Molecular marker linked with rice cold-tolerant gene qSF12 and application of molecular marker
A technology of rice and cold tolerance, applied in the field of agricultural biotechnology engineering, can solve the problems of time-consuming, low breeding efficiency, difficult phenotypic identification, etc., and achieve the effect of overcoming long time period and strong reliability
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Embodiment 1
[0051] Example 1. Acquisition of rice cold tolerance gene qSF12 at booting stage and its closely linked molecular marker STS12b 1. Mapping of rice cold tolerance QTL at booting stage and acquisition of cold tolerance gene qSF12 at booting stage
[0052] 1. Test materials
[0053] The strong cold-tolerant japonica rice variety Jilen 1 from the Rice Research Institute of the Jilin Academy of Agricultural Sciences was crossed with the cold-sensitive indica rice variety Miryang 23 from the Academy of Crop Science of the Rural Development Agency of Korea, and 253 lines of F 10 The recombinant inbred line population (RIL) was used as the targeting population.
[0054] 2. Genotype identification
[0055] The test materials were planted in the Beijing Changping Experimental Base, and the young rice leaves were clipped, and the genome-wide DNA extraction was carried out according to Doyle and Dickson (1987) with a slightly improved method of cetyltriethylammonium bromide (CTAB). A to...
Embodiment 2
[0072] Embodiment 2, the identification method of the cold tolerance of rice booting stage
[0073] The method provided by the invention to identify the cold tolerance of the rice booting stage to be tested is as follows:
[0074] 1. Extract the genomic DNA of the rice to be tested, use the genomic DNA as a template, and use the STS12b molecular marker to perform PCR amplification to obtain a PCR product;
[0075] The PCR reaction system is as follows: 10×PCR buffer (containing Mg 2+ ) 1.0μl, 10mM dNTP Mixture 0.25μl, 10pM / μl primer 0.25μl, 0.5U / μl Taq polymerase 0.25μl, 40ng / μl genomic DNA 1.0μl, ddH 2 O to make up to 10 μl.
[0076] The PCR reaction conditions were as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 40 s, 36 cycles; extension at 72°C for 10 min.
[0077] 2. Identify the cold tolerance of the rice to be tested at the booting stage according to the size of the PCR product:
[0078...
Embodiment 3
[0083] Example 3, Application of STS12b Molecular Marker in Identification of Cold Tolerance of Rice Varieties at Booting Stage
[0084] 1. Test materials
[0085] The test materials are Heigu, Italian glutinous rice, Huatanggu, Zulu valley, Baihe winnowing valley, Manpi fragrant red glutinous rice, Maiche, Baihe Xiaomagu, Taipei No. 8, rhubarb glutinous rice, back seed glutinous rice, Yunchuan white , Diaogu, Erguo and Luzu Baigu.
[0086] 2. Identification of cold tolerance at booting stage
[0087] According to the method in step 3 of Example 1 step one, the booting stage cold resistance of the test material in step one is identified.
[0088] The results are shown in Table 3. It can be seen from the table that among the 15 rice varieties, Italian Nuo, Manpi Xianghong Glutinous Rice, and Diaogu all have a cold tolerance level of 7, all of which are rice varieties with weak cold tolerance; Zulu Valley, Maiche, and Yunchuanbai Heigu and Luzu Baigu are both cold-tolerant g...
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