Molecular marker for rice grain amylose content micro-control gene ISA and application thereof

An amylose content and molecular marker technology, applied in the field of agricultural biotechnology engineering, can solve problems such as poor accuracy and achieve the effect of overcoming a long time period

Active Publication Date: 2016-08-10
浙江禾天下种业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the effects of one cause and multiple effects, multiple causes and one effect, regulatory genes, and modified genes among genes, there are o

Method used

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  • Molecular marker for rice grain amylose content micro-control gene ISA and application thereof
  • Molecular marker for rice grain amylose content micro-control gene ISA and application thereof
  • Molecular marker for rice grain amylose content micro-control gene ISA and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1. Using the Indel marker ISA-m to identify polymorphisms in the japonica rice Nipponbare with low amylose content and the indica rice Teqing with high amylose content

[0062] The specific method is as follows: select rice materials Nipponbare and Teqing from the germplasm resource bank of the China Rice Research Institute, and use Nipponbare and Teqing to obtain the F1, and use the primer ISA-m to identify its polymorphism ( figure 1 ).

[0063] 1. DNA extraction

[0064] 1) Prepare DNA extraction buffer:

[0065] Add 1 volume of DNA extraction solution (0.35M sorbitol; 0.1M Tris, pH8.2; 0.005M EDTA; the rest is water), 1 volume of nuclear lysis solution (0.2M Tris, pH7.5; 0.05M EDTA ; 2M NaCl; 0.055M CTAB; the rest is water) and 0.4 volume of 5% (mass concentration) sarkosyl solution (i.e. the aqueous solution of sodium lauryl-N-methylglycinate); finally add sodium bisulfite to prepare DNA Extraction buffer; final concentration of sodium bisulfite in DNA e...

Embodiment 2

[0085] Example 2. Using the Indel molecular marker ISA-m to identify sequence differences between the japonica rice Nipponbare with low amylose content and the indica rice Teqing with high amylose content

[0086] The specific method is: use the Indel molecular marker ISA-m to perform PCR amplification on the genomic DNA of Nipponbare and Teqing, entrust Shanghai Yingjun Biotechnology Co., Ltd. to sequence the amplified products, and compare the differences in their sequences ( figure 2 ).

[0087] 1. DNA extraction

[0088] 1) Prepare DNA extraction buffer:

[0089] With embodiment 1.

[0090] 2), the paddy rice blade of above-mentioned Nipponbare and special green is carried out as follows respectively:

[0091] With embodiment 1.

[0092] 3) PCR amplification

[0093] With embodiment 1.

[0094] 4) Recovery of PCR products

[0095] The recovery of PCR products was carried out by using the PCR product recovery kit (spin column type, catalog number: DP1403) developed ...

Embodiment 3

[0097] Example 3. Using the Indel marker ISA-m to carry out assisted selection breeding for low amylose content

[0098] The specific method is: the gene donor parent Nipponbare with low amylose content is crossed, backcrossed and selfed with the indica rice variety Teqing with high amylose content, and the segregation population is selected by the assisted selection of the resulting offspring combined with the molecular marker ISA-m The individual plants with the same middle zone type and the Japanese sunny zone type are used for breeding and improvement.

[0099] 1. DNA extraction

[0100] 1) Prepare DNA extraction buffer:

[0101] With embodiment 1.

[0102] 2), above-mentioned Nipponbare, Teqing, and the rice blades of the gained offspring are respectively processed as follows:

[0103] With embodiment 1.

[0104] 2. Indel marker detection

[0105] 1), PCR amplification

[0106] With embodiment 1.

[0107] 2), electrophoresis detection

[0108] With embodiment 1. ...

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Abstract

The invention discloses molecular marker ISA-m for rice grain amylose content micro-control gene ISA, using rice as a species. A molecular marker primer is selected from following primer pairs, wherein nucleotide sequence is 5'->3', ISA-m forward: ATAGATGCTAATGTGATGTGGC; ISA-m reverse: TGGTATAGGCACAACCGTAGA. The molecular ISA-m is useful in rice grain amylose content identification and/or rice grain amylose progeny assisted breeding.

Description

technical field [0001] The invention belongs to agricultural biotechnology engineering, and particularly relates to a molecular marker ISA-m related to rice amylose content regulation gene ISA and a method for obtaining the same. Background technique [0002] High yield and high quality have always been the long-term goal of rice breeding. After long-term efforts, especially the use of hybrid rice technology, my country has achieved universally recognized achievements in rice production. However, in the past, how to solve people's food and clothing problems has always been the first priority, so the focus of rice breeding work is mostly concentrated on the cultivation of new high-yield varieties of rice, resulting in a serious lag in the breeding of high-quality rice, especially some high-yield hybrid rice General deviations in quality. [0003] Another main reason for the slow progress in rice quality improvement is the complexity of rice quality inheritance and the limit...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 曾大力钱前李家洋朱丽代丽萍胡江张光恒郭龙彪董国军高振宇陈光
Owner 浙江禾天下种业股份有限公司
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