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Plant and Cell Physiology Advance Access published online on August 21, 2006
Plant and Cell Physiology, doi:10.1093/pcp/pcj097
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received April 18, 2006
Accepted July 25, 2006

Regular Paper

Nightshade proteinase inhibitor IIb gene is constitutively expressed in glandular trichomes

Jin Liu 1, Kuai-Fei Xia 1, Jing-Chun Zhu 2, Yu-Ge Deng 2, Xiao-Le Huang 2, Bo-Lun Hu 2, Xinping Xu 2, and Zeng-Fu Xu 2 *

1 State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China;These authors contributed equally to this work.
2 State Key Laboratory of Biocontrol and Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China

* To whom correspondence should be addressed.
Zeng-Fu Xu, E-mail: xuzengfu{at}mail.sysu.edu.cn


  Abstract

The best-known property of plant proteinase inhibitor II (PIN2) genes is their wound-inducible expression in leaves and constitutive expression in flowers. Here we show by promoter analysis in transgenic plants and in situ RT-PCR analysis that SaPIN2b, a member of the PIN2 gene family of nightshade (Solanum americanum), is also constitutively expressed in glandular trichomes. SaPIN2b promoter and its deletions were cloned and fused to the upstream of {beta}-glucuronidase (GUS) to transform the nightshade and tobacco (Nicotiana tabacum) plants. Histochemical staining assays indicated that, SaPIN2b:GUS expressed constitutively in glandular trichomes, predominantly in the gland cells, of both transgenic nightshade and tobacco plants. Constitutive expression of SaPIN2b in glandular trichomes was further confirmed by liquid phase in situ RT-PCR analysis of nightshade leaves. Deletion analysis from the 5' end of SaPIN2b promoter revealed that separate regulatory elements control SaPIN2b expression in gland cells and stalk cells of glandular trichomes. Fluorometric GUS assays showed that SaPIN2b:GUS expression was significantly increased in transgenic plant leaves after mechanical wounding or methyl jasmonate treatment. SaPIN2b promoter sequence contains six MYB binding motifs and a L1 box that are involved in trichome differentiation and development. Overexpression of SaPIN2b in tobacco resulted in a significant increase in glandular trichome density and promoting trichome branching. These results suggest that, other than an induced defensive protein of the well-known PIN2 family, SaPIN2b could also play roles in trichome-based defense by functioning as a constitutive component of trichome chemical defense and/or by regulating the development of glandular trichomes.

Keywords: GUS; Nicotiana tabacum; Promoter; Protease inhibitor; Solanum americanum; Trichomes.

The nucleotide sequence reported in this paper has been submitted to GenBank under accession number AY749108.


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