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Primers for screening internal reference genes under low temperature stress in lotus mist and their application

A technology of low temperature stress and internal reference genes, which is applied in the field of screening the primers of internal reference genes under low temperature stress in Lianwu, to achieve the effects of improving detection efficiency, reliability and specificity

Active Publication Date: 2021-04-06
POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the screening of internal reference genes of lotus mist. The present invention selects 7 commonly used internal reference genes, and uses qRT-PCR technology to evaluate their expression stability under different experimental conditions, in order to screen out stable expression genes under low temperature stress. Internal reference genes, laying the foundation for subsequent gene expression work

Method used

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  • Primers for screening internal reference genes under low temperature stress in lotus mist and their application
  • Primers for screening internal reference genes under low temperature stress in lotus mist and their application
  • Primers for screening internal reference genes under low temperature stress in lotus mist and their application

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Embodiment 1

[0016] 1 Materials and handling

[0017] Lianwu for testing Syzygium. (BI.) Merr.et Perry〕The fruit comes from the Lianwu plantation in Dongshan County, Zhangzhou City, Fujian Province. The varieties are 'Zihong' (Tub Ting Jiang), 'Black Pearl' (Heizhenzhu) and 'Emerald' (Feicui). The lotus mist fruits were harvested 10 days, 20 days, 30 days, 40 days and 50 days after full bloom in June-July 2016, and the different sampling periods were recorded as T1-T5. 5 fruits were picked from each tree in 4 directions, a total of 20 fruits, one treatment per tree, repeated 3 times. After cleaning the fruit, separate the peel and pulp with a scraper, quickly put them into liquid nitrogen for quick freezing, and then store them in a -80°C ultra-low temperature refrigerator for later use.

[0018] The seedlings of 'Zihong', 'Black Pearl' and 'Jade' used were from Xiamen Institute of Subtropical Botany. The annual lotus mist cutting seedlings with relatively consistent growth were select...

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Abstract

The invention provides primers for screening internal reference genes under low temperature stress in lotus mist and applications thereof. The primers are: forward primer 5'-CCTGGCCTCTATGATCTTATCC-3', reverse primer 5'-CAGGCATCATGTGCTTTG-3'. The present invention uses qRT-PCR to detect the expression levels of 7 candidate internal reference genes ACT-7, GAPDH, UBQ, α-TUB, CYP20-1, EF-2 and APRT-5 under the development of lotus mist fruit and low temperature stress, through geNorm , NormFinder, BestKeeper and RefFinder software for gene stability evaluation, the results show that the expression of the most stable internal reference gene CYP20-1 under low temperature stress, this not only solves the current situation that there is no internal reference gene in the existing lotus mist quantitative PCR detection, but also provides a basis for the follow-up Laying the groundwork for gene expression work.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to primers for screening internal reference genes under low temperature stress in lotus mist and applications thereof. Background technique [0002] wax apple〔 Syzygium. (BI.) Merr.et Perry〕is Myrtaceae (Mytaceae) Carmine ( Syzygium ) evergreen tree, is a typical tropical fruit tree. Lotus mist fruit is rich in nutrients, has antidiarrheal, analgesic, antibacterial, anti-inflammatory (Tina et al. 2011) and other effects, and is bright in color, crisp and refreshing, and is loved by consumers. It has developed rapidly in suitable planting areas in my country and is an important tropical characteristic fruit. kind. In terms of production, the unstable quality of lotus mist fruit and the low temperature and cold damage in winter are two prominent problems hindering the development of the lotus mist industry, which need to be solved urgently. For a long time, optimized cult...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11C12N15/29
CPCC07K14/415C12Q1/6895C12Q2600/13C12Q2600/158C12Q2600/166
Inventor 许家辉魏秀清章希娟许玲
Owner POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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