A rice sbp-box transcription factor gene and its application
A technology of transcription factor and sbp-box, applied in the field of plant bioengineering and plant improvement genetic engineering
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Embodiment 1
[0099] Example 1: SST rice transgenic experiment
[0100] 1. High salt tolerance sst Mutant acquisition and traits
[0101] A rice mutant library containing about 1000 lines was constructed by treating rice seeds with gamma rays. Large-scale screening of the rice mutant library under the salt stress of 150 mM sodium chloride concentration, after repeated verification of the candidate mutants under the salt stress of 150 mM sodium chloride concentration, observation of salt-tolerant phenotypes, and obtaining a performance height salt-tolerant mutant sst .
[0102] with molecular markers SST The gene was initially located on the sixth chromosome of rice. use sst Mutants crossed with salt-sensitive varieties to construct large-scale F 2 Group, using molecular markers to screen the exchanged individuals from the population, combined with the genotype and phenotype of the exchanged individuals, carried out map-based cloning, and successfully cloned SST Gene. Should ...
Embodiment 2
[0113] Embodiment 2: Cultivation and salt stress test of transgenic plants
[0114] The seeds of the transgenic rice obtained in Example 1 were taken, and after dormancy was broken in an oven at 45°C for one week, they were sown in petri dishes, soaked in tap water at room temperature for 2 days, and accelerated germination at 37°C for 1 day. After germination, move them into a light incubator, cultivate at 28°C, and light for 11 hours a day. After 1 day, the temperature was gradually lowered to 26°C for another day. After the seedlings are basically out, the tap water is replaced with rice culture medium for cultivation.
[0115] Test results such as figure 1 with figure 2 shown. Such as figure 1 As shown, the rice mutant sst Compared with the wild type (R401), the salt tolerance was significantly improved.
[0116] Depend on figure 2 Visible: the wild-type (R401) SST Genome fragments are transferred by transgene sst Among the mutants, the transgenic complem...
Embodiment 3
[0118] Example 3. In different plants SST presence of homologous genes
[0119] A database search (http: / / www.ncbi.nlm.nih.gov / ) found that in Oryza short-anthers ( Oryza brachyantha ), millet ( Setaria italica ), sorghum ( Sorghum bicolor ),wheat( Triticum aestivum ),corn( Zea mays ),barley( Hordeum vulgare ), Urartu Wheat ( Triticum urartu ), Brachypodium distachydium ( Brachypodium distachyon ), Festival wheat ( Aegilops tauschii ) and other genomes have homologous genes. The similarity of proteins ranged from 81.2% to 56.5%.
[0120] These homologous genes all have a conserved SBP domain with high similarity. The sequence is RCQAEGCKADLSGAKHYHRRHKVCEYHAKASVVAASGKQQRFCQQCSRFHVLTEFDEAKRSCRKRLAEHN (positions 180-249 of SEQ ID NO.2). The SBP-box transcription factor protein is combined by the SBP domain and the cis-acting element, so these homologous genes have a corresponding relationship with the cis-acting element, which indicates that other grass c...
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