Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (6): 1241-1255.doi: 10.16420/j.issn.0513-353x.2023-0774
• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles
MA Xingyun, FAN Bingli, TANG Guangcai, JIA Zhiqi, LI Ying, XUE Dongqi, ZHANG Shiwen*()
Received:
2024-01-26
Revised:
2024-04-19
Online:
2024-12-18
Published:
2024-06-21
Contact:
ZHANG Shiwen
MA Xingyun, FAN Bingli, TANG Guangcai, JIA Zhiqi, LI Ying, XUE Dongqi, ZHANG Shiwen. Preliminary Study on the Mechanism of DXR Regulating Chloroplast Development Flower Color and Fruit Coloring in Tomato[J]. Acta Horticulturae Sinica, 2024, 51(6): 1241-1255.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2023-0774
基因序列 Primer name | 正向引物序列(5′-3′) Forward primer sequence | 反向引物序列(5′-3′) Reverse primer sequence |
---|---|---|
Actin | GTCCTCTTCCAGCCATCCAT | ACCACTGAGCACAATGTTACCG |
Solyc03g114340(DXR) | AAGGCAGAGTTGGGATGGTC | TTGTCCGGATTCTCAGCGAC |
Solyc04g012120(14-3-3) | GAGCTAGGAGAGCATCGTGG | GCAACTTGAGGATGCCGTTAC |
Solyc03g034180(14-3-3) | TGATGATGCCATCGCAGAGC | TCATCCGTGGTATCAGAAGTCC |
Solyc01g079620(MYB12) | GGGCATCAAGAGAGGCAGAT | AACGACCTCCAAGAGCCTTC |
Solyc11g013110(FLS) | AAGATTTGGCCTCCTCCTGC | TCCAAACCAAGCCCAAGTGA |
Solyc04g040190(LCYB) | GTGCTCGATGCAACTGGCTT | GGTGCTCTTCCACTTCAGCC |
Solyc12g008980(LCYE) | TGGTGTATGGGAGGACGAGT | AACTCTTCCATAGGCACGGC |
Solyc05g010320(CHI) | GAATCCTCTGGGCTCCAACA | ACACACCGATCGCAGTAAAC |
Table 1 Sequence of primers used for qRT-PCR
基因序列 Primer name | 正向引物序列(5′-3′) Forward primer sequence | 反向引物序列(5′-3′) Reverse primer sequence |
---|---|---|
Actin | GTCCTCTTCCAGCCATCCAT | ACCACTGAGCACAATGTTACCG |
Solyc03g114340(DXR) | AAGGCAGAGTTGGGATGGTC | TTGTCCGGATTCTCAGCGAC |
Solyc04g012120(14-3-3) | GAGCTAGGAGAGCATCGTGG | GCAACTTGAGGATGCCGTTAC |
Solyc03g034180(14-3-3) | TGATGATGCCATCGCAGAGC | TCATCCGTGGTATCAGAAGTCC |
Solyc01g079620(MYB12) | GGGCATCAAGAGAGGCAGAT | AACGACCTCCAAGAGCCTTC |
Solyc11g013110(FLS) | AAGATTTGGCCTCCTCCTGC | TCCAAACCAAGCCCAAGTGA |
Solyc04g040190(LCYB) | GTGCTCGATGCAACTGGCTT | GGTGCTCTTCCACTTCAGCC |
Solyc12g008980(LCYE) | TGGTGTATGGGAGGACGAGT | AACTCTTCCATAGGCACGGC |
Solyc05g010320(CHI) | GAATCCTCTGGGCTCCAACA | ACACACCGATCGCAGTAAAC |
引物名称 Primer name | 正向引物序列(5′-3′) Forward primer sequence | 反向引物序列(5′-3′) Reverse primer sequence |
---|---|---|
CP098-SlDXR | AATCTAACAGTGTAGTTTGTGAGCCAGGAAGGCAGAGTTGTTTTAGAGCTAGAAATAGC | CTATTTCTAGCTCTAAAACTGCTCCCCAGGTATAATCTCCAAACTACACTGTTAGATTC |
pHellgate8-SlDXR | CATTTGGAGAGGACACGCTCGAGATGGCCCTCAATTTGCTTTCTC | TCTCATTAAAGCAGGACTCTAGATACAAGAGCTGGACTCAAACCAGAT |
pCAMBIA1302-SlDXR-GFP | ACGGGGGACTCTTGACCATGGATGGCCCTCAATTTGCTTTCTC | AAGTTCTTCTCCTTTACTAGTTACAAGAGCTGGACTCAAACCAGAT |
pMV2-SlDXR | TGCATCCAACGCGTTGGGAGCTCGCCTGCATCACACCCGTAT | GCCTTCGCCATTCTAGACTCGAGGCTATGTGTCTCGTTGTGGC |
Table 2 Sequence of primers used for expression vectors
引物名称 Primer name | 正向引物序列(5′-3′) Forward primer sequence | 反向引物序列(5′-3′) Reverse primer sequence |
---|---|---|
CP098-SlDXR | AATCTAACAGTGTAGTTTGTGAGCCAGGAAGGCAGAGTTGTTTTAGAGCTAGAAATAGC | CTATTTCTAGCTCTAAAACTGCTCCCCAGGTATAATCTCCAAACTACACTGTTAGATTC |
pHellgate8-SlDXR | CATTTGGAGAGGACACGCTCGAGATGGCCCTCAATTTGCTTTCTC | TCTCATTAAAGCAGGACTCTAGATACAAGAGCTGGACTCAAACCAGAT |
pCAMBIA1302-SlDXR-GFP | ACGGGGGACTCTTGACCATGGATGGCCCTCAATTTGCTTTCTC | AAGTTCTTCTCCTTTACTAGTTACAAGAGCTGGACTCAAACCAGAT |
pMV2-SlDXR | TGCATCCAACGCGTTGGGAGCTCGCCTGCATCACACCCGTAT | GCCTTCGCCATTCTAGACTCGAGGCTATGTGTCTCGTTGTGGC |
Fig. 1 Relative expression of SlDXR in various tissues of tomato Comparison of relative expression levels between tissues and roots. t-test,* P < 0.05;** P < 0.01
Fig. 3 Subcellular localization of tomato SlDXR in tobacco Superposition 1 represents the superposition of GFP fluorescence and chloroplast fluorescence;Superimposed 2 generations of brighfield,GFP fluorescence and chloroplast fluorescence.
Fig. 4 Identification and gene expression detection of tomato wild type(WT),T0 generation SlDXR edited lines(CR)and overexpressed lines(OE) A:Positive detection of gene editing vector CP098;B:Positive detection of overexpression lines vector pHellsgate8;C:Target and mutation sites of CRISPR/ Cas9 gene editing SlDXR;D:SlDXR gene sequence of CR lines.
Fig. 6 Phenotypic and photosynthetic related indexes determination of tomato wild type(WT),SlDXR edited(CR)and overexpressed(OE)lines A:T0 generation positive plants;B:T2 generation leaf chlorophyll fluorescence imaging;C:The light energy conversion efficiency of the maximum PSⅡof the leaves;D:Chlorophyll content in leaves. ** P < 0.01.
Fig. 9 Relative expression levels of SlDXR-related genes in tomato wild type(WT),SlDXR edited(CR)and overexpressed(OE)lines in leaves(A,B)and red ripe fruits(C-G) ** P < 0.01;*** P < 0.001.
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