Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (12): 2619-2632.doi: 10.16420/j.issn.0513-353x.2022-1136
• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles
HUANG Jiayu, LIU Jieru, KANG Jiayin, CHI Xiufeng, SHEN Ping, CAI Ming, CHENG Tangren, WANG Jia, ZHANG Qixiang, PAN Huitang*()
Received:
2023-04-21
Revised:
2023-08-13
Online:
2023-12-25
Published:
2023-12-29
Contact:
PAN Huitang
HUANG Jiayu, LIU Jieru, KANG Jiayin, CHI Xiufeng, SHEN Ping, CAI Ming, CHENG Tangren, WANG Jia, ZHANG Qixiang, PAN Huitang. Effects of Three Auxin-Related Genes LfiPIN1,LfiPIN6 and LfiARF9 on Branching of Lagerstroemia indica[J]. Acta Horticulturae Sinica, 2023, 50(12): 2619-2632.
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URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2022-1136
用途 Use | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence | 扩增子长度/bp Amplicon length |
---|---|---|---|
基因克隆 Gene clone | LfiPIN1-F | ATGATAACAGCAGCAGACTTCTACC | 1 758 |
LfiPIN1-R | TCATATGCCCAGCGAGATGTAGTA | ||
LfiPIN6-F | ATGATCACCTCCCAAGATTTCTACAACGTC | 1 545 | |
LfiPIN6-R | CTATGAGGTAACAATGAAGTAAATCAAGGTGG | ||
LfiARF9-F | ATGGCGAATCGAGCTGAAGGTCCTG | 2 025 | |
LfiARF9-R | TCAGTTTTCAGCAGTGTCCGAGCTC | ||
qRT-PCR | LfiPIN1-F | GGAGCTGCGTACAGTAACAA | 121 |
LfiPIN1-R | GCTCACTCCTCTCCAAGTAATG | ||
LfiPIN6-F | AGTGAACACTCCAGCACATAAC | 124 | |
LfiPIN6-R | CGATGACCCATAACCGGATAAC | ||
LfiARF9-F | GAACACTCTGTGCCTTCTGT | 115 | |
LfiARF9-R | GGGCTGGTTGACTTGTAGTATC | ||
EF-α-F | GACTGTGCTGTGCTCATC | 146 | |
EF-α-R | GTGGCATCCATCTTGTTG | ||
TRV载体构建及载体检测 | LfiPIN1-F | TAAGGTTACCGAATTTGGAGTGCCTCGGGGTCG | 348 |
LfiPIN1-R | GCTCGGTACCGGATCCCATCCATGTCGACTCCG | ||
LfiPIN6-F | TAAGGTTACCGAATTCGGAAAACATTGGAGAATTCGCC | 348 | |
LfiPIN6-R | GCTCGGTACCGGATCGGGCTTCCAATCTCTCCAACG | ||
LfiARF9-F | TAAGGTTACCGAATTGGGAGAAGAAAGCCAGGG | 411 | |
LfiARF9-R | GCTCGGTACCGGATCTCTGGTACAATAATTGGGGTGG | ||
TRV1-F | TTACAGGTTATTTGGGCTAG | 648 | |
TRV1-R | CCGGGTTCAATTCCTTATC | ||
TRV2-F | TGTTTGAGGGAAAAGTAGAGAACGT | 321 | |
TRV2-R | TTACCGATCAATCAAGATCAGTCGA | ||
qRT-PCR (VIGS) | LfiPIN1-F | CAGACGAAGGTGGGAGAAATC | 142 |
LfiPIN1-R | GGACCAAGTGAGACCGAATAAG | ||
LfiPIN6-F | CGTCATGCTAAGGCTCATACTC | 82 | |
LfiPIN6-R | AGACCCAGGATGCTCGAATA | ||
LfiARF9-F | GAATCAGAGCCTTCCGTTGT | 102 | |
LfiARF9-R | GGGCGTAAACCTCATCTGTT | ||
pSuper1300 ::GFP载体构建 | LfiPIN1-F | TCGACTCTAGTCTAGATGATAACAGCAGCAGACTTCTACC | 1 788 |
