Molecular marker for rapid detection of high-yield gene of Thinopyrum elongatum and its application
A technology of wheat grass and molecular markers, which is applied in the field of molecular genetics and breeding, can solve the problems of lack of high-yield gene molecular markers, time-consuming, and difficulty in phenotypic identification.
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Embodiment 1
[0059] Example 1 Creation of Near-Isogenic Lines of High-yielding Genes of Wheat-Thinopyrum elongatum
[0060] In 2015, the hybrid combination (Liangxing 66 × SNK064) was configured in the greenhouse of the Crop Research Institute of Shandong Academy of Agricultural Sciences, and obtained F 1 ; Spring 2016, will receive F 1 Backcross with Liangxing 66 and get BC 1 f 1 , and carry out molecular marker identification on the obtained plants, on the basis of molecular marker identification, the BC carrying target gene and exogenous fragment 1 f 1 Continue to backcross with Liangxing 66 and get BC 2 f 1 . In the summer and autumn of 2016, the artificial climate chamber continued to add generations, self-crossed twice, and obtained BC 2 f 3 , with the help of molecular markers and fluorescence in situ hybridization, a wheat-Thinopyrum elongatum 7el2L heterozygous line was obtained; in the winter of 2016 and the spring of 2017, the above heterozygous line continued to be self...
Embodiment 2
[0062] Example 2 Analysis of agronomic and yield-related traits of high-yielding near-isogenic lines of Echinopsis elongatum
[0063] For in situ hybridization using Oligo-pTa535.1 and Oligo-pSc119.2 as probes, the probe synthesis, probe concentration and specific operation steps refer to the method of Tang et al. [2014]. After the in situ hybridization chromosome preparation was dried naturally, 0.25 μg / mL propidium iodide was added dropwise and covered with a cover slip, and photographed under an Olympus BX51 fluorescence microscope.
[0064] The above-mentioned near-isogenic line materials (SAAS1001, SAAS1002 and SAAS1003) were planted in the Lingcheng Experimental Base in the winter of 2017. The area of the plot was 1.5 m × 6.0 m, with 3 replicates. For the flowering stage and grain filling stage of wheat, the length and width of the flag leaf, the length of the ear, the number of grains per ear, the number of fertile spikelets, the number of sterile spikelets, etc. 6 t...
Embodiment 3
[0066] Example 3 Development of Tightly Linked Molecular Markers for High Yield Genes of Echinopsis elongatum
[0067] According to the sequence information of the published wheat genome (7DL end), SSR primers were designed using Primer 3.0, and wheat varieties Jimai 22, Liangxing 66, SAAS1001 and Jimai 22 mixed DNA, SAAS1002 and Liangxing 66 mixed DN, SAAS1003 and SAAS1001 Mixed DNA and wheat-Thinopyrum elongated near-isogenic lines SAAS1001, SAAS1002 and SAAS1003 were used as materials for PCR amplification to search for molecular markers closely linked to high-yielding genes.
[0068] In the PCR amplification reaction system, the volume of the PCR reaction system is 10.0 μL, including 10 × Buffer buffer: 1.0 μL, 2.5 mM dNTP solution: 0.8 μL, Taq enzyme 0.1 μL, sterile double distilled water: 4.1 μL, 50 ng DNA template 2.0 μL (Jimai 22, Liangxing 66, SAAS1001 and Jimai 22 mixed DNA, SAAS1002 and Liangxing 66 mixed DNA, SAAS1003 and SAAS1001 mixed DNA, and wheat-Ethiopia elon...
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