Protocol: high-efficiency in-planta Agrobacterium-mediated transgenic hairy root induction of Camellia sinensis var. sinensis
Tóm tắt
Camellia sinensis var. sinensis is widely grown for tea beverages that possess significant health promoting effects. Studies on tea plant genetics and breeding are hindered due to its recalcitrance to Agrobacterium-mediated genetic transformation. Among the possible reasons, oxidation of phenolics released from explant tissues and bactericidal effects of tea polyphenols during the process of transformation play a role in the plant recalcitrance. The aim of the present study was to alleviate the harmful effects of phenolic compounds using in-planta transformation. Two-month old seedlings of tea cultivar “Nong Kangzao” were infected at the hypocotyl with wild type Agrobacterium rhizogenes and maintained in an environment of high humidity. 88.3% of infected plants developed hairy roots at the wounded site after 2 months of infection. Our data indicated that transgenic hairy root induction of tea can be achieved using A. rhizogenes following the optimized protocol. With this method, composite tea plants containing wild-type shoots with transgenic roots can be generated for “in root” gene functional characterization and root-shoot interaction studies. Moreover, this method can be applied to improve the root system of composite tea plants for a better resistance to abiotic and biotic stresses.
Tài liệu tham khảo
Food and Agriculture Organization of the United Nations-Production FAOSTAT. http://faostat.fao.org/DesktopDefault.aspx?pageID=567&lang=en#ancor. Accessed 25 July 2012.
http://www.statisticbrain.com/tea-drinking-statistics/. Accessed 30 July 2012.
Balentine DA, Wiseman SA, Bouwens LCM. The chemistry of tea flavonoids. Crit Rev Food Sci Nutr. 1997;37:693–704.
Yokogoshi H, Kato Y, Sagesaka YM, Matsuura TT, Kakuda T, Takeuchi N. Reduction effect of theanine on blood pressure and brain 5-hydroxyindoles in spontaneously hypertensive rats. Biosci Biotechnol Biochem. 1995;59:615–8.
Yokozawa T, Dong EB. Influence of green tea and its three major components upon low-density lipoprotein oxidation. Exp Toxicol Pathol. 1997;49:329–35.
Juneja LR, Chu DC, Okubo T, Nagato Y, Yokogoshi H. l-theanine—a unique amino acid of green tea and its relaxation effect in humans. Trends Food Sci Technol. 1999;10:199–204.
Kakuda T, Nozawa A, Unno T, Okamura N, Okai O. Inhibiting effects of theanine on caffeine stimulation evaluated by EEG in the rat. Biosci Biotechnol Biochem. 2000;64:287–93.
Kim KS, Song CH, Oh HJ. Effects of theanine on the release of brain alpha-wave in adult males. FASEB J. 2004;18:541–2.
Scalbert A, Manach C, Morand C, Rémésy C. Dietary polyphenols and the prevention of diseases. Crit Rev Food Sci Nutr. 2005;45:287–306.
Staszewski MV, Pilosof AMR, Jagus RJ. Antioxidant and antimicrobial performance of different argentinean green tea varieties as affected by whey proteins. Food Chem. 2011;125:186–92.
Taylor PW, Stapleton PD, Luzio JP. New ways to treat bacterial infections. Drug Discov Today. 2002;7:1086–91.
Friedman M. Overview of antibacterial, antitoxin, antiviral, and antifungal activities of tea flavonoids and teas. Mol Nutr Food Res. 2007;51:116–34.
Almajano MP, Carbó R, Jiménez JAL, Gordon MH. Antioxidant and antimicrobial activities of tea infusions. Food Chem. 2008;108:55–63.
Graham HN. Green tea composition, consumption, and polyphenol chemistry. Prev Med. 1992;21:334–50.
Shimamura T, Zhao WH, Hu ZQ. Mechanism of action and potential for use of tea catechin as an anti-infective agent. Anti-Infect Agents Med Chem. 2007;6:57–62.
Rus AM, Bressan RA, Hasegawa PM. Unraveling salt tolerance in crops. Nat Genet. 2005;37:1029–30.
Beritognolo I, Harfouche A, et al. Comparative study of transcriptional and physiological responses to salinity stress in two contrasting Populus alba L. genotypes. Tree Physiol. 2011;31:1335–55.
Mondal TK, Bhattacharya A, Ahuja PS, Chand PK. Transgenic tea (Camellia sinensis (L.) O. Kuntze cv. Kangra Jat) plants obtained by Agrobacterium-mediated transformation of somatic embryos. Plant Cell Rep. 2001;20:712–20.
