Volume 8, Issue 3, September 2020, Page: 43-53
Ectopic Expression of WUSCHEL (AtWUS) Gene Alters Plant Growth and Development in Rice
Thiveyarajan Victorathisayam, Department of Plant Biotechnology, Madurai Kamaraj University, Madurai, India
Ganapathi Sridevi, Department of Plant Biotechnology, Madurai Kamaraj University, Madurai, India
Received: Jul. 4, 2020;       Accepted: Jul. 25, 2020;       Published: Aug. 25, 2020
DOI: 10.11648/j.plant.20200803.11      View  50      Downloads  57
Developmental genes (DG)/ morphogenic genes are involved in enhancing the transformation and regeneration of plants. One such DG is the WUSCHEL (WUS) gene, a homeodomain transcription factor, and is involved in the stem cell maintenance of shoot apical meristem (SAM). In dicots, ectopic expression of WUS induced the embryogenic calli formation and organogenesis. On the other, WUS overexpression resulted in pleiotropic effects in most of the monocots. Also, very few dicots failed to regenerate due to the overexpression of WUS. In our study, the 35S driven WUS (AtWUS) gene expressing transgenic rice plants were generated. All the transgenic plants with the WUS (W) gene along with the vector (V) and untransformed (U) lines were confirmed by detailed Southern analyses. The single-copy W and V plants with complete T-DNA were taken for detailed analyses. The W plants exhibited few phenotypic changes such as thick stem, reduction in the internode length, enclosed panicle, unopen flower, pale yellow colour of the anther, and loss of viable pollens compared to the U and V plants. Interestingly, crown root formation and small vein formation in the leaves were detected in the W plants. The expression of the WUS gene was confirmed by RT-PCR analysis in the W plants. The seeds from the hemizygous plants showed enhanced embryogenic calli formation and attained early regeneration compared to U and V plants thereby confirming the role of the WUS gene in embryogenesis and regeneration.
WUSCHEL , Embryogenic Calli, Regeneration, Pleiotropic Effects, Organogenesis
To cite this article
Thiveyarajan Victorathisayam, Ganapathi Sridevi, Ectopic Expression of WUSCHEL (AtWUS) Gene Alters Plant Growth and Development in Rice, Plant. Vol. 8, No. 3, 2020, pp. 43-53. doi: 10.11648/j.plant.20200803.11
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Nagle M, Dejardin A, Pilate G, Strauss SH (2018) Opportunities for innovation in genetic transformation of forest trees, Front. Plant Sci 9: 1443. DOI: 10.3389/fpls.2018.01443.
Ikeuchi M, Ogawa Y, Iwase A, Sugimoto K (2016) Plant regeneration: cellular origins and molecular mechanisms. Development 143: 1442–1451.
Lowe K, Wu E, Wang N, et al. (2016) Morphogenic regulators Baby boom and Wuschel improve monocot transformation. Plant Cell 28 (9): 1998–2015. DOI: 10.1105/tpc.16.00124.
Altpeter F (2016) Advancing crop transformation in the era of genome editing. Plant Cell 28: 1510–1520.
Gordon-Kamm B, Sardesai N, Arling M, Lowe K, Hoerster G, Betts S, Jones T (2019) Using morphogenic genes to improve recovery and regeneration of transgenic plants. Plants 8: 38.
Thomas T (1993) Gene expression during plant embryogenesis and germination: An overview. Plant Cell 5: 1401–1410.
Schmidt EDL, Guzzo F, Toonen MAJ, de Vries SC (1997) A leucine-rich repeat-containing receptor-like kinase marks somatic plant cells competent to form embryos. Development 124: 2049–2062.
Bratzel F, López-Torrejón G, Koch M, Del Pozo JC, Calonje M (2010) Keeping cell identity in Arabidopsis requires PRC1 RING-finger homologs that catalyze H2A monoubiquitination. Curr Biol 26: 1853-9.
