Genome-wide identification of bHLH gene family and screening of genes related to prickle development in Zanthoxylum armatum
Yuwei Yi A B , Hualin Zou A , Nuo Wang A , Yansheng Xue A , Weiwei Zhang
A
B
Abstract
Zanthoxylum armatum has edible and medicinal value but its prickles make harvesting difficult. The bHLH gene family is vital in regulating physiological and developmental processes. One hundred and ninety-five ZabHLH genes from its genome were grouped into 11 subgroups and 23 subfamilies. Members of the bHLH IIIf subfamily play an important role in trichome development, and ZabHLH22, ZabHLH110, ZabHLH161, and ZabHLH194, which belong to this subfamily, were selected as candidate genes. Chromosomal localization analysis showed that 165 of 195 ZabHLHs were unevenly distributed on 31 chromosomes, and 30 ZabHLHs were localized to unanchored scaffolds. The expansion of ZabHLHs mainly includes dispersed replication and whole-genome duplication or segmental replication. Fourty-seven cis-acting elements were predicted in the promoters of ZabHLHs, with hormone-responsive elements being the most abundant. Expression profiles of four candidate genes were analyzed in two Z. armatum cultivars. Trichome development is regulated by hormones such as methyl jasmonate, salicylic acid, and auxin. The qRT-PCR results indicate that four candidate genes respond to the stress induced by these three hormones. We predict that ZabHLH110, ZabHLH161, and ZabHLH194 are most likely involved in prickle development. The results are helpful to further explore the potential roles and mechanisms of ZabHLHs in the development of Z. armatum prickles.
Keywords: bHLH, bioinformatics, genome-wide, indole acetic acid, methyl jasmonate, prickles, qRT-PCR, salicylic acid, Zanthoxylum armatum.
References
Ahmad Khan R, Mohammad , Kumar A, Abbas N (2024) AaGL3-like is jasmonate-induced bHLH transcription factor that positively regulates trichome density in Artemisia annua. Gene 904, 148213.
| Crossref | Google Scholar | PubMed |
Atchley WR, Fitch WM (1997) A natural classification of the basic helix–loop–helix class of transcription factors. Proceedings of the National Academy of Sciences 94(10), 5172-5176.
| Crossref | Google Scholar |
Atchley WR, Terhalle W, Dress A (1999) Positional dependence, cliques, and predictive motifs in the bHLH protein domain. Journal of Molecular Evolution 48, 501-516.
| Crossref | Google Scholar | PubMed |
Balkunde R, Pesch M, Hülskamp M (2010) Chapter ten - Trichome patterning in arabidopsis thaliana: from genetic to molecular models. In ‘Current topics in developmental biology, Vol. 91’ . (Ed. MCP Timmermans) pp. 299–321. (Academic Press) 10.1016/S0070-2153(10)91010-7.
Bernhardt C, Lee MM, Gonzalez A, Zhang F, Lloyd A, Schiefelbein J (2003) The bHLH genes GLABRA3 (GL3) and ENHANCER OF GLABRA3 (EGL3) specify epidermal cell fate in the Arabidopsis root. Development 130(26), 6431-6439.
| Crossref | Google Scholar | PubMed |
Bloomer RH, Juenger TE, Symonds VV (2012) Natural variation in GL1 and its effects on trichome density in Arabidopsis thaliana. Molecular Ecology 21(14), 3501-3515.
| Crossref | Google Scholar | PubMed |
Chen C, Chen H, Zhang Y, Thomas HR, Frank MH, He Y, Xia R (2020) TBtools: an integrative toolkit developed for interactive analyses of big biological data. Molecular Plant 13(8), 1194-1202.
| Crossref | Google Scholar | PubMed |
Dong H, Chen Q, Dai Y, Hu W, Zhang S, Huang X (2021) Genome-wide identification of PbrbHLH family genes, and expression analysis in response to drought and cold stresses in pear (Pyrus bretschneideri). BMC Plant Biology 21, 86.
