棉花β-tubulin基因家族的鉴定及其在纤维发育中的表达

doi:10.3864/j.issn.0578-1752.2023.23.003

摘要/Abstract

摘要：

【目的】β-微管蛋白是棉纤维细胞形态建成的基本结构单位，在纤维发育过程中具有重要作用。通过鉴定棉花β-tubulin基因家族成员，并进行生物信息学和表达模式分析，为深入探究β-tubulin基因在棉花纤维发育中的作用提供理论依据。【方法】利用BLAST方法，在4个棉种基因组中鉴定β-tubulin基因家族成员，并结合ProtParam tool分析理化性质、MEGA7.0构建系统进化树、Mapchart2.2绘制染色体定位图、MEME分析保守基序、PlantCARE分析启动子顺式作用元件；根据39个材料发育纤维转录组数据分析陆地棉β-tubulin基因家族的表达水平，并利用Spearman相关性分析鉴定影响纤维品质性状形成的β-tubulin基因。【结果】在陆地棉（Gossypium hirsutum，AD₁）、海岛棉（Gossypium barbadense，AD₂）、亚洲棉（Gossypium arboretum，A₂）和雷蒙德氏棉（Gossypium raimondii，D₅）基因组中分别鉴定到36、37、19和18个β-tubulin基因，且在四倍体棉种中的数目约是二倍体棉种数目的二倍，系统进化将其分为ClusterⅠ—ClusterⅤ共5个亚组。系统进化与共线性分析发现，与陆地棉β-tubulin基因家族相比，海岛棉与二倍体亚洲棉和雷蒙德氏棉亲缘关系更近。保守结构域均具有典型的Tubulin和Tubulin-C。理化性质分析表明，该家族基因编码的氨基酸数目为421—508，等电点为4.68—5.09。启动子顺式作用元件分析获得生长发育响应相关元件、激素响应相关元件和胁迫响应相关元件等，表明β-tubulin基因参与细胞的生长调节。利用TM-1转录组数据对36个陆地棉β-tubulin基因表达模式进行聚类分析，发现有42%基因在纤维中优势表达；此外，利用海陆群体动态纤维转录组数据筛选到1、6和11个β-tubulin基因分别与纤维马克隆值、比强度和纤维长度显著相关，其中4个基因同时影响纤维长度和比强度性状。【结论】4个棉种中共鉴定出110个β-tubulin基因家族成员，氨基酸理化性质和序列高度保守而启动子序列调控元件多样；筛选出陆地棉纤维优势表达的β-tubulin基因家族成员，并发掘潜在调控纤维发育的候选基因。

关键词: 棉花, β-tubulin基因家族, 生物信息学, 表达分析, 纤维发育

Abstract:

【Objective】β-tubulin is the basic structural unit of cotton fiber, regulates fiber cell morphogenesis, and plays a vital role in fiber development. But there is less understood how β-tubulin gene influenced the distinct characteristic of fiber quality traits in cotton. In this study, members of the β-tubulin gene family were identified in cotton, their expression profiles were analyzed, and role of β-tubulin genes were explored for fiber quality. 【Method】BLAST method was used to identify members of the β-tubulin gene family in the genomes of four cotton species. ProtParam tool was utilized to analyze physicochemical properties, MEGA7.0 to construct phylogenetic tree, Mapchart2.2 to draw chromosomal localization map, MEME to analyze conserved motif, and PlantCARE to analyze promoter cis-acting elements. Expression levels of β-tubulin genes were characterized by using transcriptome data from 39 studies on fiber development. Spearman correlation analysis was used to identify candidate genes for fiber quality traits. 【Result】Importantly, 36, 37, 19 and 18 β-tubulin genes were identified in the genomes of Gossypium hirsutum (AD₁), Gossypium barbadense (AD₂), Gossypium arboretum (A₂) and Gossypium raimondii (D₅), respectively. The number of β-tubulin genes in tetraploid cotton species is almost double than that of diploid cotton species. Phylogenetic analysis classified these genes into 5 main clusters. Phylogenetic and collinearity analysis revealed that β-tubulin genes in Gossypium barbadense is closely related to Gossypium arboretum and Gossypium raimondii as compared to Gossypium hirsutum. Furthermore, all genes have typical conservative domains with Tubulin and Tubulin-C. The genes physicochemical properties showed amino acids range from 421 to 508 with isoelectric point of 4.68 to 5.09. The analysis of promoter cis-acting elements identified growth responsive, hormone responsive, and stress responsive elements which showed β-tubulin mediates various mechanisms of cell growth regulation. Interestingly, cluster analysis on 36 β-tubulin gene expression profiles showed 42% genes in cluster П had dominant expression in fiber. In particular, 1, 6, and 11 β-tubulin genes exhibited significant correlation with fiber micronaire value, fiber strength, and fiber length, respectively. Four genes were found to influence fiber length and fiber strength traits simultaneously. 【Conclusion】A total of 110 β-tubulin gene family members were identified in the four cotton species. Their physicochemical properties and sequences of amino acids were highly conserved and the promoter sequence had diverse regulatory elements. This study characterized the expression profiles as well as molecular function of β-tubulin gene family in cotton fiber. Further discovered the potential candidate genes that probably regulate fiber quality traits in cotton. Our results may have great potential for cotton fiber quality improvement by genetic engineering.

