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Method for creating cut flower gypsophila paniculata liriomyza sativae blandchard variety by transgene

A technology of Liriomyza sativae and Gypsophila gypsophila, applied in horticultural methods, genetic engineering, plant genetic improvement, etc., can solve the problems of chemical pesticides polluting the environment, limited effectiveness, and high cost, and achieve the effect of reducing costs and reducing the degree of disease

Inactive Publication Date: 2011-06-15
FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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  • Description
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Problems solved by technology

However, due to the pollution of the environment by chemical pesticides and the high cost, the effect is limited, and it can no longer meet the requirements of flower production.

Method used

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  • Method for creating cut flower gypsophila paniculata liriomyza sativae blandchard variety by transgene
  • Method for creating cut flower gypsophila paniculata liriomyza sativae blandchard variety by transgene

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Experimental program
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Embodiment 1

[0024] A. Establishment of the efficient transformation and regeneration system of Gypsophila gypsophila: select a single plant of Gypsophila gypsophila that grows healthy under in vitro culture conditions, take small leaves as materials under sterile conditions, and in the differentiation medium: Ms+BA 1.0mg / Cultured on L+NAA 0.3mg / L for 18 days, wherein, the culture temperature was 25°C, the light intensity was 2000lx, and small buds were differentiated from the base of Gypsophila;

[0025] B, the transformation of Bt gene to Gypsophila:

[0026] (1) Preparation of Agrobacterium Competent Cells

[0027] Agrobacterium LBA4404 was inoculated in 5ml of LB liquid medium containing 50mg / L of Km and 5mg / L of Rif, cultivated overnight at 28°C and 200rpm; Continue culturing in Rif LB liquid medium until OD600 is 0.5, put the culture in ice bath for 30min, centrifuge at 5000rpm at 4°C for 5min, discard the supernatant, and replace it with 1ml of cold CaCl with a concentration of 20...

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Abstract

The invention provides a method for creating a cut flower cultivar of Gypsophila paniculata which can resist against Liriomyza sativae, which obtains a tissue culture plantlet of genetically modified Gypsophila paniculata which can resist against Liriomyza sativae by constructing a high efficient genetic transformation system of Gypsophila paniculata, sieving the background of Gypsophila paniculata antibiotics, utilizing Bt genes to carry out genetic transformation to the G33 cultivar of Gypsophila paniculata, sieving the antibiotics of transformed plants, carrying out Pcr identification to the transformed plants and the rooting and propagation of the transformed plants. The method effectively improves the capability of Gypsophila paniculata in resisting against Liriomyza sativae; compared with the control plants inoculated in the same period, the morbidity degree of Liriomyza sativae is obviously lowered, thereby indicating that the genetically modified plants can effectively lower the harm of Liriomyza sativae done to Gypsophila paniculata, reduce cost for the culture management of Gypsophila paniculata and have practical meaning to practical planting production.

Description

technical field [0001] The invention relates to a method for breeding disease-resistant flower plants, in particular to a method for breeding cut flower gypsophila genes resistant to Liriomyza sativae pests, and belongs to the field of biotechnology. Background technique [0002] Gypsophila panicuta, also known as carnation and rosy grass, is a single-year or perennial herbaceous flower of the family Caryophyllaceae, native to northern Europe and Asia. The gypsophila plant has delicate branches and leaves, pure white flowers, and more than a thousand double petals, so it has the name Gypsophila gypsophila. It is suitable for flower beds, flower paths, and rock gardens, and it is an indispensable foil for various cut flower products. Yunnan is the main cultivation area of ​​Gypsophila gypsophila, with a cultivation area of ​​1,550 mu, an annual sales volume of 2-2.3 million plants, and an average annual sales volume of more than 50 million yuan. It is one of the main sources...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/32A01H1/00A01H4/00
Inventor 李涵张宝琼李世峰蒋亚黎霞王继华张颢杨春梅
Owner FLOWER RES INST OF YUNNAN ACAD OF AGRI SCI
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