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Molecular marker of rice seed salt tolerant germination major QTL qGR2 and its application

A technology of molecular markers and rice seeds, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of few researches on seed germination period, and achieve fast and simple identification methods, high selection efficiency, Choose a well-targeted effect

Active Publication Date: 2014-03-19
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the research on QTL mapping of rice salt tolerance mainly focuses on the seedling and maturity stages, and there are few studies on the seed germination stage.

Method used

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  • Molecular marker of rice seed salt tolerant germination major QTL qGR2 and its application
  • Molecular marker of rice seed salt tolerant germination major QTL qGR2 and its application
  • Molecular marker of rice seed salt tolerant germination major QTL qGR2 and its application

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Experimental program
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Effect test

Embodiment 1

[0031] (1) Materials and methods:

[0032] 1. Material: strong salt-tolerant variety Chive Green and IR 26 Hybrid ( figure 1 ), to obtain 150 RIL families F 2:9 , for initial positioning, and then select the background-like IR containing the qGR2 locus 26 The families were backcrossed and selfed to obtain segregation BC 2 f 2 Populations are isolated, fine-mapped, and linked markers identified.

[0033] 2. Extract individual DNA by SDS method.

[0034] 3. Screening of polymorphic markers: select 900 pairs of SSR primers, and use leek green and IR 26 As a template, carry out PCR amplification, and screen for polymorphic markers.

[0035] 4. PCR reaction system: the volume is 25 microliters, including 2.5 microliters of 10×buffer, 25mM MgCl 2 1.5 microliters, 2.5 microliters of 4 pmol / microliter primer pair, 2 microliters of 2.5mM dNTPs, 0.2 microliters of 5 units / microliter Taq enzyme, 20 nanograms of template DNA, and add water to 25 microliters. The reaction program wa...

Embodiment 2

[0045] (1) Materials and methods:

[0046] 1. Material: rice variety Leek Green and IR 26 .

[0047] 2. Extract individual DNA by SDS method.

[0048] 3. Marking: RM13441, P8, RM13443.

[0049] 4. PCR reaction system: the volume is 25 microliters, including 2.5 microliters of 10×buffer, 25mM MgCl 2 1.5 microliters, 2.5 microliters of 4 pmol / microliter primer pair, 2 microliters of 2.5mM dNTPs, 0.2 microliters of 5 units / microliter Taq enzyme, 20 nanograms of template DNA, and add water to 25 microliters. The reaction program was DNA pre-denaturation at 95°C for 5 min; pre-denaturation at 95°C for 30 s, annealing at 50°C for 30 s, extension at 72°C for 30 s, and 35 cycles; finally, extension at 72°C for 10 min. PCR amplification was carried out on a biometer amplification instrument, and the amplified products were separated by electrophoresis on 8% non-denaturing polyacrylamide gel (containing 7.6g acrylamide and 0.4g methylenebisacrylamide in 100ml polyacrylamide solution...

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Abstract

Belonging to the fields seed science and technology application, the invention relates to a molecular marker of a rice seed salt tolerant germination major QTL qGR2 and its application. One or more pairs of rice genome DNA in SSR marked primers are employed to conduct PCR amplification, and the PCR amplification product is subjected to electrophoresis detection on 8% non-denaturing polyacrylamide gel. If a DNA fragment with a corresponding size is amplified, major QTL qGR2 synergetic allele for controlling rice seed salt tolerant germination can exist. By means of a molecular marker method in coseparation and close linkage with the seed salt tolerant germination gene qGR2, the salt tolerance level during rice germination can be predicted, and the salt rice variety in a rice germination period can be screened out rapidly.

Description

technical field [0001] The invention belongs to the application field of seed science and technology, and relates to a molecular marker of a main QTL site qGR2 for salt-tolerant germination of rice seeds and an application thereof. Background technique [0002] Rice (Oryza sativa L.) is one of the most important food crops in the world. About 30% of the rice arable land in the world is affected by salt damage because improper irrigation and fertilization often lead to the accumulation of salt in the soil. Salt damage has become one of the important factors affecting rice production. [0003] Different degrees of salt damage will occur in various stages of rice growth and development, such as germination, seedling, tillering, booting and maturity, among which salt damage in the germination stage is likely to occur in direct-seeded rice fields and seedling fields. In particular, with the development of the economy, direct seeding of rice has become more and more common, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 王州飞张红生程金平王建飞黄骥鲍永美
Owner NANJING AGRICULTURAL UNIVERSITY
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