Chen D, Molitor A, Liu C, Shen WH (2010) The Arabidopsis PRC1-like ring-finger proteins are necessary for repression of embryonic traits during vegetative growth. Cell Res 20 (12): 1332-44.
Yang C, Bratzel F, Hohmann N, Koch M, Turck F, Calonje M (2013) VAL- and AtBMI1-mediated H2Aub initiate the switch from embryonic to post germinative growth in Arabidopsis. Curr Biol 23 (14): 1324-9.
Xu L, Huang H (2014). Genetic and epigenetic controls of plant regeneration. Current topics in developmental biology 108: 1-33. 10.1016/B978-0-12-391498-9.00009-7.
Ikeda M, Ohme-Takagi M (2014) TCPs, WUSs, and WINDs: families of transcription factors that regulate shoot meristem formation, stem cell maintenance, and somatic cell differentiation. Front Plant Science 5: 427.
Heyman J, Canher B, Bisht A, Christiaens F, De Veylder L (2018) Emerging role of the plant ERF transcription factors in coordinating wound defense responses and repair. J Cell Sci 131: jcs208215. https://doi.org/10.1242/jcs.208215
Ikeuchi M, Shibata M, Rymen B, Iwase A, Bågman A-M, et al. (2018) A gene regulatory network for cellular reprogramming in plant regeneration. Plant Cell Physiol 59 (4): 770–82.
Gehring WJ, Qian YQ, Billeter M (1994) Homeodomain-DNA recognition. Cell 78 (2): 211–223. DOI: 10.1016/0092-8674(94)90292-5.
Van der Graff E., Laux T., Rensing SA (2009) The WUS homeobox-containing (WOX) protein family. Genome Biology 10: 248.
Zuo J, Niu QW, Frugis G, Chua NH (2002) The WUSCHEL gene promotes vegetative-to-embryonic transition in Arabidopsis. The Plant Journal 30: 349-359.
Lenhard M, Jurgens G, Laux T (2002) The WUSCHEL and SHOOTMERISTEMLESS genes fulfill complementary roles in Arabidopsis shoot meristem regulation. Development 129: 3195-3206.
Laux T, Mayer KFX, Berger J, Jurgens G (1996) The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Development 122: 87-96.
Mayer KFX., Schoof H, Haecker A, Lenhard M, Jurgens G, Laux T (1998) Role of WUSCHEL in regulating stem cell fate in the Arabidopsis shoot meristem. Cell 95: 805–815.
Lohmann JU, Hong RL, Hobe M, et al. (2001) A molecular link between stem cell regulation and floral patterning in Arabidopsis. Cell 105 (6): 793–803. doi: 10.1016/s0092-8674(01)00384-1.
Nardmann J, Werr W (2006) The shoot stem cell niche in angiosperms: expression patterns of WUS orthologues in rice and maize imply major modifications in the course of mono- and dicot evolution. Mol Biol Evol 23 (12): 2492-504.
Lu Z, Shao G, Xiong J, Jiao Y, Wang J, Liu G, Meng X, Liang Y, Xiong G, Wang Y, Li J (2015) MONOCULM 3, an ortholog of WUSCHEL in rice, is required for tiller bud formation. Journal of Genetics and Genomics 42: 71-78.
Tanaka W, Ohmori Y, Ushijima T, Matsusaka H, Matsushita T, Kumamaru T, Kawano S, Hiranoa HY (2015) Axillary meristem formation in rice requires the WUSCHEL ortholog TILLERS ABSENT1. Plant Cell 27: 1173–1184.
Tanaka W, Hirano HY (2019) Antagonistic action of TILLERS ABSENT1 and FLORAL ORGAN NUMBER2 regulates stem cell maintenance during axillary meristem development in rice. New Phytologist DOI: 10.1111/nph.16163.
