Journal of Plant Molecular Breeding

Regeneration of pinwheel phenotype and evaluation of anthocyanin in African violet (Saintpaulia ionantha Wendl.) periclinal chimera

Document Type : Original research paper

Authors

1 Department of Plant Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran

2 Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari Agricultural Sciences and Natural Resources University, Sari, Iran

3 Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran

Abstract

The mutation in the meristem layers creates different genetic backgrounds (chimera) in the plant tissue. The mutation in L1 layer of shoot apical meristem generates a periclinal chimera. UF3GT is an effective enzyme in floral coloration, inducing anthocyanin accumulation in petals. This study investigates direct and indirect regeneration systems and different explants to propagate two cultivars of periclinal chimera (Saintpaulia ionantha), namely Taro taraneh and Aghaz, using in vitro culture. The evaluation of UF3GT gene expression pattern by Real-Time PCR revealed the role of anthocyanin accumulation in the petal coloration of chimera plants. Results pertaining to both cultivars showed that inflorescence and leaf explant had the highest and lowest percentage of pinwheel phenotype, respectively. In addition, mutant characteristics were faded in the leaf regeneration of periclinal chimera. Furthermore, the highest percentage of periclinal chimera was generated in direct regeneration. Gene expression analysis revealed that UF3GT was expressed in the colorful part of chimera petal, while UF3GT expression was significantly reduced in the muted part. HPLC chromatogram also detected that cyanidin and delphinidin components were not present in the white part of either cultivar. The anthocyanin biosynthesis pathway appears to be blocked and anthocyanin accumulation does not occur in the petals. Inflorescence is likely to induce a pinwheel pattern in regenerated plants, probably owing to its lateral bud. It seems that different meristem layers are associated with the formation of epidermis and induce pinwheel phenotype.

