Clone of gene PM1 for controlling paddy rice spike and application of gene PM1
A rice and gene technology, applied in application, genetic engineering, plant gene improvement and other directions, can solve the problem of rice cluster genes not being isolated and cloned, and achieve the effect of increasing yield
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Embodiment 1
[0037] Embodiment 1: the isolated clone of PM1 gene
[0038] 1. Obtaining of pm1 mutants
[0039] The mutants identified in the present invention are from rice T-DNA insertion mutants (for the construction method of the rice T-DNA insertion mutant library, see Wu et al., Development of enhancer trap lines for functional analysis of the rice genome. PlantJ, 2003, 35 :418-427; Zhan et al., Non-random distribution of T-DNA insertions at various levels of the genome hierarchy as revealed by analyzing13,804 T-DNA flanking sequences from an enhancer-trap mutant library. Plant J, 2007,49:947- 959). The screening and identification method of mutants is as follows: from the above-mentioned rice T-DNA insertion mutant library (see http: / / rmd.ncpgr.cn / ), the T0 generation transgenic rice seeds are planted in the field after conventional seed soaking and germination procedures. After getting each T 1 A total of 20 plants were planted in the generation family. The plants were planted in...
Embodiment 2
[0063] Embodiment 2: PM1 gene expression analysis
[0064] The expression pattern of the gene was analyzed by semi-quantitative RT-PCR (Reverse-transcript PCR). RT-PCR primer RT-P2 was designed to cover exon 4-9 of the gene. The primer sequences are: P2-F5-GATATCAAGCACTGTGAAGGGT-3, P2-R5-CGGGGATGATTTGATGAACT-3, the size of the amplified product using the reverse transcription product as a template is 773bp, and the size of the amplified product using genomic DNA as a template is 2430bp. Roots, stems, leaves, leaf sheaths, and ears (1cm, 6cm, 8cm, 11cm, 14cm) of the wild type at the booting stage and the pm1-1 mutant were taken to extract RNA to prepare reverse transcription products, and GAPDH was used as an internal standard. The RT-PCR reaction program is: 94°C denaturation, 5min; 94°C, 45s, 60°C, 45s, 72°C, 45s, 30 cycles; 72°C extension, 7min; ×GC bufferⅠ10μL replaces 10×buffer2μL and other components remain unchanged, the detection results of RT-PCR are as follows F...
Embodiment 3
[0066] Example 3: Physiological analysis of the response of pm1 mutant materials to brassinosteroid (brassinosteroid, BR)
[0067] The gene predicted by Example 1 and Example 2 is a cytochrome p450 gene. Previous studies have shown that some P450 enzymes play an important role in the synthesis and inactivation of plant brassinosteroid hormone (BR). BRs is a class of hormones with steroid structure, and there are two biosynthetic pathways: one is the early C6 oxidation pathway, and the other is the late C6 oxidation pathway. Both synthetic pathways are generally present in plant cells. Studies have shown that the intermediates of the early C6 oxidation pathway are very effective in promoting hypocotyl elongation grown in the dark, and the intermediates of the late C6 oxidation pathway are more effective than the intermediates of the early C6 pathway in rescuing the phenotype of mutants under light . It shows that under different light, the synthesis and metabolism of BR may b...
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