LfiPIN1-R | TACCGGATCCACTAGTATGCCCAGCGAGATGTAG | ||
LfiPIN6-F | TCGACTCTAGTCTAGATGATCACCTCCCAAGATTTCTAC | 1 575 | |
LfiPIN6-R | TACCGGATCCACTAGTGAGGTAACAATGAAGTAAATCAAG | ||
LfiARF9-F | TCGACTCTAGTCTAGATGGCGAATCGAGCTGAAGG | 2 055 | |
LfiARF9-R | TACCGGATCCACTAGGTTTTCAGCAGTGTCCGAGC |
Table 1 Primer sequences
用途 Use | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence | 扩增子长度/bp Amplicon length |
---|---|---|---|
基因克隆 Gene clone | LfiPIN1-F | ATGATAACAGCAGCAGACTTCTACC | 1 758 |
LfiPIN1-R | TCATATGCCCAGCGAGATGTAGTA | ||
LfiPIN6-F | ATGATCACCTCCCAAGATTTCTACAACGTC | 1 545 | |
LfiPIN6-R | CTATGAGGTAACAATGAAGTAAATCAAGGTGG | ||
LfiARF9-F | ATGGCGAATCGAGCTGAAGGTCCTG | 2 025 | |
LfiARF9-R | TCAGTTTTCAGCAGTGTCCGAGCTC | ||
qRT-PCR | LfiPIN1-F | GGAGCTGCGTACAGTAACAA | 121 |
LfiPIN1-R | GCTCACTCCTCTCCAAGTAATG | ||
LfiPIN6-F | AGTGAACACTCCAGCACATAAC | 124 | |
LfiPIN6-R | CGATGACCCATAACCGGATAAC | ||
LfiARF9-F | GAACACTCTGTGCCTTCTGT | 115 | |
LfiARF9-R | GGGCTGGTTGACTTGTAGTATC | ||
EF-α-F | GACTGTGCTGTGCTCATC | 146 | |
EF-α-R | GTGGCATCCATCTTGTTG | ||
TRV载体构建及载体检测 | LfiPIN1-F | TAAGGTTACCGAATTTGGAGTGCCTCGGGGTCG | 348 |
LfiPIN1-R | GCTCGGTACCGGATCCCATCCATGTCGACTCCG | ||
LfiPIN6-F | TAAGGTTACCGAATTCGGAAAACATTGGAGAATTCGCC | 348 | |
LfiPIN6-R | GCTCGGTACCGGATCGGGCTTCCAATCTCTCCAACG | ||
LfiARF9-F | TAAGGTTACCGAATTGGGAGAAGAAAGCCAGGG | 411 | |
LfiARF9-R | GCTCGGTACCGGATCTCTGGTACAATAATTGGGGTGG | ||
TRV1-F | TTACAGGTTATTTGGGCTAG | 648 | |
TRV1-R | CCGGGTTCAATTCCTTATC | ||
TRV2-F | TGTTTGAGGGAAAAGTAGAGAACGT | 321 | |
TRV2-R | TTACCGATCAATCAAGATCAGTCGA | ||
qRT-PCR (VIGS) | LfiPIN1-F | CAGACGAAGGTGGGAGAAATC | 142 |
LfiPIN1-R | GGACCAAGTGAGACCGAATAAG | ||
LfiPIN6-F | CGTCATGCTAAGGCTCATACTC | 82 | |
LfiPIN6-R | AGACCCAGGATGCTCGAATA | ||
LfiARF9-F | GAATCAGAGCCTTCCGTTGT | 102 | |
LfiARF9-R | GGGCGTAAACCTCATCTGTT | ||
pSuper1300 ::GFP载体构建 | LfiPIN1-F | TCGACTCTAGTCTAGATGATAACAGCAGCAGACTTCTACC | 1 788 |
LfiPIN1-R | TACCGGATCCACTAGTATGCCCAGCGAGATGTAG | ||
LfiPIN6-F | TCGACTCTAGTCTAGATGATCACCTCCCAAGATTTCTAC | 1 575 | |
LfiPIN6-R | TACCGGATCCACTAGTGAGGTAACAATGAAGTAAATCAAG | ||
LfiARF9-F | TCGACTCTAGTCTAGATGGCGAATCGAGCTGAAGG | 2 055 | |
LfiARF9-R | TACCGGATCCACTAGGTTTTCAGCAGTGTCCGAGC |
Fig. 2 Phylogenetic tree(A)and motif compositions(B)of LfiPIN1 and LfiPIN6 with PIN proteins from other species At:Arabidopsis thaliana;Pt:Populus tomentosa;Pgr:Punica granatum;Eucgr:Eucalyptus grandis.
