UBC Theses and Dissertations
Pectin methyl esterification functions in seed development and germination Levesque-Tremblay, Gabriel
Homogalacturonan pectin domains are synthesized in a highly methyl esterified form and can be de-methyl esterified by the cell wall enzyme Pectin Methyl Esterase (PME). The prevalent model for PME mode of action indicates that when PMEs act on a stretch of adjacent galacturonic acid glycosides, they may strengthen the cell wall but when PMEs act on non-adjacent galacturonic acid glycosides they may loosen the cell wall. PME activity can be regulated in planta by the proteinaceous inhibitor, PMEI. I used PME and PMEI to study the importance of methyl esterification in seed development and germination. As a means to identify PMEs involved in seed coat mucilage I identified 7 PMEs expressed in the seed coat. The PME gene HIGHLY METHYL ESTERIFIED SEED (HMS) is highly expressed at 7 Day Post Anthesis (DPA) both in the seed coat and the embryo. Using a hms-1 mutant, I showed that HMS is required for normal levels of PME activity and methyl esterification in the seed, mucilage extrusion and proper embryo cell expansion, rigidity and morphogenesis between 4 and 10 DPA. The mucilage extrusion defect is a secondary effect of the function of HMS in the embryo. I hypothesize that HMS is required for cell wall loosening in the embryo to allow for cell expansion during the accumulation of storage reserves. To evaluate the importance of methyl esterification in germination my collaborators and I first showed that PME activity changed during the different stages of germination: it first increased before testa rupture and decreased during endosperm rupture. Treatment with the hormone abscisic acid (ABA) to increase dormancy prolonged PME activity in the seeds. Inversely when we negatively regulated PME activity in the A. thaliana seed with the overexpression of a PMEI (OE PMEI5), we generated larger seeds with bigger cells. These seeds germinated faster both in presence or absence of ABA. Therefore we hypothesize that the PME(s) inhibited by PMEI5 establishes stronger cell walls that restrict germination. This thesis clearly demonstrates that PME activity is important in the regulation of seed cell wall methyl esterification impacting embryo growth and germination.
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