割手密萜烯合成酶(TPS)基因家族分析及其在生物胁迫下的表达分析

doi:10.3724/SP.J.1006.2022.14237

作物学报 ›› 2022, Vol. 48 ›› Issue (12): 3029-3044.doi: 10.3724/SP.J.1006.2022.14237

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

割手密萜烯合成酶(TPS)基因家族分析及其在生物胁迫下的表达分析

林焕泰(), 张天杰, 史梦婷, 郭燕芳, 高三基, 王锦达()

福建农林大学国家甘蔗工程技术研究中心, 福建福州 350002

收稿日期:2021-12-15 接受日期:2022-03-25 出版日期:2022-12-12 网络出版日期:2022-04-19
通讯作者: 王锦达
作者简介:E-mail: 471250594@qq.com
基金资助:
国家重点研发计划项目(2018YFD201100);财政部和农业农村部国家现代农业产业技术体系建设专项(糖料, CARS-170302);福建省大学生创新训练计划项目(S202110389051);福建农林大学科技创新专项(CXZX2020084A)

Genome-wide analysis of terpene synthase (TPS) gene family and its expression under biological stress in Saccharum spontaneum

LIN Huan-Tai(), ZHANG Tian-Jie, SHI Meng-Ting, GUO Yan-Fang, GAO San-Ji, WANG Jin-Da()

National Engineering Research Center for Sugarcane, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China

Received:2021-12-15 Accepted:2022-03-25 Published:2022-12-12 Published online:2022-04-19
Contact: WANG Jin-Da
Supported by:
National Key Research and Development Program of China(2018YFD201100);China Agricultural Systrm of MOF and MARA(Sugar, CARS-170302);Innovation Training Program for College Students of Fujian Province(S202110389051);Special Fund for Science and Technology Innovation of Fujian Agriculture and Forestry University(CXZX2020084A)

摘要/Abstract

摘要：

由萜烯合酶(TPS)合成的萜类化合物在植物生物和非生物胁迫中起重要作用。作为现代甘蔗栽培种的重要亲本材料, 割手密(Saccharum spontaneum)含有大量的抗逆基因。为调查割手密TPS基因家族的特征和功能, 通过使用HMMER搜索在割手密基因组中鉴定出39个TPS基因, 该基因含有2个保守域(PF01397和PF03936)的蛋白质, 系统进化树分析显示SsTPS蛋白可以分为TPS-a、b、e/f和g四个分支。SsTPS基因家族主要通过片段复制进行扩展, 共有12个SsTPS基因参与了片段复制事件。定量实时PCR显示, 在草地贪夜蛾(Spodoptera frugiperda)胁迫和甘蔗白条黄单胞菌(Xanthomonas albilineans)感染的割手密植株中, SsTPS基因的表达模式不同, 其中7个SsTPS基因的表达受到强烈的调节。值得注意的是, SsTPS15在草地贪夜蛾的胁迫中显著上调, 但被甘蔗白条黄单胞菌感染时表达下调, 而SsTPS26、SsTPS37和SsTPS39显示相反的结果。研究结果对于进一步了解萜烯合酶的生物学作用和基于割手密的甘蔗抗逆育种具有重要意义。

关键词: 割手密, 萜烯合酶基因, 生物胁迫, 表达分析, 抗性育种

Abstract:

Terpenoids produced by the action of terpene synthase (TPS) enzymes play important roles in plant biotic and abiotic stress. Saccharum spontaneum, an important parent material of modern sugarcane cultivars, contains a large number of stress resistance genes. To investigate the characteristics and functions of the TPS gene family in S. spontaneum, 39 SsTPS genes were identified in S. spontaneum genome that encoded proteins with two conserved domains (PF01397 and PF03936) by using an HMMER search. The SsTPS proteins were divided into TPS-a, b, e/f, and g subfamilies. The SsTPS gene family had mainly expanded through segmental duplications, and a total of 12 SsTPS genes involved in segmental duplication events. In addition, qRT-PCR showed that the expression patterns of some SsTPS genes differed in S. spontaneum between Spodoptera frugiperda-stressed and Xanthomonas albilineans-infected plants, whereas the relative expression levels of seven SsTPS genes were strongly up-regulated. Notably, SsTPS15 were up-regulated in response to Spodoptera frugiperda-stressed but were down-regulated by X. albilineans infection, while SsTPS26, SsTPS37, and SsTPS39 had the opposite results. These results will be of great significance to further understanding the biological roles of terpene synthases and to develop resistant breeding in S. spontaneum.

Key words: Saccharum spontaneum, terpene synthase gene, biotic stresses, expression analysis, resistance breeding

林焕泰, 张天杰, 史梦婷, 郭燕芳, 高三基, 王锦达. 割手密萜烯合成酶(TPS)基因家族分析及其在生物胁迫下的表达分析[J]. 作物学报, 2022, 48(12): 3029-3044.