Keywords

[1] Azadi, P., Bagheri, H., Nalousi, AM., Nazari, F., and Chandler, S.F. 2016. Current status and biotechnological advances in genetic engineering of ornamental plants. Biotechnol Adv, 34(6): 1073-1090.
[2] Becraft, P.W. 2013. Using transposons for genetic mosaic analysis of plant development. Methods Mol Biol, 1057:21-42. https://doi.org/10.1007/ 978-1-62703-568-2_3
[3] Calderwood, A., Kopriva, S., and Morris, R.J. 2016. Transcript abundance explains RNA mobility data in Arabidopsis thaliana. Plant Cell, 28:610–615.
[4] Chen, WH., Hsu, ChY., Cheng, HY., Chang, H., Chen, HH., and Ger, MJ. 2011. Down regulation of putative UDP-glucose: flavonoid 3-O-glucosyltransferase gene alters flower coloring in Phalaenopsis. Plant Cell Rep, 30:1007–1017.
[5] Falcone Ferreyra, ML., Rius, SP., and Casati, P.2012. Flavonoids: biosynthesis, biological functions, and biotechnological applications. Front Plant Sci, 3:222.
[6] Filippis, I., Lopez-Cobollo, R., Abbott, J., Butcher, S., and Bishop, GJ.2013. Using a periclinal chimera to unravel layer-specific gene expression in plants. Plant J, 75(6):1039-49.
[7] Frank, M.H., and Chitwood, D.H. 2016. Plant chimeras: The good, the bad, and the Bizzaria. Dev Biol, 419: 41–53.
[8] Ghalecahi, B., Aslanpour, M., Shoor, M., Sharifi, A., Kharaz, M. 2018. Effect of light variables treatments on growth and flowering of saintpaulia (Saintpaulia ionantha). ITJEMAST, 9(6): 597-609.
[9] Gordienko, N.Y. 2002. Clonal micropropagation with preservation of decorative characters of Saintpaulia cultivars. Byulleten’ Gosudarstvewnnogo Nikitskogo Botanicheskogo Sada, 73(86): 37-38.
[10] Harborne, JB., and Williams, CA. 2001. Anthocyanins and other flavonoids. Nat Prod Rep, 18:310–333.
[11] Hartonen, K., Parshintsev, J., Sandberg, K., Bergelin, E., Nisula, L., Riekkola, ML. 2007. Isolation of flavonoids from aspen knotwood by pressurized hot water extraction and comparison with other extraction techniques. Talanta, 74: 32–38.
[12] Jones, P., and Vogt, T. 2001. Glycosyltransferases in secondary plant metabolism: tranquilizers and stimulant controllers. Planta, 3(2):164–174.
[13] Jung, S., Venkatesh, J., Kang, M.Y., Kwon, J.K., and Kang, B.Ch. 2019. A non-LTR retrotransposon activates anthocyanin biosynthesis by regulating a MYB transcription factor in Capsicum annuum. Plant Sci, 287:1-10.
[14] Kobayashi, S., Ishimaru, M., Hiraoka, K., and Honda, C. 2002. Myb-related genes of the Kyoho grape (Vitis labruscana) regulate anthocyanin biosynthesis. Planta, 215:924–933.
[15] Kolehmainen, J., Korpelainen, H., and Mutikainen, P. 2010. Inbreeding and inbreeding depression in a threathened endemic plant, the African violet (Saintpaulia ionantha ssp. grotei), of the East Usambara Mountains, Tanzania. Afr J Ecol, 48: 576-587.
[16] Krishna, H., Alizadeh, M., Singh, D., Singh, U., Chauhan, N., Eftekhari, M., and Sadh, R.H .2016. Somaclonal variations and their applications in horticultural crops improvement. Biotech, 6(1): 54-59.
[17] Lee, H., Jun, Y.S., Cha, O.K., and Sheen, J. 2019. Mitogen-activated protein kinases MPK3 and MPK6 are required for stem cell maintenance in the Arabidopsis shoot apical meristem. Plant Cell Rep, 38(3):311–319.
[18] Liobikas, J., Skemiene, K., Trumbeckaite, S., and Borutaite, V. 2016. Anthocyanins in cardioprotection: a path through mitochondria. Pharmacol Res, 113: 808-815.
[19] Lo Piero, AR., Consoli, A., Puglisi, I., Orestano, G., Recupero, GR., and Petrone, G. 2005. Anthocyaninless cultivars of sweet orange lack to express the UDP-glucose flavonoid 3-O-glucosyl transferase. J Plant Biochem Biotechnol, 14:9–14.
[20] Mercuri, A., De Benedetti, L., and Burchi, G. 2000. Agrobacterium-mediated transformation of African violet. PCTOC, 60: 39.
[21] Murashige, T., and Skoog, F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant, 15: 473–497.
[22] Nabeshima, T., Yang, S.J., Ohno, S., Honda, K., Deguch, A., Doi, M., Tatsuzawa, F., and Hosokawa, M. 2017. Histogen Layers Contributing to Adventitious Bud Formation Are Determined by their Cell Division Activities. Front Plant Sci, 8:1749.
[23] Nishihara, M., and Nakatsuka, T .2011. Genetic engineering of flavonoid pigments to modify flower color in floricultural plants. Biotech letters, 33(3): 433-441.
[24] Norris, R., and Smith, R.H .1981. Regeneration of variegated African violet (Saintpaulia ionantha WENDL.) leaf chimeras in culture. Plant Physiol, 67: 117.
