Molecular specificity labeling primers of dendrobium strongylanthum rchb.f. and detection method thereof

Abstract

The invention relates to molecular specificity labeling primers of dendrobium strongylanthum rchb.f. and a method capable of rapid identification of the dendrobium strongylanthum rchb.f. The primers have the sequences: an upstream primer of 5'-AGCGGCCGCGTGGTACAC-3', and a downstream primer of 5'-GTGTACCACGCGGCCGCT-3'. The beneficial effects mainly comprise that the molecular specificity labeling primers can perform rapid early identification on the dendrobium strongylanthum rchb.f.; and the method is simple, fast and accurate, and is an irreplaceable molecular means for apparent characteristic identification of the variety of the dendrobium strongylanthum rchb.f.

Description

(1) Technical field [0001] The invention relates to a molecular-specific marker primer of dendrobium comb lip, and a method for identifying and detecting dendrobium comb lip using the primer. (2) Background technology [0002] Dendrobium (D.orchids), also known as Dendrobium orchid, is a plant of the genus Dendrobium (Orchidaceae), widely distributed in China, Japan, the Philippines, Thailand, India, Malaysia, Australia and New Zealand in Asia and Oceania. There are more than 1,000 species in my country, and there are 74 species and 2 varieties in China. As an excellent medicinal plant, Dendrobium has high medicinal and health value. It was first seen in "Shan Hai Jing", and its medicinal use was first recorded in "Shen Nong's Materia Medica", and was later recorded in "Compendium of Materia Medica" and other successive dynasties. , has the effects of "mainly harming, eliminating numbness, lowering qi, tonifying the five Zangs, strengthening yin, thickening the stomach, ligh...

AI Technical Summary

Problems solved by technology

[0004] The identification of Dendrobium species is mainly based on appearance characteristics, but due to the long period of identification of many morphological characters, the impact of the environment, and the increasing number of varieties, the identification of varieties is becoming more and more difficult.

Since this century, some PCR-based molecular marker technologies such as RAPD (Random Amplified Polymorphic DNA, Random Amplified Polymorphic DNA), ISSR (Inter-Simple Sequence Repeat, Simple Sequence Repeat Interval Amplified Polymorphism) and SRAP ( Sequence-related amplified polymorphism) has been successively used to study the genetic diversity and genetic distance of Dendrobium germplasm resources, but there are few studies on the linkage between molecular markers and Dendrobium traits, and molecular identification between Dendrobium varieties. Moreover, these molecular markers are either universal random primers or a random combination of designed primers, the PCR amplification pattern is not only complicated, the repeatability is poor, and the specificity is not high, so it is not suitable for variety identification

SCAR (Sequence Characterized Amplified Region) marker was proposed by Paran and Michelmore on the basis of RAPD in 1993. It is based on the sequencing of specific RAPD fragments, and a pair of 18-24 bases is designed according to the sequences at both ends. Primers are achieved by specific amplification at a higher annealing temperature, and the use of specific primers eliminates the competition between random primer binding sites, so it is a very stable molecular marker and has a wide range of applications. Rapid, simple, and low-cost characteristics, so far, there is no research report on the application of SCAR markers in the identification of Dendrobium species

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