高粱CPP基因家族鉴定及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0569

摘要/Abstract

摘要：

【目的】CPP基因家族在真核生物中广泛存在，对植物的生长发育起到重要的作用。从高粱基因组中鉴定CPP基因家族成员并分析其表达特征，为高粱CPP基因家族功能研究及遗传改良提供理论依据。【方法】通过生物信息学技术的方法对高粱CPP基因家族进行序列分析，并联合转录组和RT-qPCR技术分析SbCPP在高粱不同组织及盐碱胁迫下的表达情况。【结果】高粱中共鉴定到8个SbCPP基因，不均匀地分布在7条染色体上，家族各成员间具有相似的基因结构及蛋白理化性质；结合进化树分析与共线性分析结果表明，SbCPP基因与水稻存在密切亲缘关系，且共存在6对同源基因；启动子分析发现，SbCPP基因启动子含有光响应、激素响应、应激响应等元件。转录组数据分析结果表明，SbCPP基因可能参与高粱响应盐碱胁迫的调控过程。通过实时荧光定量PCR发现，SbCPP基因在高粱盐碱胁迫中广泛表达，但家族成员在不同时期中的表达模式存在差异，SbCPP01、SbCPP04在12 h表达较高，SbCPP02在6 h表达较高，SbCPP03在1 h表达量较高，上述基因表达显著。【结论】在高粱中获得了8个CPP基因，其中SbCPP01、SbCPP02、SbCPP04基因在盐碱胁迫处理下呈现高表达，表明这些基因在高粱抵御盐碱胁迫中发挥重要作用。

关键词: 高粱, CPP基因, 生物信息学, RT-qPCR, 表达模式分析

Abstract:

【Objective】CPP gene family exists widely in eukaryotes and plays an important role in plant growth and development. Identifying the members of the CPP gene family from the sorghum genome and analyzing their expression characteristics provides a theoretical basis for functional study and genetic improvement of the CPP gene family in sorghum.【Method】Bioinformatics techniques were used to analyze the sequences of the sorghum CPP gene family, and transcriptome and RT-qPCR techniques were used to analyze the expressions of SbCPP in different tissues of sorghum and under salt alkali stress.【Result】A total of eight SbCPP genes were identified in sorghum, and they were distributed unevenly on seven chromosomes and had similar gene structure and protein physicochemical properties among the members of the families. The SbCPP gene was closely related to rice（Oryza sativa）, and there were 6 pairs of homologous genes. The promoter analysis showed that the SbCPP gene promoter contained elements such as light response, hormone response and stress response. The transcriptome data analysis results indicated that SbCPP gene might be involved in the regulation process of sorghum under saline-alkali stress. The results of real-time quantitative PCR showed that SbCPP gene was widely expressed in sorghum under saline-alkali stress, but the expression patterns of family members varied in different periods. The expressions of SbCPP01 and SbPP04 were high at 12 h, and that of SbPP02 was high at 6 h, the expression of SbPP03 was high at 1 h, and the above genes were significantly expressed.【Conclusion】Eight CPP genes were obtained from sorghum, among which SbCPP01, SbCPP02 and SbCPP04 were highly expressed under saline-alkali stress, indicating that these genes play an important role in resisting saline-alkali stress in sorghum.

Key words: sorghum, CPP gene, bioinformatics, RT-qPCR, expression pattern analysis

杜品廷, 吴国江, 王振国, 李岩, 周伟, 周亚星. 高粱CPP基因家族鉴定及表达分析[J]. 生物技术通报, 2025, 41(1): 132-142.

DU Pin-ting, WU Guo-jiang, WANG Zhen-guo, LI Yan, ZHOU Wei, ZHOU Ya-xing. Identification and Expression Analysis of CPP Gene Family in Sorghum[J]. Biotechnology Bulletin, 2025, 41(1): 132-142.

