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SSR (simple sequence repeat) primer pair linked to fruiting rate on rice chromosome 2 and application of SSR primer pair

A primer pair and seed setting rate technology, applied in the field of plant molecular genetics, achieves the effects of low cost, clear directionality, and simple identification

Inactive Publication Date: 2017-02-01
广西壮族自治区农业科学院水稻研究所
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Problems solved by technology

[0008] In order to solve the problem of early detection of the seed setting rate of breeding materials in rice breeding, and solve the aggregation problem of QTL related to the seed setting rate, the present invention provides a pair of SSR primers linked to the seed setting rate on chromosome 2 of rice, which are used to detect breeding Does this QTL related to seed setting rate exist in the material

Method used

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  • SSR (simple sequence repeat) primer pair linked to fruiting rate on rice chromosome 2 and application of SSR primer pair
  • SSR (simple sequence repeat) primer pair linked to fruiting rate on rice chromosome 2 and application of SSR primer pair
  • SSR (simple sequence repeat) primer pair linked to fruiting rate on rice chromosome 2 and application of SSR primer pair

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1 : Primer Screening

[0023] SSR primers were downloaded from www.genomene.org; the sequences of RM341SSR primers are shown in Table 1. The sequences of the SSRs in Table 6 themselves were obtained from publicly available web resources.

[0024] Table 1 RM341 sequence

[0025]

Embodiment 2

[0026] Example 2 : The CTAB method extracts the total DNA of rice, the steps are as follows:

[0027] (1) Weigh 0.4-0.5g of fresh leaves for each material, add liquid nitrogen to the mortar, grind and pulverize quickly, and then transfer to a 2.0mL sterile centrifuge tube; before this, dry materials are pulverized with a pulverizer , and take the powder;

[0028] (2) Add 700 μL of preheated 65°C CTAB extraction buffer (pH 8.0) to each tube, bathe in 65°C water for 45 minutes, take out and shake well once every 5 minutes;

[0029] (3) Add 700 μL of chloroform / isoamyl alcohol (24:1) and shake gently for 8 minutes, then centrifuge at 12000 r / min for 10 minutes;

[0030] (4) Take the supernatant and transfer it to another 1.5 mL sterile centrifuge tube, add 600 μL of pre-cooled isopropanol, mix it upside down, and freeze it in a -20°C refrigerator for more than 30 minutes;

[0031] (5) centrifuge the centrifuge tube at 12000r / min for 10min, and take the precipitate;

[0032] ...

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Abstract

The invention belongs to the field of plant molecular genetics, and relates to an SSR (simple sequence repeat) primer pair linked to a fruiting rate on a rice chromosome 2 and an application of the SSR primer pair in breeding rice varieties. According to the SSR primer pair linked to the fruiting rate on the rice chromosome 2, the sequence of an upstream primer is shown as Seq ID NO.1 and the sequence of a downstream primer is shown as Seq ID NO.2. The SSR primer pair linked to the fruiting rate on the rice chromosome 2 provided by the invention is a linked SSR marker which is related with the fruiting rate QTL, and with the application of the SSR primer pair, specific strips of a high-fruiting-rate material and a low-fruiting-rate material can be identified relatively accurately in 31 parts of core collections; the SSR primer pair can be used for predicting the fruiting rate of breeding materials in early stage and can be used for judging whether genes related to the fruiting rate exist or not; the SSR primer pair can be used for rapidly identifying whether the breeding materials contain the QTL related to the fruiting rate or not; and the SSR primer pair is low in cost, simple in identification and clear and definite in directivity.

Description

technical field [0001] The invention belongs to the field of plant molecular genetics, and relates to a pair of SSR primers linked to the seed setting rate on the No. 2 chromosome of rice and its application in breeding rice varieties. Background technique [0002] SSR marker, also known as simple sequence repeat, is one of the most commonly used microsatellite markers. The core structure is a sequence of dozens of nucleotides consisting of 2-6 nucleotides in series as repeating units, usually a single sequence with strong conservation, flanked by relatively conserved and specific single-copy sequences . According to the specific conserved sequence, primers can be artificially synthesized for PCR amplification. Due to the variation in the number of repeating units of a single microsatellite site, the change in the length of individual amplification products will produce length polymorphism. The augmented site represents a pair of alleles at this site. Thus, the PCR produc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 潘英华梁云涛徐志健韦价报
Owner 广西壮族自治区农业科学院水稻研究所
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