Appendix C. 4.5 phytoplasma Morphology Preparation of ultra-thin sections of plant samples collected by electron microscopy Appendix phytoplasma form method D.

5 results found

5.1 presenting symptoms and A. 1.1 Description of consistent and DAPI staining showed blue fluorescence can be initially determined to be overgrown apple phytoplasma Body, subject to any program by 5.2,5.3,5.4 further testing. 5.2 4.3 test, if the PCR product gel electrophoresis test result is positive, and RFLP analysis fragment size consistent with the description, can be Apple determined that the carryover overgrown plant phytoplasma original body or 16SrX group.


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