Flavonoid 3',5'-hydroxylase gene functional marker for screening high-dihydroxyl catechin-content tea plant, as well as application and application method thereof
A high dihydroxy catechin and gene function technology, applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problems of long time-consuming and low efficiency, and speed up the pace of breeding Effect
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Embodiment 1
[0021] F3'5'H Discovery of Allelic Difference Sequence, Primer Pair and Development of Functional Marker dCAPS-F3′5′H
[0022] 1. Test materials
[0023] Choose Longjing 43 and Fenghuang tea tree as research materials.
[0024] 2. Determination of catechin content
[0025] Pick one bud and two leaves of the new shoots of tea trees in spring, dry them with hot air at 120°C for 5 minutes to fix the samples in time, and dry them at 75°C until they are fully dry and store them at low temperature. When it is to be determined, use mechanical grinding, pass through a 40-mesh sieve, and store it at low temperature for future use. Weigh 0.2g sample (accurate to 0.0001g), and place it in a 10mL centrifuge tube. Add 5 mL of 70% methanol solution preheated at 70°C, mix well, and then bathe in 70°C water bath. Water bath for 10 minutes, and stir 2-3 times in the middle. Centrifuge at 3,500r / min for 10min, and transfer the supernatant. Repeat steps 2-4, combine the supernatants coll...
Embodiment 2
[0045] Genotype Analysis of Tea Tree Resources with Functional Marker dCAPS-F3′5′H
[0046] 1. Test materials
[0047] The materials studied in this experiment are listed in Table 2, including 28 tea tree resources.
[0048] Table 2 The 28 tea tree resources studied in this experiment and their dihydroxycatechin content
[0049]
[0050] 2. Functional markers for different tea tree resources F3'5'H Genotype detection of gene SNP848
[0051] Marker genotype analysis of 28 materials using molecular marker dCAPS-F3'5'H. DNA extraction, PCR amplification system, program, enzyme digestion and agarose gel conditions are the same as in Example 1. The result is as image 3 As shown, the genotype of materials 1-10 is AA, the genotype of materials 11-20 is AG, and the genotype of materials 21-28 is GG.
[0052] 3. Identification of catechin content in 28 tea tree materials
[0053] The method for identifying catechins by high-performance liquid chromatography is the same as t...
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