Skip to main content
SpringerLink
Log in

RNAi mediated silencing of lipoxygenase gene to maintain rice grain quality and viability during storage

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

Lipoxygenase (LOX) is a common enzyme which catalyzes lipid peroxidation of seeds and consequently enhances seed quality deterioration and decreases seed viability. During seed storage, peroxidation of unsaturated fatty acids occur due to enhancement of LOX activity which directly leads to reduction in seed vigour and deterioration of grain nutritional quality. This study was undertaken to overcome these problem during rice seed storage by attenuating LOX activity using RNAi technology. To improve seed storage stability, we down regulated LOX gene activity by using a functional fragment of the LOX gene under the control of both constitutive (CaMV35S) and aleurone-specific (Oleosin-18) promoter separately. To understand the storage stability, RNAi–LOX seeds and non-transgenic control seeds were subjected to accelerated aging at 45 °C and 85 % relative humidity for 14 days. Our studies demonstrate that down regulation of LOX activity reduces the seed quality deterioration under storage condition. In addition GC–MS analysis revealed that reduction of fatty acid level in non-transgenic seeds during storage was higher when compared with that of transgenic rice seeds. Furthermore, the transgenic rice seeds with reduced LOX activity exhibited enhanced seed germination efficiency after storage than that of non-transgenic rice seeds. This study will have direct impact on nutritional stability of quality rice grains.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

Abbreviations

LOX:

Lipoxygenase

MDA:

Malondialdehyde

HNE:

Hydroxynonenal

FFA:

Free fatty acid

ROS:

Reactive oxygen species

HPOD:

Hydroperoxyoctadecadienoic acid

AA:

Accelerated aging

RH:

Relative humidity

EC:

Electric conductance

AAS:

Atomic absorption spectroscopy

References

  • Berlett BS, Stadtman ER (1997) Protein oxidation in aging, disease, and oxidative stress. J Biol Chem 272:20313–20316

    Article  CAS  PubMed  Google Scholar 

  • Brash AR (1999) Lipoxygenases: occurrence, functions, catalysis, and acquisition of substrate. J Biol Chem 274:23679–23682

    Article  CAS  PubMed  Google Scholar 

  • Datta SK, Peterhans A, Datta K, Potrykus I (1990) Genetically engineered fertile indica-rice recovered from protoplasts. Nat Biotechnol 8:736–740

    Article  CAS  Google Scholar 

  • Datta K, Vasquez A, Tu J, Torrizo L, Alam MF, Oliva N, Abrigo E, Khush GS, Datta SK (1998) Constitutive and tissue-specific differential expression of the cryIA(b) gene in transgenic rice plants conferring resistance to rice insect pest. Theor Appl Genet 97:20–30

    Article  CAS  Google Scholar 

  • Datta K, Baisakh N, Oliva N, Torrizo L, Abrigo E, Tan J, Rai M, Rehana S, Al-Babili S, Beyer P, Potrykus I, Datta SK (2003) Bioengineered ‘golden’ indica rice cultivars with beta-carotene metabolism in the endosperm with hygromycin and mannose selection systems. Plant Biotechnol J 1:81–90

    Article  CAS  PubMed  Google Scholar 

  • DeLong JM, Prange RK, Hodges DM, Forney CF, Bishop MC, Quilliam M (2002) Using a modified ferrous oxidation-xylenol orange (FOX) assay for detection of lipid hydroperoxides in plant tissue. J Agr Food Chem 50:248–254

    Article  CAS  Google Scholar 

  • Devaiah SP, Pan X, Hong Y, Roth M, Welti R, Wang X (2007) Enhancing seed quality and viability by suppressing phospholipase D in Arabidopsis. Plant J 50:950–957

    Article  CAS  PubMed  Google Scholar 

  • Doria E, Galleschi L, Calucci L, Pinzino C, Pilu R, Cassani E, Nielsen E (2009) Phytic acid prevents oxidative stress in seeds: evidence from a maize (Zea mays L.) low phytic acid mutant. J Exp Bot 60:967–978

    Article  CAS  PubMed  Google Scholar 

  • Garces R, Mancha M (1993) One-step lipid extraction and fatty acid methyl esters preparation from fresh plant tissues. Anal Biochem 211:139–143

    Article  CAS  PubMed  Google Scholar 

  • Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198

    Article  CAS  PubMed  Google Scholar 

  • Himmelbach A, Zierold U, Hensel G, Riechen J, Douchkov D, Schweizer P, Kumlehn J (2007) A set of modular binary vectors for transformation of cereals. Plant Physiol 145:1192–1200

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Huang N, Angeles ER, Domingo J, Magpantay G, Singh S, Zhang G, Kumaravadivel N, Bennett J, Khush GS (1997) Pyramiding of bacterial blight resistance genes in rice: marker-assisted selection using RFLP and PCR. Theor Appl Genet 95:313–320