Suzuki C, Tanaka W, Tsuji H, Hirano HY (2019) TILLERS ABSENT1, the WUSCHEL ortholog, is not involved in stem cell maintenance in the shoot apical meristem in rice. Plant Signal Behav 14 (9): 1640565. DOI: 10.1080/15592324.2019.1640565.
Ohmori Y, Tanaka W, Kojima M, Sakakibara H, Hirano HY (2013) WUSCHEL-RELATED HOMEOBOX4 is involved in meristem maintenance and is negatively regulated by the CLE gene FCP1 in rice. Plant Cell 25: 229-241.
Weiss J, Rodriguez RA, Toksoz T, Cortines ME (2016) Meristem maintenance, auxin, jasmonic, and abscisic acid pathways as a mechanism for phenotypic plasticity in Antirrhinum majus. Scientific Reports 6: 19807.
Ferrario S, Shchennikova AV, Franken J, Immink RGH, Angenent GC (2006) Control of floral meristem determinacy in Petunia by MADS-Box transcription factors. Plant Physiology 140: 890-898.
Kamiya N, Nagasaki H, Morikami A, Sato Y, Matsuoka M (2003) Isolation and characterization of a rice WUSCHEL-type homeobox gene that is specifically expressed in the central cells of a quiescent center in the root apical meristem. Plant J 35 (4): 429–441. DOI: 10.1046/j.1365-313x.2003.01816.x.
Ramos LYS, Estrada TG, Dzib SN, Rodriguez LCZ, Castano E (2008) Overexpression of WUSCHEL in C. chinense causes ectopic morphogenesis. Plant Cell Tissue Organ Culture 96: 279–287.
Klimaszewska K, Overton C, Stewart D, Rutledge RG (2011) Initiation of somatic embryos and regeneration of plants from primordial shoots of 10-year-old somatic white spruce and expression profiles of 11 genes followed during the tissue culture process. Planta 233 (3): 635-47.
Herrera AA, Gonzalez KA, Moo CR, Figueroa FRQ, Vargas VML, Zapata LCR, DHondt CB, Solis VMS, Castano E (2008) Expression of WUSCHEL in Coffea canephora causes ectopic morphogenesis and increases somatic embryogenesis. Plant Cell Tissue Organ Culture 94: 171. https://doi.org/10.1007/s11240-008-9401-1
Hu L, Yang X, Yuan D, Zeng F, Zhang X (2011) GhHmgB3 deficiency deregulates proliferation and differentiation of cells during somatic embryogenesis in cotton. Plant Biotechnol J 9 (9): 1038-48.
Coussa OB, Obellianne M, Linderme D, Montes E, Grondard AM, Vilaine F, Pannetier C (2013) Wuschel overexpression promotes somatic embryogenesis and induces organogenesis in cotton (Gossypium hirsutum L.) tissues cultured in vitro. Plant cell reports 32: 675–686.
Zheng W, Zhang X, Yang Z, Wu J, Li F, Duan L, Liu C, Lu L, Zhang C, Li F (2014) AtWuschel promotes the formation of the embryogenic callus in Gossypium hirsutum. PLOS ONE 9: e87502.
Gaj MD, Zhang S, Harada JJ Lemaux PG (2005) Leafy cotyledon genes are essential for the induction of somatic embryogenesis of Arabidopsis. Planta 222 (6): 977-88.
Fambrini M, Durante C, Cionini G, Geri C, Giorgetti L, Michelotti V, Salvini M, Pugliesi C (2006) Characterization of LEAFY COTYLEDON1-LIKE gene in Helianthus annuus and its relationship with zygotic and somatic embryogenesis. Dev Genes Evol 216 (5): 253-64.
Hao Q, Zhang L, Yang Y, Shan Z, Zhou XA (2019) Genome-Wide Analysis of the WOX Gene Family and Function Exploration of GmWOX18 in Soybean. Plants (Basel). 8 (7): 215, Published 2019 Jul 11. DOI: 10.3390/plants8070215.
