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Molecular marker and application of rice glue consistency control gene pul

A technology of molecular marking and gel consistency, which is applied in the field of agricultural biotechnology engineering, can solve problems such as poor accuracy, and achieve the effect of improving gel consistency

Active Publication Date: 2019-05-07
ZHEJIANG NORMAL UNIVERSITY
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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 glue consistency control gene pul
  • Molecular marker and application of rice glue consistency control gene pul
  • Molecular marker and application of rice glue consistency control gene pul

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1. Using the Indel marker PUL-m to identify the polymorphisms of the indica rice Teqing with high gel consistency and the japonica rice Nipponbare with low gel consistency

[0061] The specific method is as follows: select the rice materials Nipponbare and Teqing, cross the Nipponbare and Teqing to obtain its F1, and use the primer PUL-m to identify its polymorphism ( figure 1 ).

[0062] 1. DNA extraction

[0063] 1) Prepare DNA extraction buffer:

[0064] Add 1 volume of DNA extraction solution (0.35M sorbitol; 0.1M Tris, pH8.2; 0.005MEDTA; the rest is water), 1 volume of nuclear lysis solution (0.2M Tris, pH7.5; 0.05M EDTA; 2M NaCl; 0.055MCTAB; 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 solution; the final concentration of sodium bisulfite in DNA extraction buffer was 0.02M.

[0065] The preparat...

Embodiment 2

[0084] Example 2. Using the Indel molecular marker PUL-m to identify sequence differences between the indica rice Teqing with high gel consistency and the japonica rice Nipponbare with low gel consistency

[0085] The specific method is: use the Indel molecular marker PUL-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 ).

[0086] 1. DNA extraction

[0087] 1) Prepare DNA extraction buffer:

[0088] With embodiment 1.

[0089] 2), the paddy rice blade of above-mentioned Nipponbare and special green is carried out as follows respectively:

[0090] With embodiment 1.

[0091] 3) PCR amplification

[0092] With embodiment 1.

[0093] 4) Recovery of PCR products

[0094] The recovery of PCR products was carried out by using the PCR product recovery kit (spin column type, catalog number: DP1403) developed ...

Embodiment 3

[0096] Example 3, using Indel marker PUL-m to carry out assisted selection breeding with high gel consistency

[0097] The specific method is: Teqing, the gene donor parent with high gel consistency, is sequentially crossed, backcrossed and selfed with the japonica rice variety Nipponbare with low gel consistency, and assisted selection of the resulting offspring combined with the molecular marker PUL-m is selected for the segregation population. A single plant with the same belt type as the special green belt type is used for breeding improvement.

[0098] 1. DNA extraction

[0099] 1) Prepare DNA extraction buffer:

[0100] With embodiment 1.

[0101] 2), above-mentioned Nipponbare, Teqing, and the rice blades of the gained offspring are respectively processed as follows:

[0102] With embodiment 1.

[0103] 2. Indel marker detection

[0104] 1), PCR amplification

[0105] With embodiment 1.

[0106] 2), electrophoresis detection

[0107] With embodiment 1.

[0108]...

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Abstract

The invention discloses a molecular marker PUL-m for rice gel consistency control genes PUL. The molecular marker is characterized in that paddy rice is used as a species, primers of the molecular marker are selectively primer pairs, nucleotide sequences are 5'->3', the molecular marker PUL-m in a forward direction is shown as ATCTGATTGAGGCTAGTTTCTGA, and the molecular marker PUL-m in a reverse direction is shown as ATCAACGATCACCAAACATCA. The invention further discloses application of the molecular marker PUL-m. The molecular marker PUL-m can be used for identifying different rice gel consistencies of the paddy rice and / or assisting in selectively breeding progenies of the paddy rice; single plants, with banding patterns consistent with Teqing banding patterns, of the progenies are selected for breeding when the progenies are Nipponbare and Teqing progenies.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology engineering, and in particular relates to a molecular marker related to rice gel consistency control gene PUL and its 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 limitations of t...

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 饶玉春徐江民林晗蒋玲欢黄苗苗符巧丽沈晨辉
Owner ZHEJIANG NORMAL UNIVERSITY
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