Identification and characterization of spruce genes involved in somatic embryo development

dc.contributor.authorLaw, Derek Albert
dc.contributor.examiningcommitteeDaayf, Fouad (Plant Science) Piercey-Normore, Michele (Botany)en
dc.contributor.supervisorStasolla, Claudio (Plant Science)en
dc.date.accessioned2006-07-12T15:37:40Z
dc.date.available2006-07-12T15:37:40Z
dc.date.issued2006-07-12T15:37:40Z
dc.degree.disciplinePlant Scienceen_US
dc.degree.levelMaster of Science (M.Sc.)en_US
dc.description.abstractSomatic embryogenesis can provide researchers with an important tool to study the physiological and molecular mechanisms involved in embryo development. In spruce, few lines are able to produce fully developed embryos due to the presence of malformed meristems. Genes from two families known as KNOX (knotted-like homeobox) and ARGONAUTE (AGO) have previously been found to be involved in meristem development and maintenance. This work documents the discovery of a new member of the AGO family of proteins designated as PgAGO and the further study of a KNOX gene known as HBK2. The complete coding sequence of both PgAGO and HBK2 was obtained through screening of cDNA libraries generated from white spruce (Picea glauca) somatic embryos. RNA in-situ hybridization studies showed that PgAGO mRNAs accumulate preferentially within cells of the shoot and root apical meristems in developing spruce embryos. In addition, the expression of PgAGO was low in white spruce lines unable to produce embryos in culture. Norway spruce (Picea abies) embryogenic tissue was transformed via microprojectile bombardment with an antisense construct of PgAGO. Down-regulation of PgAGO altered proper development of the apical meristems and reduced embryo regeneration. RNA in-situ hybridization studies showed that HBK2 is specifically expressed in the sub-apical and cortical regions of developing embryos. Like PgAGO, HBK2 expression was diminished in white spruce lines unable to produce embryos in culture. Transformation experiments with antisense constructs of HBK2 completely arrested somatic embryo development. This study reveals the importance of a functional meristem during embryo development.en
dc.description.noteOctober 2006en
dc.format.extent2447809 bytes
dc.format.mimetypeapplication/pdf
dc.identifier.citationTahir, Muhammad, Law, Derek Albert and Stasolla, Claudio (2006). Molecular characterization of PgAGO, a novel conifer gene of the ARGONAUTE family expressed in the apical cells and required for somatic embryo development in spruce. Tree Physiology (in press)en
dc.identifier.urihttp://hdl.handle.net/1993/261
dc.language.isoengen_US
dc.rightsopen accessen_US
dc.subjectsomaticen
dc.subjectembryogenesisen
dc.subjecthomeoboxen
dc.subjectargonauteen
dc.subjectmeristemen
dc.titleIdentification and characterization of spruce genes involved in somatic embryo developmenten
dc.typemaster thesisen_US
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