Method for improving plant traits
A plant trait, plant technology, applied in the field of biology
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Embodiment 1
[0362] Example 1. LEAT protein uses light energy to catalyze the splitting of water
[0363] 1. LEAT protein catalyzes the reduction of 2,3,5-trimethyl-1,4-p-benzoquinone, an analogue of plastoquinone, by water under the action of light.
[0364] It has been reported that proteins such as GFP can act as electron donors to reduce small molecular compounds such as NAD in oxidation states. + , potassium ferricyanide, and some proteins such as cytochrome C, and proteins with FAD as a prosthetic group (BogdanovAM et al., 2009, Nature Chemical Biology.5:459-461). The present inventors expressed recombinant proteins using purified Escherichia coli. Although pre-illuminated YFP and mCherry can be used as electron donors, one LEAT protein molecule can only provide one electron, and cannot provide electrons continuously under light ( figure 1 ). The photoreduction of quinone molecules catalyzed by LEAT proteins was analyzed by monitoring the decrease in the absorption of quinone mole...
Embodiment 2
[0372] Example 2. Comparison of the ability of LEAT proteins from different sources to catalyze water splitting to release oxygen under light
[0373] Nine fluorescent proteins and three non-fluorescent chromogenic proteins ( Figure 10 ). These 12 proteins belong to different branches such as A, B, C, and D on the evolutionary tree ( Figure 10 ). These 12 fluorescent proteins and non-fluorescent chromogenic proteins plus GFP series and dsRED series fluorescent proteins cover most of the currently reported fluorescent proteins and non-fluorescent chromogenic proteins (Alieva et al., 2008, Diversity and evolution of coarl fluorescent proteins.PLoS One3(7):e2680.doi:10.1371 / journal.pone.0002680), their molecular evolution pathways and amino acid homology are very different from the LEAT proteins of the GFP series and dsRED series ( Figure 11 A and 11B). Using Escherichia coli to express these proteins, it was found that although their fluorescence intensity, absorption spe...
Embodiment 3
[0382]Example 3, Functional research of LEAT protein mCherry transgenic plants
[0383] 1. The expression of mCherry protein in transgenic rape increased the net photosynthetic rate, promoted plant growth, and increased the biomass of transgenic plants.
[0384] After the mCherry transgenic rape is obtained, the mCherry transgenic rape is identified. The results of Southern blot identification of mCherry transgenic rapeseed are as follows Figure 13 B; Fluorescent observation in root cells of transgenic plants Figure 13 C; The results of RT-PCR and Western blot identification of transgenic plants (L1-L6) are as follows Figure 13 d. The expression and subcellular localization of mCherry protein were detected by immunoelectron microscopy. Figure 13 e. It can be seen that the plants L1-L6 are all mCherry positive transformed plants, and the mCherry protein is mainly expressed in the cytoplasm (Cy) and nucleus (N). The present invention uses the T2 generation mCherry tran...
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