Genome-Wide Identification and Characterization of KNOTTED-Like Homeobox (KNOX) Homologs in Garlic (Allium sativum L.) and Their Expression Profilings Responding to Exogenous Cytokinin and Gibberellin
Abstract
:1. Introduction
2. Results
2.1. Identification and Characterization of KNOX Family Members
2.2. Chromosomal Distribution and Gene Structures of AsKNOX Genes
2.3. Cis-Element and Conserved Motif Analysis of AsKNOX Genes
2.4. Phylogenetic Analysis of AsKNOX Homologs
2.5. Expression Patterns of AsKNOX Genes in Different Garlic Tissues
2.6. Responses of AsKNOX Genes to Exogenous GA3 and 6-BA Treatments
2.7. Subcellular Localization of AsKNOX8 and AsKNOX6 Proteins
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Phytohormone Treatments
4.2. Identification and Sequence Analysis of AsKNOX Family Members
4.3. Chromosome Distribution and Gene Structure Analysis
4.4. Phylogenetic Analysis and Multiple Sequence Alignment
4.5. RNA Isolation and Gene Expression Analysis
4.6. Verification of Subcellular Localization
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Etoh, T.; Watanabe, H.; Iwai, S. RAPD variation of garlic clones in the center of origin and the westernmost area of distribution. Mem. Fac. Agric. 2001, 37, 21–27. [Google Scholar]
- Sun, X.; Zhu, S.; Li, N.; Cheng, Y.; Liu, T. A chromosome-level genome assembly of garlic (Allium sativum L.) provides insights into genome evolution and allicin biosynthesis. Mol. Plant 2020, 13, 1328–1339. [Google Scholar] [CrossRef] [PubMed]
- Kamenetsky, R. Garlic: Botany and Horticulture; John Wiley & Sons: Hoboken, NJ, USA, 2007. [Google Scholar]
- Ni, J.; Gao, C.; Chen, M.S.; Pan, B.Z.; Ye, K.; Xu, Z.F. Gibberellin promotes shoot branching in the perennial woody plant Jatropha curcas. Plant Cell Physiol. 2015, 8, 1655–1666. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Alexopoulos, A.A.; Akoumianakis, K.A.; Passam, H.C. The effect of the time and mode of application of gibberellic acid on the growth and yield of potato plants derived from true potato seed. Russ. J. Appl. Chem. 2010, 86, 2189–2195. [Google Scholar] [CrossRef]
- Yamazaki, H.; Shiraiwa, N.; Itai, A.; Honda, I. Involvement of gibberellins in the regulation of tillering in welsh onion (Allium fistulosum L.). Hortic. J. 2015, 84, MI-050. [Google Scholar] [CrossRef] [Green Version]
- Liu, H.; Deng, R.; Huang, C.; Cheng, Z.; Meng, H. Exogenous gibberellins alter morphology and nutritional traits of garlic (Allium sativum L.) bulb. Sci. Hortic. 2019, 246, 298–306. [Google Scholar] [CrossRef]
- Liu, H.J.; Huang, C.P.; Tong, P.J.; Yang, X.; Cheng, Z.H. Response of axillary bud development in garlic (Allium sativum L.) to seed cloves soaked in gibberellic acid (GA3) solution. J. Integr. Agric. 2020, 19, 1044–1054. [Google Scholar] [CrossRef]
- Wang, Y.; Jiao, Y. Axillary meristem initiation-a way to branch out. Curr. Opin. Plant Biol. 2018, 41, 61–66. [Google Scholar] [CrossRef]
- Giulio, T.; Emiliano, C.; Ignazio, V.; Chiara, N.; Emilia, C.; Teresa, D.M.; Elisa, V.; Leonardo, B.; Beatrice, B.M.; Giovanni, M. The peach (Prunus persica L. Batsch) genome harbours 10 KNOX genes, which are differentially expressed in stem development, and the class 1 KNOPE1 regulates elongation and lignification during primary growth. J. Exp. Bot. 2012, 63, 5417–5435. [Google Scholar]
