Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (3): 193-201.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0957
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WU Xing-xing(), HONG Hai-bo, GAN Zhi-cheng, LI Rui-ning, HUANG Xian-zhong()
Received:
2023-10-10
Online:
2024-03-26
Published:
2024-04-08
Contact:
HUANG Xian-zhong
E-mail:wuxingxing2019@163.com;huangxz@ahstu.edu.cn
WU Xing-xing, HONG Hai-bo, GAN Zhi-cheng, LI Rui-ning, HUANG Xian-zhong. Cloning and Preliminary Functional Analysis of CaPI Gene in Capsicum annuum L.[J]. Biotechnology Bulletin, 2024, 40(3): 193-201.
引物名称 Primer name | 引物序列 Primer sequence(5'-3') | 引物用途 Primer purpose |
---|---|---|
CaPI-R | 35S:CaPI载体35S:CaPI Vector | |
CaPI-F | ||
qCaPI-R | CTTCTGGGAGGAGGCTATGG | RT-qPCR |
qCaPI-F | AAGATCTCAGACTGCTTGGC | |
qCaUBI3-R | TGTCCATCTGCTCTCTGTTG | RT-qPCR |
qCaUBI3-F | CACCCCAAGCACAATAAGAC | |
qAtactin2-R | CCTGGACCTGCCTCATCATAC | RT-qPCR |
qAtactin2-F | CTGGATCGGTGGTTCCATTC |
Table 1 Primer information used in this study
引物名称 Primer name | 引物序列 Primer sequence(5'-3') | 引物用途 Primer purpose |
---|---|---|
CaPI-R | 35S:CaPI载体35S:CaPI Vector | |
CaPI-F | ||
qCaPI-R | CTTCTGGGAGGAGGCTATGG | RT-qPCR |
qCaPI-F | AAGATCTCAGACTGCTTGGC | |
qCaUBI3-R | TGTCCATCTGCTCTCTGTTG | RT-qPCR |
qCaUBI3-F | CACCCCAAGCACAATAAGAC | |
qAtactin2-R | CCTGGACCTGCCTCATCATAC | RT-qPCR |
qAtactin2-F | CTGGATCGGTGGTTCCATTC |
Fig. 2 Structural analysis of CaPI protein A: Secondary structure of CaPI protein. B: Comparison of spatial structures between CaPI(left)and AtPI(right)proteins
Fig. 4 Phylogenetic tree of the CaPI proteins in C. annuum L. and other plant species AT: Arabidopsis thaliana, AT5G20240.1; Ca: Capsicum annuum, CaPI, CaMADS74 and CaMADS53; SMEL: Solanum melongena, SMEL4.1_08g015360.1.01, SMEL4.1_02g021760.1.01; Solyc: Solanum lycopersicum, Solyc06g059970.4.1, Solyc02g084630.3.1; Peaxi: Petunia hybrida, Peaxi162Scf00922g00026.1, Peaxi162Scf00591g00074.1; PGSC: Solanum tuberosum, PGSC0003DMG401007392; TRAES: Triticum aestivum, TRAES3BF021600020CFD t1, TRAES3BF048900050CFD t1, TRAES3BF068500020CFD t1, TRAES3BF009000010CFD t1; Os: Oryza sativa, Os01t0883100-01, Os05t0423400-0
Fig. 5 Tissue expression pattern of the CaPI gene using RT-qPCR A: Expression profiles of the CaPI gene in different tissues of C. annuum L.. B: Expression profiles of the CaPI gene in four tissues: sepals, petals, stamens, and pistils. Lowercase letters indicate significant differences at P=0.01 level. The same below
Fig. 6 Phenotype survey of the 35S:CaPI transgenic plants in Arabidopsis thaliana A: Comparisons of phenotypes between the 35S:CaPI transgenic lines and wild type(Col-0)plants grown for 30 d. #1 and #2 represent two independent transgenic plants, respectively. B: Statistics on the number of rosette leaves during flowering stages. C: Statistics of flowering times. D: Statistics of branch numbers during flowering stages. E: RT-qPCR analysis of the relative expression level of the CaPI gene
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