A rubber tree flowering regulatory protein hbtfl1-1 and its coding gene and application
A technique for regulating flowering and encoding genes, which can be used in applications, genetic engineering, plant genetic improvement, etc., and can solve problems such as no TFL1/CEN-like gene reports yet.
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Embodiment 1
[0031] Embodiment 1 Obtaining of 5 target genes of Hevea brasiliensis
[0032] According to the Rubber Tree Genome Database and Leaf Transcriptome Database of the Rubber Research Institute of the Chinese Academy of Tropical Agricultural Sciences, contigs obtained through homologous alignment are then spliced, and then the open reads are predicted by NCBI online software and specific primers are designed to obtain contigs containing complete open reads. DNA sequence and cDNA sequence of the reading frame.
[0033] The specific method is as follows:
[0034] Acquisition of open reading frames for 5 genes
[0035] The five gene-specific primers are as follows:
[0036] HbTFL1-1OF (EcoRI):G ATGTCAAGGATCATGGAGCC
[0037] HbTFL1-1OR(XbaI):GC TCATTCTTCTTCTTGCAGCAGTTTT
[0038] HbTFL1-2OF(EcoRI):G ATGGCAAGAATAATAGAACCTCT
[0039] HbTFL1-2OR(XbaI):GC TTAGCGCCTTCTTGCAGCAGTT
[0040] HbTFL1-3OF(EcoRI):G ATGGCAAGAATAATAGAACCTCT
[0041] HbTFL1-3OR(XbaI):GC TTAGCGTCTTCTT...
Embodiment 2
[0102] Example 2 Analysis of tissue-specific expression of 5 target genes
[0103] In this study, 3-month-old rubber tissue culture seedlings were selected as experimental materials. The roots, stems, buds, bronze leaves, discolored leaves, light green leaves and stable leaves were collected respectively, and their RNAs were extracted for future use.
[0104] The RNA obtained above was digested with DnaseI and then reverse transcribed according to the reverse transcription kit. The tissue-specific expression analysis of five target genes was performed by fluorescence quantitative technology.
[0105] The five gene fluorescent quantitative primers are:
[0106] HbTFL1-1QF:CCTCCTTAATCCCTGGCTACG,
[0107] HbTFL1-1QR:ACATCGCCTACAACTCTCCCT,
[0108] HbTFL1-2QF:GCCACATTTGGAAGGGAGATAG,
[0109] HbTFL1-2QR: TAACACTTATTTATTGAAGGCCACTTG,
[0110] HbTFL1-3QF: ATACACCTCTTAGCCAGACGCTC,
[0111] HbTFL1-3QR:TGTCGCCTCCTTGGACCTCA,
[0112] HbCEN1QF:CTAATGGCGAAGACAACAGAT,
[0113] HbCE...
Embodiment 3
[0117] Example 3 Analysis of spatiotemporal expression patterns of 5 target genes
[0118] Taking 3 months, 2 years and 10 years as the research objects, the roots, stems, buds, bronze leaves, color-changing leaves, light green leaves and stable leaves of each age group were collected in the flowering season (March) respectively, and 10 years old. RNA was extracted from the male and female flowers of the annual tree when they bloomed.
[0119] The RNA obtained above was digested with DnaseI and then reverse transcribed according to the reverse transcription kit. The temporal and spatial expression patterns of the five target genes were analyzed by fluorescence quantitative technology.
[0120] The results showed that: in the flowering season, for TFL1-like genes, the expressions of HbTFL1-1, HbTFL1-2 and HbTFL1-33 genes in their respective vegetative organs in 10-year-old trees were significantly lower than those in 3-month-old and 2-year-old trees . In the newly differenti...
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