Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (7): 123-130.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1266
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LIU Zhen-yin1(), DUAN Zhi-zhen1, PENG Ting2, WANG Tong-xin1(), WANG Jian1()
Received:
2022-10-06
Online:
2023-07-26
Published:
2023-08-17
Contact:
WANG Tong-xin, WANG Jian
E-mail:20196705310033@hainanu.edu.cn;jennifer_wang@hainanu.edu.cn;wjhainu@hainanu.edu.cn
LIU Zhen-yin, DUAN Zhi-zhen, PENG Ting, WANG Tong-xin, WANG Jian. Establishment and Optimization of Virus-induced Gene Silencing System in Bougainvillea peruviana ‘Thimma’[J]. Biotechnology Bulletin, 2023, 39(7): 123-130.
引物 Primer | 引物序列 Primer sequence(5'-3') | 长度 Length/bp |
---|---|---|
BpPDS336-EcoRI-F | TAAGGTTACCGAATTCTGCATTTTGATTGCTTTG | 368 |
BpPDS336-EcoRI-R | GCCTTCTAGAGAATTCAATGACTGGAACCCCTACTA | |
BpPDS457-EcoRI-F | TAAGGTTACCGAATTCGGTTTGAGTGTCCAAGAGTG | 489 |
BpPDS457-EcoRI-R | GCCTTCTAGAGAATTCTAATGACTGGAACCCCTACT | |
pTRV1-F | CCTGGTGCTCAGTACATAACTTTCC | 257 |
pTRV1-R | TCTTTGCTTACATCGTCCTCTTTCA | |
pTRV2-F | ACGCTGTTTGAGGGAAAAGTAG | 321 |
pTRV2-R | CCGATCAATCAAGATCAGTCGAGA | |
BpActin-RT-F | TATGAACTTCCTGATGGG | 125 |
BpActin-RT-R | GAATTGTAGGTTGTCTCG | |
BpPDS-RT-F | ATTCGCCTCGTCCTGATA | 103 |
BpPDS-RT-R | ATGGGTTTGTGACCTGCA |
Table 1 Primers used in this study
引物 Primer | 引物序列 Primer sequence(5'-3') | 长度 Length/bp |
---|---|---|
BpPDS336-EcoRI-F | TAAGGTTACCGAATTCTGCATTTTGATTGCTTTG | 368 |
BpPDS336-EcoRI-R | GCCTTCTAGAGAATTCAATGACTGGAACCCCTACTA | |
BpPDS457-EcoRI-F | TAAGGTTACCGAATTCGGTTTGAGTGTCCAAGAGTG | 489 |
BpPDS457-EcoRI-R | GCCTTCTAGAGAATTCTAATGACTGGAACCCCTACT | |
pTRV1-F | CCTGGTGCTCAGTACATAACTTTCC | 257 |
pTRV1-R | TCTTTGCTTACATCGTCCTCTTTCA | |
pTRV2-F | ACGCTGTTTGAGGGAAAAGTAG | 321 |
pTRV2-R | CCGATCAATCAAGATCAGTCGAGA | |
BpActin-RT-F | TATGAACTTCCTGATGGG | 125 |
BpActin-RT-R | GAATTGTAGGTTGTCTCG | |
BpPDS-RT-F | ATTCGCCTCGTCCTGATA | 103 |
BpPDS-RT-R | ATGGGTTTGTGACCTGCA |
Fig. 4 Phenotype of BpPDS silenced B. peruviana ‘Thimma’ A: Control check; B: negative control; C, D: pTRV2-BpPDS336 silencing treatment; E, F: pTRV2-BpPDS457 silencing treatment. The circle indicates infection treatment part
Fig. 5 PCR amplification of virus veetor pTRV1 and pTRV2 pTRV1 detection: 1, 3, 5, 7, 9, 11, 13, 15 and 17; pTRV2 detection: 2, 4, 6, 8, 10, 12, 14, 16 and 18
处理 Treatment | 叶绿素a含量 Chlorophyll a content/(mg·g-1) | 叶绿素b含量 Chlorophyll b content/(mg·g-1) | 叶绿素含量 Chlorophyll content/(mg·g-1) | 类胡萝卜素含量 Carotenoid content/(mg·g-1) |
---|---|---|---|---|
空白对照 | 0.327 ± 0.021 a | 0.214 ± 0.030 a | 0.541 ± 0.052 a | 0.114 ± 0.004 a |
阴性对照 | 0.300 ± 0.007 a | 0.153 ± 0.010 b | 0.470 ± 0.025 a | 0.114 ± 0.010 a |
pTRV2-BpPDS336 | 0.230 ± 0.023 b | 0.090 ± 0.010 c | 0.310 ± 0.060 b | 0.087 ± 0.003 b |
pTRV2-BpPDS457 | 0.190 ± 0.016 c | 0.053 ± 0.010 d | 0.260 ± 0.035 b | 0.086 ± 0.020 b |
Table 2 Content of photosynthetic pigments in the leaves of B. peruviana ‘Thimma’ under different treatments
处理 Treatment | 叶绿素a含量 Chlorophyll a content/(mg·g-1) | 叶绿素b含量 Chlorophyll b content/(mg·g-1) | 叶绿素含量 Chlorophyll content/(mg·g-1) | 类胡萝卜素含量 Carotenoid content/(mg·g-1) |
---|---|---|---|---|
空白对照 | 0.327 ± 0.021 a | 0.214 ± 0.030 a | 0.541 ± 0.052 a | 0.114 ± 0.004 a |
阴性对照 | 0.300 ± 0.007 a | 0.153 ± 0.010 b | 0.470 ± 0.025 a | 0.114 ± 0.010 a |
pTRV2-BpPDS336 | 0.230 ± 0.023 b | 0.090 ± 0.010 c | 0.310 ± 0.060 b | 0.087 ± 0.003 b |
pTRV2-BpPDS457 | 0.190 ± 0.016 c | 0.053 ± 0.010 d | 0.260 ± 0.035 b | 0.086 ± 0.020 b |
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