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Kunitz-type trypsin inhibitor gene family in Arabidopsis and Populus trichocarpa and its expression response to wounding and herbivore in Populus nigra

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Abstract

Trypsin inhibitors (TIs) play important roles in plant defense against biotic stresses. In this work, we first characterized the KTI gene families in the herbaceous model system, Arabidopsis thaliana, and the woody model system, Populus trichocarpa. Genomic analysis of AtKTIs and PtKTIs including phylogenetic relationship, gene structure, and motif preservation was presented. The temporal and spatial expression pattern of AtKTI genes under biotic and abiotic stresses has been performed by mining the publicly available microarray data. Unlike Populus, the absence of AtKTI induction under herbivore attack suggested that AtKTIs may not be closely related to herbivore defense in this plant species. In order to assess the potential of PtKTI as target genes for genetic improvement of the biotic resistance in plant species of high economic impact, we isolated KTI complementary DNAs from Populus nigra on a genome-wide scale and analyzed their respective response to Apocheima cinerarius Erschoff and mechanical wounding. A basically similar subset of PnKTIs was shown to be rapidly induced by both treatments in this study, though quantitatively distinct. This study revealed a different collection of wound- and herbivore-induced P. nigra KTI genes from those reported previously for hybrid poplar infested with Malacosoma disstri. Our data demonstrated that Populus could deploy KTI genes actively and selectively in an insect-specific manner. While KTI genes constitute good candidates for genetic engineering to improve biotic resistance in transgenic plant, their selection should be herbivore-oriented to obtain sufficient effects.

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 30800875), Tianjin Research Program of Application Foundation and Advanced Technology (10JCYBJC08600), and the Ph.D. Programs Foundation of the Ministry of Education of China (No. 200800561034).

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Authors and Affiliations

  1. School of Agriculture and Bioengineering, Tianjin University, Tianjin, 300072, China

    Yuan Ma, Qing Zhao & Jiehua Wang

  2. Lab of Biotechnology, Research Institute of Forestry, Chinese Academy of Forestry, Beijing, 100091, China

    Meng-Zhu Lu

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  1. Yuan Ma
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  2. Qing Zhao
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  3. Meng-Zhu Lu
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  4. Jiehua Wang
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Corresponding author

Correspondence to Jiehua Wang.

Additional information

Communicated by W. Boerjan

Yuan Ma and Qing Zhao contributed to this paper equally.

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Supplementary Table 1

Primers used in genome cloning and quantifying PCR analysis (DOC 61 kb)

Supplementary Table 2

Kunitz-type trypsin inhibitor genes in Populus trichocarpa (DOC 91 kb)

Supplementary Table 3

Stress treatment method used in microarray analysis of AtKTIs (DOC 92 kb)

Supplementary Figure 1

Phylogenetic relationships of Arabidopsis thaliana, Populus trichocarpa, Oryza sativa, and Sorghum vulgare Kunitz-type trypsin inhibitor genes. Sequence phylogram was constructed by neighborhood-joining bootstrap method using ClustalX1.8 software based on the multiple alignments of the protein amino acid sequences of all the KTIs, and the radial tree was generated using TreeView software (Win 32, Version 1.6.6) (DOC 1544 kb)

Supplementary Figure 2

Distribution of KTI genes on P. trichocarpa chromosomes. The names of the chromosomes are indicated below each chromosome. Segmental duplicated homologous blocks on Populus (Kalluri et al. 2007) were indicated with the same color. The position of genes was indicated with a line. Chomosomal duplication gene was connected with a single line. Tandem duplication was marked by a vertical bar (DOC 491 kb)

Supplementary Figure 3

Clustal X alignments of the amino acid sequences of KTIs from Populus nigra. The highly conserved motif was indicated by gray shadow or uplined (DOC 3344 kb)

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Ma, Y., Zhao, Q., Lu, MZ. et al. Kunitz-type trypsin inhibitor gene family in Arabidopsis and Populus trichocarpa and its expression response to wounding and herbivore in Populus nigra . Tree Genetics & Genomes 7, 431–441 (2011). https://doi.org/10.1007/s11295-010-0345-3

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