多花黄精 <i>PLT</i> 基因家族的鉴定与表达分析

朱田圆; 李小利; 张慧; 班景洁; 李丹; 罗彬彬; 朱建明; 林玉玲; 杜迎刚; 赖钟雄

doi:10.19303/j.issn.1008-0384.2023.06.006

多花黄精 PLT 基因家族的鉴定与表达分析

doi: 10.19303/j.issn.1008-0384.2023.06.006

朱田圆^1,,
李小利¹,
张慧¹,
班景洁¹,
李丹¹,
罗彬彬¹,
朱建明²,
林玉玲¹,
杜迎刚^{1, 3, ,},
赖钟雄^1, ,

1.
福建农林大学园艺植物生物工程研究所，福建　福州　350002
2.
福建金湖源生态农业有限公司，福建　邵武　354099
3.
潍坊科技学院/山东省高校设施园艺实验室，山东　潍坊　261000

基金项目: 福建农林大学横向科技项目（KH200373A）；福建省高原学科建设项目（102/71201801101）

详细信息

作者简介:
朱田圆（1997 —），女，硕士研究生，研究方向：园艺植物生物技术（E-mail：2414298958@qq.com）

通讯作者:
杜迎刚（1977 —），男，博士，教授，研究方向：害虫生物防治、园艺害虫生物技术（E-mail：eduyingang@163.com）

赖钟雄（1966 —），男，博士，研究员，研究方向：园艺植物的生物技术与遗传资源（E-mail：laizx01@163.com）

中图分类号: S567.239
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出版历程
- 收稿日期: 2022-12-02
- 修回日期: 2023-03-27
- 网络出版日期: 2023-06-02
- 刊出日期: 2023-06-28

Identification and Expressions of PLT Family of Polygonatum cyrtonema

1.
Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
Fujian Jinhuyuan Eco-agriculture Co., Ltd., Shaowu, Fujian　354099, China
3.
Weifang University of Science and Technology/Facility Horticulture Laboratory of Universities in Shandong, Weifang, Shandong　261000, China

摘要

摘要: 目的 PLT（Plethora）属于AP2/ERF型转录因子，在植物生长发育及抗胁迫等过程中发挥重要作用。本文旨在探究多花黄精PLT（Polygonatum cyrtonema Hua PLT，PcPLT）基因在根状茎器官生长发育中及盐胁迫下的调控作用，为深入研究PcPLT基因的功能提供理论依据。方法基于多花黄精转录组数据库进行PcPLT基因家族鉴定及生物信息学分析，通过qRT-PCR技术检测其在多花黄精不同组织部位及不同浓度NaCl处理下的相对表达量，利用RT-PCR技术构建PcPLT2-2、PcPLT2-7基因融合表达载体，观察其荧光信号验证亚细胞定位。结果共鉴定获得15个PcPLT家族成员，该家族无内含子结构，蛋白编码长度为159～601个氨基酸，均为亲水蛋白；进化树分析显示PcPLT与百合科石刁柏（Asparagus officinalis L.）亲缘关系最近；亚细胞定位预测及验证结果显示PcPLT2-2定位于细胞质、细胞核中，PcPLT2-7定位于细胞核中。qRT-PCR结果显示PcPLT2-3和PcPLT2-7在根状茎中表达最高，PcPLT1-3、PcPLT1-4、PcPLT2-7响应盐胁迫调控。结论 PcPLT不仅具有组织特异性还能提高抗逆性，提示可能作为器官发育调节因子参与多花黄精根状茎膨大的形态建成，为深入研究PLT调控植物根状茎器官生长发育的生物学功能和多花黄精遗传改良奠定基础。
- 多花黄精 /
- PLT /
- 基因家族 /
- 表达分析
Abstract: Objective Regulatory functions of Polygonatum cyrtonema Hua PLT (PcPLT) in the growth and development of rhizomes of the plant were investigated. Methods Based on the P. cyrtonema transcriptome database, the identity and bioinformatics of PcPLT family were obtained. Relative expressions in tissues of the plant and that under NaCl stress were detected using the qRT-PCR technique. The fusion expression vectors of PcPLT2-2 and PcPLT2-7 were constructed, and their fluorescence signals examined to determine subcellular localization. Results Fifteen PcPLT family members were identified. They were hydrophilic proteins, absent of intron structure, coded 159-601 amino acids, and evolutionarily closely related to Liliaceae Asparagus officinalis L. The predicted subcellular localization of PcPLT2-2 was in cytoplasm and nucleus, while PcPLT2-7 in nucleus only. PcPLT2-3 and PcPLT2-7 mostly in the rhizomes; and PcPLT1-3, PcPLT1-4, and PcPLT2-7 responsive to salt stress. Conclusion PcPLT were tissue-specific and capable of enhancing the stress resistance of P. cyrtonema. They might act as an organ development regulator associated with the morphogenesis of rhizome expansion. If so, the result obtained in this study would be of value for the in-depth understanding of the biological functions of PLT.
- Polygonatum cyrtonema Hua /
- PLT /
- gene family /
- expression analysis