Lowe K, La Rota M, Hoerster G, et al. (2018) Rapid genotype "independent" Zea mays L. (maize) transformation via direct somatic embryogenesis. In Vitro Cell Dev Biol Plant. 54 (3): 240–252. DOI: 10.1007/s11627-018-9905-2.
Jones T, Lowe K, Hoerster G, Anand A, Wu E, Wang N, Arling M, Lenderts B, Gordon-Kamm W (2018) Maize transformation using the morphogenic genes Baby Boom and Wuschel2, in Sandeep Kumar et al., (eds) Transgenic Plants: Methods and Protocols, Methods in Molecular Biology, vol. 1864. https://doi.org/10.1007/978-1-4939-8778-8_6
Mookkan M, Nelson-Vasilchik K, Hague J, Zhang ZJ, Kausch AP (2017) Selectable marker independent transformation of recalcitrant maize inbred B73 and sorghum P898012 mediated by morphogenic regulators BABY BOOM and WUSCHEL2. Plant Cell Rep 36 (9): 1477–1491. DOI: 10.1007/s00299-017-2169-1.
Kyo M, Maida K, Nishioka Y, Matsui K (2018) Coexpression of WUSCHEL related homeobox (WOX)2 with WOX8 or WOX9 promotes regeneration from leaf segments and free cells in Nicotiana tabacum L. Plant Biotechnology 35: 23–30.
Vijayachandra K, Palanichelvam K, Veluthambi K (1995) Rice scutellum induces Agrobacterium tumefaciens vir genes and T-strand generation. Plant Molecular Biology 29: 125-133.
Sridevi G, Sabapathi N, Meena P, Nandakumar R, Samiyappan R, Muthukrishnan S, Veluthambi K (2003) Transgenic indica rice variety Pusa Basmati 1 constitutively expressing a rice chitinase gene exhibits enhanced resistance to Rhizoctonia solani. Journal of Plant Biochemistry and Biotechnology 12: 93–101.
Hiei Y, Ohta, S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant Journal 6: 271–282.
Rogers SO, Bendich AJ (1988) Extraction of DNA from plant tissues. In: Gelvin S. B., Schilperoort R. A., Verma D. P. S. (eds) Plant Molecular Biology Manual. Kluwer Academic Publishers, Dordrecht, USA: 1–10.
Brunck CF, Jones KC, James TW (1979) Assay for nanogram quantities of DNA in cellular homogenates. Analytical Biochemistry 92: 497–500.
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98: 503–517.
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15: 473–497.
Pedersen JF, Bean SR, Funell DL, Graybosch RA (2004) “Rapid Iodine Staining Techniques for Identifying the Waxy Phenotype in Sorghum Grain and Waxy Genotype in Sorghum Pollen.” Crop Science 44 (3): 764-767.
Umesh PP, Ansari MA, Sridevi G (2017). An efficient method for high-quality RNA extraction from Moringa oleifera. Journal of Plant Sciences 5 (2): 68-74.
Shin H, Braendle C, Monahan KB, Kaplan REW, Zand TP, Mote FS, et al. (2019) Developmental fidelity is imposed by genetically separable RalGEF activities that mediate opposing signals. PLoS Genet 15 (5): e1008056. https://doi.org/10.1371/journal.pgen.1008056.
Liu J, Zheng Q, Ma Q, Gadidasu KK, Zhang P (2011) Cassava genetic transformation and its application in breeding. J Integr Plant Biol 53: 552–569.
Ma QX, Zhou WZ, Zhang P (2015) Transition from somatic embryo to friable embryogenic callus in cassava: dynamic changes in cellular structure, physiological status, and gene expression profiles. Front Plant Sc, 6: 1–14.
Mordhorst AP, Voerman KJ, Hartog MV, Meijer EA, van Went J, Koornneef M, de Vries SC (1998) Somatic embryogenesis in Arabidopsis thaliana is facilitated by mutations in genes repressing meristematic cell divisions. Genetics 149: 549–563.
