A kind of hybrid Liriodendron lhrgl1 gene and its application
A technology for hybridization of Liriodendron tulipifera and genes, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of unknown structure and function, slow progress, late start of research, etc. Effects of developmental delay
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Embodiment 1
[0024] The material used in this example is the hybrid Liriodendron tulipifera leaves on the campus of Nanjing Forestry University, which were quickly frozen in liquid nitrogen after harvest and stored in an ultra-low temperature refrigerator (-80°C).
[0025] 1) Extraction of total RNA from hybrid tulip tree leaves
[0026] According to the instructions of the NORGEN Plant Total RNA Extraction Kit, the specific operation is as follows: add appropriate amount of materials and liquid nitrogen to the treated mortar, after fully grinding, add 600 μL LysisSolution; transfer the lysed liquid in the mortar to In a new centrifuge tube; after tightly closing the cap, invert and mix several times, let it stand at room temperature for 2 min to fully lyse, centrifuge at 12000 rpm for 2 min, and transfer the supernatant to a new centrifuge tube; add an equal volume 70% ethanol, vortex and mix; transfer the mixture to the column provided in the kit (connected to the 2mL collection tube), c...
Embodiment 2
[0050] Research indicates LhRGL1 and wxya It plays an important role in various tissues and organs of Liriodendron hybrida. However, the expression levels of the two genes were the highest in the stem, indicating that the GA signaling pathway was active during the growth and development of the stem, and DELLA protein played an important regulatory role in the elongation and growth of the stem.
[0051] The plant material used in this embodiment is Arabidopsis ( Arabidopsis thaliana )Col (Columbia) wild-type seeds, kindly donated by Professor Thomas Laux from Germany.
[0052] The Escherichia coli strain used in this embodiment is E. coli JM109; Agrobacterium strains GV3101 and EHA105 were used to transform Arabidopsis thaliana; the plant expression vector used in the experiment was pBI121. All strains were purchased from Related Biological Company.
[0053] 1) LhRGL1 Construction of gene pBI121 vector
[0054] With embodiment 1 obtaining LhRGL1 The full-length O...
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