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Tomato microRNA Sly-mir156α and Sly-mir169c functional identification

Author: ZouZhe
Tutor: YeZhiBiao
School: Huazhong Agricultural University
Course: Olericulture
Keywords: microRNA mir156 mir169 Development Abiotic Tomato
CLC: S641.2
Type: Master's thesis
Year: 2010
Downloads: 290
Quote: 0
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Abstract


Tomato is an important vegetable crop genetic improvement of its important way is to find and use to regulate plant development, including plant height, leaf number and other important horticultural traits genes and adapt to the environment, including drought, cold and other abiotic stresses adaptive gene , microRNA part of this gene. microRNA widespread in eukaryotes is an important regulatory factor in the plant and animal studies have found its function is diverse, and in vivo has a very important regulatory functions including growth, stress response, hormone response and Virus-induced and many other aspects. In this study, tomato as a material, according to previous studies, the database is selected from 1nicroRNA tomato microRNA family Sly-mir156a and Sly-mir169c these two miRNA, expression vectors were constructed and transferred its excess tomato, analyze them in tomato The features and leveraging bioinformatics tools to predict their target genes in tomato, analyzes predicted target genes resulting in the expression of the corresponding conditions, expect to be able to obtain information about application value microRNA. The main research results obtained are as follows: 1. Clones selected from the database of tomato two microRNA precursor gene and overexpression construct the corresponding carrier; 2 by Agrobacterium-mediated transgenic technology, the above constructed two an expression vector into tomato, transgenic tomato plants, and through the PCR and Southern blot verification showed that the target gene into the tomato genome; 3 by RT-PCR technique validated target gene overexpression; 4. Through Sly-mir156a transgenic T1 progeny of observation, to get control of the corresponding phenotype distinct from transgenic plants showed small, leaves became volatile small, internode shortening and other features, the use of real-time PCR technique analyzes the different target genes In the case of different tissues, especially for the horticultural characteristics of tomato genetic improvement of morphogenesis provides a basis; 5 would Sly-mir169c transgenic T1 progeny of drought and salt stress for stress treatment, transgenic and control exhibited different responses, the former showing the above abiotic good resistance. Transgenic and control plants were observed stomata found transgenic lines pore size and stomatal conductance were significantly reduced compared to the control, indicating Sly-mir169c by regulating stomatal switching reduces transpiration of water thus improving plant drought tolerance. Also make use of real-time PCR technique analyzes the different target genes in different tissues and related expression under stress, as Sly-mir169c under stress study provides molecular regulatory networks preliminary evidence.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Solanaceous > Tomatoes ( tomatoes )
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