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A new method for rapid visualization of defects in leaf cuticle reveals five intrinsic patterns of surface defects in Arabidopsis
Author(s) -
Tanaka Toshihiro,
Tanaka Hirokazu,
Machida Chiyoko,
Watanabe Masaru,
Machida Yasunori
Publication year - 2004
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.2003.01946.x
Subject(s) - cuticle (hair) , plant cuticle , epidermis (zoology) , mutant , biology , arabidopsis , microbiology and biotechnology , biophysics , arabidopsis thaliana , botany , anatomy , biochemistry , gene , wax
Summary The epidermis of higher plants generates the cuticle layer that covers the outer surface of each plant. The cuticle plays a crucial role in plant development, and some mutants with defective cuticle exhibit morphological abnormalities, such as the fusion of organs. The way in which the cuticle forms and its contribution to morphogenesis are poorly understood. Conventional detection of the cuticle by transmission electron microscopy (TEM) requires laborious procedures, which include fixation, staining with osmium, and preparation of ultra‐thin sections. It is also difficult to survey entire surfaces of expanded leaves because of the limited size of specimens that can be examined. Thus, TEM is unsuitable for large‐scale screening for mutants with defective cuticle. We describe here a rapid and inexpensive method, designated the toluidine‐blue (TB) test, for detection of cuticular defects in whole leaves. We demonstrated the validity of the TB test using mutants of Arabidopsis thaliana , including abnormal leaf shape1 ( ale1 ), fiddlehead ( fdh ), and five e cer iferum ( cer ) mutants, in which the structure and/or function of the cuticle is abnormal. Genetic screening for mutants using the TB test allowed us to identify seven loci. The cuticle‐defective regions of leaves of the mutants revealed five intrinsic patterns of surface defects (classes I through V), suggesting that formation of functional cuticle on leaves involves various spatially regulated factors.