Key words: cotton, β-tubulin gene family, bioinformatics, expression analysis, fiber development

党媛玥, 马建江, 杨书贤, 宋吉坤, 贾冰, 冯盼, 陈全家, 于霁雯. 棉花β-tubulin基因家族的鉴定及其在纤维发育中的表达[J]. 中国农业科学, 2023, 56(23): 4585-4601.

DANG YuanYue, MA JianJiang, YANG ShuXian, SONG JiKun, JIA Bing, FENG Pan, CHEN QuanJia, YU JiWen. Genome-Wide Identification and Expression Analysis of β-tubulin Family in Cotton Fiber Development[J]. Scientia Agricultura Sinica, 2023, 56(23): 4585-4601.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2023.23.003

https://www.chinaagrisci.com/CN/Y2023/V56/I23/4585

图/表 8

图1

表1

棉花中β-tubulin基因家族的基本理化性质"

物种 Species	基因ID Gene ID	等电点 pI	分子量 Molecular weight (kDa)	蛋白长度 <BOLD>P</BOLD>rotein length (aa)	亚细胞定位 Subcellular localization
陆地棉 Gossypium hirsutum	GH_A01G1087	4.74	50.65	452	细胞质Cytoplasm
	GH_A03G0257	4.76	50.13	445	细胞核Nucleus
	GH_A03G0258	4.76	50.07	445	细胞核Nucleus
	GH_A03G0260	5.09	55.66	495	叶绿体Chloroplast
	GH_A03G0596	4.75	50.00	444	细胞核Nucleus
	GH_A03G0911	4.73	50.21	447	细胞质Cytoplasm
	GH_A05G0919	4.74	50.30	446	细胞核Nucleus
	GH_A05G1119	4.76	50.01	445	细胞质Cytoplasm
	GH_A05G1860	4.70	50.50	450	细胞核Nucleus
	GH_A05G2570	4.80	50.66	449	细胞核Nucleus
	GH_A06G0029	4.79	50.39	449	细胞核Nucleus
	GH_A08G2076	4.76	50.00	445	细胞核Nucleus
	GH_A08G2085	4.79	49.85	444	细胞核Nucleus
	GH_A09G0513	4.68	50.59	448	细胞质Cytoplasm
	GH_A09G1787	4.86	53.27	472	内质网Endoplasmic reticulum
	GH_A09G1903	4.76	49.73	445	细胞核Nucleus
	GH_A11G3495	4.75	50.30	447	细胞核Nucleus
	GH_A13G2534	4.75	49.97	444	细胞核Nucleus
	GH_D01G1129	4.68	50.71	452	细胞核Nucleus
	GH_D02G1082	4.72	50.21	447	细胞质Cytoplasm
	GH_D03G1367	4.75	50.00	444	细胞核Nucleus
	GH_D03G1707	4.86	49.98	444	细胞核Nucleus
	GH_D03G1709	4.76	50.10	445	细胞核Nucleus
	GH_D03G1710	4.76	50.13	445	细胞核Nucleus
	GH_D05G0911	4.70	50.21	446	细胞核Nucleus
	GH_D05G1114	4.74	50.14	446	细胞质Cytoplasm
	GH_D05G1898	4.70	50.52	450	细胞核Nucleus
	GH_D06G0016	4.73	50.67	452	细胞核Nucleus
	GH_D07G1281	4.77	50.26	447	细胞核Nucleus
	GH_D08G2085	4.76	50.00	445	细胞核Nucleus
	GH_D08G2099	4.79	49.83	444	细胞核Nucleus
	GH_D09G0508	4.71	50.36	447	细胞核Nucleus
	GH_D09G1737	4.89	53.36	472	细胞质Cytoplasm
	GH_D09G1854	4.80	49.82	445	细胞核Nucleus
	GH_D11G3501	4.75	50.30	447	细胞核Nucleus
	GH_D13G2529	4.75	49.95	444	细胞核Nucleus
海岛棉 Gossypium barbadense	GB_A01G1091	4.72	50.65	452	细胞质Cytoplasm
	GB_A03G0251	4.76	50.13	445	细胞核Nucleus
	GB_A03G0252	4.76	50.07	445	细胞核Nucleus
	GB_A03G0254	4.85	49.88	444	细胞核Nucleus
	GB_A03G0586	4.75	50.01	444	细胞核Nucleus
	GB_A03G0919	4.75	50.18	447	叶绿体Chloroplast
	GB_A05G0927	4.70	50.24	446	细胞核Nucleus
	GB_A05G1127	4.76	50.01	445	细胞质Cytoplasm
	GB_A05G1887	4.70	50.50	450	细胞核Nucleus
	GB_A05G2602	4.77	50.66	449	细胞核Nucleus