- Bürglin, T. Analysis of TALE superclass homeobox genes (MEIS, PBC, KNOX, Iroquois, TGIF) reveals a novel domain conserved between plants and animals. Nucleic Acids Res. 1997, 25, 4173. [Google Scholar] [CrossRef] [Green Version]
- Mukherjee, K.; Brocchieri, L.; Bürglin, T.R. A comprehensive classification and evolutionary analysis of plant homeobox genes. Mol. Biol. Evol. 2009, 26, 2775–2794. [Google Scholar] [CrossRef] [Green Version]
- Hay, A.; Tsiantis, M. KNOX genes: Versatile regulators of plant development and diversity. Development 2010, 137, 3153–3165. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Furumizu, C.; Alvarez, J.P.; Sakakibara, K.; Bowman, J.L.; Qu, L.J. Antagonistic roles for KNOX1 and KNOX2 genes in patterning the land plant body plan following an ancient gene duplication. PLoS Genet. 2015, 11, e1004980. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Truernit, E.; Haseloff, J. A role for KNAT class II genes in root development. Plant Signal. Behav. 2007, 2, 10–12. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhong, R.; Lee, C.; Zhou, J.; Ye, M. A battery of transcription factors involved in the regulation of secondary cell wall biosynthesis in Arabidopsis. Plant Cell 2008, 20, 2763–2782. [Google Scholar] [CrossRef] [Green Version]
- Jia, P.; Xing, L.; Zhang, C.; Zhang, D.; An, N. MdKNOX19, a class II knotted-like transcription factor of apple, plays roles in ABA signalling /sensitivity by targeting ABI5 during organ development. Plant Sci. 2021, 302, 110701. [Google Scholar] [CrossRef]
- Byrne, M.E.; Simorowski, J.; Martienssen, R.A. Asymmetric leaves1 reveals knox gene redundancy in Arabidopsis. Development 2002, 129, 1957–1965. [Google Scholar] [CrossRef]
- Venglat, S.P.; Dumonceaux, T.; Rozwadowski, K.; Parnell, L.; Datla, R. The homeobox gene BREVIPEDICELLUS is a key regulator of inflorescence architecture in Arabidopsis. Proc. Natl. Acad. Sci. USA 2002, 99, 4730–4735. [Google Scholar] [CrossRef] [Green Version]
- Ragni, L.; Belles-Boix, E.; Gunl, M.; Pautot, V. Interaction of KNAT6 and KNAT2 with BREVIPEDICELLUS and PENNYWISE in Arabidopsis inflorescences. Plant Cell 2008, 20, 888–900. [Google Scholar] [CrossRef] [Green Version]
- Yu, L.; Patibanda, V.; Smith, H. A novel role of BELL1-like homeobox genes, PENNYWISE and POUND-FOOLISH, in floral patterning. Planta 2009, 229, 693–707. [Google Scholar] [CrossRef] [PubMed]
- Jasinski, S.; Piazza, P.; Craft, J.; Hay, A.; Tsiantis, M. KNOX action in Arabidopsis is mediated by coordinate regulation of cytokinin and gibberellin activities. Curr. Biol. 2005, 15, 1560–1565. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Du, J.; Mansfield, S.D.; Groover, A.T. The Populus homeobox gene Arborknox2 regulates cell differentiation during secondary growth. Plant J. 2010, 60, 1000–1014. [Google Scholar] [CrossRef]
