Abstract
In general, stress caused by high light, high temperature and UV irradiation is unified and leads to the production of reactive oxygen species (ROS) having detrimental consequences for plants. Carotenoids are present in all photosynthetic plants as accessory pigments and have been implicated in protection against photooxidative damage. One major enzyme in the carotenoid biosynthetic pathway, β-carotene hydroxylase has been maneuvered for increasing stress tolerance of plants. Overexpression of β-carotene hydroxylase1 (BCH1) in Morus indica cv. K2 under the influence of CaMV 35S promoter was undertaken by Agrobacterium-mediated transformation. This gene when over expressed shows higher levels of carotenoids and improved oxidative stress tolerance as compared with the untransformed wild type under non-stressed and stressed conditions. The carotenoid and chlorophyll content was found to be enhanced in the overexpression lines under different stresses. The amount of ROS was found to be significantly lower in the overexpression line. Also, a corresponding increase in membrane stability suggests a positive role of the over expression of BCH1 on maintaining membrane integrity under induced oxidative stress conditions. Enhanced tolerance to high light, heat and UV irradiation was achieved in M. indica cv. K2 indicating the potential of this gene to suit the changing climatic conditions.
Similar content being viewed by others
References
Baroli I, Niyogi KK (2000) Molecular genetics of xanthophyll-dependent photoprotection in green algae and plants. Philos Trans R Soc Lond B Biol Sci 355:1385–1394
Bhatnagar S, Khurana P (2003) Agrobacterium tumefaciens-mediated transformation of Indian mulberry, Morus indica cv. K2: a time-phased screening strategy. Plant Cell Rep 21:669–675
Campos JL, Figueras X, Pinol MT, Boronat A, Tiburcio F (1991) Carotenoid and conjugated polyamine levels as indicators of ultraviolet-C induced stress in Arabidopsis thaliana. Photochem Photobiol 53:689–693
Chaitanya KV, Sundar D, Reddy AR (2001) Mulberry leaf metabolism under high temperature stress. Biol Plant 44:379–384
Chaudhary N, Nijhawan A, Khurana JP, Khurana P (2010) Carotenoid biosynthesis genes in rice: structural analysis, genome-wide expression profiling and phylogenetic analysis. Mol Genet Genomics 283:13–33
Checker VG, Chhibbar AK, Khurana P (2011) Stress-inducible expression of barley Hva1 gene in transgenic mulberry displays enhanced tolerance against drought, salinity and cold stress. Transgenic Res 21:939–957
Checker VG, Saeed B, Khurana P (2012) Analysis of expressed sequence tags from mulberry (Morus indica) roots and implications for comparative transcriptomics and marker identification. Tree Genet Gen 8:1437–1450
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal Biochem 162:156–159
D’Ambrosio C, Stigliani AL, Giorio G (2011) Overexpression of CrtR-b2 (carotene beta hydroxylase2) from S. lycopersicum L. differentially affects xanthophyll synthesis and accumulation in transgenic tomato plants. Transgenic Res 20:47–60
Das M, Chauhan H, Chibbar A, Haq QMR, Khurana P (2011) High-efficiency transformation and selective tolerance against biotic and abiotic stress in mulberry, Morus indica cv. K2, by constitutive and inducible expression of tobacco osmotin. Transgenic Res 20:231–246
Davison PA, Hunter CN, Horton P (2002) Overexpression of beta-carotene hydroxylase enhances stress tolerance in Arabidopsis. Nature 418:203–206
DellaPenna D, Pogson BJ (2006) Vitamin synthesis in plants: tocopherols and carotenoids. Annu Rev Plant Biol 57:711–738
Demmig B, Winter K, Kruger A, Czygan FC (1988) Zeaxanthin and the heat dissipation of excess light energy in Nerium oleander exposed to a combination of high light and water stress. Plant Physiol 87:17–24
