Abstract
The aim of the study is to develop transgenic rice with annexin genes (AnnBj2) to determine its salinity and water stress tolerance through Agrobacterium-mediated genetic transformation. The transgenic somatic embryos were developed on Murashige and Skoog medium fortified with 0.5 mg L−1 kinetin and 3.0 mg L−1 2,4-D. About 80% of the transformed somatic embryos germinated and was successfully established in the transgenic greenhouse. The transformed plantlets (T0 and T1 generation) were identified through PCR using NPT-II and AnnBj2 gene–specific primers as well as positive uidA reporter gene expression. Compared with non-transformed plants, the transgenic rice plants overexpressing AnnBj2 gene exhibited salt tolerance at the seedling stage. Seeds generated at T0 and T1 generations were further studied to determine their salinity tolerance through nutrient culture and pot culture experiments for water stress. When compared with the non-transgenic control, transgenic seeds (T0 and T1) had increased germination on a nutrient medium having 200 mM NaCl. The data showed that the shoot and root lengths were longer at 200 mM NaCl when compared with those of the non-transformed plants. Further, the biochemical assessment (photosynthetic pigment analysis, protein, proline content, and oxidative enzyme activity) was performed between transformed (T0 and T1) and non-transformed plantlets under stress. It was noted that the transformed plantlets were higher in chlorophyll and proline content as well as oxidative enzyme activity as compared with the non-transformed plants.
Similar content being viewed by others
References
Aananthi N, Anandakumar CR, Ushakumari R, Shanthi P (2010) Regeneration study of some indica rice cultivars followed by Agrobacterium-mediated transformation of highly regenerable cultivar Pusa Basmati. Electron J Plant Breed 1:1249–1256
Aebi H (1984) Catalase in vitro. In: Methods in enzymology, 105. Academic, New York, pp 121–126
Ahmed I, Yadav D, Shukla P, Kirti PB (2018) Heterologous expression of Brassica juncea annexin, AnnBj2 confers salt tolerance and ABA insensitivity in transgenic tobacco seedlings. Funct Integr Genomic 18:569–579
Aldemita RR, Hodges TK (1996) Agrobacterium tumefaciens-mediated transformation of japonica and indica rice varieties. Planta 199:612–617
Ashraf M, Foolad MR (2007) Roles of glycine, betaine and proline in improving plant abiotic stress resistance. Environ Exp Bot 59:206–212
Bajguz A, Piotrowska-Niczyporuk A (2013) Synergistic effect of auxins and brassinosteroids on the growth and regulation of metabolite content in the green alga Chlorella vulgaris (Trebouxiophyceae). Plant Physiol Biochem 71:290–301
Baskaran P, Dasgupta I (2012) Gene delivery using micro injection of Agrobacterium to embryonic shoot apical meristem of elite indica rice cultivars. J Plant Biochem Biotech 21:268–274
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–211
Bohnert HJ, Gong Q, Li P, Ma S (2006) Unraveling abiotic stress tolerance mechanisms – getting genomics going. Curr Opin Plant Biol 9:180–188
Bzour MI, Zuki FM, Mispan MS (2018) Introduction of imidazolinone herbicide and clearfield rice between weedy rice control efficiency and environmental concerns. Environ Rev 18:49–53
Chan MT, Chang HH, Ho SL, Tong WF, Yu SM (1993) Agrobacterium-mediated production of transgenic rice plants expressing a chimeric alpha-amylase promoter/beta-glucuronidase gene. Plant Mol Biol 22:491–506
Chan MT, Lee TM, Chang HH (1992) Transformation of indica rice (Oryza sativa L.) mediated by Agrobacterium tumefaciens. Plant Cell Physiol 33:577–583
Divya K, Jami S, Kirti PB (2010) Constitutive expression of mustard annexin, AnnBj1 enhances abiotic stress tolerance and fiber quality in cotton under stress. Plant Mol Biol 73:293–308
