Method for enhancing aphis resistance of chrysanthemum by using transgenosis

A transgenic, aphid-resistant technology, applied in horticultural methods, genetic engineering, plant genetic improvement and other directions, can solve the problems of difficult to control chrysanthemum aphids

Inactive Publication Date: 2010-09-15
ZHEJIANG SUB TROPICS CROP INST
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Problems solved by technology

[0005] The object of the invention is to solve the problem that chrysanthemum aphid is difficult to prevent and treat in actual production, and a kind of turning hpal is provided Xoo The Method of Gene Improving Aphid Resistance of Chrysanthemum

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  • Method for enhancing aphis resistance of chrysanthemum by using transgenosis
  • Method for enhancing aphis resistance of chrysanthemum by using transgenosis

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Embodiment Construction

[0040] Specific embodiments of the invention are described in detail below: this embodiment is implemented under the premise of the technical solution of the present invention, and detailed implementation methods and specific operating procedures are provided, but the protection scope of the present invention is not limited to the following the described embodiment.

[0041] The test method that does not indicate specific conditions in the following examples usually follows conventional conditions.

[0042] The main reagents used: Bacteria Genomic DNA Extraction Kit, TRIZOL Reagent, ThermoScript RT-PCR System were purchased from Invitrogen Company. Ex Taq DNA polymerase, endonuclease BamH I and Sac I, dNTPs, Goxigenin-labeled dNTPs, nucleic acid molecular mass standards, and DNA gel recovery kit are produced by TaKaRa. Genome southen hybridization was performed using kits from Roche, Switzerland. Plasmid extraction kit, product of Shanghai Shenergy Biotechnology Co., Ltd. T...

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Abstract

The invention relates to a method for enhancing the aphis resistance of chrysanthemum by using transgenosis, which belongs to the technical field of biology. The method comprises the following steps of: adopting binary plant expression vectors comprising hpalXoo genes coding harpinXoo proteins and glufosinate-resistant bar genes; integrating the hpalXoo genes and the bar genes on chrysanthemum genomic DNA; performing glufosinate resistance-based sieving to obtain resistant transformed plants; testing positive by using PCR and genomic southern blot to obtain transgenic chrysanthemum plants; and performing a biological aphis-resistance test on leaves and the whole plants to obtain aphis-resistant transgenic plants. Through the method, the aphis resistance of the chrysanthemum is improved by transforming foreign hpalXoo genes, performing normal transcriptome expression and inserting the foreign hpalXoo genes into chrysanthemum to induce a broad spectrum aphis-resistant signal to conduct relative gene NPR1 expression. The novel and practical method is provided for gene engineering technology to select and breed aphis-resistant chrysanthemum varieties.

Description

1. Technical field [0001] The invention relates to a method for improving the aphid resistance of chrysanthemum through transgene, and belongs to the field of biotechnology. 2. Background technology [0002] Chrysanthemum [Dendranthema morifolium Tzvel. (Chrysanthemum morifolium Ramat.)] is a perennial herb, which can be used for ornamental, health care, medical treatment and the preparation of natural pigments. Widely cultivated all over the world. Chrysanthemum is one of China's traditional ten famous flowers, ranking first in the world's "four major cut flowers". hpal Xoo (formerly known as hrf xoo or hrf1) is cloned from Xanthomonas oryzae pv. oryzae, the pathogen of rice bacterial blight, encoding harpin Xoo The gene can induce disease resistance and allergic cell death (Wen Weigang, Wang Jinsheng. Cloning and expression of harpin gene from Xanthomonas oryzae [J]. Acta Phytopathology, 2001, 31(4): 295-300) . hpal Xoo Coded Harpin Xoo Tobacco (Nicotiana tabacum L...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H4/00A01H5/00
Inventor 柴一秋王金生李明
Owner ZHEJIANG SUB TROPICS CROP INST
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