Hybrid Chinese tuliptree rapid molecular detection specific primer and use method thereof

A technology of hybrid Liriodendron and molecular detection, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems that cannot achieve rapid and accurate identification, affect the reliability of detection results, and RAPD Large impact and other problems, to achieve the effect of simple and fast operation, convenient material collection and high sensitivity

Inactive Publication Date: 2009-01-21
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the molecular detection and variety identification based on RAPD technology has the following main defects: (1) The detection of RAPD is greatly affected by the reaction conditions, and its repeatability and stability are poor, which seriously affects the credibility of the detection results
(2) Multiple pairs of primers need to be used, the operation is cumbersome and time-consuming, and the goal of rapid and accurate identification cannot be achieved

Method used

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  • Hybrid Chinese tuliptree rapid molecular detection specific primer and use method thereof
  • Hybrid Chinese tuliptree rapid molecular detection specific primer and use method thereof
  • Hybrid Chinese tuliptree rapid molecular detection specific primer and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Primer LT008L / R was used for species identification of Liriodendron tulipifera, Liriodendron tulipifera and hybrid Liriodendron tulipifera.

[0034] The primer pair LT008L / R was used to identify 15 samples each of Liriodendron tulipifera, Liriodendron tulipifera and Hybrid tulipwood. Results The 190bp product was specifically amplified in the Liriodendron tulipifera population, the 180bp product was specifically amplified in the Liriodendron tulipifera population, and both 180bp and 190bp products were amplified in the hybrid Liriodendron tulipifera population. 10 μL of the reaction system for PCR, including 1 μL of template DNA, 1 μL of 10×Buffer, 0.2 μL of 10 mM dNTPs (Shanghai Jierui Biological Co., Ltd.), 25 mM MgCl 2 1 μL, F-primer 0.25 μL, R-primer 0.25 μL, Ampli TaqGold 0.05 μL (Shanghai Jierui Biological Co., Ltd.), ddH2O 6.25 μL. The PCR reaction program is: pre-denaturation at 94°C for 4 minutes; gradient cooling amplification: 15 cycles of 94°C fo...

Embodiment 2

[0038] Example 2: The primers LT008L / R were used to identify the species of Liriodendron tulipifera, Liriodendron tulipifera and Hybrid tulipwood different from Example 1.

[0039] The primer pair LT008L / R was used to identify 15 samples of Liriodendron tulipifolia, Liriodendron tulipifera and hybrid Liriodendron tulipifera different from those in Example 1. Results The 190bp product was specifically amplified in the Liriodendron tulipifera population, the 180bp product was specifically amplified in the Liriodendron tulipifera population, and both 180bp and 190bp products were amplified in the hybrid Liriodendron tulipifera population. kind of product. The PCR prescription system and PCR reaction procedure are all the same as the detection and analysis steps of Liriodendron tulipifera in Example 1. The details of the samples used are shown in Table 2, and the test results are shown in figure 2

[0040] Table 2. The specific amplification results of 15 samples of Liriodendr...

Embodiment 3

[0042] Embodiment 3: Using the specific primer LT008L / R of the present invention to control the pollination progeny identification of Liriodendron tulipifera multi-paternal mixing, using the primer pair LT008L / R, the female parents are Nanka, Lewis and Lushan, and the range of the male parents is known ( Lushan, Fuyang, Missouri, South Carolina, Lewis) 86 samples of the control pollination progeny for species identification. The PCR prescription system and PCR reaction procedure are all the same as the detection and analysis steps of Liriodendron tulipifera in Example 1. The specific results of the experiment are shown in Table 3.

[0043] Table 3. The identification results of the primer LT008L / R on the pollination progeny of Liriodendron tulipiflora mixed with multiple male parents

[0044]

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Abstract

The invention discloses primers for detecting hybrid trembling poplar molecule and a usage thereof, being specially used in quick molecular detection and identification of trembling poplar, liriodendron tulipifera and hybrid trembling poplar, and belonging to the molecular identification field for new plant variety. A pair of SSR primers, i.e. LT008L/R is designed, and can realize specific amplification of a 190bp product inside the trembling poplar, a 180bp product inside liriodendron tulipifera and a 180bp product, and a 190bp product inside hybrid trembling poplar. The primer adopts a pair of specific SSR primers to carry out once PCR amplification reaction, and carries out electrophoresis by use of 8 percent polyacrylamide gel; moreover, judgment can be carried out directly according to product fragment. The primers can be directly used for the species level identification of hybrid trembling poplar, trembling poplar and liriodendron tulipifera.

Description

technical field [0001] The invention relates to molecular detection primers of Liriodendron tulipifera and usage thereof, which are specially used for rapid molecular detection and identification of Liriodendron tulipifera, Liriodendron tulipifera and hybrid Liriodendron tulipifera, and belong to the field of identification of new plant varieties. Background technique [0002] There are 2 existing species of Liriodendron in Magnoliaceae, one is Liriodendron (L.tulipiferaLinn.); the other is Liriodendron [L.chinense (Hemsl.) Sarg.]. Liriodendron grows rapidly, with a round and straight trunk, excellent material, few pests and diseases, peculiar leaf shape, elegant corolla color, and high ornamental value. Therefore, Liriodendron is an excellent landscaping and industrial timber tree species (Wang Zhangrong, et al. Hybrid breeding and utilization of Liriodendron species [M]. Beijing: China Forestry Press, 2005). [0003] Since Professor Ye Peizhong, a late well-known tree gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 李火根张红莲胥猛冯源恒
Owner NANJING FORESTRY UNIV
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