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Functional Analysis of HbFCA Gene in Rubber Tree (Hevea Brasiliensis)

Author: LuXu
Tutor: HuangHuaSun
School: Hainan University
Course:
Keywords: Rubber tree HbFCA Normalized cDNA library Yeast one-hybrid
CLC: S794.1
Type: Master's thesis
Year: 2014
Downloads: 1
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Abstract


Rubber trees are perennial tree,Children of up to5-6years,so Select breeding efficiency is greatly restricted.Therefore, the use of genetic engineering techniques to shorten the breeding cycle is important.FCA gene is an important gene autonomous flowering pathway,Has been shown to promote the importance of early flowering in Arabidopsis.In this study, the gene of Arabidopsis turn rubber tree HbFCA be obtained preliminary PCR, Southern and semi-quantitative RT-PCR to identify.Proved HbFCA gene was successfully integrated into the Arabidopsis genome and accessed to expression.By phenotype observed in transgenic Arabidopsis, HbFCA promote transgeniced wild-type Arabidopsis early flowering.And transgenic Arabidopsis seed germination also had been some impacted.Through the integration of plant expression vectors HbFCA gene promoter and GUS gene transformation of rubber trees.GUS gene was observed in embryoid bodies、 leaves and rootshave strongly expressed.but the expression is weak in the stem segments.Prove HbFCA genes involved not only in the regulation of flowering, may also be involved in the regulation of a variety of developmental processes.The results also confirmed the diversity of gene function HbFCA.In this study, the rubber tree Southern hybridization techniques were optimized.And methods for different probes labeled comparative study,proved by PCR-labeled probe is the best choice rubber tree Southern hybridization probes labeled,this method is not only more high labeling efficiency and specificity.imaging also has the advantages of low background.Rubber trees of different varieties HbFCA genes Southern Bloting verification,H.N.V8was founded HbFCA gene exists in two copies of the form,in the rest of the species studied is based on the existence of a single copy of the form.For further research HbFCA gene and promoter polymorphism in different species of rubber tree.Analyzed the promoter of rubber tree by the search for care software of Plant Care, some other responsive element except TATA-box and CAAT-box like light responsive element, cis-acting regulatory element involved in light responsiveness, cis-acting regulatory element involved in circadian control, protein binding site et al. all found in. So, the author think, these regulatory elements show HbFCA gene is highly sensitivity for light reaction. Otherwise, there are other regulatory elements in the HbFCA promoter, like cis-acting regulatory element involved in the MeJA-responsiveness, cis-acting element involved in heat stress responsiveness, cis-acting element involved in defense and stress responsiveness, cis-acting element involved in salicylic acid responsiveness. It shows that HbFCA maybe also sensitive to MeJA, orthohydroxybenzoic acid, high temperature and so on. HbFCA sequence was constructed phylogenetic tree.This is the first use of uniform length cDNA library construction strategy,successfully constructed rubber tree leaves early flowering varieties homogenization cDNA library.It has a full length cDNA sequences and gene expression is uniformly.Validated,library constructed in line with the quality of this study the basic requirements of the yeast one-hybrid.To further yeast one-hybrid screening and HbFCA promoter interactions of trans-acting factors to provide a more reliable material foundation.

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CLC: > Agricultural Sciences > Forestry > Forest tree species > Special use of broad-leaved class > Rubber tree
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