Fig. 3 Phylogenetic tree(A)and motif compositions(B)of LfiARF9 with ARF proteins from other species At:Arabidopsis thaliana;Pt:Populus tomentosa;Pgr:Punica granatum;Eucgr:Eucalyptus grandis.
Fig. 4 Expression levels of LfiPIN1,LfiPIN6 and LfiARF9 in different parts of Lagerstroemia indica standard cultivar S126 and dwarf compact cultivar D071 Different lowercase letters indicate significant difference at 0.05 level.
株系 Lines | 取样部位 Segments | 生长素 IAA | 独脚金内酯 Strigolactone | 油菜素内酯 Brassinolide |
---|---|---|---|---|
S126 | 上部 Upper | 100.93 ± 9.21 a | 64.49 ± 0.22 a | 118.60 ± 13.06 b |
中部 Middle | 94.95 ± 10.83 a | 50.97 ± 0.33 b | 164.54 ± 7.91 a | |
下部 Lower | 69.73 ± 6.01 b | 49.12 ± 0.24 c | 123.62 ± 1.79 b | |
D071 | 上部 Upper | 94.47 ± 7.20 a | 66.62 ± 0.46 a | 125.65 ± 13.00 a |
中部 Middle | 76.90 ± 3.18 b | 55.20 ± 0.34 c | 143.26 ± 0.95 a | |
下部 Lower | 77.29 ± 3.26 b | 58.88 ± 0.65 b | 131.57 ± 3.57 a |
Table 2 Contents of endogenous hormones in different parts of stem segments of two types of crape myrtleng · g-1
株系 Lines | 取样部位 Segments | 生长素 IAA | 独脚金内酯 Strigolactone | 油菜素内酯 Brassinolide |
---|---|---|---|---|
S126 | 上部 Upper | 100.93 ± 9.21 a | 64.49 ± 0.22 a | 118.60 ± 13.06 b |
中部 Middle | 94.95 ± 10.83 a | 50.97 ± 0.33 b | 164.54 ± 7.91 a | |
下部 Lower | 69.73 ± 6.01 b | 49.12 ± 0.24 c | 123.62 ± 1.79 b | |
D071 | 上部 Upper | 94.47 ± 7.20 a | 66.62 ± 0.46 a | 125.65 ± 13.00 a |
中部 Middle | 76.90 ± 3.18 b | 55.20 ± 0.34 c | 143.26 ± 0.95 a | |
下部 Lower | 77.29 ± 3.26 b | 58.88 ± 0.65 b | 131.57 ± 3.57 a |
Fig. 6 Axillary bud sprouting status in Lagerstroemia indica with silenced LfiPIN1,LfiPIN6 and LfiARF9 after seven days The arrow points to the axillary bud status after decapitation,and the area inside the dashed line indicates the location of injection for the infiltration solution.
