Skip to main content
SpringerLink
Log in

OsLOX2, a rice type I lipoxygenase, confers opposite effects on seed germination and longevity

  • Original Paper
  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

Rice production and seed storage are confronted with grain deterioration and loss of seed viability. Some members of the lipoxygenase (LOX) family function in degradation of storage lipids during the seed germination, but little is known about their influence on seed longevity during storage. We characterized the role of rice OsLOX2 gene in seed germination and longevity via over-expression and knock-down approaches. Abundant expression of OsLOX2 was detected in panicles, roots, and stems, but not in leaves. Moreover, OsLOX2 was highly induced during germination. OsLOX2 protein, located in the cytoplasm, showed a wide range of temperature adaptation (20–50 °C) and a substrate preference to linoleic acid. Lines over-expressing OsLOX2 showed accelerated seed germination under normal condition and lower seed viability after accelerated aging. RNA interference (RNAi) of OsLOX2 caused delayed germination and enhanced seed longevity. RNAi lines with strongly repressed OsLOX2 activity completely lost the capability of germination after accelerated aging. More lipid hydroperoxide were found in OE15 than the control, but less in RNAi lines than in the WT Nipponbare. Therefore, OsLOX2 acts in opposite directions during seed germination and longevity during storage. Appropriate repression of the OsLOX2 gene may delay the aging process during the storage without compromising germination under normal conditions.

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

Similar content being viewed by others

Abbreviations

LOXs:

Lipoxygenase

LA:

α-Linoleic acid

LeA:

Linolenic acid

GFP:

Green fluorescent protein

OE:

Over-expression

RNAi:

RNA interference

HPOD:

Hydroperoxide

LHP:

Lipid hydroperoxide

References

  • Axelrod B (1981) Lipoxygenase from soybeans. Methods enzymol 71:441–451

  • Beckman K, Ingram D (1994) The inhibition of the hypersensitive response of potato-tuber tissues by cytokinins-similarities between senescence and plant defense responses. Physiol Mol Plant Pathol 44:33–50

    Article  CAS  Google Scholar 

  • Blee E (2002) Impact of phyto-oxylipins in plant defense. Trends Plant Sci 7:315–321

    Article  CAS  PubMed  Google Scholar 

  • Blee E, Joyard J (1996) Envelope membranes from spinach chloroplasts are a site of metabolism of fatty acid hydroperoxides. Plant Physiol 110:445–454

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bou R, Codony R, Tres A, Decker EA, Guardiola F (2008) Determination of hydroperoxides in foods and biological samples by the ferrous oxidation–xylenol orange method: a review of the factors that influence the method’s performance. Anal Biochem 377:1–15

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

    Article  CAS  PubMed  Google Scholar 

  • Feussner I, Kindl H (1994) Particulate and soluble lipoxygenase isoenzymes-comparison of molecular and enzymatic properties. Planta 194:22–28

    Article  CAS  Google Scholar 

  • Feussner I, Wasternack C (2002) The lipoxygenase pathway. Annu Rev Plant Biol 53:275–297

    Article  CAS  PubMed  Google Scholar 

  • Feussner I, Hause B, Voros K, Parthier B, Wasternack C (1995a) Jasmonate-induced lipoxygenase forms are localized in chloroplasts of barley leaves (Hordeum-vulgare cv salome). Plant J 7:949–957

    Article  CAS  Google Scholar 

  • Feussner I, Wasternack C, Kindl H, Kühn H (1995b) Lipoxygenase-catalyzed oxygenation of storage lipids is implicated in lipid mobilization during germination. Proc Natl Acad Sci 92:11849–11853

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Feussner I, Hause B, Nellen A, Wasternack C, Kindl H (1996) Lipid-body lipoxygenase is expressed in cotyledons during germination prior to other lipoxygenase forms. Planta 198:288–293

    Article  CAS  Google Scholar 

  • Feussner I, Balkenhohl T, Porzel A, Kuhn H, Wasternack C (1997a) Structural elucidation of oxygenated storage lipids in cucumber cotyledons—implication of lipid body lipoxygenase in lipid mobilization during germination. J Biol Chem 272:21635–21641

    Article  CAS  PubMed  Google Scholar 

  • Feussner I, Kühn H, Wasternack C (1997b) Do specific linoleate 13-lipoxygenases initiate β-oxidation? FEBS Lett 406:1–5

    Article  CAS  PubMed  Google Scholar 

  • Feussner I, Kuhn H, Wasternack C (2001) Lipoxygenase-dependent degradation of storage lipids. Trends Plant Sci 6:268–273

    Article  CAS  PubMed  Google Scholar 

  • Fuse T, Sasaki T, Yano M (2001) Ti-plasmid vectors useful for functional analysis of rice genes. Plant Biotechnol 18:219–222