	GB_A06G0033	4.77	50.78	452	细胞核Nucleus
	GB_A08G2182	4.76	50.00	445	细胞核Nucleus
	GB_A08G2191	4.79	49.85	444	细胞核Nucleus
	GB_A09G0575	4.68	50.59	448	细胞质Cytoplasm
	GB_A09G1907	4.86	53.27	472	内质网Endoplasmic reticulum
	GB_A09G2018	4.76	49.72	445	细胞核Nucleus
	GB_A11G3569	4.75	50.30	447	细胞核Nucleus
	GB_A13G2683	4.75	49.97	444	细胞核Nucleus
	GB_D01G1171	4.68	50.71	452	细胞核Nucleus
	GB_D02G1129	4.75	50.80	452	细胞质Cytoplasm
	GB_D03G1389	4.75	50.00	444	细胞核Nucleus
	GB_D03G1742	4.86	49.98	444	细胞核Nucleus
	GB_D03G1744	4.76	50.10	445	细胞核Nucleus
	GB_D03G1745	4.76	50.13	445	细胞核Nucleus
	GB_D05G0908	4.70	50.21	446	细胞核Nucleus
	GB_D05G1112	4.74	50.14	446	细胞质Cytoplasm
	GB_D05G1913	4.70	50.52	450	细胞核Nucleus
	GB_D05G2606	4.76	47.37	421	细胞核Nucleus
	GB_D06G0034	4.73	50.67	452	细胞核Nucleus
	GB_D07G1286	4.81	50.33	447	细胞核Nucleus
	GB_D08G2163	4.76	50.00	445	细胞核Nucleus
	GB_D08G2175	4.79	49.83	444	细胞核Nucleus
	GB_D09G0514	4.70	50.44	447	细胞核Nucleus
	GB_D09G1750	4.86	53.38	472	细胞质Cytoplasm
	GB_D09G1866	4.80	49.82	445	细胞核Nucleus
	GB_D11G3544	5.08	57.42	508	叶绿体Chloroplast
	GB_D13G2621	4.75	49.97	444	细胞核Nucleus
亚洲棉 Gossypium arboretum	Ga01G1198	4.72	48.89	436	细胞核Nucleus
	Ga01G2180	4.75	50.00	444	细胞核Nucleus
	Ga01G2554	4.76	50.13	445	细胞核Nucleus
	Ga01G2555	4.76	50.08	445	细胞核Nucleus
	Ga01G2557	4.85	49.86	444	细胞核Nucleus
	Ga03G1117	4.73	50.21	447	细胞质Cytoplasm
	Ga05G0964	4.70	48.91	433	细胞质Cytoplasm
	Ga05G1174	4.76	50.01	445	细胞质Cytoplasm
	Ga05G1974	4.70	50.52	450	细胞核Nucleus
	Ga05G2720	4.79	50.70	449	细胞核Nucleus
	Ga06G0039	4.77	50.78	452	细胞核Nucleus
	Ga07G1306	4.81	50.32	447	细胞核Nucleus
	Ga08G2131	4.76	50.00	445	细胞核Nucleus
	Ga08G2143	4.79	49.85	444	细胞核Nucleus
	Ga09G0571	4.68	50.58	448	细胞核Nucleus
	Ga09G1894	4.76	49.71	445	细胞核Nucleus
	Ga09G1948	4.76	50.19	445	细胞核Nucleus
	Ga11G0323	4.75	50.30	447	细胞核Nucleus
	Ga13G2741	4.75	49.97	444	细胞核Nucleus
雷蒙德氏棉 Gossypium raimondii	Gorai.001G132500	4.81	50.32	447	细胞核Nucleus
	Gorai.002G123400	4.68	50.71	452	细胞核Nucleus
	Gorai.003G126300	4.75	50.00	444	细胞核Nucleus
	Gorai.003G159100	4.86	49.98	444	细胞核Nucleus
	Gorai.003G159300	4.76	50.07	445	细胞核Nucleus
	Gorai.003G159400	4.76	50.12	445	细胞核Nucleus
	Gorai.004G210600	4.76	50.00	445	细胞核Nucleus
	Gorai.004G211800	4.79	49.83	444	细胞核Nucleus
	Gorai.005G118100	4.72	50.20	447	细胞质Cytoplasm
	Gorai.006G049000	4.70	50.51	448	细胞核Nucleus
	Gorai.006G179900	4.86	53.38	472	细胞质Cytoplasm
	Gorai.006G191900	4.80	49.74	445	细胞核Nucleus
	Gorai.007G351900	4.75	50.30	447	细胞核Nucleus
	Gorai.009G094600	4.70	50.21	446	细胞核Nucleus
	Gorai.009G115600	4.74	50.14	446	细胞质Cytoplasm
	Gorai.009G194500	4.70	50.52	450	细胞核Nucleus
	Gorai.010G003800	4.73	50.67	452	细胞核Nucleus
	Gorai.013G261300	4.75	49.95	444	细胞核Nucleus