Dong J, Teng W, Buchholz WG (1996) Agrobacterium-mediated transformation of Javanica rice. Mol Breed 2:267–276
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19:11–15
Ge X, Chu Z, Lin Y, Wang S (2006) A tissue culture system for different germplasms of indica rice. Plant Cell Rep 25:392–402
Ghorbanli M, Gafarabad M, Amirkian T, Mamaghani BA (2013) Investigation of proline, total protein, chlorophyll, ascorbate and dehydroascorbate changes under drought stress in Akria and Mobil tomato cultivars. Iran J Plant Physiol 3:651–568
Giannopolitis CN, Ries SK (1977) Superoxide dismutases: I. Occurrence in higher plants. Plant Physiol 59:309–314
Gu M, Zhang J, Li H (2017) Maintenance of phosphate homeostasis and root development are coordinately regulated by MYB1, an R2R3-type MYB transcription factor in rice. J Exp Bot 68:3603–3615
Hiei Y, Komari T, Kubo T (1997) Transformation of rice mediated by Agrobacterium tumefaciens. Plant Mol Biol 35:205–218
Hiei Y, Ishida Y, Komari T (2015) Rice, indica (Oryza sativa L.). Methods Mol Biol 1223:155–167
Hossain MR, Hassan L, Patwary AK, Ferdous MJ (2009) Optimization of Agrobacterium tumefaciens mediated genetic transformation protocol for aromatic rice. J Bangladesh Agril Univ 7:235–240
International Rice Research Institute (IRRI) (2001) Website at http://www.irri.org
James D, Borphukan B, Fartyal D, Ram B, Yadav R, Singh J (2018) Concurrent overexpression of OsGS1;1 and OsGS2 genes in transgenic rice (Oryza sativa L.): impact on tolerance to abiotic stresses. Front Plant Sci 9:786
Jami SK, Clark GB, Ayele BT, Roux SJ, Kirti PB (2012) Identification and characterization of annexin gene family in rice. Plant Cell Rep 31:813–825
Jami SK, Dalal A, Divya K, Kirti PB (2009) Molecular cloning and characterization of five annexin genes from Indian mustard (Brassica juncea L. Czern and Coss). Plant Physiol Biochem 47:977–990
Jami SK, Hill RD, Kirti PB (2010) Transcriptional regulation of annexins in Indian mustard, Brassica juncea and detoxification of ROS in transgenic tobacco plants constitutively expressing AnnBj1. Plant Signal Behaviour 5:618–621
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucorodinase as sensitive and versatile gene fusion marker in higher plants. EMBO 6:3901–3907
Kamanga RM, Mbega E, Ndakidemi P (2018) Drought tolerance mechanisms in plants: physiological responses associated with water deficit stress in Solanum lycopersicum. Adv Crop Sci Tech 6:362–372
Kamoshita A, Zhang J, Siopongco J, Sarkarung S, Nguyen HT, Wade LJ (2002) Effects of phenotyping environment on identification of quantitative trait loci for rice root morphology under anaerobic conditions. Crop Sci 42:255–265
Kant T, Kothari SL, Hodges HK, Hodges TK (2001) Agrobacterium tumefaciens — mediated transformation of rice using coleoptiles and mature seed-derived callus. J Plant Biochem Biotech 10:121–126
Kathuria H, Giri J, Tyagi H, Tyagi AK (2007) Advances in transgenic rice biotechnology. Crit Rev Plant Sci 26:65–103
Khanna HK, Raina SK (1999) Agrobacterium-mediated transformation of indica rice cultivars using binary and superbinary vectors. Aust J Plant Physiol 26:311–324
Konopka-Postupolska D, Clark G, Goch G, Debski J, Floras K, Cantero A, Fijolek B, Roux S, Hennig J (2009) The role of annexin 1 in drought stress in Arabidopsis. Plant Physiol 150:1394–1410
Kumar S (2017) Phytochemistry and functional food: the needs of healthy life. J Phytochem Biochem 1:2–10
Lee S, Jeon JS, Jung KH, An G (1999) Binary vector for efficient transformation of rice. J Plant Biol 42:310–316
Lee K, Jeon H, Kim M (2002) Optimization of mature embryo-based in vitro culture system for high-frequency somatic embryogenic callus induction and plant regeneration from Japonica rice cultivars. Plant Cell Tiss Org Cult 71:237–244