    Article  CAS  Google Scholar 

  • Gardner H (1995) Biological roles and biochemistry of the lipoxygenase pathway. HortScience 30:197–205

    CAS  Google Scholar 

  • Gerhardt B (1992) Fatty acid degradation in plants. Prog Lipid Res 31:417–446

  • Grayburn W, Schneider G, Hamiltonkemp T, Bookjans G, Ali K, HildebrandI D (1991) Soybean leaves contain multiple lipoxygenases. Plant Physiol 95:1214–1218

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Heitz T, Bergey D, Ryan C (1997) A gene encoding a chloroplast-targeted lipoxygenase in tomato leaves is transiently induced by wounding, systemin, and methyl jasmonate. Plant Physiol 114:1085–1093

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271–282

    Article  CAS  PubMed  Google Scholar 

  • Höhne M, Nellen A, Schwennesen K, Kindl H (1996) Lipid body lipoxygenase characterized by protein fragmentation, cDNA sequence and very early expression of the enzyme during germination of cucumber seeds. Eur J Biochem 241:6–11

    Article  PubMed  Google Scholar 

  • Huang A (1996) Oleosins and oil bodies in seeds and other organs. Plant Physiol 110:1055

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Kindl H (1997) The oxygen-dependent modification of triacylglycerols and phospholipids, the different way of initiating lipid body mobilization. Zeitschrift für Naturforschung C A journal of biosciences 52:1–8

    CAS  Google Scholar 

  • Kuhn H, Thiele B (1999) The diversity of the lipoxygenase family—many sequence data but little information on biological significance. FEBS Lett 449:7–11

    Article  CAS  PubMed  Google Scholar 

  • Li H, Jiang L, Youn JH, Sun W, Cheng Z, Jin T, Ma X, Guo X, Wang J, Zhang X, Wu F, Wu C, Kim SK, Wan J (2013) A comprehensive genetic study reveals a crucial role of CYP90D2/D2 in regulating plant architecture in rice (Oryza sativa). New Phytol 200:1076–1088

    Article  CAS  PubMed  Google Scholar 

  • Long Q, Zhang W, Wang P, Shen W, Zhou T, Liu N, Wang R, Jiang L, Huang J, Wang Y (2013) Molecular genetic characterization of rice seed lipoxygenase 3 and assessment of its effects on seed longevity. J Plant Biol 56:232–242

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

    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 

  • Moore R (1973) Tetrazolium staining for assessing seed quality. Seed Ecol 11–23

  • Nellen A, Rojahn B, Kindl H (1995) Lipoxygenase forms located at the plant plasma-membrane. Zeitschrift Fur Naturforschung C A Journal of Biosciences 50:29–36

    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 

  • Oliw E (2002) Plant and fungal lipoxygenases. Prostaglandins Other Lipid Mediators 68–9:313–323

    Article  Google Scholar 

  • Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res 29:e45–e45

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rosahl S (1996) Lipoxygenases in plants—their role in development and stress response. Zeitschrift Fur Naturforschung C A Journal of Biosciences 51:123–138

    CAS  Google Scholar 

  • Royo J, Vancanneyt G, Perez A, Sanz C, Stormann K, Rosahl S, SanchezSerrano J (1996) Characterization of three potato lipoxygenases with distinct enzymatic activities and different organ-specific and wound-regulated expression patterns. J Biol Chem 271:21012–21019

    Article  CAS  PubMed  Google Scholar 

  • Salama AM, Pearce RS (1993) Ageing of cucumber and onion seeds: phospholipase D, lipoxygenase activity and changes in phospholipid content. J Exp Bot 44:1253–1265

    Article  CAS  Google Scholar 

  • Shibata D, Slusarenko A, Casey R, Hildebrand D, Bell E (1994) Lipoxygenases. Plant Mol Biol Report 12:S41–S42

    Article  CAS  Google Scholar 

  • Shin J-H, Kim S-R, An G (2009) Rice aldehyde dehydrogenase7 is needed for seed maturation and viability. Plant Physiol 149:905–915

    Article  CAS  PubMed  PubMed Central  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 

  • Siddique SB, Seshu D, Pardee WD (1988) Rice cultivars variability in tolerance for accelerated aging of seed [Philippines]. IRRI research paper series

  • Siedow J (1991) Plant lipoxygenase-structure and function. Annu Rev Plant Physiol Plant Mol Biol 42:145–188

    Article  CAS  Google Scholar 

  • Stephenson L, Bunker T, Dubbs W, Grimes H (1998) Specific soybean lipoxygenases localize to discrete subcellular compartments and their mRNAs are differentially regulated by source-sink status. Plant Physiol 116:923–933