表1

图2

图3

图4

图5

图6

图7

参考文献 38

[1]	肖水平, 宋国立, 余进祥. 棉花纤维品质相关基因挖掘及功能基因研究进展. 棉花科学, 2020, 42(2): 3-14.
	XIAO S P, SONG G L, YU J X. Research progress of cotton fiber quality related gene mining and functional genes. Cotton Sciences, 2020, 42(2): 3-14. (in Chinese)
[2]	杨君, 马峙英, 王省芬. 棉花纤维品质改良相关基因研究进展. 中国农业科学, 2016, 49(22): 4310-4322. doi: 10.3864/j.issn.0578-1752.2016.22.005
	YANG J, MA Z Y, WANG X F. Progress in studies on genes related to fiber quality improvement of cotton. Scientia Agricultura Sinica, 2016, 49(22): 4310-4322. (in Chinese) doi: 10.3864/j.issn.0578-1752.2016.22.005
[3]	DIXON D C, SEAGULL R W, TRIPLETT B A. Changes in the accumulation of [alpha]- and [beta]-tubulin isotypes during cotton fiber development. Plant Physiology, 1994, 105(4): 1347-1353. pmid: 12232289
[4]	KOST B, CHUA N H. The plant cytoskeleton: Vacuoles and cell walls make the difference. Cell, 2002, 108(1): 9-12. pmid: 11792316
[5]	HE X C, QIN Y M, XU Y, HU C Y, ZHU Y X. Molecular cloning, expression profiling, and yeast complementation of 19 β-tubulin cDNAs from developing cotton ovules. Journal of Experimental Botany, 2008, 59(10): 2687-2695. doi: 10.1093/jxb/ern127
[6]	NIEUWENHUIS J, BRUMMELKAMP T R. The tubulin detyrosination cycle: Function and enzymes. Trends in Cell Biology, 2019, 29(1): 80-92. doi: S0962-8924(18)30141-7 pmid: 30213517
[7]	MEIRING J C M, SHNEYER B I, AKHMANOVA A. Generation and regulation of microtubule network asymmetry to drive cell polarity. Current Opinion in Cell Biology, 2020, 62: 86-95. doi: S0955-0674(19)30091-2 pmid: 31739264
[8]	MCKEAN P G, VAUGHAN S, GULL K. The extended tubulin superfamily. Journal of Cell Science, 2001, 114(15): 2723-2733. doi: 10.1242/jcs.114.15.2723
[9]	SEAGULL R W. Cytoskeletal stability affects cotton fiber initiation. International Journal of Plant Sciences, 1998, 159(4): 590-598. doi: 10.1086/297577
[10]	OAKLEY R V, WANG Y S, RAMAKRISHNA W, HARDING S A, TSAI C J. Differential expansion and expression of α- and β-tubulin gene families in Populus. Plant Physiology, 2007, 145(3): 961-973. doi: 10.1104/pp.107.107086
[11]	MUBEEN H, SHOAIB M W, RAZA S. In silico analysis and comparison of alpha tubulin gene with other tubulin families. Journal of Advance in Biology & Biotecnology, 2016, 7(4): 1-7.
[12]	LI Y L, SUN J, LI C H, ZHU Y Q, XIA G X. Specific expression of a β-tubulin gene (GhTub1) in developing cotton fibers. Science in China Series C: Life Sciences, 2003, 46(3): 235-242. doi: 10.1360/03yc9025