Lee YP, Kim SH, Bang JW, Lee HS, Kwak SS, Kwon SY (2007) Enhanced tolerance to oxidative stress in transgenic tobacco plants expressing three antioxidant enzymes in chloroplasts. Plant Cell Rep 26:591–598
Liang X, Zhang L, Natarajan SK, Becker DF (2013) Proline mechanisms of stress survival. Antioxid Redox Signal 19:998–1011
Lin Y, Zhang Q (2005) Optimizing the tissue culture conditions for high efficiency transformation of indica rice. Plant Cell Rep 23:540–547
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408
Lowry OH, Rosebrough NI, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Mahendranath G, KrishnakanthYadav T, Jagadish P, Raghurami Reddy M, Balachandran SM, Siddiq EA, Yamini KN, Anuradha G (2018) Standardization of Agrobacterium mediated genetic transformation in Indica rice cv BPT-5204. Ann Plant Sci 7:2037–2041
Mao K, Dong Q, Li C, Liu C, Ma F (2017) Genome wide identification and characterization of apple bHLH transcription factors and expression analysis in response to drought and salt stress. Front Plant Sci 8:480
Matsuoka K, Nakamura K (1999) The large alkyl side chains of isoleucine and leucine in the NPIRL region constitute the core of the vacuolar sorting determinant of sporamin precursor. Plant Mol Biol 41:825–835
Mohanty A, Sarma NP, Tyagi AK (1999) Agrobacterium-mediated high frequency transformation of an elite indica rice variety Pusa Basmati 1 and transmission of the transgenes to R2 progeny. Plant Sci 147:127–137
Moss SE, Morgan RO (2004) The annexins. Genome Biol 5:219–224
Mostafiz SB, Wagiran A (2018) Efficient callus induction and regeneration in selected indica rice. Agronomy 8:77
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco cultures. Plant Physiol 15:473–493
Perez-Alfocea F, Estan MT, Caro M, Bolarin MC (1993) Response of tomato cultivars to salinity. Plant Soil 150:203–211
Prabhavati V, Yadav S, Kumar PA, Rajam MV (2002) Abiotic stress tolerance in transgenic egg plant (Solanum melongena L.) by introduction of bacterial mannitol phosphor dehydrogenase gene. Mol Breed 9:137–147
Proust J, Houlne G, Schantz ML, Shen WH, Schantz R (1999) Regulation of biosynthesis and cellular localization of Sp32 annexins in tobacco BY2 cells. Plant Mol Biol 39:361–372
Qiao B, Zhang Q, Liu DL, Wang HQ, Yin JY, Wang R, He ML, Cui M, Shang ZL, Wang DK, Zhu ZG (2015) A calcium-binding protein, rice annexin OsANN1, enhances heat stress tolerance by modulating the production of H2O2. J Exp Bot 66:5853–5866
Rahimian Boogar A, Salehi H, Jowkar A (2014) Exogenous nitric oxide alleviates oxidative damage in turfgrasses under drought stress. South Afr J Bot 92:78–82
Raineri DM, Bottino P, Gordon MP, Nester EW (1990) Agrobacterium-mediated transformation of rice (Oryza sativa L.). Bio/Technology 8:33–38
Rashid H, Yokoi S, Toriyama K, Kokichi H (1996) Transgenic plant production mediated by Agrobacterium in Indica rice. Plant Cell Rep 15:727–730
Saha D, Bhavya C, Ashok TH (2017) Effect of genotypes and different concentration of growth regulator on callus induction and plant regeneration through anther culture of rice. J Pharm Phytochem 6:1354–1358
Sahi C, Singh A, Kumar K, Blumwald E, Grover A (2006) Salt stress response in rice: genetics, molecular biology, and comparative genomics. Funct Integr Genomics 6:263–284
Sarangi S, Ghosh J, Bora A, Das S, Mandal AB (2011) Agrobacterium mediated genetic transformation of indica rice varieties involving Am-SOD gene. Ind J Biotech 10:9–18
Shrawat AK, Lorz H (2006) Agrobacterium-mediated transformation of cereals: a promising approach crossing barriers. Plant Biotech J 4:575–603
