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Dendrobium chrysotoxum SSR molecular marker primer composition and application thereof

A primer composition, the technology of Dendrobium drumsticks, applied in the field of molecular biology

Active Publication Date: 2021-04-30
ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As an ornamental and medicinal plant, Dendrobium chrysalis has not yet been approved and widely cultivated in production. Its wild resources or seedlings or cloned seedlings obtained by tissue culture and sterile seeding are mainly used in production. The identification, conservation, development and utilization of these Dendrobium chrysalis germplasm resources have also attracted more and more attention from all walks of life.

Method used

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  • Dendrobium chrysotoxum SSR molecular marker primer composition and application thereof
  • Dendrobium chrysotoxum SSR molecular marker primer composition and application thereof
  • Dendrobium chrysotoxum SSR molecular marker primer composition and application thereof

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Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1 Dendrobium drumstick SSR primer screening

[0036] 1. The present invention utilizes the transcriptome sequence of Dendrobium chrysalis, designs and screens 100 pairs of SSR primers, which are synthesized in Sangon Bioengineering (Shanghai) Co., Ltd. Modified with FAM (blue) and HEX (green) two kinds of biological fluorescent groups.

[0037] 2. Randomly select 7 Dendrobium chrysanthemum plants (seedlings) as test materials, cut their young leaves, use DP305 kit (China Tiangen) to extract their genomic DNA, and use 1.2% agarose gel electrophoresis and NanoDrop spectrophotometry respectively A photometer measures the quality and concentration of the DNA.

[0038] 3. Use the sample DNA as a template, and perform PCR amplification with fluorescently modified primers and downstream primers. The PCR amplification reaction system is: the total volume of the reaction system is 10 μL, including 1.0 μL DNA template (50ng / μL), 1.0 μL 10×BufferΙ Buffer, 0.1 μL TAKAR...

Embodiment 2

[0044] Application of embodiment 2 SSR primer composition in Dendrobium drumstick population genetic diversity analysis

[0045]1. The test materials are 8 populations of Dendrobium pachyrhiza from different sources (Table 3), each population samples 8-14 strains, and each plant is individually numbered, respectively marked as CA1-CA12, CB1-CB12, CC1-CC10, CD1 -CD14, CE1-CE8, CF1-CF8, CG1-CG12, CH1-CH8, a total of 84 individual plants were sampled.

[0046] Table 3. Eight groups of Dendrobium drumstick test materials

[0047] Dendrobium drumstick group number Number of sampling plants (trees) Group origin CA 12 Bhamo CB 12 Lashio, Myanmar CC 10 Menghai County, Xishuangbanna, Yunnan cd 14 Yunnan Baoshan Tengchong CE 8 Guizhou Xingyi CF 8 Menglian County, Pu'er, Yunnan CG 12 Longchuan County, Dehong, Yunnan CH 8 Cangyuan County, Yunnan Total 84 -

[0048] 2. Cut the young leaves of each pla...

Embodiment 3

[0063] Example 3 Identification and genetic relationship analysis of different Dendrobium chrysalis clones

[0064] 1. Since there is no widely cultivated Dendrobium chrysalis variety that has passed the examination and approval on the market at present, the production of Dendrobium chrysalis is mainly propagated by sowing (seedlings), branching or tissue culture to breed seedlings. This application obtained 12 excellent individual plants of Dendrobium chrysalis through screening, and constructed 12 clones of Dendrobium chrysalis by means of ramets and tissue culture, numbered Line 1-Line 12 respectively, and their main morphological characteristics are shown in Table 8. The 12 above-mentioned clones were used as test materials, and the young leaves of each clone were clipped, and the genomic DNA of Dendrobium chrysalis was extracted using the DP305 kit (Tiangen, China), and 1.2% agarose gel electrophoresis and NanoDrop spectroscopic A photometer measures the quality and conce...

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Abstract

The invention provides a dendrobium chrysotoxum SSR molecular marker primer composition and application thereof, and belongs to the technical field of molecular biology. According to the invention, 25 pairs of SSR molecular marker primers with good amplification effect and high polymorphism are developed and screened from a dendrobium chrysotoxum transcriptome sequence, the genetic diversity and genetic relationship of dendrobium chrysotoxum groups can be analyzed by adopting the SSR molecular marker primers, and germplasm identification and genetic relationship analysis can also be carried out on dendrobium chrysotoxum clones or varieties; and the method has the advantages of high efficiency, accuracy, practicability, no seasonal influence and the like, and lays a foundation for subsequent dendrobium chrysotoxum germplasm resource management, breeding, genetic protection, genetic diversity analysis among groups and in the groups and the like.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a Dendrobium drumstick SSR molecular marker primer composition and application thereof. Background technique [0002] Dendrobium chrysotoxum (Dendrobium chrysotoxum) is a perennial epiphytic herb of the genus Dendrobium orchidaceae. It is one of the widely cultivated Dendrobium species integrating ornamental, food and medicine. It is included in the 2020 edition of "Chinese Pharmacopoeia". Its stems and flowers are dried and soaked in water to drink. It has the functions of beautifying eyesight, calming the nerves and nourishing blood, anti-oxidation and enhancing immunity. At the same time, the stem of Dendrobium chrysalis is spindle-shaped, the leaves are evergreen in four seasons, the flowers are golden yellow, the flowers are large, slightly fragrant, and the flowering period is as long as one month. It can be cultivated and applied in various ways such as on raw s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 李杰王再花叶广英刘海林
Owner ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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