图/表 11

表1 高粱CPP基因家族表达分析的实时荧光定量引物

Table 1 RT-qPCR primers for the expression analysis of CPP gene family in sorghum

Gene	Forward primer（5'-3'）	Reverse primer（5'-3'）
GAPDH	CCTGTACCGTCCCTCGACTT	ATGCTTGCACCCTGTACTGC
SbCPP01	CAAGAACTACGAAGGGAGTGAGGAG	GGCACCATTCAGAGCAACATTAGC
SbCPP02	TGCTTCGCTTCTGGTACTCACTG	TCACGCCTGGCAACCTCATTC
SbCPP03	CCAACAAGAGGAACAGCAATGACAG	AGGATCTGGACGCACTTCGATATG
SbCPP04	ACAGGAAGAGGAGGAGGAAGGC	AGGTTCGCATCCACATGACTCAG
SbCPP05	CACCAAACAAGAAGCGAATCTCTCC	CCACCAAGGGAAGGGAAAGAAGG
SbCPP06	GGCTTCCATTGGTCTGAAACTTCG	TTAGTGCTTGTGTTGTTGGAGATGC
SbCPP07	CGCAAGCAGACTGAATCTCGTAAC	CGGAAGCAGGAGTGTTGTTAGAATC
SbCPP08	GTTCTCTCCACTCGGAAGCAGATAG	TTCCAGACCAGCATCAGACATACG

表2 高粱8个CPP基因家族成员的理化性质

Table 2 Physicochemical characteristics of 8 members of the CPP gene family in sorghum

基因 Gene	基因号 Gene ID	氨基酸数目Number of amino acids/aa	分子量Molecular weight/kD	等电点pI	不稳定性系数Instability index	脂肪系数Aliphatic index	亲水性平均值Grand average of hydropathicity	亚细胞定位预测 Prediction of subcellular location
SbCPP01	Sobic.010G147000.v3.2	509	56 257.23	7.11	51.64	64.24	-0.758	细胞核 Nucleus
SbCPP02	Sobic.003G303800.v3.2	613	67 297.16	8.84	57.48	62.07	-0.747	细胞核 Nucleus
SbCPP03	Sobic.009G191600.v3.2	359	40 038.23	8.5	52.7	56.55	-0.735	细胞核 Nucleus
SbCPP04	Sobic.008G160500.v3.2	409	44 239.48	6.82	80.47	64.72	-0.543	细胞核 Nucleus
SbCPP05	Sobic.008G160600.v3.2	767	82 644.05	6.54	59.91	66.4	-0.579	细胞核 Nucleus
SbCPP06	Sobic.004G124300.v3.2	543	59 446.12	7.17	58.52	53.22	-0.779	细胞核 Nucleus
SbCPP07	Sobic.001G159500.v3.2	747	81 124.88	8.1	54.62	68.3	-0.579	细胞核 Nucleus
SbCPP08	Sobic.002G050700.v3.2	776	83 360.06	6.04	53.58	64.86	-0.599	细胞核 Nucleus

表3 高粱CPP家族二级蛋白结构预测

Table 3 Prediction of secondary protein structure of sorghum CPP family

蛋白Protein	α-螺旋α-helix/%	延长链 Extended strand/%	β-转角β-turn/%	无规则卷曲Random coil/%
SbCPP01	27.11	6.48	3.34	63.06
SbCPP02	27.41	5.38	3.92	63.30
SbCPP03	21.17	9.75	3.62	65.46
SbCPP04	19.56	11.00	3.67	65.77
SbCPP05	16.95	9.91	1.96	71.19
SbCPP06	26.15	5.34	3.50	65.01
SbCPP07	18.74	9.64	2.41	69.21
SbCPP08	21.13	6.70	2.45	69.72

图1 高粱CPP基因家族染色体定位

Fig. 1 Chromosomal mapping of the CPP gene family in sorghum

图2 高粱、拟南芥和水稻基因家族的系统进化树 Sb：高粱；Os：水稻；At：拟南芥

Fig. 2 Phylogenetic tree of gene families in sorghum, Ara-bidopsis, and rice Sb: Sorghum; Os: rice; At: Arabidopsis

