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DNA barcode standard detection gene for distinguishing different elephant grass species and its application

A standard detection and barcode technology, which is applied in the field of DNA barcode standard detection of genes, can solve problems such as lack, and achieve the effects of improving efficiency, high accuracy, and short time-consuming

Active Publication Date: 2022-05-27
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Judging from the existing morphological research, there is still a lack of intuitive and obvious differences to distinguish different elephant grass species and their hybrids

Method used

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  • DNA barcode standard detection gene for distinguishing different elephant grass species and its application

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Effect test

Embodiment 1

[0032] DNA barcode standard detection genes for differentiating different elephant grass varieties, and the nucleotide sequences of the DNA barcode standard detection genes are shown in SEQ ID NO.1-SEQ ID NO.5.

[0033] The DNA barcode standard detection gene acquisition method of the present embodiment is as follows:

[0034] (1) Preliminary identification of Guimin Elephant Grass, Dwarf Elephant Grass, Purple Elephant Grass, Giant Juncao and Hybrid Elephant Grass by different biological characteristics;

[0035] (2) Germination of Elephant Grass seeds: 4 different Elephant Grass varieties (Guimin Elephant Grass, Dwarf Elephant Grass, Purple Elephant Grass and Giant Juncao) and 2 Elephant Grass Hybrid Seeds (Reyan No. 4 Elephant Grass) were taken respectively. and / or Guimu No. 1 Elephant Grass) 20 capsules, placed in a petri dish covered with moist filter paper, the petri dish was placed in a 24°C incubator (16h day, 8h night) for 14 days, and the filter paper was replaced re...

Embodiment 2

[0040] The standard detection genes of the above-mentioned SEQ ID NO.1-SEQ ID NO.5 can distinguish the following 6 varieties of elephant grass: Guimin Yinxiangcao, Dwarf elephant grass, Purple elephant grass, Jujuncao and hybrid elephant grass:

[0041] Among them, the nucleic acid sequence of the DNA barcode standard detection gene corresponding to Guimin Yinxiangcao is shown in SEQ ID NO.1;

[0042] The nucleic acid sequence of the DNA barcode standard detection gene corresponding to Dwarf Elephant Grass is shown in SEQ ID NO.2;

[0043] The nucleic acid sequence of the DNA barcode standard detection gene corresponding to the purple elephant grass is shown in SEQ ID NO.3;

[0044] The nucleic acid sequence of the DNA barcode standard detection gene corresponding to Jujuncao is shown in SEQ ID NO.4;

[0045] The DNA barcode standard detection genes of the hybrids Guimu No. 1 and Reyan No. 4 are the same, and the corresponding nucleic acid sequences are shown in SEQ ID NO.5. ...

Embodiment 3

[0047] Based on the high degree of conservation and stability of the above-mentioned gene sequences, the above-mentioned DNA barcode standard detection genes can be used to distinguish the above-mentioned 6 different elephant grass varieties.

[0048] The method of using the above DNA barcoding standard to detect genes to distinguish different elephant grass varieties is as follows:

[0049] (1) Elephant grass seed germination: Take the seeds of unknown elephant grass species, put them in a petri dish covered with moist filter paper, place the petri dish in a 24°C incubator (16h in the day, 8h in the night) for 14 days, and Replace the filter paper regularly;

[0050] (2) Extraction of seedling DNA: extract by SDS method, put the whole seedling in a 1.5ml centrifuge tube, put it in a steel ball and shake it well; add 600μl DNA buffer and mix well, water bath at 65°C for 10 minutes, and shake for 1- 2 times; put it to room temperature, add 195 μl l5mol of potassium acetate (PH...

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Abstract

The invention relates to the field of biotechnology, in particular to DNA barcode standard detection genes for distinguishing different elephant grass varieties and their applications. The DNA barcode standard detection genes of the present invention can quickly distinguish and identify different elephant grass varieties and their hybrids. The accuracy of the method is high, the operation process is simple, and the characteristics of short time-consuming, especially, adopt the present invention to accurately identify different elephant grass varieties and hybrids thereof, and can be applied to identification and variety protection of elephant grass varieties, which is very important for elephant grass varieties. The scientific protection of germplasm resources is of great significance in order to efficiently, quickly and accurately identify different Elephant grass species and their hybrids. Compared with the traditional morphological identification method, the method of the invention can be well completed in a relatively short period of time under laboratory conditions, and the efficiency can be greatly improved. It has good promotion value.

Description

【Technical field】 [0001] The invention relates to the field of biotechnology, in particular to a DNA barcode standard detection gene for distinguishing different elephant grass varieties and its application. 【Background technique】 [0002] Elephant grass (Cenchrus purpureus) is named after elephants love to eat. It is a large perennial herb of the family Poaceae and millet, native to Africa. Elephant grass is widely used as an important forage crop in the tropics and subtropics around the world because of its large biomass, rapid growth and strong adaptability. Become a potential energy grass. At present, the main cultivated elephant grass products in China include: Guimin Yinxiangcao, Dwarf elephant grass, Purple elephant grass, Reyan No. 4 elephant grass, Guimu No. 1 elephant grass, and Jujuncao; among them, Reyan No. 4 elephant grass, Guimu No. 1 Elephant Grass is a hybrid, and the shape of Elephant Grass varieties and their hybrids is very similar. In addition, the re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 张吉宇闫启易显凤李洁张正社卢丽燕
Owner LANZHOU UNIVERSITY
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