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Gene Clone and Genetic Transformation of FLC Gene in Betula Platyphylla

Author: ZuoLiNa
Tutor: QuGuanZheng
School: Northeast Forestry University
Course: Tree Genetics and Breeding
Keywords: FLC Betula platyphylla gene clone genetic transformation
CLC: S792.153
Type: Master's thesis
Year: 2013
Downloads: 27
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Abstract


Flowering, transition from vegetative to reproductive phase in plants, is regulated by both endogenous and environmental signals. FLOWERING LOCUS C (FLC) is a key repressor of the floral transition and is targeted by several regulatory pathways.We have isolated a FLC-related sequence from Betula platyphylla and named it as BpFLC. In this study, we did some researeh about BpFLC and the result are as follows:This study served to isolate a FLC gene from Betula platyphylla cDNA library. The length of the BpFLC gene was1304bp, The gene open reading frame (ORF) was627bp, The gene encodes208amino acids, with a molecular weight of50.419kD and a pI of5.17.The results revealed that BpFLC was expressed in all the tissues. The results show that the stem apex and young leaf presented the higher expression, the root exhibited the lowest expression levels.The fusion protein vector GFP-BpFLC for subcellular localization was constructed.The fusion protein vector and GFP vector were transformed into onion cells. This result was demonstrating that BpFLC protein is localized in the cell nucleus and cytoplasm.1647bp long sequence of BpFLC promoter was cloned by genome walking method. Sequence analysis showed that BpFLC’s promoter had typical characteristics of the promoter sequence, for example, and contains many cis-acting elements, such as, WRKY71OS and GARE which may be involved in the regulation of flowering time.The plant expression vector of pROKⅡ-BpFLC was constructed and transformed into Betula platyphylia via Agrobacterium infestation. The Transformed lines were tested by PCR, Q-PCR, Nouthern blot confirming that BpFLC gene have been integration into the genome of Betula platyphylla and successfully expressed, the transgenic lines had stronger expressed than wide-type ones.The plant expression vector of pROKⅡ-BpFLC was transformed into Betula platyphylla via floral dip protocols. The Transformed lines were examined by PCR, RT-PCR, Nouthern blot confirming that BpFLC gene have been transformed into Arabidopsis thaliana and expressed on the level of transcription. Q-PCR indicate that overexpression of BpFLC resulted in down-regulation of some flowering-related genes, such as FT, FD, SOC1and API.

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CLC: > Agricultural Sciences > Forestry > Forest tree species > Broad-leaved trees > Birch > Birch
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