图3 高粱CPP基因家族保守基序与基因结构分析 A：SbCPP家族成员系统进化树；B：SbCPP家族成员保守基序图；C：SbCPP家族成员基因结构图

Fig. 3 Conservative motifs and gene structure analysis of the CPP gene family in sorghum members A: Phylogenetic tree of SbCPP family members. B: Mapping of conserved motifs for SbCPP family members. C: Mapping of gene structure for SbCPP family

图4 高粱CPP基因家族种间共线性分析

Fig. 4 Interspecific collinearity analysis of the CPP gene family in sorghum

图5 高粱CPP基因家族启动子顺式作用元件

Fig. 5 Cis-acting element of CPP gene family promoter in sorghum

图6 高粱CPP基因组织表达模式 1：幼叶；2：幼根；3：幼茎；4：芽；5：穗花芽分化；6：花序梗花芽分化；7：种子；8：幼嫩根尖

Fig. 6 Analysis of tissue expression of CPP gene in sorghum 1: Young leaves. 2: Young roots. 3: Young stem. 4: Bud. 5: Panicle floral initiation. 6: Peduncle loral initiation.7: Seed. 8: Root top juvenile

图7 高粱CPP基因不同材料表达模式分析 M：盐碱敏感材料T501；N：耐盐碱材料T84：CK，2，4分别代表对照，胁迫2 d，4 d

Fig. 7 Analysis of expression patterns of CPP gene in different materials of sorghum M: Saline-alkali sensitive material T501, N: saline-alkali resistant material T84, CK, 2,4 refers to control, stress 2 d, and 4 d, respectively

图8 高粱CPP基因家族盐碱胁迫表达模式数据为3个独立生物学重复的平均值（±SE）。柱状图上方的不同字母表示 P<0.05 时显著差异

Fig. 8 Salt alkali stress expression patterns of the CPP gene family in sorghum Data are the mean of 3 independent biological replicates（±SE）. Different letters above the bar chart indicate significant differences at P<0.05

