Identification of BTB/POZ protein family in <i>Nicotiana tabacum</i> and its role in resistance to potato virus Y (PVY)

doi:10.3785/j.issn.1008-9209.2023.10.081

Journal of Zhejiang University (Agriculture and Life Sciences)

2024, Vol. 50

Issue (2): 295-307 DOI: 10.3785/j.issn.1008-9209.2023.10.081

Research Articles

Identification of BTB/POZ protein family in Nicotiana tabacum and its role in resistance to potato virus Y (PVY)

Shaowen ZHANG¹(

),Tianlun ZHAO^1,^2,³,Qinzhi XIAO⁴(

),Shuijin ZHU^1,^2,³,Jinhong CHEN^1,^2,³(

)

^1.College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, Zhejiang, China
^2.Hainan Institute of Zhejiang University, Sanya 572025, Hainan, China
^3.Zhejiang Key Laboratory of Crop Germplasm Resources, Hangzhou 310058, Zhejiang, China
^4.Yongzhou Branch of Hunan Tobacco Company, Yongzhou 425000, Hunan, China

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Abstract

The infection of Nicotiana tabacum with potato virus Y (PVY) can cause tobacco vein disease, leading to a decrease in the tobacco leaf quality. The broad complex, tramtrack, and bric-à-brac/pox virus and zinc finger (BTB/POZ) family exists widely in plants and animals. It plays a very important role in various stages of plant growth and development. In this study, 90 proteins in the BTB/POZ family of N. tabacum were identified, corresponding to 71 genes. Among these protein sequences, nine conserved motifs were identified, and the members of the BTB/POZ family were divided into six subfamilies according to the order of their appearance. Domain analysis showed that the domains of members of the same subfamily in BTB/POZ are consistent. The three-dimensional (3D) conformation of the BTB/POZ protein was predicted based on the amino acid sequence. All the members of the subfamily are mainly structured with α helixes, and the 3D conformations of members of the same subfamily are similar. Analysis of the BTB/POZ family gene expression patterns of N. tabacum K326 and mutant M867 (anti-PVY) showed that the expression levels of NtBTB2K, NtBTB2L, NtBTB2M and NtBTB6E increased significantly after inoculation with PVY, which may be related to the strong resistance of N. tabacum M867 to PVY. Cis-acting element analysis showed that the 2 000 bp upstream region of NtBTB2K, NtBTB2L, NtBTB2M and NtBTB6E contained several stress responsive elements, including TCA-element and MYB-binding site, which may be related to the upregulated expression of these genes. This study provides a theoretical basis for the study of BTB/POZ protein function, and provides a reference for disease resistance of tobacco breeding.

Key words： Nicotiana tabacum BTB/POZ family conserved motif cis-acting element

Received: 08 October 2023 Published: 25 April 2024

CLC:

S330

Corresponding Authors: Qinzhi XIAO,Jinhong CHEN E-mail: 22116179@zju.edu.cn;xwiao@126.com;jinhongchen@zju.edu.cn

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	Shaowen ZHANG
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Cite this article:

Shaowen ZHANG,Tianlun ZHAO,Qinzhi XIAO,Shuijin ZHU,Jinhong CHEN. Identification of BTB/POZ protein family in Nicotiana tabacum and its role in resistance to potato virus Y (PVY). Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(2): 295-307.

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https://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2023.10.081 OR https://www.zjujournals.com/agr/Y2024/V50/I2/295

烟草BTB/POZ蛋白家族的鉴定及其在抗马铃薯Y病毒中的作用

马铃薯Y病毒（potato virus Y, PVY）侵染烟草（Nicotiana tabacum）会引起烟草脉带病，造成烟叶品质下降。broad complex, tramtrack, and bric-à-brac/pox virus and zinc finger（BTB/POZ）是动植物体内广泛存在的蛋白质家族，在植物生长和发育的各个环节都起着非常重要的作用。本研究鉴定到90个烟草BTB/POZ家族蛋白，对应71个基因。在这些蛋白质的序列中，识别到9种保守基序（motif），并根据其出现顺序将BTB/POZ家族成员划分至6个亚家族。结构域分析表明，BTB/POZ同一亚家族成员的结构域具有一致性。对BTB/POZ蛋白的三维构象进行预测发现，所有亚家族成员结构都以α螺旋为主，同一亚家族成员的三维构象相似。烟草品种K326和突变体M867（抗PVY）的BTB/POZ家族基因表达模式分析显示，叶片接种PVY后，NtBTB2M、NtBTB2K、NtBTB2L、NtBTB6E基因的表达丰度显著增加，这可能与烟草M867对PVY的抗性较强有关。顺式作用元件分析显示，NtBTB2K、NtBTB2L、NtBTB2M和NtBTB6E基因的上游2 000 bp区域内含有若干逆境响应相关元件，包括水杨酸响应元件、MYB结合位点等，可能和这些基因的上调表达有关。本研究为BTB/POZ蛋白的功能研究提供了理论依据，为烟草抗病育种提供了一定参考。

