CabuAG gene related to pistil and stamen development of catalpa bungei and protein and application thereof

A protein, pbi121-cabuag technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem that there is no AGAMOUS homologous gene research.

Inactive Publication Date: 2015-12-09
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, catalpa is an important timber and ornamental tree species in my country, and there is no research on its AGAMOUS homologous gene

Method used

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  • CabuAG gene related to pistil and stamen development of catalpa bungei and protein and application thereof
  • CabuAG gene related to pistil and stamen development of catalpa bungei and protein and application thereof
  • CabuAG gene related to pistil and stamen development of catalpa bungei and protein and application thereof

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Experimental program
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Effect test

Embodiment 1

[0025] The molecular cloning of embodiment 1 catalpa tree CabuAG gene cDNA sequence

[0026] (1) Extraction of total RNA from flower buds of Chinese catalpa

[0027] Get 300 mg of fresh catalpa flower buds with a length of about 5 mm, put them into a cryopreservation tube, and freeze them in liquid nitrogen immediately.

[0028]Use the RNA extraction kit to extract total RNA: take the required flower buds out of the -80°C ultra-low temperature refrigerator, put them in a mortar with 2mL RLT and 100μL PLAN Taid, and grind them thoroughly at room temperature; transfer the above grinding solution into a 2mL EP tube , after centrifuging at 13000rpm for 10min, take 500μL supernatant and transfer it to a new 2mL centrifuge tube; add 250μL absolute alcohol to the supernatant, blow and mix well; add the above liquid into the adsorption column, and put the adsorption column into the collection tube Add 500 μL of washing solution to the adsorption column, centrifuge at 13,000 rpm for 1...

Embodiment 2

[0038] The protein sequence of embodiment 2 Chinese catalpa CabuAG gene code

[0039] Using the primer5 software, the full-length cDNA sequence of the CabuAG gene was translated into protein sequence. The full-length cDNA sequence was 1225bp, and the CDS was 729bp. The stop codon was removed, and a total of 242 amino acids were edited. Get SEQ ID NO: 2, and according to the characteristics of the MADS-box gene, the Catalpa CabuAG protein sequence is divided into M region, I region, K region, C region and two motifs AGmotifI and AGmotifII, see for details figure 2 .

Embodiment 3

[0040] The tissue specificity of embodiment 3 Chinese catalpa CabuAG gene expression

[0041] According to the cDNA sequence full-length of Chinese catalpa tree CabuAG gene, utilize oligo6.0 software to design the primer RTAGF of semiquantitative test: 5'-CCTTCAACTCCTACTTATTACCA-3' (as shown in SEQIDNo.11) and RTAGF: 5'-CCATTCTGCTCGTTCTTACTTAT-3' ( As shown in SEQ ID No.12). Use PCR to detect its specificity, and it can only be used under the premise of ensuring that the primers only specifically amplify the corresponding genes. Taking the catalpa actin gene as the positive control of the semi-quantitative test, the primers of the actin gene are CabuactinF: 5'-TCACACTGGTGTGATGGTTG-3' (as shown in SEQ ID No.13) and CabuactinR: 5'-AGTTCTTCTCCACAGCAGAG-3' (as shown in SEQ ID No.14 shown). And adjust the detection band brightness by the proportion of the first strand cDNA template amount of young leaves, sepals, petals, stamens and pistils. Semi-quantitative PCR reaction progra...

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Abstract

The invention relates to the field of plant molecular biology, in particular to a CabuAG gene related to the pistil and stamen development of catalpa bungei and encoded protein and application thereof. The CabuAG gene related to the pistil and stamen development of the catalpa bungei belongs to one of the following two kinds of protein, wherein the first kind of the protein is composed of amino acid shown in SEQ ID No.2, and the second kind of the protein is derived from the first kind of the protein and obtained by the fact that the amino acid sequence shown in SEQ ID No.2 is replaced by or lack of or added with one or several amino acids and has the same activity. The invention provides the gene for encoding the protein and application thereof in plant flower organ improvement. The invention further provides an express carrier containing the CabuAG gene. Compared with a wild plant, the CabuAG-transgenic plant can show the characteristics of special development of the flower organs.

Description

technical field [0001] The invention belongs to the field of plant molecular biology, and in particular relates to a CabuAG gene related to the development of pistils and stamens of Chinese catalpa, its encoded protein and its application. Background technique [0002] There are about 13 species of catalpa plants in the world, mainly distributed in eastern Asia and America. There are 6 species mainly distributed in my country, all of which are important timber and ornamental tree species, including: catalpa, catalpa, catalpa, catalpa, Tibetan catalpa and golden tree, among which catalpa is a variant of catalpa, and golden tree is an introduced species. From the analysis of kinship, morphological structure, and characteristics of flowering and fruiting, these six species of Catalpa were divided into two groups, of which Catalpa, Ash and Diana belonged to the Catalpa group, while Catalpa, Tibetan Catalpa and Golden Tree Belongs to the catalpa group. [0003] Catalpa (Catalpa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N1/21A01H5/02
CPCC07K14/415C12N15/827
Inventor 王军辉景丹龙夏燕张守攻
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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