[13]	SNUSTAD D P, HAAS N A, KOPCZAK S D, SILFLOW C D. The small genome of Arabidopsis contains at least nine expressed beta-tubulin genes. The Plant Cell, 1992, 4(5): 549-556.
[14]	CHENG Z G, CARTER J V. Temporal and spatial expression patterns of TUB9, a β-tubulin gene of Arabidopsis thaliana. Plant Molecular Biology, 2001, 47(3): 389-398. doi: 10.1023/A:1011628024798
[15]	杨建霞. TUB4对拟南芥生长发育的影响和TAO1基因新功能分析[D]. 兰州: 兰州大学, 2015.
	YANG J X. The effects of TUB4 on Arabidopsis growth and development and the analysis of the new role of TAO1[D]. Lanzhou: Lanzhou University, 2015. (in Chinese)
[16]	GAVAZZI F, PIGNA G, BRAGLIA L, GIANÌ S, BREVIARIO D, MORELLO L. Evolutionary characterization and transcript profiling of β-tubulin genes in flax (Linum usitatissimum L.) during plant development. BMC Plant Biology, 2017, 17(1): 237. doi: 10.1186/s12870-017-1186-0
[17]	睢金凯, 饶国栋, 张建国. 柳树β微管蛋白基因家族的克隆和序列分析. 西北植物学报, 2016, 36(5): 902-909.
	SUI J K, RAO G D, ZHANG J G. Cloning and sequence analysis of β-tubulin gene families in willows. Acta Botanica Boreali- Occidentalia Sinica, 2016, 36(5): 902-909. (in Chinese)
[18]	黄胜雄, 胡尚连, 孙霞, 蒋瑶. 慈竹β-tubulin基因片段的克隆及序列分析. 福建林业科技, 2009, 36(1): 7-10, 42.
	HUANG S X, HU S L, SUN X, JIANG Y. Cloning and sequence analysis of β-tubulin gene fragments of Neosinocalamus affinis. Fujian Forestry Science and Technology, 2009, 36(1): 7-10, 42. (in Chinese)
[19]	王亚婷, 林玉玲, 曾丽兰, 赖钟雄. 龙眼胚性愈伤组织微管蛋白基因β-tubulin克隆及序列分析. 福建农林大学学报(自然科学版), 2014, 43(1): 49-54.
	WANG Y T, LIN Y L, ZENG L L, LAI Z X. Cloning and sequence analysis of β-tubulin gene from embryogenic callus in Dimocarpus longan Lour. Journal of Fujian Agriculture and Forestry University (Natural Science Edition), 2014, 43(1): 49-54. (in Chinese)
[20]	LI X B, CAI L, CHENG N H, LIU J W. Molecular characterization of the cotton GhTUB1 gene that is preferentially expressed in fiber. Plant Physiology, 2002, 130(2): 666-674. doi: 10.1104/pp.005538
[21]	QIN Y, SUN H R, HAO P B, WANG H T, WANG C C, MA L, WEI H L, YU S X. Transcriptome analysis reveals differences in the mechanisms of fiber initiation and elongation between long- and short-fiber cotton (Gossypium hirsutum L.) lines. BMC Genomics, 2019, 20(1): 633. doi: 10.1186/s12864-019-5986-5
[22]	曾志锋. GhTUB7在棉纤维发育中的功能[D]. 重庆: 西南大学, 2016.
	ZENG Z F. The function of GhTUB7 in cotton fiber development[D]. Chongqing: Southwest University, 2016. (in Chinese)