关键词： 烟草, BTB/POZ家族, 保守基序, 顺式作用元件

基因名称

Gene name

引物序列（5 $′$ →3 $′$ ）

Primer sequence (5 $′$ →3 $′$ )

NtActin-7

F: TCAGGAAGGACCTCTACGGT

R: ATGTGCTAAGGGATGCGAGG

NtBTB2L

F: ACTTTGCCACCGATTTTAATG

R: TAGAATGTGCTGGAATGCG

NtBTB2K/NtBTB2M

F: ATGCACTGAAGGATGCACCA

R: ATAAGGAGCTGGACGCCTTG

NtBTB6E

F: ATGGAGGCCAGCAGCAGC

R: TCATGTGGAAGCAACCATTTCC

Table 1 Primer sequences for qRT-PCR

Fig. 1 Evolutionary tree of BTB/POZ family members in Arabidopsis thaliana, Solanum lycopersicum and N. tabacum

Table 2 Nine conserved motifs identified among BTB/POZ family in N. tabacum

Fig. 2 Conservation of nine motifs of BTB/POZ family in N. tabacumThe total height of the letter stacking at each position indicates the degree of conservation of the motif at that site. The height of a single letter in the letter stack indicates the relative frequency of the corresponding amino acid at that site.

Fig. 3 Distribution of conserved motifs of BTB/POZ family in N. tabacum

Fig. 4 Distribution of domains of the BTB/POZ family in N. tabacum

Fig. 5 Clustering heat map of expression patterns of BTB/POZ family genes in N. tabacumK326_PVY and M867_PVY indicate N. tabacum K326 and mutant M867 inoculated with PVY, respectively.

Fig. 6 Relative expression levels of differential genes in BTB/POZ familyDifferent lowercase letters above bars indicate significant differences at the 0.05 probability level.

Fig. 7 Predicted 3D conformation of BTB/POZ proteins in N. tabacumThe red part indicates the N-terminal of the protein, while the blue purple part indicates the C-terminal of the protein. Among some proteins whose 3D conformations are almost identical, only the three-dimensional (3D) conformation of the first protein is displayed.

Table 3 Cis-acting elements of highly expressed BTB/POZ family genes in N. tabacum


[1]	吴雪慧,吕若辰,王克敏,等.贵州烟区产业综合体发展模式研究[J].农业与技术,2022,42(17):166-171. DOI:10.19754/j.nyyjs.20220915040 WU X H, LÜ R C, WANG K M, et al. Research on the development model of industrial complex in Guizhou tobacco areas[J]. Agriculture and Technology, 2022, 42(17): 166-171. (in Chinese) doi: 10.19754/j.nyyjs.20220915040

[2]	NAIDU S K. Tobacco: production, chemistry and technology[J]. Crop Science, 2001, 41(1): 255. DOI: 10.2135/cropsci2001.411255x doi: 10.2135/cropsci2001.411255x

[3]	STOGIOS P J, DOWNS G S, JAUHAL J J S, et al. Sequence and structural analysis of BTB domain proteins[J]. Genome Biology, 2005, 6(10): R82. DOI: 10.1186/gb-2005-6-10-r82 doi: 10.1186/gb-2005-6-10-r82

[4]	ZOLLMAN S, GODT D, PRIVÉ G G, et al. The BTB domain, found primarily in zinc finger proteins, defines an evolutionarily conserved family that includes several developmentally regulated genes in Drosophila [J]. PNAS, 1994, 91(22): 10717-10721.

[5]	HARRISON S D, TRAVERS A A. The tramtrack gene encodes a Drosophila finger protein that interacts with the ftz trans-criptional regulatory region and shows a novel embryonic expression pattern[J]. The EMBO Journal, 1990, 9: 207-216.

[6]	GODT D, COUDERC J L, CRAMTON S E, et al. Pattern formation in the limbs of Drosophila: bric à brac is expressed in both a gradient and a wave-like pattern and is required for specification and proper segmentation of the tarsus[J]. Development, 1993, 119(3): 799-812.

