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A molecular marker, primer and application thereof tightly linked to rice high resistant starch content gene sbe3-rs

A high-resistant starch, molecular marker technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of long breeding cycle and low selection efficiency, reduce labor space, The effect of shortening the breeding cycle and improving the breeding efficiency

Inactive Publication Date: 2020-12-11
安徽丰大种业股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Conventional breeding methods based on phenotypic selection have disadvantages such as low selection efficiency and long breeding cycle. It is urgent to inject modern molecular technology, supplemented by high-efficiency genotype-directed selection, in order to quickly and efficiently breed excellent new rice varieties.

Method used

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  • A molecular marker, primer and application thereof tightly linked to rice high resistant starch content gene sbe3-rs
  • A molecular marker, primer and application thereof tightly linked to rice high resistant starch content gene sbe3-rs
  • A molecular marker, primer and application thereof tightly linked to rice high resistant starch content gene sbe3-rs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Obtaining the nucleotide sequence shown in SEQ ID NO: 3 and the nucleotide sequence shown in SEQ ID NO: 4 by the special primer pair for character identification of the highly resistant starch mutant gene sbe3-rs

[0025] 1.1 Parental Genome Amplification

[0026] The parents used to create the rice recombinant inbred line (RIL) population were Jiangtangdao 1 No. 4 and FD1710 bred by Anhui Fengda Seed Industry Co., Ltd., respectively. FD1710 is the female parent, which does not contain the high resistant starch mutant gene sbe3-rs; Jiangtangdao 1 is the male parent, which contains the high resistant starch mutant gene sbe3-rs.

[0027] Using the CTAB method to extract the genomic DNA of the leaves of the parents, the primer pair was designed with the sbe3-rs sequence (web site) published on the NCBI, and the nucleotide sequence shown in SEQ ID NO: 5 and the nucleotide shown in SEQ ID NO: 6 Composed of acid sequences, entrusted to Beijing Zixi Biological Company to synt...

Embodiment 2

[0035] Correlation verification between the amplified product of QK1 and the traits of high resistant starch content in rice

[0036] Using Jiangtangdao 1 containing the high resistant starch mutant gene sbe3-rs and FD1710 without the resistant starch mutant gene sbe3-rs as parents, 285 F 2 The substitute material is the test object. Genomic DNA was extracted from the rice to be tested by the alkaline cooking method, and the specific steps were the same as in Example 1. PCR amplification experiments were performed with QK1. The reaction system of PCR amplification is: 10X Buffer 2μl (containing Mg 2+ ), dNTP 0.4 μ l (10 mM), the nucleotide sequence shown in SEQ ID NO: 3 and the nucleotide sequence shown in SEQ ID NO: 4 each 2 μ l (5 μ M), the genomic DNA 1 μ l that the rice to be tested extracts, taq enzyme 0.5μl (5U / μl), the reaction volume is 20μl, drop a drop of mineral oil to cover; the reaction program of PCR amplification is: 94°C 5min; 94°C 60s, 53.5°C 60s, 72°C 60s,...

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Abstract

The invention discloses a molecular marker closely linked to a high resistant starch-content gene sbe3-rs of rice, a primer for amplifying the molecular marker and a method for detecting the high resistant starch-content gene sbe3-rs of the rice. The nucleotide sequence of the molecular marker is shown as SEQ ID NO: 1 or SEQ ID NO: 2. The molecular marker disclosed by the invention is located in the high resistant starch-content gene sbe3-rs and is closely linked to the high resistant starch-content gene sbe3-rs of the rice; and compared with a non-genomic inner marker, the molecular marker disclosed by the invention cannot be separated and is capable of increasing the breeding efficiency more effectively.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a molecular marker closely linked with rice high resistant starch content gene sbe3-rs, a primer and application thereof. Background technique [0002] Starch is the main source of human food and energy. After it enters the human body, most of it is digested and absorbed after being enzymatically hydrolyzed in the small intestine. A small part can escape enzymatic hydrolysis and enter the small intestine. The former is digestible starch (digestible starch, DS), which is resistant starch (resistant starch, RS). RS has important physiological functions such as lowering blood sugar, lowering blood fat, promoting intestinal health, and promoting the digestion and absorption of minerals. High-RS diets have less insulin response than those with low-RS diets, which is of great benefit to diabetic patients in controlling postprandial blood glucose levels, especially for non-insulin-dependen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 朱先飞李婧婧何立斌付求来张志清赵峰潘华柏瑞
Owner 安徽丰大种业股份有限公司
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