Plant tillering related protein and coding gene thereof
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A protein and gene technology, applied in the field of plant tiller-related proteins and their encoding genes
Inactive Publication Date: 2012-09-26
INST OF BOTANY CHINESE ACAD OF SCI
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The two genes MAX3 and MAX4 are involved in the biosynthesis of the plant branching hormone (strigolactone). In this hormone synthesis pathway, there are other key
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Embodiment 1
[0040] Embodiment 1, the acquisition of OsMADS57 protein and its coding gene
[0041] Design primers according to the results of database analysis:
[0042] Overexpression vector construction using primer: 5' end primer: 5'-CG GGATCC ATGGGGAGGGGGAAGATAGT-3' (the underlined sequence is the BamHI site), 3' end primer: 5'-GG GGTACC TTAAGGCAGATGAAGTCCCAGT-3' (the underlined sequence is the KpnI site).
[0043] Antisense expression vector construction using primer: 5' end primer: 5'-CG GGATCC TTAAGGCAGATGAAGTCCCAGT-3' (the underlined sequence is the BamHI site), 3' end primer: 5'-GG GGTACC ATGGGGAGGGGGAAGATAGT-3' (the underlined sequence is the KpnI site).
[0044] Total RNA was extracted from the three-leaf stage seedlings of japonica rice Zhonghua No. 10, and the 726bp full-length cDNA of OsMADS57 was amplified by RT-PCR. The specific operation process is as follows:
[0045] 1) Extraction of total plant RNA: select 100 mg of Oryza sativa L.cvZhonghua 10 seedlings at th...
Embodiment 2
[0054] Embodiment 2, the application of OsMAOS57 protein and its coding gene
[0055] 1. Acquisition of overexpression vector pUN-MADS57 (sense) and antisense expression vector pUN-MADS57 (antisense)
[0056] 1. Acquisition of pUN1301 plasmid
[0057] Step 1: Cut about 0.2 g of corn seedlings and grind them in liquid nitrogen; then add 800 μL of freshly prepared extraction buffer (containing 0.1M Tris-HCl pH8.0, 50mM EDTA, 0.5M NaCl, 1% SDS and 1% β-mercaptoethanol), shake vigorously to suspend it completely; bathe in water at 65°C for 30 minutes, mix by inverting every 5 minutes; then add 250 μL of pre-cooled 5M potassium acetate aqueous solution, mix by inverting immediately, and bathe in ice for 5 minutes; Add an equal amount of phenol / chloroform, extract once, and centrifuge at 12,000rpm for 5 minutes; collect the supernatant, add 0.6 times the volume of isopropanol to precipitate DNA, and let it stand at room temperature for 40 minutes; centrifuge at 12,000rpm for 15 min...
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Abstract
The invention discloses plant tillering related protein and a coding gene thereof. The protein is protein (a) or protein (b), wherein the protein (a) has amino acid sequences shown as a sequence 2 in a sequence table; and the protein (b) has an amino acid residue sequence which is derived from the sequence 2 in the sequence table by substituting and/or losing and/or adding one or more amino acid residues and has the same function. The experiment proves that expression vectors are inserted into an OsMADS57 gene which is cloned in japonica rice zhonghua 10 in forward and reverse directions so as to obtain plus sense recombinant plasmid and antisense recombinant plasmid; the plus sense recombinant plasmid and the antisense recombinant plasmid are imported into the japonica rice zhonghua 10 so as to obtain plus sense transgenic rice strains and antisense transgenic rice strains; and compared with that of wild-type rice, the tillering of the plus sense transgenic rice strains is increased, and the tillering of the plus sense transgenic rice strains is decreased.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a protein related to plant tillering and its coding gene. Background technique [0002] Plant architecture is one of the important factors affecting crop yield, especially for rice crops. At present, it is known that the tiller number, tiller angle, plant height, leaf angle, the size of rice panicle and the number of branches of rice spikelets jointly determine the rice plant type, and the tiller number and the shape of rice panicle are the most important factors determining rice yield. Key factor. For rice tiller development, the tillering stage is the most important period of rice vegetative growth, including from the differentiation of tiller buds to the formation of young panicles. General rice varieties, growth environmental conditions (temperature, water, nutritional status) and their planting density will also affect the development of tillers. Rice varieties with long grow...
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