生长素相关基因LfiPIN1、LfiPIN6和LfiARF9对紫薇分枝的影响

doi:10.16420/j.issn.0513-353x.2022-1136

摘要/Abstract

摘要：

为探究紫薇（Lagerstroemia indica）分枝性状的形成机制，以紫薇标准型和低矮紧凑型株系为材料，以分枝相关的生长素转运基因LfiPIN1、LfiPIN6和响应因子LfiARF9为对象，利用VIGS沉默（紫薇）和过表达（烟草 Nicotiana tabacum）技术对3个基因进行功能分析。结果表明，LfiPIN6和LfiARF9通过抑制侧芽的萌发而负调控分枝的数量;LfiPIN1和LfiPIN6通过调节新生侧枝的生长素含量正调控侧枝的长度，而LfiARF9并不影响侧枝长度。

关键词: 紫薇, 分枝, 生长素, 功能验证

Abstract:

To investigate the mechanism underlying branching in Lagerstroemia indica，both standard and dwarf compact L. indica strains were used as material，the function of three genes involved in auxin transport and response，namely LfiPIN1，LfiPIN6 and LfiARF9，were analyzed using virus-induced gene silencing（VIGS）in L. indica and overexpression techniques in Nicotiana tabacum. The effects of these genes on lateral bud initiation and lateral branch length in L. indica were examined. The results showed that LfiPIN6 and LfiARF9 negatively regulate the number of lateral branches in L. indica by inhibiting the sprouting of lateral buds. LfiPIN1 and LfiPIN6 positively regulate the length of lateral branches by regulating the auxin content in newly formed lateral branches. LfiARF9 was found to have no effect on lateral branch length.

Key words: Lagerstroemia indica, branching, auxin, function verification

黄佳玉, 刘洁茹, 康佳音, 池秀凤, 申萍, 蔡明, 程堂仁, 王佳, 张启翔, 潘会堂. 生长素相关基因LfiPIN1、LfiPIN6和LfiARF9对紫薇分枝的影响[J]. 园艺学报, 2023, 50(12): 2619-2632.

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.

图/表 14

参考文献 40

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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	1 758
	LfiPIN6-F	ATGATCACCTCCCAAGATTTCTACAACGTC	1 545
	LfiPIN6-R	CTATGAGGTAACAATGAAGTAAATCAAGGTGG	1 545
	LfiARF9-F	ATGGCGAATCGAGCTGAAGGTCCTG	2 025
	LfiARF9-R	TCAGTTTTCAGCAGTGTCCGAGCTC	2 025
qRT-PCR	LfiPIN1-F	GGAGCTGCGTACAGTAACAA	121
	LfiPIN1-R	GCTCACTCCTCTCCAAGTAATG	121
	LfiPIN6-F	AGTGAACACTCCAGCACATAAC	124
	LfiPIN6-R	CGATGACCCATAACCGGATAAC	124
	LfiARF9-F	GAACACTCTGTGCCTTCTGT	115
	LfiARF9-R	GGGCTGGTTGACTTGTAGTATC	115
	EF-α-F	GACTGTGCTGTGCTCATC	146
	EF-α-R	GTGGCATCCATCTTGTTG	146
TRV载体构建及载体检测	LfiPIN1-F	TAAGGTTACCGAATTTGGAGTGCCTCGGGGTCG	348
	LfiPIN1-R	GCTCGGTACCGGATCCCATCCATGTCGACTCCG	348
	LfiPIN6-F	TAAGGTTACCGAATTCGGAAAACATTGGAGAATTCGCC	348
	LfiPIN6-R	GCTCGGTACCGGATCGGGCTTCCAATCTCTCCAACG	348
	LfiARF9-F	TAAGGTTACCGAATTGGGAGAAGAAAGCCAGGG	411
	LfiARF9-R	GCTCGGTACCGGATCTCTGGTACAATAATTGGGGTGG	411
	TRV1-F	TTACAGGTTATTTGGGCTAG	648
	TRV1-R	CCGGGTTCAATTCCTTATC	648
	TRV2-F	TGTTTGAGGGAAAAGTAGAGAACGT	321
	TRV2-R	TTACCGATCAATCAAGATCAGTCGA	321
qRT-PCR (VIGS)	LfiPIN1-F	CAGACGAAGGTGGGAGAAATC	142
	LfiPIN1-R	GGACCAAGTGAGACCGAATAAG	142
	LfiPIN6-F	CGTCATGCTAAGGCTCATACTC	82
	LfiPIN6-R	AGACCCAGGATGCTCGAATA	82
	LfiARF9-F	GAATCAGAGCCTTCCGTTGT	102
	LfiARF9-R	GGGCGTAAACCTCATCTGTT	102
pSuper1300 ::GFP载体构建	LfiPIN1-F	TCGACTCTAGTCTAGATGATAACAGCAGCAGACTTCTACC	1 788
	LfiPIN1-R	TACCGGATCCACTAGTATGCCCAGCGAGATGTAG	1 788
	LfiPIN6-F	TCGACTCTAGTCTAGATGATCACCTCCCAAGATTTCTAC	1 575
	LfiPIN6-R	TACCGGATCCACTAGTGAGGTAACAATGAAGTAAATCAAG	1 575
	LfiARF9-F	TCGACTCTAGTCTAGATGGCGAATCGAGCTGAAGG	2 055
	LfiARF9-R	TACCGGATCCACTAGGTTTTCAGCAGTGTCCGAGC	2 055

