Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (10): 1945-1955.doi: 10.16420/j.issn.0513-353x.2021-0581
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ZHANG Qian1, YANG Nan1,2, SANG Haiyu1, ZHAO Rong1, SONG Xiaoxi1, CHEN Longqing2, XIANG Lin1, ZHAO Kaige1,*()
Received:
2021-06-18
Revised:
2021-09-09
Online:
2021-10-25
Published:
2021-11-01
Contact:
ZHAO Kaige
E-mail:zhaokaige@mail.hzau.edu.cn
CLC Number:
ZHANG Qian, YANG Nan, SANG Haiyu, ZHAO Rong, SONG Xiaoxi, CHEN Longqing, XIANG Lin, ZHAO Kaige. Cloning and Functional Analysis of CpTT8 Related to Anthocyanin Synthesis in Wintersweet(Chimonanthus praecox)[J]. Acta Horticulturae Sinica, 2021, 48(10): 1945-1955.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2021-0581
作用 Funciton | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence |
---|---|---|
qPCR | CpTT8qF | TCTGACTGAGTCCGAGTGGTTC |
CpTT8qR | TTGCTGTCAGCCTCATTTGC | |
内参Internal reference | CpRPL8qF | AAGATTGTACCAAGTGGTTGC |
CpRPL8qR | TGCTCTACAGGGTTCATAGC | |
克隆Cloning | CpTT8-S1300F | CAAATCGACTCTAGAAAGCTTATGGCCTGCCCACCGAGCAG |
CpTT8-S1300R | GCCCTTGCTCACCATGGTACCGTTCTGGGAAATGATTTGGT | |
酵母双杂交载体构建 Constructing yeast two-hybrid vectors | CpTT8-ADF | GCCATGGAGGCCAGT GAATTCATGGCCTGCCCACCGAGCAG |
CpTT8-ADR | AGCTCGAGCTCGATGGATCCCGTTCTGGGAAATGATTTGGT | |
CpTT8-BDF | ATGGCCATGGAGGCCGAATTCATGGCCTGCCCACCGAGCAG | |
CpTT8-BDR | CGCTGCAGGTCGACGGATCCCGTTCTGGGAAATGATTTGGT |
Table 1 Primers used in this study
作用 Funciton | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence |
---|---|---|
qPCR | CpTT8qF | TCTGACTGAGTCCGAGTGGTTC |
CpTT8qR | TTGCTGTCAGCCTCATTTGC | |
内参Internal reference | CpRPL8qF | AAGATTGTACCAAGTGGTTGC |
CpRPL8qR | TGCTCTACAGGGTTCATAGC | |
克隆Cloning | CpTT8-S1300F | CAAATCGACTCTAGAAAGCTTATGGCCTGCCCACCGAGCAG |
CpTT8-S1300R | GCCCTTGCTCACCATGGTACCGTTCTGGGAAATGATTTGGT | |
酵母双杂交载体构建 Constructing yeast two-hybrid vectors | CpTT8-ADF | GCCATGGAGGCCAGT GAATTCATGGCCTGCCCACCGAGCAG |
CpTT8-ADR | AGCTCGAGCTCGATGGATCCCGTTCTGGGAAATGATTTGGT | |
CpTT8-BDF | ATGGCCATGGAGGCCGAATTCATGGCCTGCCCACCGAGCAG | |
CpTT8-BDR | CGCTGCAGGTCGACGGATCCCGTTCTGGGAAATGATTTGGT |
Fig. 2 Protein sequence alignment of CpTT8 and other bHLH transcription factors that regulate anthocyanin synthesis At: Arabidopsis thaliana;DK:Diospyros kaki;Aa:Artemisia annua;Lc:Litchi chinensis;Md:Malus × domestica;Nt:Nicotiana tabacum;Pa:Prunus avium;Ph:Petunia hybrida;Pp:Prunus persica;Vv:Vitis vinifera;Zm:Zea mays;Os:Oryza sativa.
Fig. 4 The phenotype of transgenic Arabidopsis thaliana overexpressing CpTT8 induced by MeJA for 5 days(A),total anthocyanins content(B)and semi-quantitative detection of gene expression(C) **:Represents a significant difference between transgenic lines and wild type(** P ≤ 0.01).
Fig. 6 The activation function of CpTT8 on the promoter of ANS A:The effect of CpTT8 on the promoter of CpANS;B:The effect of CpTT8 on the promoter of AtANS. Different letters indicate a significant difference(P ≤ 0.05).
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