    Article  CAS  Google Scholar 

  • Ida S, Masaki Y, Morita Y (1983) The isolation of multiple forms and product specificity of rice lipoxygenase. Agr Biol Chem 47:637–641

    Article  CAS  Google Scholar 

  • ISTA (1993) International rules for seed testing. Seed Sci Technol 21:1–228

    Google Scholar 

  • Jiang J, Wu J, Wu Y, Song M, Wang X, Liu B, Yu Z (2009) Storage stability reversion mutation of a rice line devoid of LOX-1, 2 acquired by ion beam irradiation. Plasma Sci Technol 1:116–121

    Google Scholar 

  • Kaewnaree P, Vichitphan S, Klanrit P, Siri B, Vichitphan K (2011) Effect of accelerated aging process on seed quality and biochemical changes in sweet pepper (Capsicum annuum Linn.) seeds. Biotechnol 10:175–182

    Article  CAS  Google Scholar 

  • Kapoor N, Arya A, Siddiqui MA, Kumar H, Amir A (2011) Physiological and biochemical changes during seed deterioration in aged seeds of rice (Oryza sativa L.) Am. J Plant Physiol 6:28–35

    Article  Google Scholar 

  • Kim D, Han S, Song J (2011) Evaluation of the inorganic compound leakage and carbohydrates as indicator of physiological potential of Ulmus parvifolia seeds. New Forest 41:3–11

    Article  Google Scholar 

  • Kwon KC, Verma D, Jin S, Singh ND, Daniell H (2013) Release of proteins from intact chloroplasts induced by reactive oxygen species during biotic and abiotic stress. PLoS ONE 8:e67106

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Li J, Zhang Y, Yu Z, Wang Y, Yang Y, Liu Z, Jiang J, Song M, Wu Y (2007) Superior storage stability in low lipoxygenase maize varieties. J Stored Prod Res 43:530–534

    Article  CAS  Google Scholar 

  • McDonald MB (1999) Seed deterioration: physiology, repair and assessment. Seed Sci Technol 27:177–237

    Google Scholar 

  • Meng L, Feldman L (2010) A rapid TRIzol-based two-step method for DNA-free RNA extraction from Arabidopsis siliques and dry seeds. Biotechnol J 5:183–186

    Article  CAS  PubMed  Google Scholar 

  • Mizuno K, Iida T, Takano A, Yokoyama M, Fujimura T (2003) A new 9-lipoxygenase cDNA from developing rice seeds. Plant Cell Physiol 44:1168–1175

    Article  CAS  PubMed  Google Scholar 

  • Murthy UM, Kumar PP, Sun WQ (2003) Mechanisms of seed ageing under different storage conditions for Vigna radiata (L.) Wilczek: lipid peroxidation, sugar hydrolysis, Maillard reactions and their relationship to glass state transition. J Exp Bot 54:1057–1067

    Article  CAS  PubMed  Google Scholar 

  • Nishida I (2004) Molecular biology and biotechnology of plant organelles. In: Daniell H, Chase C (eds) Plastid metabolic pathways for fatty acid metabolism. Springer, Netherland, pp 543–564

  • Ohta H, Ida S, Mikami B, Morita Y (1986) Purification and characterization of rice lipoxygenase component 3 from embryos (biological chemistry). Agri Biol Chem 50:3165–3171

    Article  CAS  Google Scholar 

  • Ohta H, Shirano Y, Tanaka K, Morita Y, Shibata D (1992) cDNA cloning of rice lipoxygenase L-2 and characterization using an active enzyme expressed from the cDNA in Escherichia coli. Eur J Biochem 206:331–336

    Article  CAS  PubMed  Google Scholar 

  • Orak H, Başer I, Korkut KZ, Bilgin O, Orak A (2010) Investigations of peroxidase and lipoxygenase enzyme activities and their relationships with colour and chemical characteristics in commercial dent corn (Zea Mays L.) hybrids. J Agro Crop Sci 1:33–40

    Google Scholar 

  • Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Wright SY, Hinchliffe E, Adams JL, Silverstone AL, Drake R (2005) Improving the nutritional value of golden rice through increased pro-vitamin A content. Nat Biotechnol 23:482–487

    Article  CAS  PubMed  Google Scholar 

  • Peng YL, Shirano Y, Ohta H, Hibino T, Tanaka K, Shibata D (1994) A novel lipoxygenase from rice. Primary structure and specific expression upon incompatible infection with rice blast fungus. J Biol Chem 269:3755–3761

    CAS  PubMed  Google Scholar 

  • Qu LQ, Takaiwa F (2004) Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice. Plant Biotechnol J 2:113–125

    Article  CAS  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning, a laboratory manual. Ed.3. (1, 2, 3). Cold Spring Harbour Laboratory Press, Cold spring Harbour

    Google Scholar 

  • Schaffrath U, Zabbai F, Dudler R (2000) Characterization of RCI-1, a chloroplastic rice lipoxygenase whose synthesis is induced by chemical plant resistance activators. Eur J Biochem 267:5935–5942

