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Internal reference genes suitable for the study of water stress gene expression in lotus and their application

A water stress and gene technology, applied in the field of agricultural biology, can solve the problem of no internal reference gene, and achieve the effect of improving stability

Active Publication Date: 2021-07-09
SUZHOU ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the above problems existing in the prior art, and provide an internal reference gene suitable for the study of gene expression of lotus flooding stress. The present invention solves the current situation that there is no internal reference gene in the study of gene expression of lotus flooding stress

Method used

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  • Internal reference genes suitable for the study of water stress gene expression in lotus and their application
  • Internal reference genes suitable for the study of water stress gene expression in lotus and their application
  • Internal reference genes suitable for the study of water stress gene expression in lotus and their application

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Embodiment

[0028] 1. Material treatment: take lotus seeds as the material, soak the seeds under the conditions of 25°C and 16h / day light after the tail is broken, and select the seeds with the same bud length for the flooding stress test after 1-2 weeks after the emergence of hypocotyls. The flooding stress time was 1h, 2h, 4h, 12h.

[0029] 2. Fluorescence quantitative test: RNA extraction uses leaf bud material, the extraction method refers to the Shanghai Sangon column type plant total RNA extraction and purification kit manual (product number: B518661-0100), cDNA first strand synthesis refers to Shanghai Sangon M-MuLV First-Strand cDNA Synthesis Kit Instructions (Product No.: B532435-0020).

[0030] According to the five internal reference candidate sequences, primers were designed using primer premier5.0 software, and the primer sequences and estimated lengths of amplified fragments are shown in Table 1.

[0031] For the qPCR method, refer to the manual of Shanghai Sangon 2xSG Fast...

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Abstract

The invention discloses internal reference genes suitable for the expression research of lotus flood stress genes. The internal reference genes include: lotus actin gene and ribosomal protein 18sRNA gene. The present invention discloses two internal reference genes for studying the expression of water-flooding stress genes in lotus, and discloses real-time fluorescent quantitative PCR primers designed based on these two genes, solving the problem that there is no internal reference gene in the study of water-flooding stress gene expression in lotus The current situation of the current situation, the internal reference gene of the present invention can be stably expressed when the lotus responds to flooding stress, and the designed real-time fluorescent quantitative PCR primers are used for gene expression analysis when the lotus responds to flooding stress, which can improve the stability, reliability and reliability of the research. repeatability.

Description

technical field [0001] The invention relates to the field of agricultural biotechnology, in particular to the field of lotus flooding stress gene expression, in particular to an internal reference gene suitable for the study of lotus flooding stress gene expression and application thereof. Background technique [0002] The lotus is a perennial emergent plant and the only aquatic flower among the top ten famous flowers in my country. Because of its rich flower colors, long flowering period, and green leaves, it is deeply loved by people, plus it has profound meaning and cultural value. It is an excellent waterscape garden. Landscaping plants and commonly used plants for home gardening are widely used and distributed in our country. Flooding stress: The lotus is a kind of aquatic flower, and the change of the environment makes the lotus often face the impact of the flooding stress. Therefore, mining resources and genes for tolerance to flooding stress is one of the goals of lo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/158C12Q2600/166
Inventor 徐君姜红卫李欣江君蒋华伟
Owner SUZHOU ACADEMY OF AGRI SCI
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