Lopez SJ, Kumar RR, Pius PK, Muraleedharan N. Agrobacterium tumefaciens-mediated genetic transformation in tea (Camellia sinensis (L.) O. Kuntze). Plant Mol Biol Rep. 2004;22:201–2.
Sandal I, Saini U, Lacroix B, Bhattacharya A, Ahuja PS, Citovsky V. Agrobacterium-mediated genetic transformation of tea leaf explants: effects of counteracting bactericidity of leaf polyphenols without loss of bacterial virulence. Plant Cell Rep. 2007;26:169–76.
Yam TS, Shah S, Hamilton-Miller JM. Microbiological activity of whole and fractionated crude extracts of tea (Camellia sinensis), and of tea components. FEMS Microbiol Lett. 1997;152:169–74.
Ru Z, Lai Y, Xu C, Li L. Polyphenol oxidase (PPO) in early stage of browning of Phalaenopsis leaf explants. J Agric Sci. 2013;5:57–64.
Song DP, Feng L, Rana MM, Gao MJ, Wei S. Effects of catechins on Agrobacterium- mediated genetic transformation of Camellia sinensis. Plant Cell, Tissue Organ Cult. 2014;119:27–37.
Naz S, Ali A, Iqbal J. Phenolic content in vitro cultures of chick pea (Cicer arietinum L.) during callogenesis and organogenesis. Pak J Bot. 2008;40:2525–39.
Ngomuo M, Mneney E, Ndakidemi P. Control of lethal browning by using ascorbic acid on shoot tip cultures of a local Musa spp. (Banana) cv. Mzuzu in Tanzania. Afr J Biotechnol. 2014;13:1721–5.
Rana MM, Han ZH, Song DP, Liu GF, Li DX, Wan XC, Karthikeyan A, Wei S. Effect of medium supplements on Agrobacterium rhizogenes mediated hairy root induction from the callus tissues of Camellia sinensis var. Sinensis. Int J Mol Sci. 2016;17:1132–49.
Ali M, Kiani BH, Mannan A, Ismail T, Mirza B. Enhanced production of artemisinin by hairy root cultures of Artemisia dubia. J Med Plant Res. 2012;6:1619–22.
Sujatha G, Zdravković-Korać S, Ćalić D, Flamini G, Ranjitha Kumaria BD. High-efficiency Agrobacterium rhizogenes-mediated genetic transformation in Artemisia vulgaris: Hairy root production and essential oil analysis. Ind Crops Prod. 2013;44:643–52.
Christey MC. Use of Ri-mediated transformation for production of transgenic plants. In Vitro Cell Dev Biol Plant. 2001;37:687–700.
Georgiev MI, Agostini E, Ludwig-Muller J, Xu J. Genetically transformed roots: from plant disease to biotechnological resource. Trends Biotechnol. 2012;30:528–37.
Guillon S, Tremouillaux-Guiller J, Pati PK, Rideau M, Gantet P. Hairy root research: recent scenario and exciting prospects. Curr Opin Plant Biol. 2006;9:341–6.
Taylor CG, Fuchs B, Collier R, Lutke WK. Generation of composite plants using Agrobacterium rhizogenes. Methods Mol Biol. 2006;343:155–67.
Chilton MD, et al. Agrobacterium rhizogenes inserts T-DNA into the genome of the host plant root cells. Nature. 1982;295:432–4.
Nagel R, Elliott A, Masel A, Birch RG, Manners JM. Electroporation of binary Ti plasmid vector into Agrobacterium tumefaciens and Agrobacterium rhizogenes. FEMS Microbiol Lett. 1990;67:325–8.
Jefferson RA. Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep. 1987;5:387–405.
Collier R, Fuchs B, Walter N, Kevin Lutke W, Taylor CG. Ex vitro composite plants: an inexpensive, rapid method for root biology. Plant J. 2005;43:449–57.
Colpaert N. Composite Phaseolus vulgaris plants with transgenic roots as research tool. Afr J Biotechnol. 2008;7:404–8.
Clemow SR, Clairmont L, Madsen LH, Guinel FC. Reproducible hairy root transformation and spot-inoculation methods to study root symbioses of pea. Plant Methods. 2011;7:46.
Estrada-Navarrete G, et al. Agrobacterium rhizogenes transformation of the Phaseolus spp.: a tool for functional genomics. Mol Plant-Microbe Interact. 2006;19:1385–93.
Vieweg MF, et al. The promoter of the Vicia faba L. gene VfLb29 is specifically activated in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots from different legume and non-legume plants. Mol Plant-Microbe Interact. 2004;17:62–9.