Molecular marker for dwarfing wheat marker gene, and primer and application thereof
A technology of molecular markers and dwarfing genes, which is applied in the field of molecular genetic breeding, can solve the problems of difficult phenotype identification, low breeding efficiency, and laboriousness, and achieve the effects of improving breeding efficiency, reducing breeding costs, and reducing operating procedures
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Embodiment 1
[0045] 1. Dwarf wheat obtains:
[0046] In 2007, Nannong 9918 was treated with 0.35% EMS (ethyl methyl sulfonate) at the Institute of Cytogenetics, Nanjing Agricultural University, and in 2008 M 1 Harvest according to single ear. Seed M in November 2008 2 Dai Suixing, in 2009 observed its agronomic traits in the field during the whole growth period, and found a dwarf mutant, which was harvested as a single plant. In 2009, this single plant was planted in M 3 Generation strains, and in 2010 phenotype identification, found that M 3 The mutant traits of the generation strain can be inherited stably, named NM9, and preserved in the China Center for Type Culture Collection with the preservation date of 2014.5.22 and the preservation number CCTCC P201408.
[0047] 2. Population construction and phenotypic identification of hybrid offspring of NM9×Nannong 9918 and NM9×Sumai 3:
[0048] (1) Population construction of hybrid offspring: F 1 , selfing to obtain F 2 , each F 2 F ...
Embodiment 2
[0061] Using Chinese spring and its 21 missing four-body systems to map Rht‐x linkage markers WMC296 and GWM122 for missing four bodies. 21 common wheat Chinese spring deficient‐tetrasome materials (N1AT1B, N1BT1A, N1DT1A, N2AT2D, N2BT2D, N2DT2A, N3AT3D, N3BT3D, N3DT3B, N4AT4D, N4BT4D, N4DT4B, N5AT5D, N5BT5A, N5DT5B, N6AT6D, N6BT6D), from the United States Introduced from Kansas State University and preserved by the Institute of Cytogenetics, Nanjing Agricultural University.
[0062]Reaction system for PCR amplification: PCR reagent composition: 1 μL DNA template containing 20‐100ng DNA, 1.0 μL 10×PCR buffer, 0.8 μL MgCl2, 0.8 μL dNTP, 0.2 μL each of upstream and downstream primers, 0.15 μL Taq DNA polymerase, 4.85 μL ddH2O; PCR program: 94°C pre-denaturation for 3 minutes; then 94°C denaturation for 30 seconds, 55°C renaturation for 45 seconds, 72°C extension for 50 seconds, 36 cycles; final extension at 72°C for 10 minutes; storage at 10°C; amplification after PCR amplificat...
Embodiment 3
[0065] Using the SSR marker GWM122 closely linked to Rht-x, the wheat materials NM9, Sumai 3 (Rht8), Chinese spring (without wheat dwarf gene), Zheng 9405 (Rht-B1b, Rht-D1b) containing different Rht genes ), Yannong 19 (Rht‐D1b), Aisumai 3 (Rht‐B1c), Yangmai 5 (Rht8), Xinong 04 (Rht‐D1c), Yannong 23 (Rht‐B1b, Rht‐D1b), PCR amplification was performed on the DNA of XN0004(Rht21).
[0066] Reaction system for PCR amplification: PCR reagent composition: 1 μL DNA template containing 20‐100ng DNA, 1.0 μL 10×PCR buffer, 0.8 μL MgCl2, 0.8 μL dNTP, 0.2 μL each of upstream and downstream primers, 0.15 μL Taq DNA polymerase, 4.85 μL ddH2O; PCR program: 94°C pre-denaturation for 3 minutes; then 94°C denaturation for 30 seconds, 55°C renaturation for 45 seconds, 72°C extension for 50 seconds, 36 cycles; final extension at 72°C for 10 minutes; storage at 10°C; amplification after PCR amplification reaction The product was separated by electrophoresis on a non-denaturing polyacrylamide gel...
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