HTML全文

图 1 PcPLT 蛋白保守基序Motif、保守结构域及基因结构

Figure 1. Conserved motif, conserved domains, and structure of PcPLT

下载: 全尺寸图片幻灯片

图 2 多花黄精PLT 家族系统进化树

Pc：多花黄精， At：拟南芥， Os：水稻， Ao：石刁柏， Eg：油棕， Vv：葡萄。

Figure 2. Phylogenetic trees of P. cyrtonema PLT families

Pc: P. cyrtonema Hua; At: Arabidopsis thaliana L.; Os: Oryza sativa L.; Ao: A. officinalis L.; Eg: Elaeis guineensis; Vv: Vitis vinifera.

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图 3 PcPLT在不同组织部位及盐胁迫下的表达模式分析

Figure 3. Expressions of PcPLTs in various tissues and under NaCl stress

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图 4 PcPLT在不同组织部位及盐胁迫下的qRT-PCR表达分析

A：PcPLT在不同组织部位的表达；B：不同NaCl浓度处理下PcPLT的表达。

Figure 4. qRT-PCR expressions of PcPLTs in various tissues and under NaCl stress

A: Expressions of PcPLT in tissues; B: PcPLT expression under stresses of varied NaCl concentrations.

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图 5 PcPLT蛋白在洋葱表皮中的亚细胞定位

A：pCAMBIA1302-GFP荧光信号；B：pCAMBIA1302-PcPLT2-2-GFP荧光信号；C：pCAMBIA1302-PcPLT2-7-GFP荧光信号。

Figure 5. Subcellular localization of PcPLT in onion epidermal cells

A: pCAMBIA1302-GFP fluorescence signal; B: pCAMBIA1302-PcPLT2-2-GFP fluorescence signal; C: pCAMBIA1302-PcPLT2-7-GFP fluorescence signal.

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表 1 引物序列

Table 1. Sequences of primers used

引物名称 Primer name	引物序列 Primer sequence (5′-3′)	退火温度Tm/℃	用途 Usage
PLT2-2-Q-F PLT2-2-Q-R	CGTCATCTTATCGTGGAGTTAC CATCTTTCCGCCTTGTTACT	55	qRT-PCR引物
PLT2-7-Q-F PLT2-7-Q-R	TCAAGTGTAATGGGAGGGA AGAGAATAGCGGCAATGTTG	55	qRT-PCR引物
β-tubulin-Q-F β-tubulin-Q-R	TTGTCGAAAATGCTGACGAG CAAGCTTCCGGAGATCAGAG	60	内参基因引物
PLT2-2-GFP-F PLT2-2-GFP-R	CATGCCATGGATGAAGAAGTTTGCGGTGCC GGACTAGTTAACTCAAGTTTGCAGGGTT	60	载体构建、亚细胞定位
PLT2-7-GFP-F PLT2-7-GFP-R	CATGCCATGGATGGGGAACTTGACCAAG GGACTAGTGCTCCTGGGGTAGAAATAGG	64	载体构建、亚细胞定位
PLT1-3-Q-F PLT1-3-Q-R	TACCTCCTCCACCACTTCTC TCTTCAAATCAGAGTCCGC	60	qRT-PCR引物
PLT1-4-Q-F PLT1-4-Q-R	ATGGTGAGTGCTTGGCTT TGCCTCCCTTTCCTTGTT	50	qRT-PCR引物

