Related protein HTDF (high tillering and dwarf) for tillering and plant length of rice, and encoding gene and application thereof
A coding and rice technology, applied in the field of rice genetics and breeding, to achieve the effect of improving rice yield
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Embodiment 1
[0028] Example 1. Acquisition of Rice Tiller and Dwarf-related Proteins and Their Encoding Genes
[0029] 1. Genetic analysis of rice tiller and dwarf related gene HTDF
[0030] We found a new rice mutant with short stems and multiple tillers in the field. Compared with the wild type Taipei 309 (TP309), this mutant mainly exhibits traits such as more tillers and dwarfing plants, so we named it htdf (high tillering and dwarf)( figure 1 ). At the rice maturity stage, the average number of tillers in the mutant htdf (35.43±5.39) was significantly higher than that of the wild-type variety Taipei 309 (19.3±1.21) ( figure 1 ); the plant height of the mutant was also significantly reduced, about 73.8% of the wild-type variety ( figure 2 ).
[0031] Orthogonal and reciprocal F of mutant htdf to Taipei 3091 The plant height and tillering traits of the generation plants were similar to those of Taipei 309, indicating that F 1 The plant height and tillering traits of the generati...
Embodiment 2
[0045] The expression analysis of embodiment 2, HTDF
[0046] In order to study the expression pattern of the HTDF gene, it was analyzed by quantitative RT-PCR in the roots, seedlings, nodes, internodes, leaves, ears, calli, etc. of the japonica rice variety Taipei 309 (Oryza sativa L. subsp. expression in the organization. The fluorescence signal (expressed in Ct value) of HTDF gene and the fluorescence signal (expressed in Ct value) of Actin1 gene (internal reference gene) are according to the formula (relative expression amount=2 -ΔCt , where ACt=Ct 目的基因 -Ct 内参基因 ) calculated value as the relative expression level of HTDF gene, the results are shown in Figure 6 . The results showed that the HTDF gene was expressed in all the tested tissues, and the expression level in ear and callus was relatively high.
Embodiment 3
[0047] Embodiment 3, cultivating the rice that dwarf stalk and / or tiller number increase
[0048] Using the KOME full-length cDNA clone (No. AK108569) as a template, PCR amplification, enzyme digestion, and connection to the expression vector pTCK303 with Ubiquitin promoter, the interference vector RNAi-HDTF was constructed, and the RNAi-HDTF Transformed into Agrobacterium tumefaciens strain LBA4404 (Cat. No. 18313015, Invitrogen Company), and screened to obtain a recombinant Agrobacterium strain containing the interference vector RNAi-HDTF, using wild-type rice variety TP309 mature embryos to induce callus. Agrobacterium transformation, a total of 8 independent T0 strains were obtained.
[0049] Statistical analysis of the plant type of T1 transgenic plants transgenic for RNAi-HDTF gene and control TP309 plants showed that the number of tillers and plant height of T1 transgenic plants transgenic for RNAi-HDTF gene increased significantly. It shows that using the method of ...
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