The Transcriptomic Sequencing And Salt Resistance Related Gene Cloning And Identification Of The Enriched Salt Glandes Of Spartina Alterniflor

With the help of salt glands, the halophyte can secret excess salt and effectively reduce the salt concentration in plant to adapt to the salt environment. Salt gland is a specialized structure of the halophyte in order to adapt to the salt environment. For salt of physiological processes glandular secrete salt research indicates that, in high salt stress condition, salt gland can bring Na+ out from the plant body with a higher rate. Based on what's mentioned above, We can predict that there will be expression and regulation of relevant special genes which controls its development and the mechanism of the ion efficient transport. Therefore, the study of the development of salt gland can help to discover the genes that are related to the development of the salt gland and the efficient ion transportion, as well as the mechanism of it.Salt glands contains lots of informations which are related to the secretion salt, Transcriptomic analysis can be used to research of cells regulation of gene transcription and information of space-time in overall level, which is an effective method to study the molecular mechanisms of biological (organization) life process in regulating gene expression. In the salt induced cDNA library of Spartina alterniflor, there are many salt resistance related EST sequences which obviously differed in abundance and type, when compared with other species. The analysis of plant of sequences in the library shows that there exists inevitably connections between the expression mode and the salt secretion activities of its salt gland. However, in order to study the genetic mechanism of salt secretion by salt gland more accurately and on all sides, especially the mechanism of the genes which regulate ion transportion efficienttly. It is necessary to research the gene expression profile of salt gland, to analysis and research the expression and regulation of the genes, get the genetics information of the genes'composition and regulation.In recent years, the technique of Laser capture microscopic cutting (LCM) appears to greatly promote the study of gene expression profile of specific type of cell in the plant .This subject by using laser capture microscopic cutting (LCM) technology, use Spartina alterniflor which is highly resistant to salt as materials. Under the condition of salt induction, collect salt glands to research the gene expression profile, furtherly research the character of gene expression of salt gland in ion transportion and the vesicle-associated membrane protein , explore the role of them in the process of salt secreteion. Overexpressed the VAMP gene which is related to vesicles transport, studies the correlation of salt-tolerant of Spartina alterniflor, and analysis the preliminary function of the transgenic plants showed that the gene may participate in the response through regulate the related processes of glucose transporter under the salt stress. In this subject, we get pure transgenic straines, under the stresses of salt and glucose which have showed some patiences and sensitivity, indicating that, under the stress condition, VAMP genes regulate the plant's resist to adversity reaction.Innovation points1, This subject first attempt to use LCM technology to capture and separate precisely the salt gland of Spartina alterniflor; through the transcriptomic sequencing and gene expression profile analysis , discusses the molecular mechanism and expression of related-genes, salt refusion in plant and the ion compartmentalization, when different types of cells are under salt stress.2, This subject to optimize technical conditions for collect salt glands, such as the paraffin technology, scanning electron microscopic technique, laser cutting technology experiment condition, to observe cellular forms and distribution characteristics.3, This subject to optimize and explore the method and process of enrich salt gland and RNA extract , such as the optimization design of the method of high quality RNA extract, and the factors influencing the RNA quality in material pretreatment processes.4,This subject first attempt to transcriptomic sequencing and gene expression profile analysis of the halophyte, research the expression of the salt secretion related gene of salt gland, molecular mechanism, and explore the ion efficient transport mechanism of the salt secretion in salt gland molecular level.5, First cloning the Vamp gene of Spartina alterniflor which is related to vesicles transport, and preliminary analyze its functions through excessively expressed in Arabidopsis.