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A marker gene for identifying heat-tolerant blueberry varieties and its application

A technology for marking genes and blueberries, applied in the field of plant biology, can solve the problems of consuming manpower and material resources, long experiment cycle, etc., and achieve the effect of low cost, high repeatability and simple operation

Active Publication Date: 2017-12-26
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the conventional field natural identification method has the characteristics of easy operation, it needs to be cultivated for several years before it can be evaluated, and only large-scale cultivation can ensure the accuracy of the results, so it often requires a lot of manpower and material resources, and there is still an experimental period. long-term disadvantages

Method used

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  • A marker gene for identifying heat-tolerant blueberry varieties and its application

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Experimental program
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Effect test

Embodiment 1

[0018] A marker gene for identifying blueberry heat-resistant varieties, o-type thioredoxin (TRXo), its nucleotide sequence SEQID No.1.

Embodiment 2

[0020] For the nucleotide sequence of the above-mentioned genes, Primer Premier 5.0 software (a professional software developed by Premier Company in Canada for the design and evaluation of PCR or sequencing primers and hybridization probes) was used to design qRT-PCR primers, SEQ ID No. 2 and SEQ ID No.3.

Embodiment 3

[0022] Using the CTAB method to extract the total RNA of the blueberry leaves to be tested under the control and 45°C high temperature stress treatment for 6 hours, and reverse-transcribe it into cDNA as a template, and use GAPDH as an internal reference gene (SEQ ID No.4 and SEQ ID No.5) for qRT -PCR identification (Taking Dalian Bao Bioluminescent Quantitative Kit RR82LR as an example):

[0023] The qRT-PCR reaction system is as follows:

[0024]

[0025] Reaction conditions: 95°C for 30s; 95°C for 5s, 58°C for 35s, a total of 40 cycles, for qRT-PCR detection, and using 2 -ΔΔCt method for computational analysis.

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Abstract

The invention relates to a marker gene for identifying blueberry heat-resistant varieties and application thereof, belonging to the field of plant biotechnology. Utilizing the marker gene, thioredoxin o-type (TRXo), was able to up-regulate expression to different degrees in different heat-tolerant blueberry varieties under high temperature stress. Primers were designed using the CDS sequence of the gene as a template, and the detection method was to artificially simulate the climate and heat stress at 45°C for 6 h, extract total RNA, reverse transcribe it into cDNA as a template, perform qRT‑PCR detection, and use the 2‑ΔΔCt method to calculate analyze. By analyzing the ratio of the expression of the gene under heat stress and the control, if the ratio is greater than or equal to 5, the blueberry variety can be identified as a heat-tolerant variety. Compared with the field natural identification method, the method is faster, more stable and more accurate, and is the basis for rapid screening of heat-resistant varieties for popularization and planting and breeding of new heat-resistant varieties.

Description

technical field [0001] The invention relates to a marker gene o-type thioredoxin (TRXo) for identifying blueberry heat-resistant varieties and an application thereof, belonging to the field of plant biotechnology. Background technique [0002] As a new fruit, blueberry (Vaccinium spp.) has high nutritional value and economic benefits. Due to the unique health care function of blueberry, it is one of the most popular small fruits in developed countries such as the United States and Japan, and is listed by the International Food and Agriculture Organization. One of the top five healthy foods for human beings, it can be called "the king of the third generation fruit in the world". Blueberries are originally produced in cold northern regions. Planting blueberries in subtropical regions can not only prolong the growth period, but also greatly advance the fruit maturity period. Blueberries are currently at a high price in the domestic market. However, due to the hot and dry summe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/53C12N15/11
Inventor 陈文荣施洁余柯达朱心怡吴洁慧陈晓青
Owner ZHEJIANG NORMAL UNIVERSITY
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