| Crossref | Google Scholar | PubMed |
Fan Y, Yang H, Lai D, He A, Xue G, Feng L, Chen L, Cheng X-B, Ruan J, Yan J, Cheng J (2021) Genome-wide identification and expression analysis of the bHLH transcription factor family and its response to abiotic stress in sorghum [Sorghum bicolor (L.) Moench]. BMC Genomics 22, 415.
| Crossref | Google Scholar | PubMed |
Feller A, Machemer K, Braun EL, Grotewold E (2011) Evolutionary and comparative analysis of MYB and bHLH plant transcription factors. The Plant Journal 66(1), 94-116.
| Crossref | Google Scholar | PubMed |
Feng S, Liu Z, Hu Y, Tian J, Yang T, Wei A (2020) Genomic analysis reveals the genetic diversity, population structure, evolutionary history and relationships of Chinese pepper. Horticulture Research 7, 158.
| Crossref | Google Scholar | PubMed |
Feng Z, Sun L, Dong M, Fan S, Shi K, Qu Y, Zhu L, Shi J, Wang W, Liu Y, Chen X, Weng Y, Liu X, Ren H (2023) Identification and functional characterization of CsMYCs in cucumber glandular trichome development. International Journal of Molecular Sciences 24(7), 6435.
| Crossref | Google Scholar | PubMed |
Gao F, Dubos C (2024) The arabidopsis bHLH transcription factor family. Trends in Plant Science 29(6), 668-680.
| Crossref | Google Scholar | PubMed |
Guo X-J, Wang J-R (2017) Global identification, structural analysis and expression characterization of bHLH transcription factors in wheat. BMC Plant Biology 17, 90.
| Crossref | Google Scholar | PubMed |
Guo Z, Zhang Q, Zhang Y, Wu C, Zheng Y, Tong F, Zhang L, Lu R, Pan X, Tan H, Lv Z (2023) Effects of exogenous indole-3-acetic acid on the density of trichomes, expression of artemisinin biosynthetic genes, and artemisinin biosynthesis in Artemisia annua. Biotechnology and Applied Biochemistry 70(6), 1870-1880.
| Crossref | Google Scholar | PubMed |
Han W, Zhang Q, Suo Y, Li H, Diao S, Sun P, Huang L, Fu J (2023) Identification and expression analysis of the bHLH gene family members in Diospyros kaki. Horticulturae 9(3), 380.
| Crossref | Google Scholar |
Heim MA, Jakoby M, Werber M, Martin C, Weisshaar B, Bailey PC (2003) The basic helix–loop–helix transcription factor family in plants: a genome-wide study of protein structure and functional diversity. Molecular Biology and Evolution 20(5), 735-747.
| Crossref | Google Scholar | PubMed |
Huang X, Su L, Xian B, Yu Q, Zhang M, Fan J, Zhang C, Liu Y, He H, Zhong X, Li M, Chen S, He Y, Li Q (2024) Genome-wide identification and characterization of the sweet orange (Citrus sinensis) basic helix-loop-helix (bHLH) family reveals a role for CsbHLH085 as a regulator of citrus bacterial canker resistance. International Journal of Biological Macromolecules 267, 131442.
| Crossref | Google Scholar | PubMed |
Inthima P, Nakano M, Otani M, Niki T, Nishijima T, Koshioka M, Supaibulwatana K (2017) Overexpression of the gibberellin 20-oxidase gene from Torenia fournieri resulted in modified trichome formation and terpenoid metabolities of Artemisia annua L. Plant Cell, Tissue and Organ Culture (PCTOC) 129, 223-236.
| Crossref | Google Scholar |
Koornneeff M, Dellaert LWM, Van der Veen JH (1982) EMS- and relation-induced mutation frequencies at individual loci in Arabidopsis thaliana (L.) Heynh. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 93(1), 109-123.
| Crossref | Google Scholar |
Li X, Duan X, Jiang H, Sun Y, Tang Y, Yuan Z, Guo J, Liang W, Chen L, Yin J, Ma H, Wang J, Zhang D (2006) Genome-wide analysis of basic/helix-loop-helix transcription factor family in rice and Arabidopsis. Plant Physiology 141(4), 1167-1184.
