Primers and their application for screening wheat leaf rust resistance qtl
A technology for leaf rust and wheat, applied in primers for screening wheat leaf rust resistance QTL and its application field, can solve problems such as yield loss, and achieve the effect of prolonging disease resistance
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[0034] A new QTL for wheat leaf rust resistance- QLr.hebau-2DS The SSR markers gwm261 and gwm484 are obtained:
[0035] Fundulea 900, Thatcher and 188 Fundulea 900 / ThatcherF 2:3 The populations were planted in Baoding Experimental Field, Hebei during the wheat growing season for three consecutive years in 2010-2011, 2011-2012 and 2012-2013 respectively. A completely randomized block design was adopted for field planting, and a total of 2 repetitions were set up. The row length was 1.2 m and the row spacing 0.25 m, about 50 grains per row. One row of high-sensitivity variety Zhengzhou 5389 was planted in every 10 rows as a control. The induced row (Zhengzhou 5389) was planted vertically with the test material.
[0036] The rows were inoculated with mixed leaf rust strains, and when the leaves of the induced rows were fully infected, the rust resistance of the population was identified, and the field maximum disease severity (MDS) was statistically analyzed.
[0037] The DN...
Embodiment 1
[0043] 1. Extract Thatcher genomic DNA as a template;
[0044] 2. Using wms261 and wms484 as primers and Thatcher genomic DNA as a template, carry out PCR amplification. 10 μL PCR reaction system: 10×buffer 1 μL, 10 mmol L -1 dNTPs 0.2 μL, 4 μmol L -1 Primer 1 μL, 30 ngμL -1 DNA template 1 μL, Taq Enzyme 0.5 U, ddH 2 O 6.7 μL.
[0045] The reaction program was pre-denaturation at 94°C for 5 min; denaturation at 94°C for 45 sec, annealing at 55°C for 45 sec, extension at 72°C for 45 sec, 35 cycles; final extension at 72°C for 10 min, and storage at 10°C.
[0046] 3. Separation and detection of the amplified products: add 2.5 μL 6×Loading buffer to the amplified products, separate them by electrophoresis in a non-denaturing polyacrylamide gel with a concentration of 12% (w / v), and develop the color by silver staining. Such as figure 1 and figure 2 shown ( figure 1 Non-denaturing polyacrylamide gel electrophoresis pictures of wms261 amplification products, figure...
Embodiment 2
[0048] 1. Extract Fundulea 900 genomic DNA as a template;
[0049] 2. Using wms261 and wms484 as primers and Fundulea 900 genomic DNA as a template, carry out PCR amplification. 10 μL PCR reaction system is: 10×buffer 1 μL, 10 mmol L -1 dNTPs 0.2 μL, 4 μmol L -1 Primer 1 μL, 30ng μL -1 DNA template 1 μL, Taq Enzyme 0.5 U, ddH 2 O 6.7 μL.
[0050] The reaction program was pre-denaturation at 94°C for 5 min; denaturation at 94°C for 45 sec, annealing at 55°C for 45 sec, extension at 72°C for 45 sec, 35 cycles; final extension at 72°C for 10 min, and storage at 10°C.
[0051] 3. Separation and detection of the amplified products: add 2.5 μL 6×Loading buffer to the amplified products, separate them by electrophoresis in a non-denaturing polyacrylamide gel with a concentration of 12% (w / v), and develop the color by silver staining. Such as figure 1 and figure 2 shown ( figure 1 Non-denaturing polyacrylamide gel electrophoresis pictures of wms261 amplification product...
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