Overexpression of SlARG2 or SlTD2 in Arabidopsis enhances resistance against Plutella xylostella L.
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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.
KeywordsArabidopsis Insect-resistance Overexpression Diamondback moth SlARG2 SlTD2
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).
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.
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