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Chive internal reference gene and its screening method and application

An internal reference gene and chive technology, applied in the field of molecular biology, can solve the problems that restrict the research process of chive molecular breeding, achieve the effects of shortening the detection time, improving detection efficiency, and optimizing the PCR amplification program

Active Publication Date: 2021-06-25
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This seriously restricts the research progress of chive molecular breeding

Method used

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  • Chive internal reference gene and its screening method and application
  • Chive internal reference gene and its screening method and application
  • Chive internal reference gene and its screening method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] An internal reference gene of chives, the internal reference gene is AfActin, and the nucleotide sequence of the internal reference gene AfActin is shown in SEQ ID NO.1.

Embodiment 2

[0030] A primer for PCR amplification of an internal reference gene AfActin, the nucleotide sequence of the PCR primer is: the forward sequence is shown in SEQ ID NO.2; the reverse sequence is shown in SEQ ID NO.3.

Embodiment 3

[0032] A method for screening internal reference genes of chives, comprising the following steps:

[0033] 1. Materials and methods

[0034] 1. Test material

[0035] The heat-resistant "Guixiangcong No. 1" potted plant and the heat-labile "AF35" potted plant were used as research materials.

[0036] Place the chive potted plants in a light incubator. The conditions of the control group are: 6:00-20:00 during the day: 23°C, humidity 70%, light 3. 20:00-6:00 the next day, 20°C, humidity 65%, light 0; high temperature stress treatment group conditions 6:00-20:00 during the day: 38°C, humidity 70%, light 3. 20:00-6:00 the next day, 35°C, humidity 65%, light 0; The shallot leaves were collected at 12h, 24h, 48h, and 72h, and 10 plants were treated for each treatment, and each stress treatment was biologically repeated three times. The materials were quick-frozen in liquid nitrogen and stored at –80°C for later use.

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Abstract

The invention provides an internal reference gene of chives and its screening method and application, belonging to the field of molecular biology. Select 13 candidate internal reference genes Ac18S, Af18s, Af28s, AcActin, AfActin, AfTUA, AfTUB, AsSAND, AfMYH, AfGAPDH, AfUBQ, AfEF1a, AfCYC, and use real-time fluorescent quantitative RT-qPCR technology to express their expression levels under high temperature stress The stability of the 13 internal reference genes was evaluated in combination with GeNorm, NormFinder and BestKeeper software, and finally a comprehensive analysis was conducted to obtain the most suitable internal reference gene for chives under high temperature stress as AfActin, which provided a basis for subsequent scientific research on gene expression in chives. refer to.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to an internal reference gene of chives and a screening method and application thereof. Background technique [0002] Chives, also known as: green onions, chives, northern onions, fire onions. Latin name: Allium fistulosumL.var.caespitosum Makino. Liliaceae, Allium, bulbs clustered, oblong-ovate, narrowly ovate or ovate-cylindrical; bulb skin reddish-brown, purple-red to yellow-white, membranous Textured or thin leathery, not cracked. The leaves are hollow cylindrical, acuminate to the top, dark green, often slightly white powder. The plant is small, the leaves are very thin, the texture is soft, the taste is fragrant, slightly spicy, and it is mainly used for seasoning and removing fishy smell. It is native to western Asia and is widely cultivated in southern my country, as well as in some parts of Europe and Asia. Chives are not only an important spicy seasoning that is indisp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/11C12Q1/6809C12Q1/6895
CPCC07K14/415C12Q1/6809C12Q1/6895C12Q2600/158C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/101C12Q2537/165
Inventor 郭元元陈振东张力陈琴蒋月喜李洋宋焕忠车江旅秦龙妹
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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