参考文献 28

[1]	李顺国, 刘猛, 刘斐, 等. 中国高粱产业和种业发展现状与未来展望[J]. 中国农业科学, 2021, 54(3): 471-482. doi: 10.3864/j.issn.0578-1752.2021.03.002
	Li SG, Liu M, Liu F, et al. Current status and future prospective of sorghum production and seed industry in China[J]. Sci Agric Sin, 2021, 54(3): 471-482. doi: 10.3864/j.issn.0578-1752.2021.03.002
[2]	罗洪, 张丽敏, 夏艳, 等. 能源植物高粱基因组研究进展[J]. 科技导报, 2015, 33(16): 17-26. doi: 10.3981/j.issn.1000-7857.2015.16.002
	Luo H, Zhang LM, Xia Y, et al. An update ongenome research of biofuel sorghum(Sorghum bicolour)[J]. Sci Technol Rev, 2015, 33(16): 17-26.
[3]	Kim M, Day DF. Composition of sugar cane, energy cane, and sweet sorghum suitable for ethanol production at Louisiana sugar Mills[J]. J Ind Microbiol Biotechnol, 2011, 38(7): 803-807.
[4]	Yang Z, Li JL, Liu LN, et al. Photosynthetic regulation under salt stress and salt-tolerance mechanism of sweet sorghum[J]. Front Plant Sci, 2020, 10: 1722.
[5]	吴国江, 周伟, 李艳肖, 等. 高粱ZF-HD基因家族鉴定与盐碱胁迫下的表达分析[J]. 浙江农业学报, 2024, 36(6): 1217-1231. doi: 10.3969/j.issn.1004-1524.20230861
	Wu GJ, Zhou W, Li YX, et al. Identification and expression analysis under saline-alkali stress of ZF-HD gene family in sorghum[J]. Acta Agric Zhejiangensis, 2024, 36(6): 1217-1231. doi: 10.3969/j.issn.1004-1524.20230861
[6]	吴慧琳. 高粱全基因组测序的完成揭示了未被发现的宝贵遗传资源[J]. 农业生物技术学报, 2013, 21(11): 1278.
	Wu HL. Whole-genome sequencing reveals untapped genetic potential in Africa's indigenous cereal crop sorghum[J]. J Agric Biotechnol, 2013, 21(11): 1278.
[7]	王彬, 陈敏氡, 林亮, 等. 植物干旱胁迫的信号通路及相关转录因子研究进展[J]. 西北植物学报, 2020, 40(10): 1792-1806.
	Wang B, Chen MD, Lin L, et al. Signal pathways and related transcription factors of drought stress in plants[J]. Acta Bot Boreali Occidentalia Sin, 2020, 40(10): 1792-1806.
[8]	赵才美, 黄兴奇, 殷富有, 等. 水稻NAC转录因子家族的研究进展[J]. 植物科学学报, 2020, 38(2): 278-287.
	Zhao CM, Huang XQ, Yin FY, et al. Research progress on NAC transcription factor family in Oryza sativa L[J]. Plant Sci J, 2020, 38(2): 278-287.
[9]	Yang ZF, Gu SL, Wang XF, et al. Molecular evolution of the CPP-like gene family in plants: insights from comparative genomics of Arabidopsis and rice[J]. J Mol Evol, 2008, 67(3): 266-277.
[10]	王凯. 拟南芥和水稻CPP转录因子家族的生物信息学分析[J]. 生物技术通报, 2010, 26(2): 76-84.
	Wang K. Bioinformatic analysis of the CPP transcription factors family in Arabidopsis and rice[J]. Biotechnol Bull, 2010, 26(2): 76-84.
[11]	Lu T, Dou YC, Zhang C. Fuzzy clustering of CPP family in plants with evolution and interaction analyses[J]. BMC Bioinformatics, 2013, 14(Suppl 13): S10.
[12]	Song XY, Zhang YY, Wu FC, et al. Genome-wide analysis of the maize(Zea may L.)CPP-like gene family and expression profiling under abiotic stress[J]. Genet Mol Res, 2016, 15(3). DOI: 10.4238/gmr.15038023.
[13]	Zhou Y, Hu LF, Ye SF, et al. Genome-wide identification and characterization of cysteine-rich polycomb-like protein(CPP)family genes in cucumber(Cucumis sativus)and their roles in stress responses[J]. Biologia, 2018, 73(4): 425-435.
[14]	杨如兴, 王鹏杰, 陈芝芝, 等. 茶树CPP转录因子家族的全基因组鉴定及分析[J]. 西北植物学报, 2019, 39(6): 1024-1032.
	Yang RX, Wang PJ, Chen ZZ, et al. Genome-wide identification and analysis of CPP transcription factor family in tea plants[J]. Acta Bot Boreali Occidentalia Sin, 2019, 39(6): 1024-1032.
[15]	Hauser BA, He JQ, Park SO, et al. TSO1 is a novel protein that modulates cytokinesis and cell expansion in Arabidopsis[J]. Development, 2000, 127(10): 2219-2226. doi: 10.1242/dev.127.10.2219 pmid: 10769245
