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Primer pairs and their application for screening internal reference genes in the development of Lianwu fruit

A developmental process and primer pair technology, applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., to achieve the effects of improving detection efficiency, reliability, and specificity

Active Publication Date: 2020-09-11
POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report about 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 test conditions, in order to screen out the genes that are stably expressed in the fruit development process. Internal reference genes, laying the foundation for subsequent gene expression work

Method used

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  • Primer pairs and their application for screening internal reference genes in the development of Lianwu fruit
  • Primer pairs and their application for screening internal reference genes in the development of Lianwu fruit
  • Primer pairs and their application for screening internal reference genes in the development of Lianwu fruit

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Embodiment 1

[0019] 1 Materials and handling

[0020] Lianwu for testing Syzygium. (BI.) Merr.et Perry〕Fruits come from the Lianwu plantation in Dongshan County, Zhangzhou City, Fujian Province, and the varieties are 'Zihong' (Tub Ting Jiang), 'Black Pearl' (Black Pearl) and 'Jade' (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.

[0021] 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 a primer pair for screening reference genes of wax apple fruit development process and application thereof. The primers comprise an ACT-7 forward primer 5'-CGTTTGGCTCACTCCATACA-3', a reverse primer 5'-AGGTCTTTGTTGGTGAGCTATT-3'; an alpha-TUB forward primer 5'-TCCTTTAAGCCTATGTCGTAACC-3', and a reverse primer 5'-R: TCCGCAGCTTTCTTCTTT-3'. With the utilization of a qRT-PCR detection technology and through geNorm, NormFinder, BestKeeper and RefFinder software for gene stability evaluation, results show that reference genes which are most stably expressed during pulp and pericarp development process are respectively ACT-7 and alpha-TUB. The present situation that existing wax apple quantitative PCR detection has no reference gene is solved, and a foundation is also laid for subsequent gene expression work.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a pair of primers for screening internal reference genes in the development process of lotus mist fruit and its application. 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, optim...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/158C12Q2600/166C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 魏秀清许玲许家辉章希娟
Owner POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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