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Expression detection method for sweet sorghum sucrose phosphate synthase gene and amplification primer

A technology for phosphate synthase and gene amplification, which is applied in the field of amplification primers, sweet sorghum sucrose phosphate synthase gene, and expression detection of sweet sorghum sucrose phosphate synthase gene, which can solve the problems of no comprehensive reports and achieve band The effects of clearness, shortening the breeding cycle, and overcoming difficulties in phenotypic identification

Inactive Publication Date: 2018-08-24
SHENYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no comprehensive report on the detection method of SPS gene expression in sweet sorghum

Method used

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  • Expression detection method for sweet sorghum sucrose phosphate synthase gene and amplification primer
  • Expression detection method for sweet sorghum sucrose phosphate synthase gene and amplification primer
  • Expression detection method for sweet sorghum sucrose phosphate synthase gene and amplification primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1: Correlation analysis between sweet sorghum sucrose phosphate synthase (SPS) activity and sucrose content

[0076] The present invention is by measuring the F of male parent, female parent and their hybrid 5 The activity of SPS in leaves of high-sugar group and low-sugar group, to understand the variation of SPS between different groups in different periods. Parents and F 5 The activity of SPS in leaves of high-sugar group and low-sugar group increased gradually. At the maturity stage, the activities of the three sweet sorghum varieties, the female parent, the high-sugar population and the low-sugar population, all reached the maximum values ​​of 14.86, 17.63, and 13.76 μmol / h.g.FW; dropped. There was no significant difference in the activities of SPS among the four varieties at the seedling stage, but there was a difference in the activities of SPS among the four varieties from the jointing stage. The SPS activity of the high-sugar group and the low-sugar...

Embodiment 2

[0077] Implementation example 2: Analysis of extraction results of sweet sorghum RNA

[0078] Agarose gel electrophoresis detection showed that the RNA extracted by the modified Trizol method had three bands, and the brightness of the 28S band in the total RNA was about twice that of the 18S band, indicating that it was less degraded during the extraction process. The integrity of the extracted RNA is good. The 5S band is relatively light, indicating that there is no contamination of RNA, which can be used as a template in the next experiment.

Embodiment 3

[0079] Implementation example 3: sucrose phosphate synthase (SPS) primer design

[0080] The primers in Table 1 were designed based on a known cDNA sequence of the coding region of sweet sorghum SPS3-1 gene, using Oligo6 software and Primer primer software. Named as SPS-1, β-Actin is the housekeeping gene, and the lengths of the target fragments amplified by the two pairs of primers are 121bp and 210bp respectively.

[0081] Table 1 Primers used for SPS gene amplification

[0082]

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Abstract

The invention discloses an expression detection method for a sweet sorghum sucrose phosphate synthase gene and an amplification primer. Sweet sorghum LTR102 (female parent), sorghum 654 (male parent)and a high sugar group and a low sugar group of a generation F5 obtained by hybridation of the sweet sorghum LTR102 and the sorghum 654 are used as test materials; the sucrose contents and the activities of the sucrose phosphate synthase (SPS) in leaves of the sweet sorghum LTR102, the sorghum 654 and the high sugar group and the low sugar group of the generation F5 in different periods are measured in the seedling stage, the elongation stage, the heading period and the maturation period, so as to determine the relevance between the activity of the SPS of the sweet sorghum and the sucrose content. The SPS gene expressions of the test materials in different developing periods are qualitatively analyzed, so as to determine a difference between the SPS gene expressions in different periods. The difference between SPS gene expression quantities of the test materials in different growing periods are quantitatively analyzed, and the relative concentration of the SPS gene expression and the sucrose content are subjected to relevance analysis; a result shows that the relative concentration of the SPS gene expression and the sucrose content are obviously relevant; and a quick detection method for the sweet sorghum SPS gene is established according to the result of the invention, so as to achieve the aims of quickly detecting and determining the best harvest time and screening novel sweet sorghum materials with high sucrose content.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a sweet sorghum sucrose phosphate synthase (SPS) gene, in particular to an expression detection method and an amplification primer for the sweet sorghum sucrose phosphate synthase gene. Background technique [0002] Sucrose is the main product of photosynthesis in higher plants, the main form of carbon transport, the main form of carbohydrate accumulation and storage, and the main substrate of "sink metabolism". It has a special position in plant sugar metabolism (Farrar et al. ,2000). Especially for sweet sorghum, as a good raw material for sugar industry, production of alcohol, fiber and feed mainly depends on the rich sugar in the stalk, of which 85% is sucrose, 9% glucose, 6% fructose (Gnansounou et al.,2005 ). The use of sweet sorghum sugar to produce ethanol as an alternative energy source has aroused great attention in recent years. To explore the mechanism of sweet sorghum su...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851C12Q1/6895C12N15/11
CPCC12Q1/6851C12Q1/6895C12Q2600/13C12Q2600/158C12Q2531/113C12Q2563/107
Inventor 李玥莹逄洪波李雪梅马莲菊邹剑秋王兰兰张颖魏鑫高华晨钱美玲
Owner SHENYANG NORMAL UNIV
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