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Phytoparasitica

, Volume 45, Issue 5, pp 695–705 | Cite as

Overexpression of SlARG2 or SlTD2 in Arabidopsis enhances resistance against Plutella xylostella L.

  • Huixin Geng
  • Qiujie Zhang
  • Xiao Yang
  • Shuzhen MenEmail author
Article
  • 94 Downloads

Abstract

Tomato (Solanum lycopersicum L.) ARGINASE2 (ARG2) and THREONINE DEAMINASE2 (TD2) are involved in plant defense. These enzymes act in the midgut of herbivores fed on tomato plants to degrade the essential amino acids Arg and Thr, respectively. Although it has been demonstrated that overexpression of the SlARG2 gene in tomato enhanced its resistance against M. sexta larvae, knock-down the expression of SlTD2 reduced the resistance of tomato to lepidopteran herbivores; it remains unclear whether overexpression of SlTD2 could enhance the resistance of the host plants to herbivores, or whether combined overexpression of SlARG2 and SlTD2 could lead to synergistically enhanced resistance to insects. Here, we generated transgenic Arabidopsis plants overexpressing SlARG2 (SlARG2 OE) and SlTD2 (SlTD2 OE) individually as well as in combination (SlARG2-SlTD2 OE). Overexpression of these genes did not affect Arabidopsis development, seed yield, or Arg and Thr content. Insect-feeding bioassay was performed by feeding diamondback moth (Plutella xylostella L.) larvae on detached leaves of wild-type, SlARG2 OE, SlTD2 OE, and SlARG2-SlTD2 OE plants. Larvae fed on SlARG2 OE leaves showed approximately 31% to 35% reduction in weight and 6% to 10% reduction in survival rate compared to those fed on wild-type leaves. Although larvae fed on SlTD2 OE leaves showed no reduction in survival rate, they gained less weight. Whereas larvae fed on SlARG2-SlTD2 OE leaves showed neither reduction in weight nor reduction in survival rate. We further investigated the arginase enzymatic activity of the SlARG2 OE and SlARG2-SlTD2 OE transgenic plants. The SlARG2 OE line most resistant to diamondback moth larvae displayed the highest arginase activity. Our data indicate that overexpression of SlARG2 or SlTD2 in Arabidopsis can enhance its resistance against diamondback moth, whereas combined overexpression of SlARG2 and SlTD2 did not generate synergistically increased resistance to diamondback moth.

Keywords

Arabidopsis Insect-resistance Overexpression Diamondback moth SlARG2 SlTD2 

Notes

Acknowledgements

We thank Gregg A. Howe for providing the pB2GW7-ARG2 and pBI121-TD2 plasmids, Nam-Hai Chua for providing the pBA002 plasmid, Shaohui Yang and Jiehua Wang for providing diamondback moth eggs, Tongmei Xia for helping with amino acid measurement. This work was supported by the National Natural Science Foundation of China (31570247 and 31460453); the Major Technological Program on Cultivation of New Varieties of Genetically Modified Organisms (NO. 2011ZX08004-004); and the Natural Science Foundation of Tianjin (No. 14JCYBJC41200).

Author contributions

SM conceived and designed the experiments. QZ constructed the SlARG2-SlTD2 OE vector; generated SlARG2 OE, SlTD2 OE, and SlARG2-SlTD2 OE transgenic Arabidopsis plants; and performed the phenotypic analysis of these plants. HG performed the insect bioassay and arginase activity assay; and analyzed the amino acid content. HG, QZ, XY, and SM analyzed the data. XY and HG drafted the manuscript; SM rewrote most part of the manuscript.

Supplementary material

12600_2017_626_MOESM1_ESM.doc (46 kb)
ESM 1 (DOC 45 kb)
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Fig. S1 Life cycle of diamondback moth (Plutella xylostella L.). (A) Eggs. (B-D) Larvae at first (B), third (C), second (arrow in D), and fourth instar (arrowhead in D). (E) Pupa. (F-G) Adult in back (F) and bottom view (G). (TIFF 4796 kb)
12600_2017_626_Fig5_ESM.gif (176 kb)

High resolution image (GIF 176 kb)

12600_2017_626_MOESM3_ESM.tif (11.6 mb)
Fig. S2 Phenotypes of SlARG2 and SlTD2 OE seedlings. (A) Five-day-old seedling grown under 16-h light/8-h dark growth conditions. (B-D) Root and hypocotyl length of 5-day-old seedlings grown under 16-h light/8-h dark growth conditions (B, C) or in the dark (D, E). Data from three experiments are presented as the means ± SD (n = 20 for each experiment). Differences between the wild-type (WT) and transgenic lines were significant at P < 0.05 (*) or P < 0.01 (**) by Student’s t test. (TIFF 11846 kb)
12600_2017_626_Fig6_ESM.gif (182 kb)

High resolution image (GIF 181 kb)

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Plant Biology and Ecology, College of Life SciencesNankai UniversityTianjinChina

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