Demmig-Adams B (1990) Carotenoids and photoprotection: a role for the xanthophyll zeaxanthin. Biochim Biophys Acta 1020:1–24
Demmig-Adams B, Adams WW III (1996) The role of xanthophyll cycle carotenoids in the protection of photosynthesis. Trends Plant Sci 1:21–26
Diretto G, Welsch R, Tavazza R, Mourgues F, Pizzichini D, Beyer P, Giuliano G (2007) Silencing of beta-carotene hydroxylase increases total carotenoid and beta-carotene levels in potato tubers. BMC Plant Biol 7:11
Du H, Wang N, Cui F, Li X, Xiao J, Xiong L (2010) Characterization of the beta-carotene hydroxylase gene DSM2 conferring drought and oxidative stress resistance by increasing xanthophylls and abscisic acid synthesis in rice. Plant Physiol 154:1304–1318
Götz T, Sandmann G, Römer S (2002) Expression of a bacterial carotene hydroxylase gene (crtZ) enhances UV tolerance in tobacco. Plant Mol Biol 50:129–142
Guzman I, Hamby S, Romero J, Bosland PW, O’Connell MA (2010) Variability of carotenoid biosynthesis in orange colored Capsicum spp. Plant Sci 179:49–59
Havaux M (1998) Carotenoids as membrane stabilizers in chloroplasts. Trends Plant Sci 3:147–151
Havaux M, Niyogi KK (1999) The violaxanthin cycle protects plants from photooxidative damage by more than one mechanism. Proc Natl Acad Sci USA 96:8762–8767
Hiscox JD, Israelstam GF (1979) A method for the extraction of chlorophyll from leaf tissue without maceration. Can J Bot 57:1332–1334
Jahns P, Holzwarth AR (2011) The role of the xanthophyll cycle and of lutein in photoprotection of photosystem II. Biochim Biophys Acta 1817:182–193
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Khurana P, Checker VG (2011) The advent of genomics in mulberry and perspectives for productivity enhancement. Plant Cell Rep 30:825–838
Kim CS, Lee CH, Shin JS, Chung YS, Hyung NI (1997) A simple and rapid method for isolation of high quality genomic DNA from fruit trees and conifers using PVP. Nucleic Acids Res 25:1085–1086
Kim IJ, Ko KC, Kim CS, Chung WII (2001) Isolation and characterization of cDNAs encoding β-carotene hydroxylase in Citrus. Plant Sci 161:1005–1010
Krinsky N (1998) The antioxidant and biological properties of the carotenoids. Ann NY Acad Sci 854:443–447
Lal S, Bhatnagar S, Khurana P (2006) Screening of Indian mulberry for abiotic stress tolerance and ameliorative effect of calcium on salinity stress. Physiol Mol Biol Plants 12:193–199
Lal S, Gulyani V, Khurana P (2008) Overexpression of HVA1 gene from barley generates tolerance to salinity and water stress in transgenic mulberry (Morus indica). Transgenic Res 17:651–663
Lal S, Ravi V, Khurana JP, Khurana P (2009) Repertoire of leaf expressed sequence tags (ESTs) and partial characterization of stress-related and membrane transporter genes from mulberry (Morus indica L.). Tree Genet Gen 5:359–374
Li F, Vallabhaneni R, Wurtzel ET (2008) PSY3, a new member of the phytoene synthase gene family conserved in the poaceae and regulator of abiotic stress-induced root carotenogenesis. Plant Physiol 146:1333–1345
Li Q, Farre G, Naqvi S, Breitenbach J, Sanahuja G, Bai C, Sandmann G, Capell T, Christou P, Zhu C (2010) Cloning and functional characterization of the maize carotenoid isomerase and β-carotene hydroxylase genes and their regulation during endosperm maturation. Transgenic Res 19:1053–1068
Liang YS, Bae HJ, Kang SH, Lee T, Kim MG, Kim YM, Ha SH (2009) The Arabidopsis beta-carotene hydroxylase gene promoter for a strong constitutive expression of transgene. Plant Biotechnol Rep 3:325–331
Muller P, Li XP, Niyogi KK (2001) Non-photochemical quenching: a response to excess light energy. Plant Physiol 125:1558–1566