[23]	李彩丽. 毛竹细胞骨架微管蛋基因PeTua3和PeTub3研究[D]. 北京: 中国林业科学研究院, 2012.
	LI C L. Study on PeTua3 and PeTub3 encoding micro tuhulin of cvtoskeleton in moso bamboo (Phyllostachys edulis)[D]. Beijing: Chinese Academy of Forestry, 2012. (in Chinese)
[24]	DU X M, HUANG G, HE S P, YANG Z E, SUN G F, MA X F, LI N, ZHANG X Y, SUN J L, LIU M, JIA Y H, PAN Z E, GONG W F, LIU Z H, ZHU H Q, MA L, LIU F Y, YANG D G, WANG F, FAN W, GONG Q, PENG Z, WANG L R, WANG X Y, XU S J, SHANG H H, LU C R, ZHENG H K, HUANG S W, LIN T, ZHU Y X, LI F G. Resequencing of 243 diploid cotton accessions based on an updated A genome identifies the genetic basis of key agronomic traits. Nature Genetics, 2018, 50(6): 796-802. doi: 10.1038/s41588-018-0116-x pmid: 29736014
[25]	PATERSON A H, WENDEL J F, GUNDLACH H, GUO H, JENKINS J, JIN D C, LLEWELLYN D, SHOWMAKER K C, SHU S Q, UDALL J, YOO M J, BYERS R, CHEN W, DORON-FAIGENBOIM A, DUKE M V, GONG L, GRIMWOOD J, GROVER C, GRUPP K, HU G J, LEE T H, LI J P, LIN L F, LIU T, MARLER B S, PAGE J T, ROBERTS A W, ROMANEL E, SANDERS W S, SZADKOWSKI E, TAN X, TANG H B, XU C M, WANG J P, WANG Z N, ZHANG D, ZHANG L, ASHRAFI H, BEDON F, BOWERS J E, BRUBAKER C L, CHEE P W, DAS S, GINGLE A R, HAIGLER C H, HARKER D, HOFFMANN L V, HOVAV R, JONES D C, LEMKE C, MANSOOR S, UR RAHMAN M, RAINVILLE L N, RAMBANI A, REDDY U K, RONG J K, SARANGA Y, SCHEFFLER B E, SCHEFFLER J A, STELLY D M, TRIPLETT B A, VAN DEYNZE A, VASLIN M F S, WAGHMARE V N, WALFORD S A, WRIGHT R J, ZAKI E A, ZHANG T Z, DENNIS E S, MAYER K F X, PETERSON D G, ROKHSAR D S, WANG X Y, SCHMUTZ J. Repeated polyploidization of Gossypium genomes and the evolution of spinnable cotton fibres. Nature, 2012, 492(7429): 423-427. doi: 10.1038/nature11798
[26]	HU Y, CHEN J D, FANG L, ZHANG Z Y, MA W, NIU Y C, JU L Z, DENG J Q, ZHAO T, LIAN J M, BARUCH K B, FANG D, LIU X, RUAN Y L, RAHMAN M U, HAN J L, WANG K, WANG Q, WU H T, MEI G F, ZANG Y H, HAN Z G, XU C Y, SHEN W J, YANG D F, SI Z F, DAI F, ZOU L F, HUANG F, BAI Y L, ZHANG Y G, BRODT A, BEN-HAMO H, ZHU X F, ZHOU B L, GUAN X Y, ZHU S J, CHEN X Y, ZHANG T Z. Gossypium barbadense and Gossypium hirsutum genomes provide insights into the origin and evolution of allotetraploid cotton. Nature Genetics, 2019, 51(4): 739-748. doi: 10.1038/s41588-019-0371-5
[27]	LETUNIC I, DOERKS T, BORK P. SMART: Recent updates, new developments and status in 2015. Nucleic Acids Research, 2015, 43(D1): D257-D260. doi: 10.1093/nar/gku949
