Application of E-like gene in rice in cultivating early-flowering and dwarf transgenic plants

A technology of transgenic plants and genes, which is applied in the field of application of rice E genes in the cultivation of early flowering and dwarf transgenic plants, and can solve problems such as weak research

Inactive Publication Date: 2010-01-20
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The functional role of E genes in monocotyledonous plants is far from clear in dicotyledonous plants such as Arabidopsis and petunias; however, according to gene expression patterns and phylogenetic analysis, E gene functions occur in monocotyledonous plants There is a large variation, but the research on it is still very weak

Method used

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  • Application of E-like gene in rice in cultivating early-flowering and dwarf transgenic plants
  • Application of E-like gene in rice in cultivating early-flowering and dwarf transgenic plants
  • Application of E-like gene in rice in cultivating early-flowering and dwarf transgenic plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1, Regulatory function of rice class E genes in flower organ development

[0057] 1. Gene silencing

[0058] 1. Gene cloning

[0059] The OsMADS8 gene was cloned by the RACE method, and the OsMADS7 gene was obtained by screening the rice cDNA library with the OsMADS8 gene as a probe.

[0060] 1) Construction of rice young panicle cDNA library

[0061] Total RNA was extracted from 1-5 cm young panicles of wild-type rice Zhonghua 11 (purchased from the Institute of Crop Science, Chinese Academy of Agricultural Sciences). After mRNA was isolated, a cDNA library was constructed using Stratagene's ZAP-cDNA Synthesis Kit. After titer detection: the titer of the original library was 3.7×10 6 The number of plaques / ml, the titer of the amplified library was 9.3×10 9 Number of plaques / ml. The identification of blue and white spots shows that the plaque rate is only 0.8%. 17 phage plaques were randomly selected, and plasmids were extracted after circularization, and ...

Embodiment 2

[0145] Embodiment 2, the overexpression of gene

[0146] 1. Gene overexpression

[0147] 1. Construction of overexpression vector

[0148] In order to facilitate the detection of protein levels in transgenic plants, we fused the MYC-FLAG tag in front of or behind the OsMADS8 coding region; the Ubiqutin promoter is a promoter with strong expression in monocotyledonous plants, and the Ubiqutin promoter used in Example 2 The promoter is the promoter prepared in Example 1. We chose this promoter to construct two OsMADS8 overexpression vectors: pU1301MFOs8 and pU1301Os8MPF ( Figure 24 ).

[0149] Primers H8F and H8R were designed according to the OsMADS8 sequence, and the forward primer H8F was introduced into the BamHI site in front of the start codon. The sequence is: 5'ACTA ggATCC ATggggAgAgggAgggT-3'; H8R reverse primer, mutating the stop codon TGA in the OsMADS8 sequence to ATC (Ile) and introducing a ClaI site, the sequence is: 5'-CAgg ATGAT gggTAgCCATgTCggCA-3'. The...

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Abstract

The invention discloses an OSMADS8 gene for cultivating dwarf transgenic plants; the nucleotide sequence of the OSMADS8 gene is between 109 and 852 from the 5' terminal in the Genbank, with number of U78892.1; the OSMADS8 gene can be also used for cultivating the transgenic plants with early-flowering characteristics. Besides, by RNAi technology, when OsMADS7 and OsMADS8 genes are simultaneously suppressed, transgenic rice lines are subjected to evident phenotype changes which are mainly shown as follows: the plants are dwarf and late-flowering, and the inner three-round floral organs all have the characteristics of partial palea and lose the determinacy of flower meristem.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of rice E genes in cultivating early flowering and dwarf transgenic plants. Background technique [0002] MIKC-type MADS-box genes encode a large family of transcription factors, because these genes have four structural domains similar to M (MADS), I (Intervening), K (Keratin-like) and C (C-terminal) , hence the name, these genes play an important regulatory role in flower development. Studies on the molecular genetics of flower development in the dicotyledonous model plants Arabidopsis thaliana, Antirrhinum majus and Petunia hybrida have shown that various homeotic MADS-box genes are widely involved in flower development. Regulatory processes in various stages of organ development, and established the ABCDE model of flower development. According to the model, A, B, C, D, and E represent five types of characteristic genes of floral organs, and their gene products re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82
Inventor 孟征吴凤韩加坤崔荣峰
Owner INST OF BOTANY CHINESE ACAD OF SCI
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