- Cheng, X.; Li, M.; Abdullah, M.; Li, G.; Lin, Y. In silico genome-wide analysis of the pear (Pyrus bretschneideri) KNOX family and the functional characterization of PbKNOX1, an Arabidopsis Brevipedicellus orthologue gene, involved in cell wall and lignin biosynthesis. Front. Genet. 2019, 10, 632. [Google Scholar] [CrossRef] [PubMed]
- Vollbrecht, E.; Veit, B.; Sinha, N.; Hake, S. The developmental gene Knotted-1 is a member of a maize homeobox gene family. Nature 1991, 350, 241–243. [Google Scholar] [CrossRef] [PubMed]
- Vollbrecht, E.; Reiser, L.; Hake, S. Shoot meristem size is dependent on inbred background and presence of the maize homeobox gene, knotted1. Development 2000, 127, 3161. [Google Scholar] [CrossRef]
- Jia, P.; Zhang, C.; Xing, L.; Li, Y.; Shah, K.; Zuo, X.; Zhang, D.; An, N.; Han, M.; Ren, X. Genome-wide identification of the MdKNOX gene family and characterization of its transcriptional regulation in Malus domestica. Front. Plant Sci. 2020, 11, 128. [Google Scholar] [CrossRef]
- Jia, P.; Xing, L.; Zhang, C.; Chen, H.; An, N. MdKNOX15, a class I knotted-like transcription factor of apple, controls flowering and plant height by regulating GA levels through promoting the MdGA2ox7 transcription. Environ. Exp. Bot. 2021, 185, 104411. [Google Scholar] [CrossRef]
- Hackbusch, J.; Richter, K.; Müller, J.; Salamini, F.; Uhrig, J.F. A central role of Arabidopsis thaliana ovate family proteins in networking and subcellular localization of 3-aa loop extension homeodomain proteins. Proc. Natl. Acad. Sci. USA 2005, 102, 4908–4912. [Google Scholar] [CrossRef] [Green Version]
- Gonzalez, D.H. Plant transcription factors evolutionary, structural and functional aspects. In Introduction to Transcription Factor Structure and Function; Academic Press: Cambridge, MA, USA, 2016; pp. 3–11. ISBN 978-0-12-800854-6. [Google Scholar]
- Lucas, W.J.; Bouche-Pillon, S.; Jackson, D.; Nguyen, L.; Baker, L.; Ding, B.; Hake, S. Selective trafficking of KNOTTED1 Homeodomain protein and its mRNA through plasmodesmata. Science 1995, 270, 1980–1983. [Google Scholar] [CrossRef]
- Bueno, N.; Alvarez, J.M.; Ordás, R. Characterization of the Knotted1-Like Homeobox (KNOX) gene family in Pinus pinaster Ait. Plant Sci. 2020, 301, 110691. [Google Scholar] [CrossRef] [PubMed]
- Li, E.; Bhargava, A.; Qiang, W.; Friedmann, M.; Forneris, N.; Savidge, R.; Johnson, L.; Mansfield, S.; Ellis, B.; Douglas, C. The Class II KNOX gene KNAT7 negatively regulates secondary wall formation in Arabidopsis and is functionally conserved in Populus. New Phytol. 2012, 194, 102–115. [Google Scholar] [CrossRef] [PubMed]
- Wang, S.; Yamaguchi, M.; Grienenberger, E.; Martone, P.T.; Samuels, A.L.; Mansfield, S.D. The Class II KNOX genes KNAT3 and KNAT7 work cooperatively to influence deposition of secondary cell walls that provide mechanical support to Arabidopsis stems. Plant J. 2020, 101, 293–309. [Google Scholar] [CrossRef]
- Huang, L.M. ATH1 and KNAT2 proteins act together in regulation of plant inflorescence architecture. J. Exp. Bot. 2012, 63, 1423–1433. [Google Scholar]
- Tsuda, K.; Hake, S. Diverse functions of KNOX transcription factors in the diploid body plan of plants. Curr. Opin. Plant Biol. 2015, 27, 91–96. [Google Scholar] [CrossRef] [Green Version]
