SN/T 2398-2010Identification of candidatus phytoplasma mali (English PDF)

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.

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.