[28]	LARKIN M A, BLACKSHIELDS G, BROWN N P, CHENNA R, MCGETTIGAN P A, MCWILLIAM H, VALENTIN F, WALLACE I M, WILM A, LOPEZ R, THOMPSON J D, GIBSON T J, HIGGINS D G. Clustal W and Clustal X version 2.0. Bioinformatics, 2007, 23(21): 2947-2948. doi: 10.1093/bioinformatics/btm404 pmid: 17846036
[29]	KUMAR S, STECHER G, TAMURA K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 2016, 33(7): 1870-1874. doi: 10.1093/molbev/msw054 pmid: 27004904
[30]	BJELLQVIST B, BASSE B, OLSEN E, CELIS J E. Reference points for comparisons of two-dimensional maps of proteins from different human cell types defined in a pH scale where isoelectric points correlate with polypeptide compositions. Electrophoresis, 1994, 15(3/4): 529-539. doi: 10.1002/elps.v15:1
[31]	VOORRIPS R E. MapChart: Software for the graphical presentation of linkage maps and QTLs. Journal of Heredity, 2002, 93(1): 77-78. doi: 10.1093/jhered/93.1.77 pmid: 12011185
[32]	CHEN C J, CHEN H, ZHANG Y, THOMAS H R, FRANK M H, HE Y H, XIA R. TBtools: An integrative toolkit developed for interactive analyses of big biological data. Molecular Plant, 2020, 13(8): 1194-1202. doi: S1674-2052(20)30187-8 pmid: 32585190
[33]	KRZYWINSKI M, SCHEIN J, BIROL I, CONNORS J, GASCOYNE R, HORSMAN D, JONES S J, MARRA M A. Circos: An information aesthetic for comparative genomics. Genome Research, 2009, 19(9): 1639-1645. doi: 10.1101/gr.092759.109 pmid: 19541911
[34]	BAILEY T L, BODEN M, BUSKE F A, FRITH M, GRANT C E, CLEMENTI L, REN J Y, LI W W, NOBLE W S. MEME Suite: Tools for motif discovery and searching. Nucleic Acids Research, 2009, 37(supp1_2): W202- W208. doi: 10.1093/nar/gkp335
[35]	LESCOT M, DÉHAIS P, THIJS G, MARCHAL K, MOREAU Y, VAN DE PEER Y, ROUZÉ P, ROMBAUTS S. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Research, 2002, 30(1): 325-327.
[36]	YOSHIKAWA M, YANG G X, KAWAGUCHI K, KOMATSU S. Expression analyses of β-tubulin isotype genes in rice. Plant and Cell Physiology, 2003, 44(11): 1202-1207. doi: 10.1093/pcp/pcg150
[37]	RAO G D, ZENG Y F, HE C Y, ZHANG J G. Characterization and putative post-translational regulation of α- and β-tubulin gene families in Salix arbutifolia. Scientific Reports, 2016, 6: 19258. doi: 10.1038/srep19258
[38]	CHEN B J, ZHAO J J, FU G Y, PEI X X, PAN Z E, LI H G, AHMED H, HE S P, DU X M. Identification and expression analysis of Tubulin gene family in upland cotton. Journal of Cotton Research, 2021, 4(3): 229-238.