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Genetic transformation method of PttKN1 gene in cardamine

A genetic transformation method, a technology of broken rice chestnut, which can be applied in the field of biological genetics and can solve problems such as undetectable

Inactive Publication Date: 2014-07-30
NORTHWEST A & F UNIV
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Problems solved by technology

[0025] PttKN1 gene expression analysis showed that PttKN1 gene was highly expressed in stem apical meristem, and also expressed in phloem, cambium and cork cambium, meristem and undifferentiated cells, but not in young and undifferentiated cells of poplar. detected in mature leaves

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  • Genetic transformation method of PttKN1 gene in cardamine

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Embodiment

[0041] The genetic transformation method of the PttKN1 gene of the present embodiment in broken rice chestnuts specifically includes:

[0042] (1) Preparation of plant material and carrier: plant vernalized Cardamine hirsuta seeds in a mixed substrate of vermiculite: peat soil: perlite (1:1:1), cover with plastic wrap, and after two days Expose to light, and remove the film after three days. Choose vigorously growing and more unopened flower buds for later use. Culture conditions: relative humidity 80%, constant temperature 20-24°C, light intensity 80-200μmol / M 2 S, 16h light culture. Agrobacterium strain GV3101 containing the pPCV702 plasmid vector was kindly provided by Dr. Olf Olsson, University of Gothenburg, Sweden. pPCV702 contains a CaMV35S promoter and a neomycin phosphotransferase gene; the PttKNI gene is fused downstream of the CaMV35S promoter.

[0043] (2) Transformation of Chrysoprina sativa by Agrobacterium-mediated cathedral dipping method: Genetic transform...

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Abstract

The invention relates to a genetic transformation method of PttKN1 gene in cardamine, and the method specifically comprises the following steps: (1) preparing a plant material and a carrier; (2) using an agrobacterium mediated inflorescence impregnation method for transformation of cardamine; (3) performing resistant plant screening and phenotypic observation; (4) performing RT-PCR (reverse transcription-polymerase chain reaction) identification; and (5) performing analytic statistics of the results. According to the method, the agrobacterium mediated inflorescence impregnation method is used for genetic transformation of the cardamine. After screening by use of 100mg / L kanamycin, resistant plants show leaf morphological changes, abnormal growth points, ascidial leaves, incised leaves, cladodes and other morphological changes. The RT-PCR tests show that the morphological changes of the resistant plants can be caused by heterologous expression of the PttKN1 gene, and an important foundation for further study on the function of the gene in compound leaf formation and compound leaf plant development is laid.

Description

technical field [0001] The invention relates to a genetic transformation method of the PttKN1 gene in the rice chestnut, which belongs to the technical field of biological genetics. Background technique [0002] Homeobox genes (homeobox genes) encode homeobox proteins, which can bind to the cis-regulatory regions of their respective target genes, thus playing an important role as transcription factors in the development of organisms. From yeast, fungi to animals and plants, a large number of homeobox genes have been isolated and extensively studied; and it has been found that homeobox proteins play an important role in the morphogenesis of organisms. [0003] The first homeobox gene isolated in plants was the KNOTTED1 (ZmKN1) gene in maize, which functions in the shoot apical meristem (SAM). ZmKN1 gain-of-function mutant / heterologous expression plants showed that ZmKN1 inhibited leaf differentiation and formed tumor-like structures near leaf veins. Loss-of-function mutants...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00C12Q1/68
Inventor 徐全乐郭瑞军胡鑫
Owner NORTHWEST A & F UNIV
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