Primer for screening reference genes of syzygium samarangense under low-temperature stress and application thereof

A technology of low temperature stress and internal reference gene, which is applied in the field of screening primers for internal reference genes under low temperature stress of Lianwu fog, and achieves the effects of improving stability, improving detection efficiency and improving reliability.

Active Publication Date: 2017-12-15
POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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  • Abstract
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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 expr...

Method used

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  • Primer for screening reference genes of syzygium samarangense under low-temperature stress and application thereof
  • Primer for screening reference genes of syzygium samarangense under low-temperature stress and application thereof
  • Primer for screening reference genes of syzygium samarangense under low-temperature stress and application thereof

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Embodiment 1

[0016] 1 Materials and Handling

[0017] For the test lotus mist [ Syzygium. samarangense (BI.) Merr.et Perry] The fruit comes from the Lianwu plantation in Dongshan County, Zhangzhou City, Fujian Province, and the varieties are 'Purple Red' (Tub Ting Jiang), 'Black Pearl' (Heizhenzhu) and 'Feicui' (Feicui). The lotus mist fruits were collected from June to July 2016 at 10 d, 20 d, 30 d, 40 d and 50 d after full bloom, and different sampling periods were recorded as T1-T5. 5 fruits were picked from each tree in 4 directions, 20 fruits in total, and each tree was treated as 1, repeated 3 times. After cleaning the fruit, separate the peel and pulp with a peeling knife, 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 'Purple Red', 'Black Pearl' and 'Emerald' were from Xiamen Institute of Subtropical Botany. Choose the annual lotus foggy cutting seedlings with c...

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Abstract

The invention provides a primer for screening reference genes of syzygium samarangense under low-temperature stress and application thereof. The primer comprises forward primer 5'-CCTGGCCTCTATGATCTTATCC-3' and reverse primer 5'-CAGGCATCATGTGCTTTG-3'. The expression level of 7 candidate reference genes ACT-7, GAPDH, UBQ, alpha-TUB, CYP20-1, EF-2 and APRT-5 under fruit development and low-temperature stress of syzygium samarangense can be detected by utilizing qRT-PCR, and gene stability is evaluated through geNorm, NormFinder, RestKeeper and RefFinder software. Results indicate that the reference gene CYP20-1 is expressed most stably under low-temperature stress, so that the situation that no reference gene exists in existing syzygium samarangense quantitative PCR detection can be solved, and foundation is laid for subsequent gene expression work.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer used for screening an internal reference gene under low temperature stress of the lotus mist and its application. Background technique [0002] wax apple〔 Syzygium. samarangense (BI.) Merr.et Perry] is the genus of Mytaceae (Mytaceae) Syzygium ) evergreen tree, is a typical tropical fruit tree. Lianwu 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 is developing rapidly in suitable planting areas in my country and is an important tropical fruit. kind. In terms of production, unstable fruit quality and low temperature and cold damage in winter are two prominent problems hindering the development of the lotus fog industry, which need to be solved urgently. For a long time, optimizing cultivation measures a...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/29
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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