Abstract
HpaXm produced by the cotton leaf blight bacterium Xanthomonas citri subsp. malvacearum is a novel harpin elicitor of the induced hypersensitive response (HR) in tobacco. We investigated whether fragments of HpaXm, compared with fragments of Hpa1Xoo, are sufficient for HR or plant growth promotion (PGP) elicitation using four synthetic peptides (HpaXm35-51, HpaXm10-39, Hpa1Xoo36-52 and Hpa1Xoo10-40). We also heated the fragments to determine the heat tolerance of the functional fragments. HpaXm35-51 and Hpa1Xoo36-52 induced hypersensitive response (HR). Bursts of reactive oxygen intermediates (ROI) induced by HpaXm35-51 and Hpa1Xoo36-52 were earlier and stronger than those induced by HpaXm and Hpa1Xoo. In plants treated with HpaXm35-51 or Hpa1Xoo36-52, the expression of the HR marker genes Hin1 and Hsr203J and the active oxygen metabolism related gene AOX were significantly upregulated. These findings suggest that the predicted α-helical structures of the HpaXm35-51 and Hpa1Xoo36-52 fragments are crucial for HR. PGP result by soaking seeds in unheated/heated HpaXm10-39 or Hpa1Xoo10-40 solution prior to transfer, which obviously enhances root growth and the aerial parts of plants. The PGP related gene NtEXP6 was greatly enhanced when plants were sprayed with a solution of HpaXm10-39 or Hpa1Xoo10-40; heated fragment treatments induced higher levels of NtEXP6 expression than unheated HpaXm fragments. In addition, HR marker genes induced by the heated fragments had lower expression levels than when induced with unheated HpaXm fragments. Moreover, the expression levels of HR marker genes and PGP related genes induced by treatment with Hpa1Xoo fragments before or after heating were the opposite of those induced by HpaXm fragments. Different functional fragments of harpin and different harpins with the same functional region have different degrees of heat tolerance. Therefore, the heat resistance of harpin is conservative, but the degree of heat tolerance of the functional fragments is specific.
Highlights
Harpins, encoded by hrp genes, are secreted by Gram-negative bacteria during interactions with host plants to cause diseases [1]
The harpin-encoding gene hrpN of E. amylovora induces disease resistance through the systemic acquired resistance (SAR) pathway in Arabidopsis [3]; HrpN induces drought tolerance in Arabidopsis mediated by ABI2-dependent abscisic acid signaling [4]; Hpa1 of Xanthomonas axonopodis pv. glycines can elicit a typical HR in nonhost tobacco [5]; expression of hpaGXooc elicits hypersensitive response (HR), which induces disease- and insect-resistance in plants, and enhances plant growth [6]; HrpZ of Pseudomonas syringae pv. phaseolicola enhances resistance to rhizomania diseases in transgenic Nicotiana benthamiana and sugar beet [7]
The homologous position in HpaXm was threonine at the 44th amino acid site (Fig 1)
Summary
Harpins, encoded by hrp (hypersensitive response and pathogenicity) genes, are secreted by Gram-negative bacteria during interactions with host plants to cause diseases [1]. Glycines can elicit a typical HR in nonhost tobacco [5]; expression of hpaGXooc elicits hypersensitive response (HR), which induces disease- and insect-resistance in plants, and enhances plant growth [6]; HrpZ of Pseudomonas syringae pv. The HR elicitor activity of the synthetic peptide of this region is the same as that of the HpaG protein at the same concentration [8]; Hairpin XopA did not elicit HR in tobacco. Vesicatoria results in the lack of HR-eliciting activity [8] Both the P44 (aa 269–312) and P24 (aa 290–313) sequences, which represent putative α-helical fragments of HrpZ from Pseudomonas syringae pv. HpaXm of Xanthomonas citri subsp. malvacearum with two heptads from the N-terminal a-helical region of HpaXm displayed activity in inducing HR [14]
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