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Molecular Marker and Application of Rice Gene qse3 with Strong Salt Tolerance and High Vitality

A technology of molecular markers and high activity, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems that the genes related to salt-tolerant germination of rice seeds are rarely reported, and achieve rapid identification methods. Simple, clear location, and the effect of improving breeding efficiency

Active Publication Date: 2019-11-08
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A number of genes related to seed germination have been successfully cloned by map-based cloning, such as: qLTG3-1, Sdr4, OsVP1, OsGA20ox1, OsFbx352, and GD1, etc., but there are still few reports on rice seed germination-related genes

Method used

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  • Molecular Marker and Application of Rice Gene qse3 with Strong Salt Tolerance and High Vitality
  • Molecular Marker and Application of Rice Gene qse3 with Strong Salt Tolerance and High Vitality
  • Molecular Marker and Application of Rice Gene qse3 with Strong Salt Tolerance and High Vitality

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Materials and methods:

[0031] 1. Materials: The strong salt-tolerant cultivar Chicaiqing was used as the donor, and the salt-sensitive indica rice IR26 was used as the recipient, and isolated BC was obtained by backcrossing and selfing 2 f 2 Segregate the population and complete the initial positioning; further construct the high-generation backcross segregation population BC 5 f 2 , select the exchange strain, and then let the exchange strain self-fertilize to produce BC 5 f 3The population was changed from heterozygous fragments to homozygous, fine-mapped in combination with phenotypes, and molecular markers linked to qSE3 were determined.

[0032] 2. Extract individual DNA by CTAB method.

[0033] 3. Screening of polymorphic markers: Select 1000 pairs of SSR primers, and use leek green and IR26 as templates for PCR amplification to screen for polymorphic markers.

[0034] 4. PCR reaction system: the volume is 25 microliters, including 2.5 microliters of 1...

Embodiment 2

[0043] (1) Materials and methods:

[0044] 1. Materials: rice variety Chicaiqing and IR26.

[0045] 2. Extract individual DNA by CTAB method.

[0046] 3. Markers: jm3, jm6, jm31 and jm32.

[0047] 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 7.6 g of acrylamide and 0.4 g of methylenebisacrylamide in 100 ml of polyacry...

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Abstract

The invention discloses a molecular marker of a highly-salt-tolerant and high-vitality gene qSE3 of paddy rice and application of the molecular marker. PCR amplification is performed on paddy rice genome DNA by using a pair of or multiple pairs of primers in the SSR marker, electrophoresis detection is performed on the PCR amplification product on 8% non-denaturing polyacrylamide gel, and enhancing allele controlling the highly-salt-tolerant and high-vitality gene qSE3 of the paddy rice exists if the DNA fragment with the corresponding size is obtained through amplification. By the molecular marker and a molecular marking method in co-segregation with and closely linked with the highly-salt-tolerant and high-vitality gene qSE3, the salt tolerance level of the paddy rice during a germination stage can be predicted, and salt-tolerant species of paddy rice seeds at the germination stage can be screened out fast.

Description

technical field [0001] The invention belongs to the application field of seed science and technology, and relates to a molecular marker of rice strong salt-tolerant high-activity gene qSE3 and its application. Background technique [0002] Rice (Oryza sativa L.) is one of the important food crops in my country. It is a freshwater marsh plant and is sensitive to salt. Due to the accumulation of salt in the soil due to long-term irrigation and improper fertilization, the area of ​​saline-alkali rice cultivation in my country accounts for about 20% of the total area of ​​rice cultivation. At the same time, my country's existing coastal tidal flats cover an area of ​​more than 35 million mu, which is still growing at a certain speed every year. In recent years, with the development of the economy, the shortage of rural labor force has become increasingly common, and the light and simple cultivation of direct seeding and machine-transplanted rice has become more and more common,...

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 王州飞张红生程金平何永奇王建飞黄骥鲍永美
Owner NANJING AGRICULTURAL UNIVERSITY