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Plant Molecular Biology Reporter

, Volume 33, Issue 4, pp 962–974 | Cite as

The TsnsLTP4, a Nonspecific Lipid Transfer Protein Involved in Wax Deposition and Stress Tolerance

  • Wei Sun
  • Yan Li
  • Yanxiu Zhao
  • Hui ZhangEmail author
Original Paper

Abstract

Nonspecific lipid transfer proteins (nsLTPs), a group of small, basic proteins that are ubiquitously distributed throughout the plant kingdom, are thought to participate in cutin formation as well as in defense reactions against abiotic and biotic stresses. However, whether nsLTPs are involved in cuticular wax deposition remains unknown. We identified a salt-induced gene, TsnsLTP4, encoding an nsLTP from Thellungiella salsuginea. TsnsLTP4 expression was significantly induced by salt, polyethylene glycol (PEG), abscisic acid (ABA), and high (37 °C) and low (4 °C) temperatures. Transgenic onion epidermal cells transiently expressing a TsnsLTP4-DsRed fusion protein demonstrated that TsnsLTP4 was targeted to the cell wall. Overexpression of TsnsLTP4 in transgenic Arabidopsis plants resulted in increased epicuticular wax deposition, particularly of wax components with a carbon chain length of more than C27. Moreover, the amount of wax deposited was strongly reduced in RNA interference (RNAi)-knockdown TsnsLTP4 transgenic T. salsuginea lines. Cuticle permeability was inversely related to the expression level of TsnsLTP4. Further analyses indicated that overexpression of TsnsLTP4 resulted in significantly enhanced drought and salt tolerance in transgenic Arabidopsis. In summary, our studies suggested that TsnsLTP4 may play a role in wax deposition and in plant tolerance against abiotic stresses.

Keywords

Stress tolerance Thellungiella salsuginea TsnsLTP4 Wax deposition 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (30470158), the National Program on Key Basic Research Project of China (2012CB114204), Science Foundation for The Excellent Youth Scientists of Shandong Province (BS2009NY038), and funds from Shandong Normal University (Dr Foundation). We sincerely thank Dr. Chang le Ma for comments on our manuscript.

Supplementary material

11105_2014_798_Fig9_ESM.gif (98 kb)
Fig. S1

Northern blot analysis of the expression of the TsnsLTP4. Total RNA samples from leaves of plants were analyzed. The plants were untreated or treated with 4 °C, 19 mM PEG, or 100 mM NaCl for various durations, as indicated. The cDNA of TsnsLTP4 were labeled with 32P as probes. 18S rRNA was used as an internal control of characterized total RNA. The bottom panels are calculations of the relative expression. (GIF 98 kb)

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Fig. S2

TsnsLTP4 gene expression analysis by Northern blot in leaves of transgenic plants. a The TsnsLTP4 gene expression level of the overexpression lines of Arabidopsis compared with the wild-type Arabidopsis and T. salsuginea. b The transgenic lines of T. salsuginea harboring the RNAi construct compared with the wild-type T. salsuginea. 18S rRNA was used as an internal control of characterized total RNA. The bottom panels are calculations of the relative expression. (GIF 73 kb)

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Fig. S2

TsnsLTP4 gene expression analysis by Northern blot in leaves of transgenic plants. a The TsnsLTP4 gene expression level of the overexpression lines of Arabidopsis compared with the wild-type Arabidopsis and T. salsuginea. b The transgenic lines of T. salsuginea harboring the RNAi construct compared with the wild-type T. salsuginea. 18S rRNA was used as an internal control of characterized total RNA. The bottom panels are calculations of the relative expression. (GIF 73 kb)

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Fig. S3

Phenotypes of wild-type and Ts-nsLTP overexpression plants under 19 mM PEG − 6,000 and 200 mM mannitol. Similar results were observed in three independent experiments. a PEG tolerance assay of the wild type and TsnsLTP overexpression plants. The four weeks plants were grown in soil and then watered 19 mM PEG − 6,000 four days. b Fresh and c dry weight of the plants in a under PEG stress. Statistical significance of the measurements was determined using a Student’s t test (a indicate P < 0.01) by comparing control and treatment plants, (b indicate P < 0.01) by comparing wild-type and transgenic plants under same concentration PEG treatment. d The MDA contents in the wild type plants and the transgenic plants under 200 mM mannitol treatment. The data presented are the means ± SD (n = 12); **P < 0.01 (Student’s t test). (GIF 292 kb)

11105_2014_798_Fig13_ESM.gif (66 kb)
Fig. S3

Phenotypes of wild-type and Ts-nsLTP overexpression plants under 19 mM PEG − 6,000 and 200 mM mannitol. Similar results were observed in three independent experiments. a PEG tolerance assay of the wild type and TsnsLTP overexpression plants. The four weeks plants were grown in soil and then watered 19 mM PEG − 6,000 four days. b Fresh and c dry weight of the plants in a under PEG stress. Statistical significance of the measurements was determined using a Student’s t test (a indicate P < 0.01) by comparing control and treatment plants, (b indicate P < 0.01) by comparing wild-type and transgenic plants under same concentration PEG treatment. d The MDA contents in the wild type plants and the transgenic plants under 200 mM mannitol treatment. The data presented are the means ± SD (n = 12); **P < 0.01 (Student’s t test). (GIF 292 kb)

11105_2014_798_Fig14_ESM.gif (63 kb)
Fig. S3

Phenotypes of wild-type and Ts-nsLTP overexpression plants under 19 mM PEG − 6,000 and 200 mM mannitol. Similar results were observed in three independent experiments. a PEG tolerance assay of the wild type and TsnsLTP overexpression plants. The four weeks plants were grown in soil and then watered 19 mM PEG − 6,000 four days. b Fresh and c dry weight of the plants in a under PEG stress. Statistical significance of the measurements was determined using a Student’s t test (a indicate P < 0.01) by comparing control and treatment plants, (b indicate P < 0.01) by comparing wild-type and transgenic plants under same concentration PEG treatment. d The MDA contents in the wild type plants and the transgenic plants under 200 mM mannitol treatment. The data presented are the means ± SD (n = 12); **P < 0.01 (Student’s t test). (GIF 292 kb)

11105_2014_798_Fig15_ESM.gif (35 kb)
Fig. S3

Phenotypes of wild-type and Ts-nsLTP overexpression plants under 19 mM PEG − 6,000 and 200 mM mannitol. Similar results were observed in three independent experiments. a PEG tolerance assay of the wild type and TsnsLTP overexpression plants. The four weeks plants were grown in soil and then watered 19 mM PEG − 6,000 four days. b Fresh and c dry weight of the plants in a under PEG stress. Statistical significance of the measurements was determined using a Student’s t test (a indicate P < 0.01) by comparing control and treatment plants, (b indicate P < 0.01) by comparing wild-type and transgenic plants under same concentration PEG treatment. d The MDA contents in the wild type plants and the transgenic plants under 200 mM mannitol treatment. The data presented are the means ± SD (n = 12); **P < 0.01 (Student’s t test). (GIF 292 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Key Laboratory of Plant Stress Research, College of Life ScienceShandong Normal UniversityJinanChina

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