[7]	SCOFIELD M, KORUTLA L, JACKSON T, et al. Nucleus accumbens 1, a pox virus and zinc finger/bric-a-brac tramtrack broad protein binds to TAR DNA-binding protein 43 and has a potential role in amyotrophic lateral sclerosis[J]. Neuroscience, 2012, 227: 44-54. DOI: 10.1016/j.neuro-science.2012.09.043 doi: 10.1016/j.neuro-science.2012.09.043

[8]	XU L, WEI Y, REBOUL J, et al. BTB proteins are substrate-specific adaptors in an SCF-like modular ubiquitin ligase containing CUL-3[J]. Nature, 2003, 425(6955): 316-321. DOI: 10.1038/nature01985 doi: 10.1038/nature01985

[9]	PINTARD L, WILLIS J H, WILLEMS A, et al. The BTB protein MEL-26 is a substrate-specific adaptor of the CUL-3 ubiquitin-ligase[J]. Nature, 2003, 425(6955): 311-316. DOI: 10.1038/nature01959 doi: 10.1038/nature01959

[10]	AN J P, LIU Y J, ZHANG X W, et al. Dynamic regulation of anthocyanin biosynthesis at different light intensities by the BT2-TCP46-MYB1 module in apple[J]. Journal of Experi-mental Botany, 2020, 71(10): 3094-3109. DOI: 10.1093/jxb/eraa056 doi: 10.1093/jxb/eraa056

[11]	AN J P, ZHANG X W, YOU C X, et al. MdWRKY40 promotes wounding-induced anthocyanin biosynthesis in association with MdMYB1 and undergoes MdBT2-mediated degradation[J]. New Phytologist, 2019, 224(1): 380-395. DOI: 10.1111/nph.16008 doi: 10.1111/nph.16008

[12]	JULIAN J, COEGO A, LOZANO-JUSTE J, et al. The MATH-BTB BPM3 and BPM5 subunits of Cullin3-RING E3 ubiquitin ligases target PP2CA and other clade A PP2Cs for degradation[J]. PNAS, 2019, 116(31): 15725-15734. DOI: 10.1073/pnas.1908677116 doi: 10.1073/pnas.1908677116

[13]	LECHNER E, LEONHARDT N, EISLER H, et al. MATH/BTB CRL3 receptors target the homeodomain-leucine zipper ATHB6 to modulate abscisic acid signaling[J]. Developmental Cell, 2011, 21(6): 1116-1128. DOI: 10.1016/j.devcel.2011.10.018 doi: 10.1016/j.devcel.2011.10.018

[14]	庄晓峰,董海涛,李德葆.水稻抗病性反应的cDNA微阵列分析及一个新基因OsBTB的发现[J].植物病理学报,2005,35(3):221-228. DOI:10.13926/j.cnki.apps.2005.03.006 ZHUANG X F, DONG H T, LI D B. Analysis of cDNA microarrays of resistant responses and discovery of a novel gene contained BTB/POZ domain in resistant rice[J]. Acta Phytopathologica Sinica, 2005, 35(3): 221-228. (in Chinese with English abstract) doi: 10.13926/j.cnki.apps.2005.03.006

[15]	CHAHTANE H, ZHANG B, NORBERG M, et al. LEAFY activity is post-transcriptionally regulated by BLADE ON PETIOLE2 and CULLIN3 in Arabidopsis [J]. New Phytologist, 2018, 220(2): 579-592. DOI: 10.1111/nph.15329 doi: 10.1111/nph.15329

[16]	MALNOY M, JIN Q, BOREJSZA-WYSOCKA E E, et al. Overexpression of the apple MpNPR1 gene confers increased disease resistance in Malus×domestica [J]. Molecular Plant-Microbe Interactions, 2007, 20(12): 1568-1580. DOI: 10.1094/MPMI-20-12-1568 doi: 10.1094/MPMI-20-12-1568

[17]	YOCGO R E, CREISSEN G, KUNERT K, et al. Two different banana NPR1-like coding sequences confer similar protection against pathogens in Arabidopsis [J]. Tropical Plant Biology, 2012, 5(4): 309-316. DOI: 10.1007/s12042-012-9112-y doi: 10.1007/s12042-012-9112-y

[18]	STOGIOS P J, PRIVÉ G G. The BACK domain in BTB-kelch proteins[J]. Trends in Biochemical Sciences, 2004, 29(12): 634-637. DOI: 10.1016/j.tibs.2004.10.003 doi: 10.1016/j.tibs.2004.10.003

[19]	ROBERTS D, PEDMALE U V, MORROW J, et al. Modula-tion of phototropic responsiveness in Arabidopsis through ubiquitination of phototropin 1 by the CUL3-RING E3 ubiquitin ligase CRL3NPH3 [J]. The Plant Cell, 2011, 23(10): 3627-3640. DOI: 10.1105/tpc.111.087999 doi: 10.1105/tpc.111.087999

