Abstract
Tea is a widely consumed nonalcoholic beverage, made from tender tea leaves, which contain extraordinarily high amount of specialized metabolites. These specialized metabolites are beneficial to human health and crucial in tea brisk, umami taste, and pleasant scent. Agrobacterium-assisted genetic transformation studies on tea plant are largely limited due to recalcitrant nature of tea plant. Release of oxidation phenolics that are dominant from explant tissues and the antibacterial nature of polyphenols during the transformation are among the possible reasons for tea plant recalcitrance. Our main objective in this study was to create an in vitro system for selection of transgenic roots from composite plants for possible production of specialized metabolites. Three-week-old in vitro-germinated seedlings were used in generating composite plants. The hypocotyl and the stems were wounded using a sonicator and a sterilized needle and infected with Agrobacterium rhizogenes harboring pBI121 plasmid and maintained in aseptic conditions. Molecular assay of roots from antibiotic selected plants showed amplification of rol B C and gus confirming the integration of Agrobacterium rhizogenes T-DNA region into the root system. From our data, it was clear that hairy root generation in tea can be achieved using A. rhizogenes. With this method, composite tea plants containing wild-type shoots with transgenic roots can be developed for gene functional characterization as well as specialized metabolite production and root-shoot interaction research in plants.
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References
Ahmed S, Orians CM, Griffin TS et al (2014) Effects of water availability and pest pressures on tea (Camellia sinensis) growth and functional quality. AoB Plants. https://doi.org/10.1093/aobpla/plt054
Alagarsamy K, Shamala LF, Wei S (2018) Protocol: high-efficiency in-planta Agrobacterium-mediated transgenic hairy root induction of Camellia sinensis var sinensis. Plant Methods:14. https://doi.org/10.1186/s13007-018-0285-8
Almajano MP, Carbó R, Jiménez JAL, Gordon MH (2008) Antioxidant and antimicrobial activities of tea infusions. Food Chem 108:55–63. https://doi.org/10.1016/j.foodchem.2007.10.040
Alpizar E, Dechamp E, Espeout S et al (2006) Efficient production of Agrobacterium rhizogenes-transformed roots and composite plants for studying gene expression in coffee roots. Plant Cell Rep 25:959–967
Balentine DA, Wiseman SA, Bouwens LCM (1997) The chemistry of tea flavonoids. Crit Rev Food Sci Nutr 37:693–704. https://doi.org/10.1080/10408399709527797
Banerjee B (1992) Botanical classification of tea. In: Tea. Springer, Dordrecht, pp 25–51
Beritognolo I, Harfouche A, Brilli F et al (2011) Comparative study of transcriptional and physiological responses to salinity stress in two contrasting Populus alba L. genotypes. Tree Physiol 31:1335–1355. https://doi.org/10.1093/treephys/tpr083
Borzelleca JF, Peters D, Hall W (2006) A 13-week dietary toxicity and toxicokinetic study with l-theanine in rats. Food Chem Toxicol. https://doi.org/10.1016/j.fct.2006.03.014
Chacko SM, Thambi PT, Kuttan R, Nishigaki I (2010) Beneficial effects of green tea: a literature review. Chin Med 5:13. https://doi.org/10.1186/1749-8546-5-13
Chen K, Wang Y, Zhang R et al (2019) CRISPR/Cas genome editing and precision plant breeding in agriculture. Annu Rev Plant Biol. https://doi.org/10.1146/annurev-arplant-050718-100049
Chilton M-D, Tepfer DA, Petit A et al (1982) Agrobacterium rhizogenes inserts T-DNA into the genomes of the host plant root cells. Nature 295:432–434. https://doi.org/10.1038/295432a0
Da Silva PM (2013) Tea: a new perspective on health benefits. Food Res Int
Ding Y, Kalo P, Yendrek C et al (2008) Abscisic acid coordinates nod factor and cytokinin signaling during the regulation of nodulation in Medicago truncatula. Plant Cell Online 20:2681–2695. https://doi.org/10.1105/tpc.108.061739
Graham HHN (1992) Green tea composition, consumption, and polyphenol chemistry. Prev Med (Baltim) 21:334–350. https://doi.org/10.1016/0091-7435(92)90041-F
Han ZX, Rana MM, Liu GF et al (2016) Green tea flavour determinants and their changes over manufacturing processes. Food Chem 212:739–748
Hansen J, Jorgensen J-E, Stougaard J, Marcker KA (1989) Hairy roots ? A short cut to transgenic root nodules. Plant Cell Rep 8:12–15. https://doi.org/10.1007/BF00735768
Jefferson RA (1989) The GUS reporter gene system. Nature 342:837–838. https://doi.org/10.1038/342837a0
Juneja L (1999) L-theanine—a unique amino acid of green tea and its relaxation effect in humans. Trends Food Sci Technol 10:199–204. https://doi.org/10.1016/S0924-2244(99)00044-8
Kakuda T, Nozawa A, Unno T et al (2000) Inhibiting effects of theanine on caffeine stimulation evaluated by EEG in the rat. Biosci Biotechnol Biochem 64:287–293. https://doi.org/10.1271/bbb.64.287
Koster H, Quandt H, Broer I et al (1995) The promoter of the Vicia faba L. VfENOD-GRP3 gene encoding a glycine-rich early nodulin mediates a predominant gene expression in the interzone II-III region of transgenic Vicia hirsuta root nodules. Plant Mol Biol 29:759–772. https://doi.org/10.1007/BF00041166