LIN Huan-Tai, ZHANG Tian-Jie, SHI Meng-Ting, GUO Yan-Fang, GAO San-Ji, WANG Jin-Da. Genome-wide analysis of terpene synthase (TPS) gene family and its expression under biological stress in Saccharum spontaneum[J]. Acta Agronomica Sinica, 2022, 48(12): 3029-3044.

图/表 10

表1

表2

割手密SsTPS家族成员基本信息和理化性质"

基因名称 Gene name	基因编号 Gene ID	氨基酸数 Number of amino acids	等电点 pI	分子量 Molecular weight (kD)	负电荷残基 Asp+Glu	正电荷残基Arg+Lys	不稳定系数Coefficient of instability	平均疏水性 GRAVY	脂肪系数 AI	α-螺旋 α-helix(%)	延伸链 Extended chain (%)	无规则卷曲 Random curl (%)
基因名称 Gene name	基因编号 Gene ID	氨基酸数 Number of amino acids	等电点 pI	分子量 Molecular weight (kD)	负电荷残基 Asp+Glu	正电荷残基Arg+Lys	不稳定系数Coefficient of instability	平均疏水性 GRAVY	脂肪系数 AI	α-螺旋 α-helix(%)	延伸链 Extended chain (%)	无规则卷曲 Random curl (%)	SsTPS1	Sspon.01G0009730-1A	514	5.90	59,392.66	72	62	36.28	-0.306	88.29	57.39	11.67	30.93
SsTPS2	Sspon.01G0010010-1A	496	5.29	57,214.41	78	57	37.65	-0.288	94.92	54.03	14.72	31.25
SsTPS3	Sspon.01G0044880-1B	497	5.75	56,761.45	69	54	40.04	-0.312	83.84	49.90	17.51	32.60
SsTPS4	Sspon.04G0011840-1A	735	9.77	81,962.81	74	104	60.66	-0.460	75.17	43.67	8.03	48.30
SsTPS5	Sspon.04G0011840-2B	353	5.78	41,253.51	51	43	47.03	-0.333	87.00	64.31	3.68	32.01
SsTPS6	Sspon.04G0011840-3C	501	6.43	57,711.12	62	58	45.18	-0.311	87.96	55.09	6.99	37.92
SsTPS7	Sspon.05G0015000-1A	581	5.53	66,604.19	78	61	39.07	-0.216	91.02	51.29	12.22	36.49
SsTPS8	Sspon.06G0010490-1P	528	5.33	61,308.56	76	59	39.80	-0.173	98.26	52.65	12.12	35.23
SsTPS9	Sspon.06G0010500-1A	376	5.10	43,465.54	59	41	52.71	-0.328	87.95	52.39	11.17	36.44
SsTPS10	Sspon.06G0011520-1A	433	5.84	49,561.84	62	51	40.69	-0.242	83.44	55.43	9.70	34.87
SsTPS11	Sspon.06G0011680-1A	468	5.32	53,911.93	71	58	49.94	-0.247	97.67	60.47	7.69	31.84
SsTPS12	Sspon.06G0010490-1A	538	5.34	62,470.81	78	61	40.28	-0.201	96.43	50.74	12.64	36.62
SsTPS13	Sspon.06G0021690-1B	325	5.52	37,701.75	51	39	46.74	-0.380	89.14	55.38	12.62	32.00
SsTPS14	Sspon.06G0023410-1B	397	5.47	46,236.32	64	52	38.72	-0.254	96.93	50.13	12.85	37.03
SsTPS15	Sspon.06G0010490-2B	538	5.34	62,468.83	78	61	40.74	-0.192	97.16	50.93	12.64	36.43
SsTPS16	Sspon.06G0011680-2B	607	5.96	70,631.16	81	70	49.39	-0.222	94.23	50.74	13.67	35.58
SsTPS17	Sspon.06G0024130-1B	606	5.97	70,317.44	86	74	53.41	-0.363	83.83	55.61	13.53	30.86
SsTPS18	Sspon.06G0015570-2B	533	5.29	61,439.38	76	57	45.23	-0.220	83.49	50.66	14.82	34.52
SsTPS19	Sspon.06G0023520-2C	491	5.08	57,333.63	76	54	49.64	-0.302	84.83	46.84	13.65	39.51
SsTPS20	Sspon.06G0011660-2C	482	5.04	55,885.74	75	53	48.91	-0.274	86.72	47.51	18.67	33.82
SsTPS21	Sspon.06G0011680-1P	414	5.47	47,542.37	62	48	48.39	-0.301	93.16	53.86	10.63	35.51
SsTPS22	Sspon.06G0030650-1C	358	9.40	41,189.25	38	51	38.63	-0.391	79.55	46.37	13.69	39.94
SsTPS23	Sspon.06G0011680-3C	468	5.15	53,761.70	72	55	51.78	-0.219	98.93	58.55	9.83	31.62
SsTPS24	Sspon.06G0034360-1D	317	4.93	36,898.43	50	32	39.19	-0.134	91.29	48.26	17.98	33.75
SsTPS25	Sspon.06G0011680-2P	333	8.65	38,083.58	38	42	35.22	-0.328	83.75	38.44	21.32	40.24
SsTPS26	Sspon.06G0024130-2D	545	5.56	63,780.15	81	68	48.69	-0.286	90.11	53.39	13.58	33.03
SsTPS27	Sspon.06G0024130-1P	497	5.71	57,610.39	75	65	43.03	-0.181	93.00	51.71	17.30	30.99
SsTPS28	Sspon.06G0011680-4D	328	8.47	37,598.00	37	40	33.44	-0.324	82.65	42.38	17.38	40.24
SsTPS29	Sspon.07G0027750-1B	509	5.55	59,706.37	75	59	55.08	-0.414	90.84	52.65	9.23	38.11
SsTPS30	Sspon.08G0029890-1D	629	5.83	71,361.82	80	67	49.40	-0.160	86.38	46.26	13.67	40.06
SsTPS31	Sspon.01G0039100-1B	348	5.62	39,581.24	49	39	48.50	-0.288	84.74	47.14	12.93	39.66
SsTPS32	Sspon.01G0039100-2C	550	8.25	62,827.75	67	70	54.38	-0.376	82.91	50.36	9.45	40.18
SsTPS33	Sspon.04G0000830-1A	388	5.42	44,015.32	47	38	54.09	-0.088	93.84	55.41	7.47	37.11
SsTPS34	Sspon.05G0030980-1C	545	6.04	62,692.00	73	67	49.04	-0.309	84.02	41.47	12.29	46.24
SsTPS35	Sspon.05G0002480-3D	783	6.03	87,936.45	90	18	46.73	-0.168	89.13	47.51	11.11	41.38
SsTPS36	Sspon.04G0020000-1A	550	5.33	62,580.79	74	53	52.94	-0.116	92.62	49.27	16.55	34.18
SsTPS37	Sspon.04G0019960-2C	515	5.28	59,084.06	71	52	47.41	-0.090	96.37	53.59	14.17	32.23
SsTPS38	Sspon.04G0020000-4P	563	6.13	63,167.56	68	58	50.45	-0.145	89.68	52.75	11.90	35.35
SsTPS39	Sspon.04G0020000-3D	552	5.25	63,021.29	78	56	51.02	-0.123	95.27	54.53	11.78	33.70