处理 Treatment | 枝条长度/cm Branch lenghth | 节间长度/cm Internode length | 腋芽数 Number of axillary bud | 腋芽萌发数 Number of sprouted axillary bud |
---|---|---|---|---|
未处理植株(WT) | 13.73 ± 0.91 a | 2.32 ± 0.14 a | 10.75 ± 0.24 ab | 0 |
pTRV2对照植株pTRV2 control plant | 13.25 ± 1.48 a | 2.30 ± 0.29 ab | 9.38 ± 0.80 ab | 0 |
LfiPIN1沉默植株LfiPIN1 silenced plant | 8.66 ± 1.85 b | 1.72 ± 0.15 d | 9.98 ± 1.94 ab | 0 |
LfiPIN6沉默植株LfiPIN6 silenced plant | 8.70 ± 1.82 b | 1.98 ± 0.22 c | 8.72 ± 1.78 b | 1.31 ± 0.68 |
LfiARF9沉默植株LfiARF9 silenced plant | 12.12 ± 2.45 a | 2.07 ± 0.08 bc | 11.55 ± 2.04 a | 1.64 ± 0.78 |
Table 3 Phenotype of new shoots in Lagerstroemia indica with silenced LfiPIN1、LfiPIN6、and LfiARF9
处理 Treatment | 枝条长度/cm Branch lenghth | 节间长度/cm Internode length | 腋芽数 Number of axillary bud | 腋芽萌发数 Number of sprouted axillary bud |
---|---|---|---|---|
未处理植株(WT) | 13.73 ± 0.91 a | 2.32 ± 0.14 a | 10.75 ± 0.24 ab | 0 |
pTRV2对照植株pTRV2 control plant | 13.25 ± 1.48 a | 2.30 ± 0.29 ab | 9.38 ± 0.80 ab | 0 |
LfiPIN1沉默植株LfiPIN1 silenced plant | 8.66 ± 1.85 b | 1.72 ± 0.15 d | 9.98 ± 1.94 ab | 0 |
LfiPIN6沉默植株LfiPIN6 silenced plant | 8.70 ± 1.82 b | 1.98 ± 0.22 c | 8.72 ± 1.78 b | 1.31 ± 0.68 |
LfiARF9沉默植株LfiARF9 silenced plant | 12.12 ± 2.45 a | 2.07 ± 0.08 bc | 11.55 ± 2.04 a | 1.64 ± 0.78 |
处理 | 生长素/(ng · g-1) IAA | 独脚金内酯/(ng · g-1) Strigolactone | 油菜素内酯/(ng · g-1) Brassinolide |
---|---|---|---|
未处理植株(WT) | 90.02 ± 2.54 b | 62.45 ± 2.31 b | 125.89 ± 3.78 bc |
pTRV2对照植株 pTRV2 control plant | 99.81 ± 1.75 a | 68.67 ± 0.83 a | 129.79 ± 2.34 b |
LfiPIN1沉默植株 LfiPIN1 silenced plant | 83.13 ± 2.63 c | 64.82 ± 0.48 b | 122.49 ± 2.70 c |
LfiPIN6沉默植株 LfiPIN6 silenced plant | 82.86 ± 2.83 c | 55.05 ± 2.15 c | 146.08 ± 4.11 a |
LfiARF9沉默植株LfiARF9 silenced plant | 93.90 ± 3.19 b | 56.63 ± 1.58 c | 126.15 ± 2.82 bc |
Table 4 Hormone content in stem segments of plants after gene silencing hormone content in stem segments of Lagerstroemia indica with silenced LfiPIN1,LfiPIN6,and LfiARF9
处理 | 生长素/(ng · g-1) IAA | 独脚金内酯/(ng · g-1) Strigolactone | 油菜素内酯/(ng · g-1) Brassinolide |
---|---|---|---|
未处理植株(WT) | 90.02 ± 2.54 b | 62.45 ± 2.31 b | 125.89 ± 3.78 bc |
pTRV2对照植株 pTRV2 control plant | 99.81 ± 1.75 a | 68.67 ± 0.83 a | 129.79 ± 2.34 b |
LfiPIN1沉默植株 LfiPIN1 silenced plant | 83.13 ± 2.63 c | 64.82 ± 0.48 b | 122.49 ± 2.70 c |
LfiPIN6沉默植株 LfiPIN6 silenced plant | 82.86 ± 2.83 c | 55.05 ± 2.15 c | 146.08 ± 4.11 a |
LfiARF9沉默植株LfiARF9 silenced plant | 93.90 ± 3.19 b | 56.63 ± 1.58 c | 126.15 ± 2.82 bc |
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