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Identification and application of sugarcane SPSB gene isotype 2

An isotype, gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem of molecular assisted breeding yet to be developed and so on

Pending Publication Date: 2021-01-12
SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, my country's sugarcane breeding is basically based on traditional hybridization, and molecular assisted breeding has yet to be developed.

Method used

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  • Identification and application of sugarcane SPSB gene isotype 2
  • Identification and application of sugarcane SPSB gene isotype 2
  • Identification and application of sugarcane SPSB gene isotype 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Cloning and sequencing analysis of the cDNA sequence of sugarcane SPSB gene of embodiment 1

[0042] 1. Sample RNA extraction and cDNA synthesis

[0043]In the present invention, sugarcane YZ14-401 leaves at the mature stage are used as detection objects, and the total RNA is extracted with the EasyPurePlant RNA Kit provided by Quanshijin Company, and finally dissolved in sterilized ultrapure water, and stored at 4°C after detection. The OD value of the RNA sample at 260nm and 280nm was measured with a UV photometer. Calculate the RNA content and the ratio of OD260 / OD280 to evaluate the quality of RNA extraction; after the RNA detection is completed, take out a certain amount and dilute it to 10ng / μL for the synthesis template of cDNA. First-Strand cDNA Synthesis SuperMix kit was used to synthesize cDNA.

[0044] 2. Primer design

[0045] The published SPSB gene sequence was obtained from NCBI, and the GenBank ID was JN584485. Primer5.0 software was used to design p...

Embodiment 2

[0055] Cloning and sequencing analysis of the cDNA sequence of the sugarcane SPSB gene of embodiment 2

[0056] 1. Sample RNA extraction and cDNA synthesis

[0057] In the present invention, sugarcane YZ14-401 leaves at the mature stage are used as detection objects, and the total RNA is extracted with the EasyPurePlant RNA Kit provided by Quanshijin Company, and finally dissolved in sterilized ultrapure water, and stored at 4°C after detection. The OD value of the RNA sample at 260nm and 280nm was measured with a UV photometer. Calculate the RNA content and the ratio of OD260 / OD280 to evaluate the quality of RNA extraction; after the RNA detection is completed, take out a certain amount and dilute it to 10ng / μL for the synthesis template of cDNA. First-Strand cDNA Synthesis SuperMix kit was used to synthesize cDNA.

[0058] 2. Primer design

[0059] The published SPSB gene sequence was obtained from NCBI, and the GenBank ID was JN584485. Primer5.0 software was used to des...

Embodiment 3

[0066] Example 3 Cloning and sequencing analysis of sugarcane SPSB gene cDNA sequence

[0067] 1. Sample RNA extraction and cDNA synthesis

[0068] In the present invention, sugarcane YZ14-401 leaves at the mature stage are used as detection objects, and the total RNA is extracted with the EasyPurePlant RNA Kit provided by Quanshijin Company, and finally dissolved in sterilized ultrapure water, and stored at 4°C after detection. The OD value of the RNA sample at 260nm and 280nm was measured with a UV photometer. Calculate the RNA content and the ratio of OD260 / OD280 to evaluate the quality of RNA extraction; after the RNA detection is completed, take out a certain amount and dilute it to 10ng / μL for the synthesis template of cDNA. First-Strand cDNA Synthesis SuperMix kit was used to synthesize cDNA.

[0069] 2. Primer design

[0070] The published SPSB gene sequence was obtained from NCBI, and the GenBank ID was JN584485. Primer5.0 software was used to design primers in th...

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Abstract

The invention relates to identification and application of sugarcane SPSB gene isotype 2, and belongs to the technical field of plant molecular breeding. A cDNA sequence of high-sugar sugarcane YZ14-401 is used as a template, SPSB2351 is used as a primer for PCR amplification, a haplotype of an SPSB gene is screened through sequencing, a fifth intron part retention sequence exists between a fifthexon and a sixth exon, and sequences at two ends of the primer are completely the same as a CDS sequence of the SPSB gene. The SPSB gene isotype 2 can be used for designing a specific primer in a fifth intron retention section, identifying the expression of the isotype 2 and carrying out molecular marker-assisted selection. The sequence required for identifying the gene expression level of the SPSB gene isotype 2 is provided for sugarcane sugar trait marker-assisted breeding; tracking, identification and expression abundance detection of different haplotypes or isotypes of the SPSB gene in sugarcane filial generations are facilitated; and the application has important significance in rapid establishment of sugarcane germplasm with excellent heredity.

Description

technical field [0001] The invention belongs to the technical field of plant molecular breeding, and in particular relates to a sugarcane SPSB gene (JN584485) isoform 2 and its identification method, and the application of the isoform gene-specific sequence in the identification and expression of SPSB alleles to develop molecular markers. Background technique [0002] With the development and progress of gene sequencing technology and bioinformatics, it has brought hope to the research of polyploid plants such as sugarcane, which can solve the problem of unclear positioning and tracking of sugarcane hybridization purposes and functional traits. The study of genes has brought possibility, so the traits of hybrid offspring are seriously separated. A gene has incomplete expression or different copies in controlling a certain trait. After mRNA is translated into cDNA, there are multiple isotypes of genes. The difference in the efficacy of the gene controlling related traits will...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/11C12Q1/686C12Q1/6895
CPCC07K14/415C12Q1/686C12Q1/6895C12Q2600/13C12Q2600/158C12Q2600/172C12Q2565/125C12Q2535/122
Inventor 刘洪博陆鑫李纯佳毛钧李旭娟林秀琴徐超华孔春艳刘新龙
Owner SUGARCANE RES INST OF YUNNAN ACADEMY OF AGRI SCI