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Molecular marker and application of rice amylose content micro-controlling gene agps2a

A technology of amylose content and molecular markers, applied in the field of agricultural biotechnology engineering, can solve the problems of poor accuracy and achieve the effect of overcoming the long time period

Active Publication Date: 2015-10-28
CHINA NAT RICE RES INST
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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 often large differences between individual phenotypes and genotypes, so the accuracy of individual selection through field phenotypic traits is poor

Method used

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  • Molecular marker and application of rice amylose content micro-controlling gene agps2a
  • Molecular marker and application of rice amylose content micro-controlling gene agps2a
  • Molecular marker and application of rice amylose content micro-controlling gene agps2a

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Using the Indel marker AGPS2a-m to identify polymorphisms in the japonica rice Nipponbare with low amylose content and the indica rice Teqing with high amylose content

[0058] The specific method is: 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 primers AGPS2a-m to identify its polymorphism ( figure 1 ).

[0059] 1. DNA extraction

[0060] 1) Prepare DNA extraction buffer:

[0061] 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 (that is, the aqueous solution of sodium lauryl-N-methylglycinate); finally add sodium bisulfite to prepare DNA Extraction buffer; final concentration of sodium bisulfite in DNA extra...

Embodiment 2

[0081] Example 2. Using the Indel molecular marker AGPS2a-m to identify sequence differences between the japonica rice Nipponbare with low amylose content and the indica rice Teqing with high amylose content

[0082] The specific method is: use the Indel molecular marker AGPS2a-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 ).

[0083] 1. DNA extraction

[0084] 1) Prepare DNA extraction buffer:

[0085] With embodiment 1.

[0086] 2), the rice leaves of the above-mentioned Nipponbare and Teqing were respectively treated as follows:

[0087] With embodiment 1.

[0088] 3) PCR amplification

[0089] With embodiment 1.

[0090] 4) Recovery of PCR products

[0091] The recovery of PCR products was carried out by using the PCR product recovery kit (centrifugal column type, catalog number: DP1403) develop...

Embodiment 3

[0093] Example 3. Using the Indel marker AGPS2a-m to carry out assisted selection breeding for low amylose content

[0094] The specific method is: the gene donor parent Nipponbare with a low amylose content, and the indica rice variety Teqing with a high amylose content are sequentially crossed, backcrossed and selfed, and the resulting progeny are assisted by selection of the molecular marker AGPS2a-m, and selected to segregate Individual plants with the same band type as the Japan Sunshine band type in the population were used for breeding improvement.

[0095] 1. DNA extraction

[0096] 1) Prepare DNA extraction buffer:

[0097] With embodiment 1.

[0098] 2), the paddy leaves of the above-mentioned Nipponbare, Teqing, and the resulting offspring were respectively treated as follows:

[0099] With embodiment 1.

[0100] 2. Indel marker detection

[0101] 1), PCR amplification

[0102] With embodiment 1.

[0103] 2), electrophoresis detection

[0104] With embodimen...

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Abstract

The invention discloses a molecular marker AGPS2a-m of a rice amylose content micro-control gene AGPS2a. According to the molecular marker AGPS2a-m, paddy is taken as a species, primers of the molecular marker are selected from a primer pair shown in a drawing, the nucleotide sequence of the molecular marker is 5'-3', the forward sequence of the AGPS2a-m is TCTTCTTTTAGATCTTAAATTTCAGA, and the reverse sequence of the AGPS2a-m is CACATCAAAGTTGTAGTTTTGTGA. The molecular marker AGPS2a-m can be applied to the assay of paddy rice amylose content and / or the assisted selective breeding of descendants thereof; for the descendants of Nipponbare and Teqing, single plants with banding patterns consistent with those of Nipponbare are selected from the descendants and are applied to breeding improvement.

Description

technical field [0001] The invention belongs to agricultural biotechnology engineering, and particularly relates to a molecular marker related to rice amylose content regulation gene AGPS2a and its obtaining method and application. 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 lim...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 曾大力钱前李家洋田志喜杨窑龙张光恒郭龙彪高振宇朱丽胡江董国军胡兴民颜美仙
Owner CHINA NAT RICE RES INST
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