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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (12): 3029-3044.doi: 10.3724/SP.J.1006.2022.14237

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

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()   

  1. National Engineering Research Center for Sugarcane, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
  • Received:2021-12-15 Accepted:2022-03-25 Online:2022-12-12 Published:2022-04-19
  • Contact: WANG Jin-Da E-mail:471250594@qq.com;jdwang@fafu.edu.cn
  • 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)

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

Table 1

Primer sequence used to detect SsTPS gene expression level"

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

Table 2

Basin information and physiochemical properties of SsTPS genes family in S. spontaneum"

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

Fig. 1

Phylogenetic tree of TPS gene family in S. spontaneum (Ss), Oryza sativa (Os), and Sorghum bicolor (Sb) TPS-a, b, e/f, and g represent different branches of TPS gene family, respectively."

Fig. 2

Genetic structures of gene family members of S. spontaneum A: the phylogenetic tree of SsTPSs. B: the prediction of intron-exon structure of SsTPSs. C: the motifs prediction of SsTPSs. D: the motif of base sequence structure."

Fig. 3

Chromosomal localization of the terpene synthase genes family in S. spontaneum"

Fig. 4

Collinear analysis of the terpene synthase genes family in S. spontaneum"

Fig. 5

Types and numbers of cis-acting elements in promoter regions of each member of the TPS gene family in S. spontaneum 1-39 represent the S. spontaneum TPS gene family member from SsTPS1 to SsTPS39, respectively."

Fig. 6

Heat map of the expression of TPS genes in different tissues of S. spontaneum"

Fig. 7

Relative expression levels of 12 candidate SsTPS genes in S. spontaneum under S. frugiperda stress *, **, and *** mean significant difference at the 0.05, 0.01, and 0.001 probability levels, respectively."

Fig. 8

Expression analysis of 12 SsTPS genes under X. albilineans infection in S. spontaneum Different lowercase letters above the bars mean significant difference at the 0.05 probability level among the treatments."

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