| Crossref | Google Scholar | PubMed |
Li X, Xiang F, Han W, Qie B, Zhai R, Yang C, Wang Z, Xu L (2021) The MIR-domain of PbbHLH2 Is involved in regulation of the anthocyanin biosynthetic pathway in ”Red Zaosu” (PyrusBretschneideri Rehd.) pear fruit. International Journal of Molecular Sciences 22(6), 3026.
| Crossref | Google Scholar | PubMed |
Liu X, Bartholomew E, Cai Y, Ren H (2016) Trichome-related mutants provide a new perspective on multicellular trichome initiation and development in cucumber (Cucumis sativus L). Frontiers in Plant Science 7, 1187.
| Crossref | Google Scholar |
Liu Y, Li Q, Yang W, Sun B, Zhou Y, Zheng Y, Huang M, Yang W (2020) Characterization of the potent odorants in Zanthoxylum armatum DC Prodr. pericarp oil by application of gas chromatography–mass spectrometry–olfactometry and odor activity value. Food Chemistry 319, 126564.
| Crossref | Google Scholar | PubMed |
Liu R, Wang Y, Tang S, Cai J, Liu S, Zheng P, Sun B (2021) Genome-wide identification of the tea plant bHLH transcription factor family and discovery of candidate regulators of trichome formation. Scientific Reports 11, 10764.
| Crossref | Google Scholar | PubMed |
Liu X, Tang N, Xu F, Chen Z, Zhang X, Ye J, Liao Y, Zhang W, Kim S-U, Wu P, Cao Z (2022a) SMRT and Illumina RNA sequencing reveal the complexity of terpenoid biosynthesis in Zanthoxylum armatum. Tree Physiology 42(3), 664-683.
| Crossref | Google Scholar |
Liu X, He X, Liu Z, Wu P, Tang N, Chen Z, Zhang W, Rao S, Cheng S, Luo C, Xu F (2022b) Transcriptome mining of genes in Zanthoxylum armatum revealed ZaMYB86 as a negative regulator of prickly development. Genomics 114(3), 110374.
| Crossref | Google Scholar |
Liu Q, Wen J, Wang S, Chen J, Sun Y, Liu Q, Li X, Dong S (2023) Genome-wide identification, expression analysis, and potential roles under low-temperature stress of bHLH gene family in Prunus sibirica. Frontiers in Plant Science 14, 1267107.
| Crossref | Google Scholar | PubMed |
Ma X, Li K, Wang Z, Wei D, Tang Q (2020) Research progress on regulatory models of different types of epidermal hairs in plants. Chinese Journal of Biotechnology 36(10), 2051-2065.
| Crossref | Google Scholar | PubMed |
Moon J, Skibbe D, Timofejeva L, Rachel Wang C-JR, Kelliher T, Kremling K, Walbot V, Zacheus Cande W (2013) Regulation of cell divisions and differentiation by MALE STERILITY32 is required for anther development in maize. The Plant Journal 76(4), 592-602.
| Crossref | Google Scholar | PubMed |
Morohashi K, Grotewold E (2009) A systems approach reveals regulatory circuitry for Arabidopsis Trichome initiation by the GL3 and GL1 selectors. PLoS Genetics 5(2), e1000396.
| Crossref | Google Scholar | PubMed |
Morohashi K, Zhao M, Yang M, Read B, Lloyd A, Lamb R, Grotewold E (2007) Participation of the Arabidopsis bHLH Factor GL3 in trichome initiation regulatory events. Plant Physiology 145(3), 736-746.
| Crossref | Google Scholar | PubMed |
Oppenheimer DG, Herman PL, Sivakumaran S, Esch J, Marks MD (1991) A myb gene required for leaf trichome differentiation in Arabidopsis is expressed in stipules. Cell 67(3), 483-493.
| Crossref | Google Scholar | PubMed |
Pattanaik S, Patra B, Singh SK, Yuan L (2014) An overview of the gene regulatory network controlling trichome development in the model plant, Arabidopsis. Frontiers in Plant Science 5, 259.
| Crossref | Google Scholar |
Payne CT, Zhang F, Lloyd AM (2000) GL3 encodes a bHLH protein that regulates trichome development in arabidopsis through interaction with GL1 and TTG1. Genetics 156(3), 1349-1362.
