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Transgenic method of flame nandina
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A flame nantian bamboo and genetically modified technology, applied in the field of genetic engineering, can solve the problems of long time-consuming, labor-intensive, and cumbersome operations, and achieve the effects of avoiding tedious work, improving infection efficiency, and reducing pollution
Active Publication Date: 2019-03-26
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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Abstract
Description
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Problems solved by technology
Although there are relatively mature transgenic systems in other plants, the traditional method is laborious, time-consuming, cumbersome and inefficient, and too few transgenic plants are obtained, which brings obstacles to the later identification of transgenic plants.
And because the transgenic methods of different species are also different, the success rate of transgenic methods borrowed from other plants is very low, so how to obtain an efficient and fast stable transformation system of flaming nandina is a problem to be solved urgently in the transgenic of flaming nandina
Method used
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Examples
Experimental program
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Effect test
Embodiment 1
[0041] The genetically modified method of Nan Tianzhu includes the following steps:
[0042] 1. Strain activation
[0043] ① Transform the plasmid pCAMBIA2301 into Agrobacterium LBA4404 to obtain the recombinant Agrobacterium LBA4404 containing plasmid pCAMBIA2301.
[0044] ②The recombinant Agrobacterium LBA4404 containing plasmid pCAMBIA2301 was streaked on YEB solid medium supplemented with Km 50mg / L (kanamycin) and Rif 50mg / L (rifampicin), and cultured at 28°C for 2 days.
[0045] ②Pick 2-3 single colonies grown on YEB solid medium, and inoculate them into 50mL YEB liquid medium supplemented with Km 50mg / L and Rif50mg / L, and shake culture on a shaker at 28°C and 220rpm for about 12-16h The OD600 value of the bacterial solution is 0.8.
[0046] ③At room temperature, centrifuge the bacterial solution at 5000rpm for 8 minutes, remove the supernatant, invert the centrifuge tube on the sterile filter paper, remove the supernatant as much as possible, resuspend the bacteria with 100mL MS ...
Embodiment 2
[0057] Change the infection time, and set the infection time to 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 70 minutes, and 80 minutes. The rest of the steps are the same as in Example 1. Investigate the effect of the infection time on the conversion rate of Nantian bamboo influences.
[0058] The results are shown in Table 1.
[0059] Table 1 The influence of infection time on the conversion rate of Nantian bamboo
[0060] deal with
[0061] As shown in Table 1, when the time is shorter, the conversion rate of Tianzhu flame is lower. As the infection time increases, the conversion rate increases. When the infection time reaches 80 minutes, the conversion rate decreases, and the pollution of Agrobacterium and other bacteria is also serious, with a pollution rate of 7.8%.
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Abstract
The invention discloses a transgene method for Nandina domestica Fire power. The method comprises the following steps: an exogenous plasmid vector is transferred into agrobacterium tumefaciens LBA4404, and recombinant agrobacterium tumefaciens is obtained; the recombinant agrobacterium tumefaciens is subjected to activated culture until OD (optical density) 600 is 0.4-0.6, and then leaf explants of the Nandina domestica Fire power are infected for 50-70 min; the infected leaf explants are subjected to bud inducing culture and rooting culture sequentially, Nandina domestica Fire power plants are obtained, and transgenic Nandina domestica Fire power is screened from the Nandina domestica Fire power plants. An efficient, rapid and stable Nandina domestica Fire power genetic transformation system is established with the method, the transformation rate can be higher than 10%, practice foundation is provided for further improvement of Nandina domestica Fire power varieties with application of the genetic transformation method, and development of future molecular breeding work of the Nandina domestica Fire power is facilitated.
Description
Technical field [0001] The present invention relates to the technical field of genetic engineering, in particular to a genetically modified method of Nantian bamboo. Background technique [0002] Nandina domestica Fire power (Nandina domestica Fire power) is an emerging landscaping tree species, which is now widely used in gardens and landscapes. Nantian bamboo is cold-resistant and evergreen, with elliptical or ovoid leaves, which are obviously different from the original species with lanceolate leaves, which is more important than the original species. Flame Nantian bamboo encounters cold air every autumn, and the leaf color changes from green to bright red. The leaf color can last until the beginning of the second spring, so it has an excellent viewing effect in the south. In landscaping, the flaming Nantian bamboo can be cultivated in the tree altar at the same time as the camellia, and it is also suitable for planting in the lower layer of the plant community to enrich the ...
Claims
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Application Information
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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00A01H6/00
Inventor 贾思振张立伟王媛花颜志明
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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