Peach polypeptide hormone synthesis gene pprgf1 and its application
A technology of polypeptide hormones and synthetic genes, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as fruit softening and unclear interaction mechanism, and achieve the effect of promoting ripening
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Embodiment 1
[0033] Example 1 Peach polypeptide hormone synthesis gene PpRGF1 isolation and identification of
[0034] experiment method
[0035] 1. Gene PpRGF1 separation
[0036] The peach pulp RNA was extracted with a plant polysaccharide and polyphenol kit (DP441, Tiangen Biochemical Technology (Beijing) Co., Ltd.), and single-stranded cDNA was obtained by a reverse transcription kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.). cDNA was used as the template, and the following sequences were used as primers, and the gene was obtained by PCR (the kit FastStart SYBR Green Master was purchased from Roche Biotechnology Co., Ltd.) PpRGF1 The full-length sequence of the PCR amplification electropherogram is shown in figure 1 shown. Gene PpRGF1 The full-length sequence is shown in SEQ ID NO: 1 in the sequence table, with a total of 489 bp, and the amino acid sequence of the co-encoded protein is shown in SEQ ID NO: 2 in the sequence table, with a total of 162. Peach PpRGF1 wi...
Embodiment 2
[0067] Example 2 Gene PpRGF1 The role of transgenic tomatoes
[0068] 2.1 Genes PpRGF1 Effects on tomato fruit development
[0069] Image 6 Observation diagrams of wild-type tomato and transgenic tomato fruits at different developmental stages. from Image 6 It can be seen that the transgenic tomato began to change color at 41d after flowering, and the wild-type tomato began to change color at 45d after flowering, that is, the transgenic tomato matured about 4d earlier than the wild-type tomato. same.
[0070] 2.3 Genes PpRGF1 Effects on tomato fruit firmness and ethylene release
[0071] The ethylene levels of transgenic and wild-type tomato fruits at different days after flowering (37dpa, 41dpa, 45dpa, 50dpa, 53dpa, 59dpa) were measured. Figure 7 Comparison of ethylene release changes of wild-type tomato and transgenic tomato fruits at different ripening stages. from Figure 7 It can be seen that the ethylene release of wild-type and transgenic tomato showed a tr...
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