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stewart RR, Bewley JD (1980) Lipid peroxidation associated with accelerated aging of soybean axes. Plant Physiol 65:245–248

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Takano K (1993) Advances in cereal chemistry and technology in Japan. Cereal Foods World 38:695–698

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vertucci CW, Roos EE (1990) Theoretical basis of protocols for seed storage. Plant Physiol 94:1019–1023

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang R, Shen W, Liu L, Jiang L, Liu Y, Su N, Wan J (2008) A novel lipoxygenase gene from developing rice seeds confers dual position specificity and responds to wounding and insect attack. Plant Mol Biol 66:401–414

    Article  CAS  PubMed  Google Scholar 

  • Wang F, Wang R, Jing W, Zhang W (2012) Quantitative dissection of lipid degradation in rice seeds during accelerated aging. Plant Growth Regul 66:49–58

    Article  CAS  Google Scholar 

  • Warthmann N, Chen H, Ossowski S, Weigel D, Hervé P (2008) Highly specific gene silencing by artificial miRNAs in rice. PLoS ONE 3:e1829

    Article  PubMed  PubMed Central  Google Scholar 

  • Wilson D Jr, McDonald M Jr (1986) The lipid peroxidation model of seed ageing. Seed Sci Technol 14:269–300

    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 

  • Zhang Y, Su J, Duan S, Ao Y, Dai J, Liu J, Wang P, Li Y, Liu B, Feng D, Wang J, Wang H (2011) A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods 7:30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zou Y, Kim AR, Kim JE, Choi JS, Chung HY (2002) Peroxynitrite scavenging activity of sinapic acid (3, 5-dimethoxy-4-hydroxycinnamic acid) isolated from Brassica juncea. J Agric Food Chem 50:5884–5890

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We wish to thank Prof. Mingliang Xu for critical review of the draft manuscript. We also gratefully acknowledge Xi Liu for excellent field work. This project was financially supported by the National Transformation Science and Technology Program (2013ZX08001-006).

Author information

Authors and Affiliations

  1. State Key Laboratory for Crop Genetics and Germplasm Enhancement, Jiangsu Provincial Center of Plant Gene Engineering, Nanjing Agricultural University, Weigang 1, Nanjing, 210095, China

    Jiexue Huang, Maohong Cai, Qizhang Long, Linglong Liu, Qiuyun Lin, Ling Jiang, Saihua Chen & Jianmin Wan

  2. Institute of Crop Science, The National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, Beijing, 10081, China

    Jianmin Wan

Authors
  1. Jiexue Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maohong Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qizhang Long
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Linglong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qiuyun Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ling Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Saihua Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jianmin Wan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Saihua Chen or Jianmin Wan.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 11 kb)

11248_2014_9803_MOESM2_ESM.pdf

Fig. 1 Purification and SDS-PAGE of the HIS and HIS-LOX2 proteins. HIS and HIS-LOX2 fusion proteins induced by 0.1 mM IPTG at 18 °C for 16 h were purified by His•Bind® kits (Novagen). The purified proteins were electrophoresed on 15 % SDS–polyacrylamide gel (SDS-PAGE). (M) High molecular weight marker; (1) Soluble purified HIS protein from E.coli. BL21 (DE3) harboring pET30a; (2) Soluble purified HIS-LOX2 fusion protein from E.coli. BL21 (DE3) harboring pET30a-OsLOX2; Arrows indicate the positions of target proteins. (PDF 15 kb)

11248_2014_9803_MOESM3_ESM.pdf

Fig. 2 Seed survival curve of Wuyunjing 7 and Nipponbare. The survival curve was made by the germination rate of Wuyunjing 7 (upper) and Nipponbare (lower) seeds after different days of accelerated aging. Values are mean ± SD of three replicates. The LD50 of Wuyunjing 7 and Nipponbare were about 16 days and 26 days after accelerated aging, respectively. (PDF 13 kb)

11248_2014_9803_MOESM4_ESM.pdf

Fig. 3 Seed viability test by Tetrazolium assay. (a) Staining pattern in seeds with or without vigor. After Tetrazolium assay, the embryo of the viable seeds can be stained deeply and totally as uppermost panel indicated, whereas, seed without vigor shows a light stain pattern in partial or stainless (lower three pannels). (b-c) After 25 days accelerated aging, seed viability of Nipponbare (b) and hRNAi9 (c) were identified by TTC and calculated by the percentage of deeply stained seeds. (PDF 60 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huang, J., Cai, M., Long, Q. et al. OsLOX2, a rice type I lipoxygenase, confers opposite effects on seed germination and longevity. Transgenic Res 23, 643–655 (2014). https://doi.org/10.1007/s11248-014-9803-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-014-9803-2

Keywords

Search

Navigation