A fluorescent molecular marker and its primers for rice grain width regulation gene gw8
A technology for regulating genes and fluorescent molecules, applied in the field of biomolecules, can solve problems such as low efficiency, use of toxic substances, etc., and achieve the effects of simple and reliable method, rapid inspection, and improvement of grain shape agronomic traits
Active Publication Date: 2021-08-13
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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[0004] Aiming at the above technical problems, the present invention discloses a fluorescent molecular marker of the rice grain width regulating gene GW8 and its primers, which realizes rapid detection of whether a rice variety contains a GW8 allele with two base deletions; the method is simple and solves the problem of The currently used method has the disadvantage of low efficiency and the use of toxic substances
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[0026] The preferred embodiments of the present invention will be further described in detail below.
[0027] A fluorescent molecular marker of rice grain width regulation gene GW8, which adopts the following steps:
[0028] (1) get rice leaves, obtain the genomic DNA of rice by CTAB extraction method (cetyltrimethylammonium bromide method);
[0029] (2) PCR amplification: three primers Allele-AT, Allele-GC and Common-R1 were simultaneously added to the PCR reaction system, and the GW8 gene in the rice plant was amplified to obtain a PCR product.
[0030] The sequences of the primers are as follows:
[0031] Allele-AT: GAAGGTCGGAGTCAACGGATTACTAATTTGTGCTGATATATCAAACATCAT;
[0032] Allele-GC: GAAGGTGACCAAGTTCATGCTACTAATTTGTGCTGATATATCAAAACATCGC;
[0033] Common-R1: TGAGAATCTTGCCCCTGAATTTTG.
[0034] Among them, the PCR reaction program is: 95°C, 5min; then 10 cycles of 95°C, 20sec, 65°C (-0.8°C per cycle), 1min; then 32 cycles of 95°C, 20sec, 57°C, 1min;
[0035] The PCR re...
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The present invention provides a fluorescent molecular marker of rice grain width regulation gene GW8 and primers thereof, the primers include Allele-AT primers, Allele-GC primers and Common-R1 primers, the sequence of the Allele-AT primers is as SEQ ID NO : 1, the sequence of the Allele-GC primer is shown in SEQ ID NO: 2, and the sequence of the Common-R1 primer is shown in SEQ ID NO: 3. By adopting the technical scheme of the present invention, it is realized to quickly detect whether the rice variety contains GW8 alleles with two base deletions; the method is simple, and solves the disadvantages of low efficiency and the use of toxic substances in the currently used methods; it can be applied to molecular markers Assisted selection of GW8 alleles to improve rice grain shape agronomic traits.
Description
technical field [0001] The invention belongs to the technical field of biomolecules, and in particular relates to a fluorescent molecular marker of rice grain width regulating gene GW8 and a primer thereof. Background technique [0002] As one of the important food crops, rice feeds about 50% of the world's population. Simultaneously improving rice yield and quality remains a challenging problem for rice breeders since rice yield and quality are negatively correlated. Grain size is an important factor affecting yield and quality, and is one of the main goals of rice breeding. In high-temperature areas, grain filling is difficult due to wide grain size, resulting in a decline in rice quality. However, the loss of function of the grain width negative regulator gene GW8 resulted in thinner grains and improved rice quality. Therefore, the development of fluorescent molecular markers of GW8 gene for molecular marker-assisted selection breeding has important research significan...
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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 邓国富高利军卿冬进戴高兴黄娟高菊梁海福伍豪周维永周萌陈韦韦
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI