Application of WRKY55 transcription factor in plant salt resistance
A stress-resistant, sequence-listed technology, applied in the biological field, can solve the problems of little research on WRKY transcription factors
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Embodiment 1
[0060] Example 1: Bioinformatics analysis of sweet sorghum WRKY55 gene
[0061] 1 test material
[0062] Sweet sorghum (Sb02g011050), and wild rice with short anthers (XM_015839135.1), Brachypodium distachyda (XM_014901792.1), zucchini (XM_023659186.1), corn (XM_020540231.2), cassava ( WRKY protein sequences of XM_021761582.1), Anthurium (KY597634.1), Vitis vinifera (KY411919.1), Arabidopsis (AT2G40740).
[0063] 2 test method
[0064] Firstly, the primary structure and physicochemical properties of SbWRKY55 protein were analyzed by software ExPASy. The detection of the signal peptide and the prediction of the transmembrane region use the software Signal4.1 and TMHMM. SMART online software can predict the structure and functional domains of SbWRKY55. The homologous sequences of SbWRKY55 were obtained from NCBI BLASTp on the Internet, and then the homologous sequences were compared in terms of proteins with software such as MegAlign and a phylogenetic tree reflecting the ph...
Embodiment 2
[0078] Example 2 Cloning, Vector Construction and Subcellular Localization of Sweet Sorghum WRKY55 Gene
[0079] 1 Test materials and material handling
[0080] Seeds of sweet sorghum salt-tolerant and high-sugar inbred line M-81E (gifted by Shandong Academy of Agricultural Sciences), tobacco (Nicotiana benthamiana) seeds, Escherichia coli strain (DH5α), Agrobacterium strain (GV3101), cloning vector pEASY-Blunt3 simple, pROKII-GFP expression vector driven by CaMV35S promoter, etc.
[0081]Select sweet sorghum seeds with similar sizes and full grains, and wash them in a mesh bag with running water for 12 hours. The sand was washed with tap water and divided into pots. Then plant the seeds evenly in the sand, 8 plants per pot, with a depth of about 2cm, not too shallow, to prevent them from being washed out when watering. Water tap water once a day until the hole at the bottom of the flower pot can flow out, which is counted as thoroughly watered. After emergence, pour 1 / 2 H...
Embodiment 3
[0114] Example 3: Overexpression of Arabidopsis SbWRKY55 and the acquisition of AtWRKY55 mutant lines
[0115] 1 test material
[0116] Columbia ecotype (Col-0) Arabidopsis plants were purchased from Tair website. Arabidopsis WRKY55 (AT2G40740) mutants were wrky55-1 (SALK_084192), wrky55-2 (SALK_021677), wrky55-3 (SALK_121437), wrky55 -4 (SALK_084288), wrky55-5 (SALK_082916), wrky55-6 (CS838667).
[0117] 2 test method
[0118] 2.1 Obtaining Arabidopsis SbWRKY55 overexpression lines
[0119] After the inflorescence of Arabidopsis thaliana was infiltrated with Agrobacterium pROKⅡ-GFP-SbWRKY55, it was covered with a black plastic film and cultured in the dark for about 24 hours, and then the black plastic wrap was removed for normal culture. After about 7 days, the second infection is carried out, and the culture is continued in an artificial climate incubator (22°C-16h light / 18°C-8h dark). After about 24 hours, the black plastic film was removed, and the cultivation was con...
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