Unidirectional movement of cellulose synthase complexes in Arabidopsis seed coat epidermal cells deposit cellulose involved in mucilage extrusion, adherence, and ray formation.
Cellulose synthase5 (CESA5) synthesizes cellulose necessary for seed mucilage adherence to seed coat epidermal cells of Arabidopsis (Arabidopsis thaliana). The involvement of additional CESA proteins in this process and details concerning the manner in which cellulose is deposited in the mucilage pocket are unknown. Here, we show that both CESA3 and CESA10 are highly expressed in this cell type at the time of mucilage synthesis and localize to the plasma membrane adjacent to the mucilage pocket. The isoxaben resistant1-1 and isoxaben resistant1-2 mutants affecting CESA3 show defects consistent with altered mucilage cellulose biosynthesis. CESA3 can interact with CESA5 in vitro, and green fluorescent protein-tagged CESA5, CESA3, and CESA10 proteins move in a linear, unidirectional fashion around the cytoplasmic column of the cell, parallel with the surface of the seed, in a pattern similar to that of cortical microtubules. Consistent with this movement, cytological evidence suggests that the mucilage is coiled around the columella and unwinds during mucilage extrusion to form a linear ray. Mutations in CESA5 and CESA3 affect the speed of mucilage extrusion and mucilage adherence. These findings imply that cellulose fibrils are synthesized in an ordered helical array around the columella, providing a distinct structure to the mucilage that is important for both mucilage extrusion and adherence.
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Jonathan S
http://orcid.org/0000-0003-4512-7374
JS
Griffiths
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Krešimir
http://orcid.org/0000-0003-3030-7129
K
Šola
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Rekha
http://orcid.org/0000-0002-1910-9802
R
Kushwaha
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Patricia
P
Lam
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Mizuki
http://orcid.org/0000-0002-2206-190X
M
Tateno
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Robin
R
Young
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Cătălin
http://orcid.org/0000-0001-9105-014X
C
Voiniciuc
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Gillian
http://orcid.org/0000-0003-1126-8355
G
Dean
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Shawn D
http://orcid.org/0000-0002-0175-554X
SD
Mansfield
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546.
Seth
S
DeBolt
Department of Botany (J.S.G., K.Š., P.L., R.Y., C.V., G.D., G.W.H.) and Department of Wood Science (S.D.M.), University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4; andDepartment of Horticulture Plant Physiology/Biochemistry/Molecular Biology Program (R.K., M.T., S.D.) and University of Kentucky Seed Biology Group (R.K., M.T., S.D.), University of Kentucky, Lexington, Kentucky 40546 george.haughn@ubc.ca.
George W
http://orcid.org/0000-0001-8164-8826
GW
Haughn
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D010368:Pectins / Q000378:metabolism; D019441:Plant Epidermis / Q000166:cytology*; D062087:Plant Mucilage / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012639:Seeds / Q000166:cytology*; D016335:Zinc Fingers
D029681:Arabidopsis Proteins; D009097:Multienzyme Complexes; D062087:Plant Mucilage; D049452:Green Fluorescent Proteins; D010368:Pectins; D002482:Cellulose; D005964:Glucosyltransferases; C478648:cellulose synthase