Method for quickly culturing homozygous transgenic ornamental collard with insect resistance and herbicide resistance
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A genetically modified, cabbage technology, applied in herbicides and algaecides, pesticides, gardening methods, etc.
Inactive Publication Date: 2010-09-08
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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We have established high-frequency adventitious bud induction and plant regeneration technology in several excellent breeding materials (Zhao Xiushu et al., 2009), but there are no reports on the use of transgenic technology to cultivate insect-resistant and herbicide-resistant ornamental kale at home and abroad
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Embodiment 1
[0052] Embodiment 1, the method for cultivating recombinant ornamental kale
[0053] Experiment 1. Cultivation and identification of recombinant ornamental kale
[0054] (1) Cultivation of recombinant ornamental kale
[0055] In order to investigate whether the established in vitro culture system is effective, the control explants were set up with the same materials in the experiment, and the control explants were directly entered into the bud induction medium (Table 1) after pre-cultivation, without contact with Agrobacterium, and without screening culture (ie no steps 4, 5, 6).
[0056] 1. Obtaining sterile materials
[0057] Select seeds with plump grains, uniform size and no pests and diseases, disinfect with 75% alcohol for 30 seconds, rinse with sterile water for 3 times, disinfect with 3% sodium hypochlorite (V / V,) for 15 minutes (shake constantly, add a drop of Tween), rinse with sterile water 4 times, dry the water on the surface of the seeds with sterile filter pa...
Embodiment 2
[0209] Embodiment 2, transgenic regenerated strain obtains homozygous material rapidly by free microspore culture
[0210] In general, in the obtained transgenic plants, the exogenous gene is present in a heterozygous state. If the genetic background of the recipient material is also heterozygous, such as in cabbage crops, it usually takes 8-10 years to mature. In order to obtain completely homozygous material, it can be used for hybrid seed production. In this experiment, the haploid breeding method was adopted, combined with the characteristics of the transgenic material, to quickly breed homozygous transgenic material within 1-2 years.
[0211] Transgenic kale donor mother plants 173-2, 173-6, 173-7, 173-8, 173-9 with good insect resistance were planted in plastic greenhouses with insect-proof nets, and collected in early March to June The microspores are cultured in vitro by the method of the present invention, and homozygous transgenic plants can be obtained at one time....
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Abstract
The invention discloses a method for quickly culturing homozygous transgenic ornamental cabbages with insect resistance and herbicide resistance. The method for culturing the transgenic cabbages comprises the following steps of: immersing the cotyledons of the cabbages by using recombinant agrobacterium tumefaciens containing a target gene and then co-culturing to obtain the transgenic cabbages. In the invention, the inductivity of the adventitious buds of explants after conversion treatment reaches 48 percent in the selection of a culture medium, the number of regeneration buds on each explant is averagely 1 to 2, and the conversion frequency of the regeneration buds reaches 35 percent by the validation of molecular identification. The result of the manual feeding of the larvae of diamond back moths proves that the obtained transgenic regeneration plant has insect resistance, and the larva lethality of a best resistant material can reach 97 percent. The process from the conversion of the explants to the acquisition of the homozygous transgenic resistant material only needs a transient time of three years.
Description
technical field [0001] The invention relates to a method for rapidly cultivating homozygous insect-resistant and herbicide-resistant transgenic ornamental kale. Background technique [0002] Ornamental kale (Brassica oleracea var. acephala), a biennial herb of the Brassicaceae family, is a horticultural variety of Brassica oleracea. Focusing on foliage viewing, the leaves are beautiful and changeable, the heart leaves are as colorful as flowers, and the whole plant is shaped like a blooming peony, so people vividly call it "leaf peony". It has strong cold resistance and can withstand low temperatures of about -8°C. Recently, it has become the main landscape ground cover plant in autumn and winter to early spring in many cities, especially in the Yangtze River Basin and cities south of the Yellow River. my country's current seeds basically rely on imports, and domestic breeding work is rare. [0003] Part of the growth period of ornamental kale is in the season of severe in...
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