Kumazawa K (2006) Flavor chemistry of tea and coffee drinks. Food Sci Technol Res 12:71–84
Liu C, Xiang L, Zheng X (2015) Health benefits of theanine in green tea: a review 14:1943–1949
Lobell DB, Schlenker W, Costa-Roberts J (2011) Climate trends and global crop production since 1980. Science 80. https://doi.org/10.1126/science.1204531
Mohanpuria P, Kumar V, Ahuja PS, Yadav SK (2011) Agrobacterium-mediated silencing of caffeine synthesis through root transformation in Camellia sinensis L. Mol Biotechnol 48:235–243. https://doi.org/10.1007/s12033-010-9364-4
Mondal TK, Bhattacharya A, Sood A, Ahuja PS (1999) An efficient protocol for somatic embryogenesis and its use in developing transgenic tea (Camellia sinensis (L.) O. Kuntze) for field transfer. Curr Plant Sci Biotechnol Agric 36:181–184
Mondal TK, Bhattacharya A, Laxmikumaran M, Ahuja PS (2004) Recent advances of tea ( Camellia sinensis ) biotechnology. Plant Cell Tissue Organ Cult 76:195–254
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Plasencia A, Dupas A, Ladouce N et al (2016) Eucalyptus hairy roots, a fast, efficient and versatile tool to explore function and expression of genes involved in wood formation. Plant Biotechnol J 14:1381–1393. https://doi.org/10.1111/pbi.12502
Prabhu SA, Ndlovu B, Engelbrecht J, Van Den Berg N (2017) Generation of composite Persea americana (Mill.) (avocado) plants: a proof-of-concept-study. PLoS One 12:1–22. https://doi.org/10.1371/journal.pone.0185896
Rus AM, Bressan RA, Hasegawa PM (2005) Unraveling salt tolerance in crops. Nat Genet 37:1029–1030. https://doi.org/10.1038/ng1005-1029
Sandal I, Saini U, Lacroix B et al (2007) Agrobacterium-mediated genetic transformation of tea leaf explants: effects of counteracting bactericidity of leaf polyphenols without loss of bacterial virulence. Plant Cell Rep 26:169–176. https://doi.org/10.1007/s00299-006-0211-9
Scalbert A, Manach C, Morand C et al (2005) Dietary polyphenols and the prevention of diseases. Crit Rev Food Sci Nutr 45:287–306. https://doi.org/10.1080/1040869059096
Singh HR, Hazarika P (2020) Biotechnological approaches for tea improvement. Elsevier Inc. https://doi.org/10.1016/b978-0-12-816678-9.00004-7
Song CH, Jung JH, Oh JS, Kim KS (2003) Effects of theanine on the release of brain alpha wave in adult males. Korean J Nutr 36:918–923Â
Suzuki T, Miyoshi N, Hayakawa S et al (2016) Health benefits of tea consumption. In: Beverage impacts on health and nutrition. Humana Press, Cham
Taylor PW, Stapleton PD, Paul Luzio J (2002) New ways to treat bacterial infections. Drug Discov Today 7:1086–1091. https://doi.org/10.1016/S1359-6446(02)02498-4
Veena V, Taylor CG (2007) Agrobacterium rhizogenes: recent developments and promising applications. In Vitro Cell Dev Biol Plant 43:383–403
Villarreal D, Laffargue A, Posada H et al (2009) Genotypic and environmental effects on coffee (Coffea Arabica L.) bean fatty acid profile: impact on variety and origin chemometric determination. J Agric Food Chem. https://doi.org/10.1021/jf902441n
von Staszewski M, Pilosof AMR, Jagus RJ (2011) Antioxidant and antimicrobial performance of different Argentinean green tea varieties as affected by whey proteins. Food Chem 125:186–192. https://doi.org/10.1016/j.foodchem.2010.08.059
Webster CC (1978) Tea. By T. Eden (3rd edn). Longman Group, London (1976), pp 236, £12.00. Exp Agric 14:179. https://doi.org/10.1017/S0014479700008620
Wei C, Yang H, Wang S et al (2018) Draft genome sequence of Camellia sinensis var. sinensis provides insights into the evolution of the tea genome and tea quality. https://doi.org/10.1073/pnas.1719622115
Wollenberg E, Vermeulen SJ, Girvetz E et al (2016) Reducing risks to food security from climate change. Glob Food Sec 11:34–43
Xia EH, Bin ZH, Sheng J et al (2017) The tea tree genome provides insights into tea flavor and independent evolution of caffeine biosynthesis. Mol Plant 10:866–877. https://doi.org/10.1016/j.molp.2017.04.002
Xia EH, Tong W, Wu Q et al (2020) Tea plant genomics: achievements, challenges and perspectives. Hortic Res 7. https://doi.org/10.1038/s41438-019-0225-4
Zhu H (2008) The Tropical Flora of Southern Yunnan, China, and Its Biogeographic Affinities. Ann Mo Bot Gard 95(4): 661–680. https://doi.org/10.3417/2006081
Acknowledgments
We thank all our lab members Jing-Yi Pang, Dong-Wei Zhao, Zhen Yan, and Meng-Xian Zhang for their technical support.
Funding
This work supported by the National Natural Science Foundation of China (Grant Number 31370687 and 31770734) and funding was provided by Research Fund for the Doctoral Program of Higher Education (Grant No. 20123418110002) to Shu Wei.
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Shamala, L.F., Wei, S. (2020). An Improved In Vitro Protocol for Agrobacterium rhizogenes-Mediated Transformation of Recalcitrant Plants for Root Biology Studies: A Case Study of Tea Plants (Camellia sinensis var. sinensis). In: Srivastava, V., Mehrotra, S., Mishra, S. (eds) Hairy Root Cultures Based Applications. Rhizosphere Biology. Springer, Singapore. https://doi.org/10.1007/978-981-15-4055-4_12
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