1 <& /page/page_title.mas, title => 'The Tomato Fruit Cuticular Cell Wall Proteome' &>
3 <div class="indentedcontent">
5 The plant cuticle is a highly specialized hydrophobic cell wall that
6 covers the aerial organs of land plants and provides protection
7 against desiccation, pathogens, UV radiation and herbivory. It is
8 continuous with the outer-periclinal polysaccharide cell wall of the
9 epidermis and consists of organic soluble waxes embedded in, and
10 layered on, a non-soluble polyester matrix of ω-substituted
11 fatty acids. The waxes include both aliphatic compounds, derived from
12 very long chain fatty acids, and secondary metabolites, such as
13 triterpenoids and flavonoids.
17 Little is yet known about many key mechanisms of cuticle biogenesis,
18 and experimental strategies to identify new proteins that associate
19 with cutin and waxes could provide a valuable means to identify new
24 Although Arabidopsis research has greatly accelerated the discovery of
25 new cuticle-related genes, its cuticle poses some experimental
26 limitations since it is relatively thin, fragile and difficult to
27 isolate in substantial quantities. Conversely, tomato fruit cuticles
28 are astomatous and large amounts of intact cuticular material can be
29 isolated for chemical and biomechanical analyses. For example, the
30 fruit accumulate on the order of 1 mg cm<sup>-2</sup> cutin,
31 compared to the stem of Arabidopsis, which has 0.5-10 μg
32 cm<sup>-2</sup>. Thus, the typical 6 week period of early tomato
33 fruit development represents a remarkably rapid and extensive phase of
37 <div style="width: 420px" class="captioned_image img_float_left caption_right">
39 <img src="/documents/img/secretom/tomato_cuticle_stain_400.jpg" />
42 Light microscope image of the surface of an expanding tomato fruit
43 with the cuticle stained red.
49 To identify new candidate proteins with a potential role in cuticle
50 biosynthesis, and particularly those that are secreted to the
51 apoplast, we are generating a more comprehensive inventory of proteins
52 that are associated with the outermost surface tissues of plant
53 organs, we are using a range of complementary protein fractionation
54 strategies (1D gel-based and liquid chromatography) coupled with
55 several sensitive mass spectrometry (MS) platforms.
59 We are also characterizing a number of tomato fruit cuticle mutants
60 and have developing a new 3D imaging technique using confocal scanning
61 laser microscopy for visualizing the architecture, deposition patterns
62 and micro-structure of plant cuticles. This has allowed 3-D cuticle
63 modeling based on the reconstruction of serial optical sections and
64 its use in the identification of several previously unreported
65 features of the tomato fruit cuticle. This information, together with
66 the proteome study is providing new insights into the molecular
67 processes underlying cuticle formation and restructuring.
71 <div style="width: 370px; margin-left: 0" class="captioned_image img_float_right">
73 <img src="/documents/img/secretom/tomato_cuticle_3d_350.jpg" />
76 A 3-D rendering of the tomato fruit (cv. M8) cuticle.
81 <div style="float: left; width: 345px">
82 <&| /secretom/section_templates/objectives.mas &>
84 Isolate and identify proteins from the surface of expanding tomato
85 fruits at the time of maximal cuticle biosynthesis, using solvent
86 dips followed by gel- and liquid chromatography-based separation and
89 Identify potentially secreted proteins and characterize their
90 expression patterns during fruit ontogeny
92 Incorporate this information into related studies of the tomato
93 fruit cuticle t develop a more detailed understanding of cuticle
94 deposition and restructuring
99 <&| /secretom/section_templates/data_items.mas,
100 default_ref_base => '/download/data/secretom/Tomato_fruit_cuticular_cell_wall_proteome'
102 - text: "Tomato fruit (cv. M82) surface proteins identified"
103 ref: Tomato_fruit_surface_proteins.xlsx
104 - text: "Tomato fruit (cv. M82) surface proteins: from gel bands"
105 ref: Tomato_fruit_surface_proteins_gel_bands.xlsx
106 - text: "Tomato fruit (cv. M82) surface proteins: from gel slabs"
107 ref: Tomato_fruit_surface_proteins_gel_slabs.xlsx
108 - text: "Tomato fruit (cv. M82) surface proteins: from gel free"
109 ref: Tomato_fruit_surface_proteins_gel_free.xlsx
112 <&| /secretom/section_templates/publications.mas, entitize => 1 &>
114 Yeats, T.H. and Rose, J.K.C. (2008) The biochemistry and biology of extracellular plant lipid-transfer proteins (LTPs) Protein Science 17: 191-198.
116 Buda, G.J., Isaacson, T., Matas, A.J., Paolillo, D.J. and Rose, J.K.C. (2009) Three dimensional imaging of plant cuticle architecture using confocal scanning laser microscopy. The Plant Journal 60: 378-385 (front cover).
118 Isaacson, T., Kosma, D.K., Matas, A.J., Buda, G.J., He, Y., Yu, B., Pravitasari, A., Batteas, J.D., Stark, R.E., Jenks, M.A. and Rose, J.K.C. (2009) Cutin deficiency in the tomato fruit cuticle consistently affects resistance to microbial infection and biomechanical properties, but not transpirational water loss. The Plant Journal 60: 363-377.
120 Yeats, T.H., Howe, K.J., Matas, A.J., Buda, G.J., Thannhauser, T.W. and Rose, J.K.C. (2010) Mining the tomato fruit cuticular and epidermal cell wall proteome for proteins associated with cuticle biogenesis. Journal of Experimental Botany 61: 3759-3771.