| Crossref | Google Scholar | PubMed |
Pires N, Dolan L (2010) Origin and diversification of basic-helix-loop-helix proteins in plants. Molecular Biology and Evolution 27(4), 862-874.
| Crossref | Google Scholar | PubMed |
Qi T, Song S, Ren Q, Wu D, Huang H, Chen Y, Fan M, Peng W, Ren C, Xie D (2011) The jasmonate-ZIM-domain proteins interact with the WD-Repeat/bHLH/MYB complexes to regulate jasmonate-mediated anthocyanin accumulation and trichome initiation in Arabidopsis thaliana. The Plant Cell 23(5), 1795-1814.
| Crossref | Google Scholar | PubMed |
Serna L, Martin C (2006) Trichomes: different regulatory networks lead to convergent structures. Trends in Plant Science 11(6), 274-280.
| Crossref | Google Scholar | PubMed |
Shangguan X-X, Yang C-Q, Zhang X-F, Wang L-J (2016) Functional characterization of a basic helix-loop-helix (bHLH) transcription factor GhDEL65 from cotton (Gossypium hirsutum). Physiologia Plantarum 158(2), 200-212.
| Crossref | Google Scholar | PubMed |
Sun H, Fan H-J, Ling H-Q (2015) Genome-wide identification and characterization of the bHLH gene family in tomato. BMC Genomics 16, 9.
| Crossref | Google Scholar | PubMed |
Symonds VV (2004) Genetic analyses of natural variation in the model plant Arabidopsis thaliana: neutral marker, quantitative genetic, and population genetic approaches. Doctor of Philosophy, The University of Texas at Austin. Available at http://hdl.handle.net/2152/1418.
Symonds VV, Hatlestad G, Lloyd AM (2011) Natural allelic variation defines a role for ATMYC1: trichome cell fate determination. PLoS Genetics 7, e1002069.
| Crossref | Google Scholar | PubMed |
Tang N, Cao Z, Wu P, Liu Y, Lou J, Hu Y, Sun X, Si S, Chen Z (2023) Comparative transcriptome analysis reveals hormone, transcriptional and epigenetic regulation involved in prickle formation in Zanthoxylum armatum. Gene 871, 147434.
| Crossref | Google Scholar | PubMed |
Toledo-Ortiz G, Huq E, Quail PH (2003) The Arabidopsis basic/helix-loop-helix transcription factor family. The Plant Cell 15(8), 1749-1770.
| Crossref | Google Scholar | PubMed |
Traw MB, Bergelson J (2003) Interactive effects of jasmonic acid, salicylic acid, and gibberellin on induction of trichomes in Arabidopsis. Plant Physiology 133(3), 1367-1375.
| Crossref | Google Scholar | PubMed |
Walker AR, Davison PA, Bolognesi-Winfield AC, James CM, Srinivasan N, Blundell TL, Esch JJ, Marks MD, Gray JC (1999) The TRANSPARENT TESTA GLABRA1 locus, which regulates trichome differentiation and anthocyanin biosynthesis in Arabidopsis, encodes a WD40 repeat protein. The Plant Cell 11(7), 1337-1349.
| Crossref | Google Scholar | PubMed |
Wang M, Tong S, Ma T, Xi Z, Liu J (2021) Chromosome-level genome assembly of Sichuan pepper provides insights into apomixis, drought tolerance, and alkaloid biosynthesis. Molecular Ecology Resources 21(7), 2533-2545.
| Crossref | Google Scholar | PubMed |
Wang T, Wang S, Wang Y, Li J, Yan F, Liu Y, Zhao L, Wang Q (2020) Jasmonic acid–induced inhibition of root growth and leaf senescence is reduced by GmbHLH3, a soybean bHLH transcription factor. Canadian Journal of Plant Science 100(5), 477-487.
| Crossref | Google Scholar |
Xia X-C, Hu Q-Q, Li W, Chen Y, Han L-H, Tao M, Wu W-Y, Li X-B, Huang G-Q (2018) Cotton (Gossypium hirsutum) JAZ3 and SLR1 function in jasmonate and gibberellin mediated epidermal cell differentiation and elongation. Plant Cell, Tissue and Organ Culture (PCTOC) 133, 249-262.