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表 2 多花黄精PLT 家族成员信息

Table 2. Information on PcPLT family members

基因名称 Gene name	基因ID Gene ID	注释到拟南芥成员 Annotated Arabidopsis	氨基酸数量 Amino acids/aa	分子量 Molecular weight/Da	等电点 pI	脂肪系数 Aliphatic index	二级结构 Secondary structure/%				亚细胞定位 Subcellular localization
基因名称 Gene name	基因ID Gene ID	注释到拟南芥成员 Annotated Arabidopsis	氨基酸数量 Amino acids/aa	分子量 Molecular weight/Da	等电点 pI	脂肪系数 Aliphatic index	α-螺旋 Alpha helix	β-转角 Beta turn	延伸链 Extended strand	无规卷曲 Random coil	亚细胞定位 Subcellular localization
PcPLT1-1	＞Cluster-11683.110576.p1	AtPLT1	317	35195.85	8.96	54.64	32.18	5.36	10.73	51.74	细胞核
PcPLT1-2	＞Cluster-11683.110575.p1	AtPLT1	469	51308.34	6.47	53.41	25.59	5.33	12.15	56.93	细胞核
PcPLT1-3	＞Cluster-11683.173509.p1	AtPLT1	468	51236.27	6.47	52.88	25.21	5.13	12.61	57.05	细胞核
PcPLT1-4	＞Cluster-11683.233671.p1	AtPLT1	421	47120.43	7.71	58.24	24.47	7.84	13.78	53.92	细胞质、细胞核
PcPLT2-1	＞Cluster-11683.222146.p1	AtPLT2	601	66506.19	6.51	59.63	18.47	3.99	12.48	65.06	细胞质、细胞核
PcPLT2-2	＞Cluster-11683.100274.p1	AtPLT2	277	30714.02	7.25	61.66	34.30	7.22	15.52	42.96	细胞质、细胞核
PcPLT2-3	＞Cluster-11683.77849.p1	AtPLT2	180	20995.90	9.64	70.94	33.89	7.78	26.11	32.22	细胞核
PcPLT2-4	＞Cluster-11683.163200.p1	AtPLT2	159	17827.93	6.48	67.61	42.77	5.66	16.35	35.22	细胞核
PcPLT2-5	＞Cluster-11683.105355.p1	AtPLT2	312	34191.20	6.94	73.91	26.92	6.73	13.14	53.21	细胞质、细胞核
PcPLT2-6	＞Cluster-11683.151415.p2	AtPLT2	199	22672.77	9.11	74.02	39.70	7.54	22.61	30.15	细胞核
PcPLT2-7	＞Cluster-11683.204809.p1	AtPLT2	332	36898.44	9.11	61.84	25.00	5.12	12.65	57.23	细胞核
PcPLT2-8	＞Cluster-11683.147730.p1	AtPLT2	395	44135.24	9.07	63.01	27.59	6.84	15.95	49.62	细胞质、细胞核
PcPLT2-9	＞Cluster-11683.151416.p1	AtPLT2	391	43773.69	8.89	59.64	24.55	7.42	13.81	54.22	细胞质、细胞核
PcPLT2-10	＞Cluster-11683.133871.p1	AtPLT2	380	42322.23	9.27	61.89	24.47	7.37	13.68	54.47	细胞质、细胞核
PcPLT2-11	＞Cluster-11683.182808.p2	AtPLT2	184	20988.80	9.26	69.46	39.67	7.07	18.48	34.78	细胞核

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