    Article  CAS  PubMed  Google Scholar 

  • Shin MG, Yoon SH, Rhee JS, Kwon TAIWAN (1986) Correlation between oxidative deterioration of unsaturated lipid and n-hexanal during storage of brown rice. J Food Sci 51:460–463

    Article  CAS  Google Scholar 

  • Shirasawa K, Takeuchi Y, Ebitani T, Suzuki Y (2008) Identification of gene for rice (Oryza sativa) seed lipoxygenase-3 involved in the generation of stale flavor and development of SNP markers for lipoxygenase-3 deficiency. Breed Sci 58:169–176

    Article  CAS  Google Scholar 

  • Song M, Wu Y, Zhang Y, Liu BM, Jiang JY, Xu X, Yu ZL (2007) Mutation of rice (Oryza sativa L.) LOX-1/2 near-isogenic lines with ion beam implantation and study of their storability. Nucl Instrum Methods 265:495–500

    Article  CAS  Google Scholar 

  • Steffens B, Steffen-Heins A, Sauter M (2013) Reactive oxygen species mediate growth and death in submerged plants. Front Plant Sci 4. doi: 10.3389/fpls.2013.00179

  • Suzuki Y, Ise K, Li C, Honda I, Iwaim Y, Matsukura U (1999) Volatile components in stored rice [Oryza sativa (L.)] of varieties with and without lipoxygenase-3 in seeds. J Agric Food Chem 47:1119–1124

    Article  CAS  PubMed  Google Scholar 

  • Umate P (2011) Genome-wide analysis of lipoxygenase gene family in Arabidopsis and rice. Plant Signal Behav 6:335–338

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wu Q, Lin J, Liu JZ, Wang X, Lim W, Oh M, Park J, Rajashekar CB, Whitham SA, Cheng NH, Hirschi KD, Park S (2012) Ectopic expression of Arabidopsis glutaredoxin AtGRXS17 enhances thermotolerance in tomato. Plant Biotechnol J 10:945–955

    Article  CAS  PubMed  Google Scholar 

  • Xu X, Guo R, Cheng C, Zhang H, Zhang Y, Wang X (2013) Overexpression of ALDH2B8, an aldehyde dehydrogenase gene from grapevine, sustains Arabidopsis growth upon salt stress and protects plants against oxidative stress. Plant Cell, Tissue Organ Cult 114:187–196

    Article  CAS  Google Scholar 

  • Zhang Y, Yu Z, Lu Y, Wang Y, She D, Song M, Wu Y (2007) Effect of the absence of lipoxygenase isoenzymes on the storage characteristics of rice grains. J Stored Prod Res 43:87–91

    Article  CAS  Google Scholar 

  • Zhou G, Qi J, Ren N, Cheng J, Erb M, Mao B, Lou Y (2009) Silencing OsHI-LOX makes rice more susceptible to chewing herbivores, but enhances resistance to a phloem feeder. Plant J 60:638–648

    Article  CAS  PubMed  Google Scholar 

  • Zhu C, Sanahuja G, Yuan D, Farré G, Arjó G, Berman J, Zorrilla-López U, Banakar R, Bai C, Pérez-Massot E (2013) Bio fortification of plants with altered antioxidant content and composition: genetic engineering strategies. Plant Biotechnol J 11:129–141

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thankfully acknowledge Department of Biotechnology (DBT), Government of India for financial support in the form of DBT programme support (Sanction no. BT/01/COE/06/05) that made the research possible. We are thankful to the Leibniz Institute of Plant Genetics and Crop Plant Research, D–06466 Gatersleben, Germany for providing the pIPKb-006 and pIPKb-009 destination vectors. We thank Prof. S Muthukrishnan, Kansas State University, USA for reviewing and editorial support of the MS.

Author information

Authors and Affiliations

  1. Plant Molecular Biology and Biotechnology Laboratory, Department of Botany, University of Calcutta, 35 Ballygunge Circular Road, Kolkata, 700019, WB, India

    Dipak Gayen, Nusrat Ali, Moumita Ganguly, Soumitra Paul, Karabi Datta & Swapan K. Datta

  2. Division of Crop Sciences, Indian Council of Agricultural Research (ICAR), Krishi Bhavan, Dr. Rajendra Prasad Road, New Delhi, 110001, India

    Swapan K. Datta

Authors
  1. Dipak Gayen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nusrat Ali
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Moumita Ganguly
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Soumitra Paul
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Karabi Datta
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Swapan K. Datta
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Swapan K. Datta.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 4932 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gayen, D., Ali, N., Ganguly, M. et al. RNAi mediated silencing of lipoxygenase gene to maintain rice grain quality and viability during storage. Plant Cell Tiss Organ Cult 118, 229–243 (2014). https://doi.org/10.1007/s11240-014-0476-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-014-0476-6

Keywords

Search

Navigation