PLANT GALLS: A MODEL SYSTEM TO STUDY PLANT DEVELOPMENT
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Cynipid gall formation is achieved by the intimate insect-plant interaction where by cynipid wasps redirect host plant development to form novel structures to protect and nourish the developing larva. To investigate the molecular mechanisms involved in this interaction, and extend our understanding of plant development, four approaches were taken. 1) A PCR based approach to search for genes to known signalling molecules: chitiooligosaccharides, or Nod factors, that control nodulation in the Rhizobia-legume interaction. PCR analysis was used to investigate the presence of the nodC gene in the cynipid gall wasp genome,h owever, no nodC-like sequencesw ere found. 2) SDS-PAGE analysis was carried out to compare inner-gall and non-gall protein signatures, demonstrating the variation between gall and non-gall tissue, and also that the protein signatures of inner-gall tissues vary between gall species. N-terminal sequencing and western blot analysis lead to the identification of a number of innergall proteins such as protein disulphide isomerase (PDI), formate dehydrogenase (FDH) and putative biotin carboxyl carrier protein (BCCP), involved in the synthesis of lipids in seeds. Analysis of the temporal and spatial expression of the putative BCCP revealed expression to be concentrated in the inner-gall cells throughout development, in all the gall species tested. 3) Cytological analysis of the inner-gall tissue was carried out throughout development of several gall species to investigate differences in their patterns of development and cytological characteristics of the inner-gall tissue, with many inner-gall cells being polytene. 4) A gall formation bioassay, to enable the activity of possible signals involved in gall formation to be tested, was developed. Rose callus tissue was used as a test tissue and the cynipid larval extract was exposed to this as a source of the active molecules. The induction of proteins in the callus after exposure to the larval extract was used as a molecular marker for activity. The polytene characteristic and the possible expression of seed proteins, suggest that seed developmental pathways may be used during gall formation.
AuthorsHarper, Lisa Janine
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