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	1 758
	LfiPIN6-F	ATGATCACCTCCCAAGATTTCTACAACGTC	1 545
	LfiPIN6-R	CTATGAGGTAACAATGAAGTAAATCAAGGTGG	1 545
	LfiARF9-F	ATGGCGAATCGAGCTGAAGGTCCTG	2 025
	LfiARF9-R	TCAGTTTTCAGCAGTGTCCGAGCTC	2 025
qRT-PCR	LfiPIN1-F	GGAGCTGCGTACAGTAACAA	121
	LfiPIN1-R	GCTCACTCCTCTCCAAGTAATG	121
	LfiPIN6-F	AGTGAACACTCCAGCACATAAC	124
	LfiPIN6-R	CGATGACCCATAACCGGATAAC	124
	LfiARF9-F	GAACACTCTGTGCCTTCTGT	115
	LfiARF9-R	GGGCTGGTTGACTTGTAGTATC	115
	EF-α-F	GACTGTGCTGTGCTCATC	146
	EF-α-R	GTGGCATCCATCTTGTTG	146
TRV载体构建及载体检测	LfiPIN1-F	TAAGGTTACCGAATTTGGAGTGCCTCGGGGTCG	348
	LfiPIN1-R	GCTCGGTACCGGATCCCATCCATGTCGACTCCG	348
	LfiPIN6-F	TAAGGTTACCGAATTCGGAAAACATTGGAGAATTCGCC	348
	LfiPIN6-R	GCTCGGTACCGGATCGGGCTTCCAATCTCTCCAACG	348
	LfiARF9-F	TAAGGTTACCGAATTGGGAGAAGAAAGCCAGGG	411
	LfiARF9-R	GCTCGGTACCGGATCTCTGGTACAATAATTGGGGTGG	411
	TRV1-F	TTACAGGTTATTTGGGCTAG	648
	TRV1-R	CCGGGTTCAATTCCTTATC	648
	TRV2-F	TGTTTGAGGGAAAAGTAGAGAACGT	321
	TRV2-R	TTACCGATCAATCAAGATCAGTCGA	321
qRT-PCR (VIGS)	LfiPIN1-F	CAGACGAAGGTGGGAGAAATC	142
	LfiPIN1-R	GGACCAAGTGAGACCGAATAAG	142
	LfiPIN6-F	CGTCATGCTAAGGCTCATACTC	82
	LfiPIN6-R	AGACCCAGGATGCTCGAATA	82
	LfiARF9-F	GAATCAGAGCCTTCCGTTGT	102
	LfiARF9-R	GGGCGTAAACCTCATCTGTT	102
pSuper1300 ::GFP载体构建	LfiPIN1-F	TCGACTCTAGTCTAGATGATAACAGCAGCAGACTTCTACC	1 788
	LfiPIN1-R	TACCGGATCCACTAGTATGCCCAGCGAGATGTAG	1 788
	LfiPIN6-F	TCGACTCTAGTCTAGATGATCACCTCCCAAGATTTCTAC	1 575
	LfiPIN6-R	TACCGGATCCACTAGTGAGGTAACAATGAAGTAAATCAAG	1 575
	LfiARF9-F	TCGACTCTAGTCTAGATGGCGAATCGAGCTGAAGG	2 055
	LfiARF9-R	TACCGGATCCACTAGGTTTTCAGCAGTGTCCGAGC	2 055

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

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

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