Sreenivasulu N, Grimm B, Wobus U, Weschke W (2000) Differential response of antioxidant components to salinity stress in salt-tolerant and salt-sensitive seedlings of foxtail millet (Setaria italica). Physiol Plant 109:435–442
Sridevi G, Dhandapani M, Veluthambi K (2005) Agrobacterium-mediated transformation of White Ponni, a non-basmati variety of indica rice (Oryza sativa L.). Curr Sci 88:128–132
Tanaka Y, Hibino T, Hayashi Y, Tnaka A, Kishitani S (1999) Salt tolerance of transgenic rice over-expressing yeast mitochondrial Mn-SOD in chloroplasts. Plant Sci 14:131–138
Tang Y, Liu K, Zhang J, Li X, Xu K, Zhang Y (2017) JcDREB2, a physic nut AP2/ERF gene, alters plant growth and salinity stress responses in transgenic rice. Front Plant Sci 8:306. https://doi.org/10.3389/fpls.2017.00306
Toki S, Hara N, Ono K, Onodera H, Tagiri A, Oka S, Tanaka H (2006) Early infection of scutellum tissue with Agrobacterium allows high-speed transformation of rice. The Plant J 47:969–976
Tyagi AK, Mohanty A (2000) Rice transformation for crop improvement and functional genomics. Plant Sci 158:1–18
Vendruscolo EC, Schuster I, Pileggi M, Scapim CA, Molinari HB, Marur CJ, Vieira LG (2007) Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat. J Plant Physiol 164:136–146
Vernon LP (1960) Spectrophotometric determination of chlorophylls and heophytins in plant extracts. Anal Chem 32:1144–1150
Wahab AG (2017) Grain and feed annual Kuala Lumpur Malaysia. Global Agricultural Information Network, Kuala Lumpur
Wang FZ, Wang QB, Kwon SY, Kwak SS, Su WA (2005) Enhanced drought tolerance of transgenic rice plants expressing a pea manganese superoxide dismutase. J Plant Physiol 162:465–472
Wang H, Zhang H, Gao F, Li J, Li Z (2007) Comparison of gene expression between upland and lowland rice cultivars under water stress using cDNA microarray. Theor Appl Genet 115:1109–1126
Wei Q, Luo Q, Wang R, Zhang F, He Y, Zhang Y (2017) A wheat R2R3-type MYB transcription factor TaODORANT1 positively regulates drought and salt stress responses in transgenic tobacco plants. Front Plant Sci 8:1374
Yadav D, Ahmed I, Shukla P, Boyidi P, Kirti PB (2016) Overexpression of Arabidopsis AnnAt8 alleviates abiotic stress in transgenic Arabidopsis and tobacco. Plants. 5:18
Yang A, Dai X, Zhang WH (2012) A R2R3-type MYB gene, OsMYB2, is involved in salt, cold, and dehydration tolerance in rice. J Exp Bot 63:2541–2556
Yang P, Liang Y, Shen S, Kuang T (2006) Proteome analysis of rice uppermost internodes at the milky stage. Proteomics 6:3330–3338
Zhang F, Li S, Yang S, Wang L, Guo W (2015) Overexpression of a cotton annexin gene, GhAnn1, enhances drought and salt stress tolerance in transgenic cotton. Plant Mol Biol 87:47–67
Zhang Y, Sompong T (2015) Optimization of certain parameters for transformation of indica rice HomKra Dang Ngah variety via Agrobacterium mediated transformation. Kasetsart J Nat Sci 49:676–686
Zhu N, Cheng S, Liu X, Du H, Dai M, Zhou DX (2015) The R2R3-type MYB gene OsMYB91 has a function in coordinating plant growth and salt stress tolerance in rice. Plant Sci 236:146–156
Acknowledgments
The authors wish to acknowledge the Department of Biotechnology, Government of India, for providing the infrastructural facility under the PG-HRD teaching program. The authors wish to acknowledge Prof. P.B. Kirti, University of Hyderabad, Hyderabad, India, for proving the gene construct.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Editor: Prakash Lakshmanan
Rights and permissions
About this article
Cite this article
Mallick, S.R., Jadhao, K.R. & Rout, G.R. Overexpression of annexin gene in rice (Oryza sativa L.) for salinity and water stress. In Vitro Cell.Dev.Biol.-Plant 57, 86–101 (2021). https://doi.org/10.1007/s11627-020-10108-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-020-10108-6