Maize gene zmlip2 and its application
A corn and gene technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as positioning research that has not yet been reported, and achieve the effects of huge potential for economic benefits, reducing toxic effects, and enhancing growth conditions.
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Embodiment 1
[0026] Example 1 Transcriptome Analysis of the Roots of Maize Inbred Line 178
[0027] 1-1. Lead stress treatment of corn inbred line materials
[0028] Select plump corn inbred line 178 seeds to be sterilized with 75% ethanol for 1 min, soak in 10℅ H2O2 solution for 15 min and shake continuously during disinfection, then rinse with deionized water for 3 to 5 times until the residual H2O2 is rinsed clean, and then use Soak in deionized water for 4 hours, and finally germinate with filter paper in the dark at 28°C. After about 2-3 days, transfer the germinated corn to a floating plate, and culture it with Hoagland nutrient solution under the conditions of light (16h) / darkness (8h) at 28°C for hydroponics, and change the culture medium twice a week. Six seedlings with consistent growth were selected for stress treatment. Remove the endosperm of these 6 seedlings and move them into plastic containers with nutrient solution. Two treatments were set up in this experiment: normal...
Embodiment 2
[0047] Example 2. The acquisition and analysis of ZmLIP2 interference strains
[0048] In this example, Ubi is used as the promoter, NOS is used as the terminator, bar is used as the selection marker gene, and kanamycin is used as the selection marker gene. The target gene fragment primers are designed according to the ZmLIP2 sequence, and the CUB vector is used as the backbone. Homologous recombination method , construct the interference vector of ZmLIP2 gene by double digestion with BamHI and SmalⅠ. Transform the constructed interference vector into Agrobacterium competent cells. The T0 generation ZmLIP2 gene interference maize was obtained by infecting wild-type maize C01 immature embryos with Agrobacterium.
[0049] The following are the specific steps:
[0050] 2-1. Extraction and purification of corn RNA
[0051] The total RNA of the maize inbred line Qi C01 (hereinafter also referred to as wild-type maize) was extracted by the Trizol method, and the specific method r...
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