Stone SL, Braybrook SA, Paula SL, Kwong LW, Meuser J, et al. (2008) Arabidopsis LEAFY COTYLEDON2 induces maturation traits and auxin activity: implications for somatic embryogenesis. Proc. Natl. Acad. Sci 105: 3151–56.
Devic M, Roscoe T (2016) Seed maturation: simplification of control networks in plants. Plant Science 252: 335–346.
Honda E. Yew CL, Yoshikawa T, Sato Y, Hibara KI, Itoh JI (2018) LEAF LATERAL SYMMETRY1, a member of the WUSCHEL-RELATED HOMEOBOX3 gene family, regulates lateral organ development differentially from other paralogs. NARROW LEAF2 and NARROW LEAF3 in rice. Plant Cell Physiol 59 (2): 376–391.
Li H, Xu YY, Chong K, Wang H. (2004) Analysis of transgenic Tobacco with overexpression of Arabidopsis WUSCHEL gene. Acta Botanica Sinica 46: 224-229.
Wei XJ, Jiang L, Xu JF, Liu X, Liu SJ, Zhai HQ, Wan JM (2009) The distribution of japonica rice cultivars in the lower region of the Yangtze River valley is determined by its photoperiod-sensitivity and heading date genotypes. Journal of Integrative Plant Biology 51: 922-32.
Park SJ, Kim SL, Lee S, Je BI, Piao HL, Park SH, Kim CM, Ryu CH, Park SH, Xuan YH, Colasanti J, An G, Han C. D. (2008) Rice Indeterminate 1 (OsId1) is necessary for the expression of Ehd1 (Early heading date 1) regardless of photoperiod. The Plant Journal 56: 1018–1029.
Zhao J, Chen H, Ren D, Tang H, Qiu R, Feng J, Long Y, Niu B, Chen D, Zhong T, Liu YG, Guo J (2015) Genetic interactions between diverged alleles of Early heading date 1 (Ehd1) and Heading date 3a (Hd3a)/ RICE FLOWERING LOCUS T1 (RFT1) control differential heading and contribute to regional adaptation in rice (Oryza sativa). New Phytologist 208: 936–948.
Xue W, Xing YZ, Weng X, Zhao Y, Tang W, Wang L, Zhou H, Yu S, Xu C, Li X, Zhang Q (2008) Natural variation in Ghd7 is an important regulator of heading date and yield potential in rice. Nature Genetics 40: 761-767.
Kobayashi M, Yamaguchi I, Murofushi N, Ota Y, Takahashi N (1988) Fluctuation and localization of endogenous gibberellins in rice. Agricultural Biological Chemistry 52: 1189–1194.
Yin C, Gan L, Ng D, Zhou X, Xia K (2007) Decreased panicle-derived indole-3-acetic acid reduces gibberellin A1 level in the uppermost internode, causing panicle enclosure in male-sterile rice Zhenshan 97A. Journal of Experimental Botany 58 (10): 2441–2449. DOI 10.1093/jxb/erm077.
Zhu L, Hu J, Zhu KM, Fang YX, Gao ZY, He YH, Zhang GH, Guo LB, Zeng DL, Dong GJ, Yan MX, Liu J, Qian Q (2011) Identification and characterization of SHORTENED UPPERMOST INTERNODE 1, a gene negatively regulating uppermost internode elongation in rice. Plant Mol Biol 77: 475–487.
Takahashi M, Nagasawa NS, Kitano H, Nagato Y (1998) panicle phytomer 1 mutation affects the panicle architecture of rice. Theoretical and Applied Genetics 96: 1050-1056.
Lu H, Dai Z, Li L, Wang J, Miao X, Shi Z (2017) OsRAMOSA2 shapes panicle architecture through regulating pedicel length. Frontiers in Plant Science 8: 1538.
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