Ramel F, Birtic S, Ginies C, Soubigou-Taconnat L, Triantaphylides C, Havaux M (2012) Carotenoid oxidation products are stress signals that mediate gene responses to singlet oxygen in plants. Proc Natl Acad Sci USA 109:5535–5540
Ren Y, Bang H, Lee EJ, Gould J, Rathore KS, Patil BS, Crosby KM (2013) Levels of phytoene and β-carotene in transgenic honeydew melon (Cucumis melo L. inodorus). Plant Cell Tissue Organ Cult 113:291–301
Sagar AD, Briggs WR (1990) Effects of high light stress on carotenoid-deficient chloroplasts in Pisum sativum. Plant Physiol 94:1663–1670
Sairam RK (1994) Effect of moisture stress on physiological activities of two contrasting wheat genotypes. Indian J Exp Biol 32:584–593
Sandmann G (2001) Genetic manipulation of carotenoid biosynthesis: strategies, problems and achievements. Trends Plant Sci 6:14–17
Tan CP, Zhao FQ, Su ZL, Liang CW, Qin S (2007) Expression of beta-carotene hydroxylase gene (crtR-B) from the green alga Haematococcus pluvialis in chloroplasts of Chlamydomonas reinhardtii. J Appl Phycol 19:347–355
Taylor M, Ramsay G (2005) Carotenoid biosynthesis in plant storage organs: recent advances and prospects for improving plant food quality. Physiol Plant 124:143–151
Thayer SS, Bjorkman O (1990) Leaf xanthophyll content and composition in sun and shade determined by HPLC. Photosynth Res 23:331–343
Tian L, DellaPenna D (2004) Progress in understanding the origin and functions of carotenoid hydroxylases in plants. Arch Biochem Biophys 430:22–29
Tuteja N (2007) Abscisic acid and abiotic stress signaling. Plant Signal Behav 2:135–138
Wang N, Fang W, Han H, Sui N, Li B, Meng QW (2008) Overexpression of zeaxanthin epoxidase gene enhances the sensitivity of tomato PSII photoinhibition to high light and chilling stress. Physiol Plant 132:384–396
Watanabe T (1958) Substances in mulberry leaves which attract silkworm larvae (Bombyx mori). Nature 182:325–326
Wellburn A (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313
Wright LA, Murphy TM, Travis RL (1981) The effect of ultraviolet radiation on wheat root vesicles enriched in plasma membrane. Photochem Photobiol 33:343–348
Xiong L, Zhu JK (2003) Regulation of abscisic acid biosynthesis. Plant Physiol 133:29–36
Yang G, Wang Y, Xia D, Gao C, Wang C, Yang C (2014) Overexpression of a GST gene (ThGSTZ1) from Tamarix hispida improves drought and salinity tolerance by enhancing the ability to scavenge reactive oxygen species. Plant Cell Tissue Organ Cult 117:99–112
Yu B, Lydiate DJ, Schafer UA, Hannoufa A (2007) Characterization of a beta-carotene hydroxylase of Adonis aestivalis and its expression in Arabidopsis thaliana. Planta 226:181–192
Zhou X, Yuan Y, Yang Y, Rutzke M, Thannhauser TW, Kochian LV, Li L (2009) Involvement of a broccoli COQ5 methyl transferase in the production of volatile selenium compounds. Plant Physiol 151:528–540
Acknowledgments
This work was financially supported by Grants received from DBT (Department of Biotechnology, Government of India). BS acknowledges the award of Junior Research Fellowship and MD acknowledges the award of Senior Research Fellowship by CSIR (Council of Scientific and Industrial Research) and DBT (Department of Biotechnology), respectively.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Additional information
Bushra Saeed and Manaswini Das have contributed equally to this work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Saeed, B., Das, M. & Khurana, P. Overexpression of β-carotene hydroxylase1 (BCH1) in Indian mulberry, Morus indica cv. K2, confers tolerance against UV, high temperature and high irradiance stress induced oxidative damage. Plant Cell Tiss Organ Cult 120, 1003–1014 (2015). https://doi.org/10.1007/s11240-014-0654-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-014-0654-6