[20]	MISRA A, MCKNIGHT T D, MANDADI K K. Bromo-domain proteins GTE9 and GTE11 are essential for specific BT2-mediated sugar and ABA responses in Arabidopsis thaliana [J]. Plant Molecular Biology, 2018, 96(4/5): 393-402. DOI: 10.1007/s11103-018-0704-2 doi: 10.1007/s11103-018-0704-2

[21]	WANG X F, AN J P, LIU X, et al. The nitrate-responsive protein MdBT2 regulates anthocyanin biosynthesis by interacting with the MdMYB1 transcription factor[J]. Plant Physiology, 2018, 178(2): 890-906. DOI: 10.1104/pp.18.00244 doi: 10.1104/pp.18.00244

[22]	RYALS J, WEYMANN K, LAWTON K, et al. The Arabidopsis NIM1 protein shows homology to the mammalian transcription factor inhibitor IκB[J]. The Plant Cell, 1997, 9(3): 425-439.

[23]	WU Y, ZHANG D, CHU J Y, et al. The Arabidopsis NPR1 protein is a receptor for the plant defense hormone salicylic acid[J]. Cell Reports, 2012, 1(6): 639-647. DOI: 10.1016/j.celrep.2012.05.008 doi: 10.1016/j.celrep.2012.05.008

[24]	WANG W, WITHERS J, LI H, et al. Structural basis of salicylic acid perception by Arabidopsis NPR proteins[J]. Nature, 2020, 586(7828): 311-316. DOI: 10.1038/s41586-020-2596-y doi: 10.1038/s41586-020-2596-y

[25]	ZHOU F, ZHANG K, ZHENG X, et al. BTB and TAZ domain protein BT4 positively regulates the resistance to Botrytis cinerea in Arabidopsis [J]. Plant Signaling & Behavior, 2022, 17(1): 2104003. DOI: 10.1080/15592324.2022.2104003 doi: 10.1080/15592324.2022.2104003

[26]	TRAN L S P, NAKASHIMA K, SAKUMA Y, et al. Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cis-element in the early responsive to dehydration stress 1 pro-moter[J]. The Plant Cell, 2004, 16(9): 2481-2498. DOI: 10.1105/tpc.104.022699 doi: 10.1105/tpc.104.022699

[27]	XIAO Q Z, CUI G X, CHEN Y R, et al. Combined analysis of mRNA and miRNA transcriptomes reveals the regulatory mechanism of PVY resistance in tobacco[J]. Industrial Crops and Products, 2022, 176: 114322. DOI: 10.1016/j.indcrop.2021.114322 doi: 10.1016/j.indcrop.2021.114322

[28]	EDWARDS K D, FERNANDEZ-POZO N, DRAKE-STOWE K, et al. A reference genome for Nicotiana tabacum enables map-based cloning of homeologous loci implicated in nitrogen utilization efficiency[J]. BMC Genomics, 2017, 18: 448. DOI: 10.1186/s12864-017-3791-6 doi: 10.1186/s12864-017-3791-6

[29]	CHERN M, FITZGERALD H A, CANLAS P E, et al. Over-expression of a rice NPR1 homolog leads to constitutive activation of defense response and hypersensitivity to light[J]. Molecular Plant-Microbe Interactions, 2005, 18(6): 511-520. DOI: 10.1094/MPMI-18-0511 doi: 10.1094/MPMI-18-0511

[30]	REN Y R, ZHAO Q, YANG Y Y, et al. Interaction of BTB-TAZ protein MdBT2 and DELLA protein MdRGL3a regulates nitrate-mediated plant growth[J]. Plant Physiology, 2021, 186(1): 750-766. DOI: 10.1093/plphys/kiab065 doi: 10.1093/plphys/kiab065

[31]	JI X L, LI H L, QIAO Z W, et al. The BTB protein MdBT2 recruits auxin signaling components to regulate adventitious root formation in apple[J]. Plant Physiology, 2022, 189(2): 1005-1020. DOI: 10.1093/plphys/kiac084 doi: 10.1093/plphys/kiac084

[32]	陈西霞.苹果BTB蛋白MdBT2与MdNAC1互作负调控铁离子稳态[D].泰安:山东农业大学,2022. CHEN X X. BTB protein MdBT2 interacts with MdNAC1 negatively regulates iron homeostasis in apple[D]. Tai’an: Shandong Agricultural University, 2022. (in Chinese with English abstract)

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