Characteristic DNA sequence for dendrobium officinale test-tube plantlet and application thereof
A technology of DNA sequence and Dendrobium candidum, applied in the field of molecular biology, can solve the problems of difficult identification, inability to determine the type of source, and small plants, and achieve the effects of good specificity, great practical value, and high accuracy
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Embodiment 1
[0054] The acquisition of the characteristic DNA sequence of embodiment 1 Dendrobium officinale tube plantlet
[0055] 1 Materials and methods
[0056] 1.1 Materials
[0057] The names and sources of the experimental materials are shown in Table 1. The wild Dendrobium officinale were collected from Yunnan, Guangxi and other production areas, and the test-tube seedlings of Dendrobium officinale were provided by the Shanghai Institute of Traditional Chinese Medicine.
[0058] Table 1. Material names and sources
[0059] Numbering
[0060] 1
D. devonianum Dendrobium I
12
D.sp good bamboo stick
23
D. trigonopus dendrobium
2
D. devonianum Dendrobium II
13
D. aphyllum dendrobium
24
D. fimbriatum Verbena officinale
3
D. strongylanthum comb-lipped Dendrobium
14
D. chrysanthum bouquet Dendrobium...
Embodiment 2
[0100] The acquisition and identification of the specific probe of embodiment 2 Dendrobium officinale test-tube plantlets
[0101] 1 method
[0102] 1.1 The design of the specific probe of Dendrobium officinale tube plantlet
[0103] For the Dendrobium candidum test-tube plantlet characteristic DNA sequence that embodiment 1 obtains, use Primer Premier 5.0 (PREMIERBiosoft International development) software design to carry out primer sequence design, have designed four pairs of primers, synthesize:
[0104] (1) AS1 group: S1 5'AGGAAGATGAACTTGCCGATAC 3' (SEQ ID NO: 8)
[0105] A1 5' TCGGTCGTCCCGTCGTAT 3' (SEQ ID NO: 9)
[0106] (2) AS2 group: S2 5'AAGTGCCAAATGTTGAGTC 3' (SEQ ID NO: 10)
[0107] A2 5'CCTGGAATCTGTCGTGCT 3' (SEQ ID NO: 11)
[0108] (3) AS3 group: S3 5'CGAGACACCGTCCATTCA 3' (SEQ ID NO: 12)
[0109] A3 5'TTCCGACATTGGGCTTTA 3' (SEQ ID NO: 13)
[0110] (4) AS4 group: S4 5'AGGAAGATGAACTTGCCGATAC 3' (SEQ ID NO: 14)
[0111] ...
Embodiment 3
[0117] The genetic analysis of embodiment 3 different algebra Dendrobium officinale test-tube plantlets
[0118] By establishing a special test-tube plantlet culture line of Dendrobium candidum, the genetic status of plants of different generations in the same line was compared longitudinally.
[0119] Take different generations 1, 2, 4, 6, 8, 10, An (Dendrobium candidum test-tube seedling samples with passage number more than 10 generations) and wild Dendrobium candidum tissue culture test-tube plantlets and wild Dendrobium, carry out DNA extraction, use specificity The probes were amplified by PCR, and the results were as follows Figure 4 shown. The results of amplification with the specific probe of Dendrobium officinale test tube plantlets showed that there was no change in each generation, and the same bands were all expanded.
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