[16]	Sijacic P, Wang WP, Liu ZC. Recessive antimorphic alleles overcome functionally redundant loci to reveal TSO1 function in Arabidopsis flowers and meristems[J]. PLoS Genet, 2011, 7(11): e1002352.
[17]	Cvitanich C, Pallisgaard N, Nielsen KA, et al. CPP1 a DNA-binding protein involved in the expression of a soybean leghemoglobin c3 gene[J]. Proc Natl Acad Sci USA, 2000, 97(14): 8163-8168. doi: 10.1073/pnas.090468497 pmid: 10859345
[18]	田骄阳, 王秋霞, 郑淑文, 等. 全基因组水平蒺藜苜蓿CPP基因家族的鉴定及表达模式分析[J]. 草业学报, 2022, 31(7): 111-121. doi: 10.11686/cyxb2021215
	Tian JY, Wang QX, Zheng SW, et al. Genome-wide identification and expression profile analysis of the CPP gene family in Medicago truncatula[J]. Acta Prataculturae Sin, 2022, 31(7): 111-121.
[19]	罗巍. 甜高粱种质资源耐苏打盐碱性鉴定及机制分析[D]. 通辽: 内蒙古民族大学, 2023.
	Luo W. Identification and mechanism analysis of alkalinity tolerance of sweet sorghum germplasm resources to soda salt[D]. Tongliao: Inner Mongolia University for the Nationalities, 2023.
[20]	黄静, 郑晶, 胡乐佳, 等. 陆地棉CPP转录因子家族全基因组鉴定及分析[J]. 分子植物育种, 2022, 20(14): 4556-4566.
	Huang J, Zheng J, Hu LJ, et al. Genome-wide identification and analysis of CPP transcription factor family in Gossypium hirsutum L[J]. Mol Plant Breed, 2022, 20(14): 4556-4566.
[21]	杨楠, 孙瑞青, 孙宇, 等. 杧果CPP转录因子家族基因的鉴定及表达分析[J]. 植物遗传资源学报, 2021, 22(5): 1452-1462.
	Yang N, Sun RQ, Sun Y, et al. Identification and expression analysis of CPP transcription factor family genes in mango[J]. J Plant Genet Resour, 2021, 22(5): 1452-1462.
[22]	崔江慧, 杨溥原, 常金华. 高粱GRF基因家族鉴定及在非生物胁迫下的表达分析[J]. 中国农业科技导报, 2021, 23(4): 37-46. doi: 10.13304/j.nykjdb.2020.0278
	Cui JH, Yang PY, Chang JH. Identification and expression analysis under abiotic stress of GRF gene family in sorghum[J]. J Agric Sci Technol, 2021, 23(4): 37-46.
[23]	吴金丽, 汤泽洋, 鲁鑫, 等. 玉米PDI基因家族鉴定和表达模式分析[J]. 植物遗传资源学报, 2024, 25(1): 97-110.
	Wu JL, Tang ZY, Lu X, et al. Identification and expression pattern analysis of PDI gene family in maize[J]. J Plant Genet Resour, 2024, 25(1): 97-110.
[24]	刘晓龙, 徐晨, 邵勤, 等. 脱落酸提高水稻抗逆性的研究进展[J]. 东北农业科学, 2022, 47(6): 29-33.
	Liu XL, Xu C, Shao Q, et al. Research progress of abscisic acid in improving rice stress resistance[J]. J Northeast Agric Sci, 2022, 47(6): 29-33.
[25]	罗丽娟. 环境因子对柑橘柠檬酸积累及其相关基因的影响[D]. 武汉: 华中农业大学, 2019.
	Luo LJ. Effects of environmental factors on citric acid accumulation and related genes in Citrus[D]. Wuhan: Huazhong Agricultural University, 2019.
[26]	杨平, 蔡芸菲, 孙悦, 等. 生长素引发对盐胁迫下棉花生长发育及产量品质的影响[J]. 分子植物育种, 2023, 21(20): 6851-6859.
	Yang P, Cai YF, Sun Y, et al. Effects of auxin priming treatment on cotton growth, yield and quality under salt stress[J]. Mol Plant Breed, 2023, 21(20): 6851-6859.
[27]	牛佳斌, 唐凯, 夏迎萌, 等. 辣椒NRT基因家族的系统鉴定、进化与表达分析[J]. 南京农业大学学报, 2024, 47(4): 653-664.
	Niu JB, Tang K, Xia YM, et al. Systematic identification, evolution and expression analysis of NRT gene family in pepper[J]. J Nanjing Agric Univ, 2024, 47(4): 653-664.
[28]	刘涛涛, 李薇, 冯甜甜, 等. 小麦CPP基因家族鉴定和TaCPP20-5B克隆分析[J]. 分子植物育种, 2023, 21(17): 5569-5575.
	Liu TT, Li W, Feng TT, et al. Identification of the CPP gene family and TaCPP20-5B cloning analysis in wheat[J]. Mol Plant Breed, 2023, 21(17): 5569-5575.

编辑推荐 0

Metrics

阅读次数

全文

155

HTML			PDF

最新录用	在线预览	正式出版	最新录用	在线预览	正式出版
0	0	29	0	0	126

来源	本网站	其他网站

次数	150	5
比例	97%	3%

摘要

170

最新录用	在线预览	正式出版

0	0	170

	来源	本网站

	次数	170
	比例	100%