[25] Peary, J., Lineberger, R., Malinich, T., and Wertz, M. 1988. Stability of leaf variegation of Saintpaulia ionantha during in vitro propagation and during chimeral separation of a pinwheel flowering form. Amer J Bot, 75(51): 603-608.
[26] Pierik, R.L.M., and Steegmans, H.H.M. 1983. Vegetative propagation of a chimerical Yucca elephantipes Regel in vitro. Sci Hort, 21: 267-272.
[27] Pourcel, L., Irani, NG., Koo, AJ., Bohorquez-Restrepo, A., Howe, GA., and Grotewold, E. 2013. A chemical complementation approach reveals genes and interactions of flavonoids with other pathways. Plant J, 74: 383–397.
[28] Qi, Y., Lou, Q., Li, H., Yue, J., Liu, Y., and Wang, Y. 2013. Anatomical and biochemical studies of bicolored flower development in Muscari latifolium. Protoplasma, 250: 1273–1281. [29] Sato, M., Kawabe, T., Hosokawa, M., Tatsuzawa, F., and Doi, M. 2011. Tissue culture-induced flower-color changes in Saintpaulia caused by excision of the transposon inserted in the flavonoid 3', 5' hydroxylase (F3'5'H) promoter. Plant Cell Rep, 30:929–939.
[30] Shajiee, K., Naderi, R., and Khalighi, A. 2006. Evaluation of Leaf Patterns of Chlorophyll Deficiency Findings from Somaclonal Variations in African violet. Iranian J agri sci, 37(1):54-61.
[31] Smith, R.H, and Norris, R. E. 1983. In vitro propagation of African violet chimeras. Hort Science 18(4): 436- 437.
[32] Stephan, L., Tilmes, V., and Hulskamp, M. 2019. Selection and validation of reference genes for quantitative Real-Time PCR in Arabis alpina. PLoS ONE 14(3): e0211172.
[33] Sun, W., Meng, X., Liang, L., Li, Y., Zhou, T., Cai, X., Wang, L., and Gao, X. 2017. Overexpression of a Freesia hybrida flavonoid 3-O-glycosyltransferase gene, Fh3GT1, enhances transcription of key anthocyanin genes and accumulation of anthocyanin and flavonol in transgenic petunia (Petunia hybrida). In Vitro Cell Dev Biol Plant, 53(5): 478–488.
[34] Sun, Y.J., Chen, X.Q., Sun, N., Chen, J., and Zhang, L. 2010. Karyotype analysis of Saintpaulia ionantha. J. Tianjin Agri Univ. 17: 5-8.
[35] Sunpui, W., and Kanchanapoom, K. 2002. Plant regeneration from petiole and leaf of African violet (Saintpaulia ionantha Wendl.) cultured in vitro. Songklanakarin J Sci Technol, 24(3): 357-364.
[36] Szymkowiak, E.J, and Sussex, I.M. 1996. What chimeras can tell us about plant development. Annu Rev Plant Physiol Plant Mol Biol 47: 351–376. http://dx.doi. org/10.1146/annurev.arplant.47.1.351
[37] Teixeira da Silva, J.A., Dewir, Y., Wicaksono, A., Kher, M., Kim, H., Hosokawa, M., and Zeng, S. 2016. Morphogenesis and developmental biology of African Violet (Saintpaulia ionantha H. WENDL.). J Plant Develop. 23: 15-25.
[38] Warschefsky, E.J., Klein, L.L., Frank, M.H., Chitwood, D.H., Londo, J.P., vonWettberg, E.J.B., and Miller, A.J. 2016. Rootstocks: diversity, domestication, and impacts on shoot phenotypes. Trends Plant Sci, 21: 418–437.
[39] Wei, S., Lingj, L., Xiangyu, M., Yueqing, L., Fengzhan, G., Xingxue, L., Shucai, W., Xiang, G., and Li, W. 2016. Biochemical and molecular characterization of a flavonoid 3-O-glycosyltransferase responsible for anthocyanins and flavonols biosynthesis in Freesia hybrida. Front Plant Sci. 7:410.
[40] Wongpiyasatid, A., Jompuk, P., Chusreeaeom, K., and Taychasinpitak, T. 2007. Effects of chronic gamma irradiation on adventitious plantlet formation of Saintpaulia ionantha (African violet) detached leaves. Kasetsart J Nat Sci, 41: 414-419.
[41] Yang, S.J., Ohno, Sh., Deguchi, A., Sato, M., Goto, M., and Doi, M. 2017. The histological study in sympetalous corolla development of pinwheel-type flowers of Saintpaulia. Sci Hortic, 223:10–18.
[42] Yoshida, K., Mori, M., and Kondo, T. 2009. Blue flower color development by anthocyanins: from chemical structure to cell physiology. Nat Prod Rep 26(7):884-915.
[43] Yunqing, Ch., Jianfeng, L., Xue, Zh., Zhiwen, Ch. 2010. Tissue Culture and Regeneration System from Leaves of Saintpaulia ionantha In Vitro. Chinese Agri Sci Bulletin, 10:1-12.
[44] Yuzhou, G., Qingbo, L., Jieqiong, L., Ming, X., and Binghui L. 2013. Callus Induction and Plant Regeneration of Saintpaulia ionantha. J Agric Eng 3:1-12.
[45] Zhou, J., Hirata, Y., Nou, I.S., Shiotani, H., and Ito, T. 2002. Interactions between different genotypic tissues in citrus graft chimeras. Euphytica1 26: 355–364.
Journal of Plant Molecular Breeding
Volume 7, Issue 2
December 2021
Pages 39-49
  • Receive Date: 09 November 2020
  • Revise Date: 15 May 2022
  • Accept Date: 16 March 2021
  • First Publish Date: 16 March 2021