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Transgenic Research

, Volume 23, Issue 4, pp 643–655 | Cite as

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

  • Jiexue Huang
  • Maohong Cai
  • Qizhang Long
  • Linglong Liu
  • Qiuyun Lin
  • Ling Jiang
  • Saihua ChenEmail author
  • Jianmin WanEmail author
Original Paper

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.

Keywords

Oryza sativa L. OsLOX2 Seed germination Seed longevity Grain storage 

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

Notes

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).

Supplementary material

11248_2014_9803_MOESM1_ESM.pdf (11 kb)
Supplementary material 1 (PDF 11 kb)
11248_2014_9803_MOESM2_ESM.pdf (15 kb)
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 (13 kb)
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 (61 kb)
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)

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Copyright information

© Springer International Publishing Switzerland 2014

Authors and Affiliations

  • Jiexue Huang
    • 1
  • Maohong Cai
    • 1
  • Qizhang Long
    • 1
  • Linglong Liu
    • 1
  • Qiuyun Lin
    • 1
  • Ling Jiang
    • 1
  • Saihua Chen
    • 1
    Email author
  • Jianmin Wan
    • 1
    • 2
    Email author
  1. 1.State Key Laboratory for Crop Genetics and Germplasm Enhancement, Jiangsu Provincial Center of Plant Gene EngineeringNanjing Agricultural UniversityNanjingChina
  2. 2.Institute of Crop Science, The National Key Facility for Crop Gene Resources and Genetic ImprovementChinese Academy of Agricultural SciencesBeijingChina

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