Method, primer pair and kit for identifying color of watermelon pulp and application thereof
A primer pair, watermelon technology, applied in the directions of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effect of improving efficiency and accuracy, easy to observe, and easy to operate
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Embodiment 1
[0039] Embodiment 1: The acquisition method of specific identification flesh color primer
[0040] (1) Acquisition of experimental materials
[0041] Select the garden female parent whose mature fruit has red pulp traits, cultivated watermelon as the male parent, select Yellow Crimson with orange pulp traits, and cultivated watermelon as the female parent, and cross the above male and female parents to obtain F 1 generation; then for the above obtained F 1 F 2 Generation groups, with F 1 Generation and F 2 The generation group was used as the test material.
[0042] Sow the materials of each generation in step (1) separately, harvest the fruit at the mature stage, cut the fruit crosswise, and judge the color of the pulp by observation. Each color observation is checked by three people separately to ensure the accuracy of the investigation results. f 1 Generation and F 2 The survey results of the pulp color of each individual plant in the generation group showed that: F ...
Embodiment 2
[0047] Embodiment 2: A kind of method for identifying the character of watermelon orange pulp
[0048] (1) extract the genomic DNA of the watermelon to be tested;
[0049] (2) Using the genomic DNA obtained in step (1) as a template, using the primer pair to perform PCR amplification to obtain a PCR product;
[0050] (3) The PCR product obtained in step (2) is digested with restriction endonuclease TaqI to obtain a digested product;
[0051] (4) Judging the pulp color of the watermelon by detecting the enzyme cleavage product.
[0052] The specific judgment method is: use agarose gel electrophoresis to detect the enzyme-cut products. If there are two enzyme-cleaved products, it is judged that the watermelon has orange pulp properties; if there are one or more enzyme-cleaved products, it is judged that the watermelon has red pulp properties.
[0053] The above PCR reaction system is: 1 μL of upstream and downstream primers (primer concentration is 2 pM), DNA template 2 μL (co...
Embodiment 3
[0057] Example 3: Utilize the obtained primers to screen watermelon varieties with different flesh colors
[0058] For judging the accuracy of the developed primer pair identification watermelon orange pulp, select 24 F described in embodiment 1 2 Substitute the sample of watermelon seedling, carry out pulp color identification to each sample according to the identification method described in embodiment 2, identification result is as follows image 3 As shown, the enzyme digestion products of samples 1-8 and 17-24 are one or three, so samples 1-8 and 17-24 are identified as watermelon seedlings with red pulp traits; the enzyme digestion products of samples 9-16 are two Therefore, samples 9-16 were identified as watermelon seedlings with orange pulp traits. Harvest F 2 Substitute fruit, cut the fruit crosswise, and judge the color of the pulp by observation. It has been verified that 24 F 2 The primer pair identification results of the generation samples were 100% consisten...
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