- Yanai, O.; Shani, E.; Dolezal, K.; Tarkowski, P.; Ori, N. Arabidopsis KNOXI proteins activate cytokinin biosynthesis. Curr. Biol. 2005, 15, 1566–1571. [Google Scholar] [CrossRef] [Green Version]
- Hay, A.; Kaur, H.; Phillips, A.; Hedden, P.; Tsiantis, M. The gibberellin pathway mediates knotted1-type homeobox function in plants with different body plans. Curr. Biol. 2002, 12, 1557–1565. [Google Scholar] [CrossRef] [Green Version]
- Ye, S.G.; Zai, W.S.; Xiong, Z.L.; Zhang, H.L.; Ma, Y.R. Genome-wide identification of gene family in tomato and their evolutionary relationship in Solanaceae. J. Nucl. Agric. Sci. 2017, 31, 1263–1271. [Google Scholar]
- Jie, G.; Yang, X.; Zhao, W.; Lang, T.; Tore, S. Evolution, diversification, and expression of KNOX proteins in plants. Front. Plant Sci. 2015, 6, 882. [Google Scholar]
- Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 2001, 25, 402–408. [Google Scholar] [CrossRef]
- Liu, M.; Wu, Z.; Jiang, F. Selection and validation of garlic reference genes for quantitative real-time PCR normalization. Plant Cell Tissue Organ Cult. 2015, 122, 435–444. [Google Scholar] [CrossRef]
Gene | Gene ID | Protein Length (aa) | pI | Mw (kDa) | GRAVY | Predicted Subcellular Localization |
---|---|---|---|---|---|---|
AsKNOX1 | Asa1G01043.1 | 354 | 7.29 | 40.21 | −0.961 | Nucleus, Chloroplast |
AsKNOX2 | Asa2G04444.1 | 160 | 4.05 | 17.25 | −0.367 | Nucleus |
AsKNOX3 | Asa2G07204.1 | 307 | 5.10 | 34.65 | −0.815 | Nucleus |
AsKNOX4 | Asa2G07205.1 | 331 | 5.33 | 37.68 | −0.684 | Nucleus |
AsKNOX5 | Asa4G01931.1 | 278 | 5.86 | 32.13 | −0.674 | Nucleus |
AsKNOX6 | Asa4G03695.1 | 322 | 5.61 | 36.62 | −0.752 | Nucleus |
AsKNOX7 | Asa6G00294.1 | 315 | 5.68 | 35.84 | −0.793 | Nucleus, Cytoplasm |
AsKNOX8 | Asa8G00804.1 | 287 | 5.08 | 32.42 | −0.592 | Nucleus |
AsKNOX9 | Asa8G04023.1 | 330 | 6.08 | 37.41 | −0.604 | Nucleus |
AsKNOX10 | Asa0G05087.1 | 324 | 6.39 | 37.27 | −0.785 | Nucleus, Cytoplasm |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Zhang, S.; Pan, Y.; Zhi, C.; Zheng, Y.; Wang, X.; Li, X.; Cheng, Z. Genome-Wide Identification and Characterization of KNOTTED-Like Homeobox (KNOX) Homologs in Garlic (Allium sativum L.) and Their Expression Profilings Responding to Exogenous Cytokinin and Gibberellin. Int. J. Mol. Sci. 2021, 22, 9237. https://doi.org/10.3390/ijms22179237
Zhang S, Pan Y, Zhi C, Zheng Y, Wang X, Li X, Cheng Z. Genome-Wide Identification and Characterization of KNOTTED-Like Homeobox (KNOX) Homologs in Garlic (Allium sativum L.) and Their Expression Profilings Responding to Exogenous Cytokinin and Gibberellin. International Journal of Molecular Sciences. 2021; 22(17):9237. https://doi.org/10.3390/ijms22179237
Chicago/Turabian StyleZhang, Siyu, Yupeng Pan, Chengchen Zhi, Yujie Zheng, Xi’ao Wang, Xiaxia Li, and Zhihui Cheng. 2021. "Genome-Wide Identification and Characterization of KNOTTED-Like Homeobox (KNOX) Homologs in Garlic (Allium sativum L.) and Their Expression Profilings Responding to Exogenous Cytokinin and Gibberellin" International Journal of Molecular Sciences 22, no. 17: 9237. https://doi.org/10.3390/ijms22179237