Application of set of SNP loci in screening of populus tomentosa with characters of high photosynthesis and quick growth and kit
A technology of photosynthesis and poplar tomentosa, applied in the field of molecular genetics, can solve the problems of not providing poplar selection methods, shorten the breeding cycle and accelerate the breeding process
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Embodiment 1
[0061] Screening of SNP functional loci significantly associated with photosynthetic and growth traits
[0062] (1) DNA extraction
[0063] Genomic DNA of Populus tomentosa was extracted with DNeasy Plant Mini Kits (Qiagen, Inc., Valencia, CA, USA).
[0064] (2) Amplify the target gene
[0065] The total RNA extraction and cDNA synthesis of Populus tomentosa leaves were carried out according to the methods described by RNeasy Plant Mini Kits (Qiagen, Inc., Valencia, CA, USA) and Clonetech kits, respectively. Apply 25 μL of DNA polymerase chain reaction (PCR) system, take Populus tomentosa leaf cDNA as template, add 2.5 μL of 10× buffer, 1.8 μL of 25mmolL -1 MgCl 2 , 1μL 10mmolL -1 dNTP, Taq DNA polymerase 1.0U (the above reagents were purchased from Promega), 100nmolL -1 Add 1 μL of each amplification primer to 25 μL by adding appropriate amount of double distilled water. Amplification primers are shown in Table 2.
[0066] Table 2 Primer sequences used for gene amplifi...
Embodiment 2
[0078] The method of association mapping was used to identify the functional SNP markers in the PetC gene of Populus tomentosa related to the photosynthetic and growth traits of poplar.
[0079] 1. Selection of associated populations: The National Populus tomentosa gene bank was collected and preserved in 1982, including 1047 genotyped individuals. The meteorological conditions in the main distribution area of Populus tomentosa were investigated, and based on 16 meteorological factors such as heat, light and water, the entire distribution area of Populus tomentosa was finally divided into the southern climate zone, the northeast climate zone and the northwest climate zone. All materials were collected in 100 counties of 10 provinces and cities (Beijing, Hebei, Henan, Shandong, Shanxi, Shaanxi, Gansu, Ningxia, Anhui and Jiangsu) covered by three climate zones, and were uniformly planted in Guan County, Shandong Province by root propagation State Nursery. The present invent...
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