Protein for controlling growth of plants as well as coding gene and application thereof
A technology for encoding genes and plants, applied in the field of plant development-related proteins and their encoding genes and applications, to achieve the effects of changing plant cell elongation, increasing plant biomass, and improving plant shape
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Embodiment 1
[0060] Example 1, the discovery of OsILI1
[0061] 1. Obtaining and Morphological Observation of Mutant Plant i1i1-D
[0062] 1. Obtaining the mutant plant i1i1-D
[0063] A Nipponbare rice T-DNA insertion mutant library was constructed, and a mutant plant i1i1-D with a significantly increased leaf angle was found.
[0064] 2. Morphological observation of mutant plants i1i1-D
[0065] The following morphological observations were carried out on the mutant plant i1i1-D.
[0066] (1) leaf angle
[0067] The most important phenotype of i1i1-D was that the leaf angle increased significantly at each growth stage. At the seedling stage, the mutant grew for about 5 days after germination, and after the second leaf was fully developed, it could be observed that the angle between the second leaf and the leaf angle was bent more than 90 degrees (see figure 1 ). At the tillering stage, each new tiller of the mutant showed a significantly increased leaf angle (see figure 2 ).
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Embodiment 2
[0081] Embodiment 2, the discovery of AtIBH1
[0082] 1. Discovery of AtIBH1
[0083] The analysis of OsILI1 protein sequence shows that OsILI1 belongs to the basic helix-loop-helix (bHLH) family of transcription factors, but it does not have a typical basic domain. Such bHLH proteins are classified as class D bHLH proteins, which cannot directly bind DNA and regulate their functions mainly by forming heterodimers with canonical bHLH transcription factors. Using the full-length OsILI1 protein as bait, yeast two-hybrid technology was used to screen for proteins interacting with OsILI1. After the coding sequence of OsILI1 was amplified from the cDNA library, it was connected into the pBD-GAL4 vector, and then the constructed vector was transformed into yeast AH109, and after being made into competent cells, the yeast two-hybrid library at the three-leaf stage of rice was co-transformed. The efficiency of the sieve library is 3×10 5 Clones / g DNA, the total amount of screenin...
Embodiment 3
[0093] Example 3, Obtaining of AtIBH1 Overexpression Arabidopsis Transgenic Plants
[0094] 1. Construction of recombinant expression vector
[0095] 1. Obtaining the 35S promoter fragment
[0096] Plasmid vector pBI221 (Clontech) was double digested with restriction endonucleases HindIII and BamHI, and a 35S promoter fragment with a length of about 0.8 kb was recovered after detection by agarose gel electrophoresis.
[0097] 2. The Noster poly A termination sequence was excised from the plasmid vector pBI221 (Clontech) with restriction enzymes Sac I and EcoR I, and connected to the corresponding site of the vector pUC19 (TaKaRa Company) to obtain a recombinant vector named as pUC19-Noster. Restriction endonucleases HindIII and BamHI were used to double digest pUC19-Noster, and after detection by agarose gel electrophoresis, the large linearized vector fragment was recovered, and the recovered fragment was connected with the 35S promoter fragment obtained in step 1 to obtain...
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