| Crossref | Google Scholar |
Xu D, Zhuo Z, Wang R, Ye M, Pu B (2019) Modeling the distribution of Zanthoxylum armatum in China with MaxEnt modeling. Global Ecology and Conservation 19, e00691.
| Crossref | Google Scholar |
Yan H, Wu Z, Liu Y, Weng Q, Yi Y, Huang X (2021) Functional divergence of RrGL3 and RrEGL3 from Rosa roxburghii in mediating trichome development. Plant Cell, Tissue and Organ Culture (PCTOC) 147, 313-324.
| Crossref | Google Scholar |
Yang J, Gao M, Huang L, Wang Y, van Nocker S, Wan R, Guo C, Wang X, Gao H (2017) Identification and expression analysis of the apple (Malus × domestica) basic helix-loop-helix transcription factor family. Scientific Reports 7, 28.
| Crossref | Google Scholar | PubMed |
Zhang F, Gonzalez A, Zhao M, Payne CT, Lloyd A (2003) A network of redundant bHLH proteins functions in all TTG1-dependent pathways of Arabidopsis. Development 130(20), 4859-4869.
| Crossref | Google Scholar | PubMed |
Zhang T, Lv W, Zhang H, Ma L, Li P, Ge L, Li G (2018) Genome-wide analysis of the basic helix-loop-helix (bHLH) transcription factor family in maize. BMC Plant Biology 18, 235.
| Crossref | Google Scholar | PubMed |
Zhang X-Y, Qiu J-Y, Hui Q-L, Xu Y-Y, He Y-Z, Peng L-Z, Fu X-Z (2020) Systematic analysis of the basic/helix-loop-helix (bHLH) transcription factor family in pummelo (Citrus grandis) and identification of the key members involved in the response to iron deficiency. BMC Genomics 21, 233.
| Crossref | Google Scholar | PubMed |
Zhao H, Wang X, Zhu D, Cui S, Li X, Cao Y, Ma L (2012) A single amino acid substitution in IIIf subfamily of basic helix-loop-helix transcription factor AtMYC1 leads to trichome and root hair patterning defects by abolishing its interaction with partner proteins in Arabidopsis. Journal of Biological Chemistry 287(17), 14109-14121.
| Crossref | Google Scholar | PubMed |
Zhao L, Bi W, Jia Y, Shi J, Chi Y, Yu M, Wang C (2023) Genome-wide characterization of bHLH family genes and expression analysis in response to osmotic stress in Betula platyphylla. Plants 12(21), 3687.
| Crossref | Google Scholar |
Zhou Z, Sun L, Zhao Y, An L, Yan A, Meng X, Gan Y (2013) Zinc Finger Protein 6 (ZFP6) regulates trichome initiation by integrating gibberellin and cytokinin signaling in Arabidopsis thaliana. New Phytologist 198(3), 699-708.
| Crossref | Google Scholar | PubMed |
Zhou NN, Tang KX, Jeauffre J, Thouroude T, Arias DCL, Foucher F, Oyant LH-S (2020a) Genetic determinism of prickles in rose. Theoretical and Applied Genetics 133, 3017-3035.
| Crossref | Google Scholar | PubMed |
Zhou X, Liao Y, Kim S-U, Chen Z, Nie G, Cheng S, Ye J, Xu F (2020b) Genome-wide identification and characterization of bHLH family genes from Ginkgo biloba. Scientific Reports 10, 13723.
| Crossref | Google Scholar | PubMed |
Zhou N, Simonneau F, Thouroude T, Oyant LH-S, Foucher F (2021) Morphological studies of rose prickles provide new insights. Horticulture Research 8, 221.
| Crossref | Google Scholar | PubMed |
Zi Y, Zhang M, Yang X, Zhao K, Yin T, Wen K, Li X, Liu X, Zhang H (2024) Identification of the sweet orange (Citrus sinensis) bHLH gene family and the role of CsbHLH55 and CsbHLH87 in regulating salt stress. The Plant Genome 17(3), e20502.
| Crossref | Google Scholar | PubMed |