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Yunnan-type hybridrice restoring line molecular auxiliary selecting method
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A molecular-assisted, restorer technology, applied in the fields of botanical equipment and methods, rice cultivation, horticulture, etc., to avoid cumbersome procedures, reduce blindness, and improve breeding efficiency
Inactive Publication Date: 2006-08-09
YUNNAN AGRICULTURAL UNIVERSITY
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Abstract
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Problems solved by technology
At present, there is no method for selecting and identifying restorer lines of Dian-type hybrid rice with relatively complete technology and effective methods.
Method used
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Embodiment 1
[0018] 15 days after transplanting, the leaves were taken to extract DNA. The composition of the PCR reaction solution and the amplification procedure are as follows:
[0019] Composition of reaction solution: ddH 2 O6ul, 10×buffer2ul, MgCl 2 (25mM) 2ul, dNTP (2.5umol / L) 2ul, primer (2umol / L) 2ul each, template DNA 1ul, Taq enzyme 0.5U, total volume 17ul
[0020] Amplification procedure: 93°C for 4min;
[0021] 93℃1min, 50℃1min, 68℃1min, 35 cycles;
[0022] 72℃4min
[0023] The test results are shown in Table 1.
[0024] OSR33
[0025] T 0.05 The value is 1.96, T 0.01 Value 2.63
Embodiment 2
[0027] Thirty days after transplanting, the leaves were taken to extract DNA. The PCR reaction solution composition and amplification procedure are as follows:
[0028] Composition of reaction solution: ddH 2 O 8ul, 10×buffer 2ul, MgCl 2 (25mM) 2ul, dNTP (2.5umol / L) 2ul, primer (2umol / L) 2ul each, template DNA 2ul, Taq enzyme 0.5U, total volume 20ul
[0029] Amplification procedure: 94℃ 4min;
[0030] 94℃1min, 55℃1min, 72℃1min, 35 cycles;
[0031] The test results at 72°C for 4 minutes are shown in Table 2.
[0032] OSR33
[0033] T 0.05 The value is 1.96, T 0.01 Value 2.63
Embodiment 3
[0035] 40 days after transplanting, the leaves were taken to extract DNA. The PCR reaction solution composition and amplification procedure are as follows:
[0036] Composition of reaction solution: ddH 2 O 12ul, 10×buffer 2ul, MgCl 2 (25mM) 2ul, dNTP (2.5umol / L) 2ul, primer (2umol / L) 2ul each, template DNA 3ul, Taq enzyme 1.5U, total volume 25ul
[0037] Amplification procedure: 95°C for 4min;
[0038] 95℃1min, 60℃1min, 75℃1min, 35 cycles;
[0039] 72℃4min
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Abstract
The present invention relates to a method for molecular auxiliary selection of Dian-type hybrid rice restoring line, belonging to the field of breeding technology. Said method includes the following steps: sowing and transplanting the material to be identified, after the material is transplanted for 15-40 days, taking leaf, extracting total DNA of rice; determining sample DNA concentration and quantitatively diluting DNA to 25 ng / ul; then using micro-satellite marker OSR33 and RM228 which are positioned in tenth chromosome of rice and tightly linked with Dian-type restoring gene as primer, making PCR reaction; making poly-acrylamidegel electrophoresis, utilizing silver staining method to detect PCR product and electrophoresis result; then judging that the tested material has restoring gene or not and can be used as restoring line of Dian-type hybrid rice or not. Said invention can quickly and effectively screen restoring line of Dian-type hybrid rice.
Description
Technical field [0001] The invention relates to a method for molecular assisted selection of Dian-type hybrid rice restorer lines, which belongs to the field of agricultural biotechnology, more specifically to the field of breeding. Background technique [0002] Rice is one of the important food crops in our country. The breeding of hybrid rice in the 1970s has made a great contribution to our country's food production and solving the problem of food and clothing. Statistics in 1998 showed that my country's hybrid rice planting area accounted for more than 50% of the rice area, and the cumulative increase in grain production was 350 billion kilograms. Dian-type japonica hybrid rice Since the realization of the three-line combination of Dian-type japonica hybrid rice in 1973, ten Dian-type sterile lines with different cytoplasmic types have been bred, and a sequence of excellent japonica hybrid combinations have been successfully assembled, which has been used for food production....
Claims
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