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Market-free Transgenic System Establishment of Citrus and Preliminary Application

Author: LiLinJie
Tutor: DengZiNiu
School: Hunan Agricultural University
Course: Pomology
Keywords: Chemically induced removal of the marker gene system Citrus canker pthA-NLS Sweet orange
CLC: S666
Type: Master's thesis
Year: 2010
Downloads: 41
Quote: 0
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Abstract


Plant resistance marker genes in transgenic research can improve the efficiency of the screening of the transformants . However, now can take advantage of the very limited types of marker genes , is not conducive to long-term research on genetically modified plants . And it is reported on the safety of genetically modified plants to suspect that hinder the development of a transgenic plant research . In removing marker genes policy , can be induced independently removing pathway showing better utilization prospect . Citrus canker is a worldwide disease quarantine , and greatly harm the industry , due to the lack of resistance resources and effective prevention and control methods , the disease still can not be eradicated . pthA-NLS gene function, inhibition of pathogen virulence factor built into this chemically induced knockout vector designed to citrus receptor genetically modified plants in pthA-NLS eliminate marker genes transferred from the anti-ulcer disease . The results of this study are as follows : the test as test materials to achieve eco- sugar orange , using a combination of chemical induction system the XVE and a bit of site-specific recombination system Cre / LoxP marker gene knockout vector citrus unmarked transgenic the research findings show that chemically induced excision marker gene in the citrus is feasible , and the initial establishment of an effective citrus unmarked transgenic system . Vitro grafting by fluorescence microscopy successful observed the expression of the green fluorescent protein , and after 30 d , extracting the scions genomic DNA for PCR amplification, to complete excision of the marker gene is detected . 2 . PthA-NLS gene was replaced gfp reporter gene construct into the the p35S-gfp carrier the XhoI and SpeI sites successfully constructed the citrus canker pthA-NLS gene unmarked transformation vector . 3 the p35S-NLS carrier using Agrobacterium-mediated transformation sweet orange seedlings Festival Room stem segments , transformed seedlings the purpose gene -specific primers XhoIF and SpeI PCR assay , 27 transgenic scarlet sweet orange . P1, P2 and P4 three primer multiplex PCR assay , does not detect the occurrence of the recombination phenomenon . Will have been identified as transgenic tubes grafted seedlings transferred to the induction of a liquid medium containing a of 6μM- estradiol , proceed induced . A month after its RT-PCR analysis found that part of the restructuring resection phenomenon occurred .

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CLC: > Agricultural Sciences > Gardening > Fruit trees gardening > Citrus
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