表2

图1

图2

图3

图4

图5

图6

图7

图8

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基因名称 Gene name	正向引物 Forward primer (5'-3')	反向引物 Reverse primer (5'-3')
SsTPS5	AGTACCGCATGCCCTACCTA	CACCGCTTCTTGCTGAAACC
SsTPS6	AGTACCGCATGCCCTACCTA	CGCTGCAAGTGTTCCTCAAC
SsTPS8	ATGATGTTCAGGGGCTGCAA	CATGTATGCACGCTTGGGTG
SsTPS10	AGGGGAAAGTGCGTCAGATG	AGTGTGGATACGGCTTAGCG
SsTPS15	AAAAGACCGGACCTTCCACC	CATGTATGCACGCTTGGGTG
SsTPS18	ACCAAGGAGGCATTCGAGTG	CGCGTTCTTCCTCCCTTTCT
SsTPS26	GTCCCATGCACTCCATCACA	GAGGTGGCAACGGTTCCTAA
SsTPS27	AATGGGCTCTCACCTTTCCG	AAGCTTGTTCACGCTCATGC
SsTPS35	TGTTCCTAGGCCATGCAAGG	TTTTGGACTCCGAGTGGCTC
SsTPS36	TGCTGCTGCGCAATGATTTT	GGTCGCTCTTCTGACGTTGA
SsTPS37	GAGCCAAGCCTTGCAGAGTA	ATGGGCAGGCTCTGTAGGTA
SsTPS39	GAAATACCTGTGGTGCGGGA	CCACAACATAGACAAGCGCG
GAPDH	CACGGCCACTGGAAGCA	TCCTCAGGGTTCCTGATGCC

割手密萜烯合成酶(TPS)基因家族分析及其在生物胁迫下的表达分析

Genome-wide analysis of terpene synthase (TPS) gene family and its expression under biological stress in Saccharum spontaneum

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