Application of soybean MYB transcription factor GmMYB181

A transcription factor and soybean technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as pollen inactivity, biological function speculation, and short plants

Inactive Publication Date: 2014-03-26
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2007, Yang et al. found that overexpression of the AtMYB24 gene in Arabidopsis thaliana would lead to short stature and poor development of floral organs, non-dehiscent anthers, and inactive pollen.
Due to the large number of MYB transcription factors, although they are generally involved

Method used

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  • Application of soybean MYB transcription factor GmMYB181
  • Application of soybean MYB transcription factor GmMYB181
  • Application of soybean MYB transcription factor GmMYB181

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Cloning and identification of soybean GmMYB181 and its coding gene

[0036] The experimental material is soybean (Glycine max(L.)Merr.) local variety Monkey Hair, provided by the Germplasm Resources Laboratory of the National Soybean Improvement Center of Nanjing Agricultural University. In this study, multiple floral organ dominantly expressed genes were identified from soybean flowers by gene chip technology, and one gene fragment (Gma.18002.1.S1_at) encoding MYB transcription factor was selected for further study. The gene fragment was spliced ​​by EST, and sequence comparison revealed that it was the named gene GmMYB181 (DQ822906) on the NCBI website. Primers were designed to further isolate and clone the full-length cDNA sequence of the GmMYB181 gene. The specific method is as follows:

[0037] Soybean flowers were taken, ground in liquid nitrogen, and total RNA was extracted using a plant total RNA extraction kit (TIANGEN DP404). Take 5 μg of total RNA...

Embodiment 2

[0038]Example 2 Expression characteristics of GmMYB181 in different organs and fruits of soybean at different stages

[0039] The expression of GmMYB181 in soybean organs and fruits at different development stages was studied by real-time fluorescent quantitative PCR technology. Soybean experimental materials were sown in Jiangpu Farm in early June, and the field management was routine. Collect roots, stems and leaves; when the third compound leaf unfolds, collect mature postflowers in full bloom; take pods and seeds of 7DAF (7 days after flowering), 15DAF, 25DAF and 40DAF. After collection, the materials were quick-frozen in liquid nitrogen and stored in a -80°C refrigerator for later use.

[0040] The extraction of total RNA was the same as in Example 1. The soybean constitutively expressed gene Tubulin (GenBank accession number: AY907703) was used as an internal reference gene, and the amplification primers were: Tubulin forward primer sequence: 5'GGAGTTCACAGAGGCAGAG3' (S...

Embodiment 3

[0042] Example 3 Functional Identification of GmMYB181 Transcription Factor

[0043] Using Invitrogen's Gateway Technology with Clonase TM II kit, recombine GmMYB181 into the plant expression vector pMDC83 (Curtis et al, 2003, Plant Physiology.133, 462-469), transform Escherichia coli DH5α, and spread the transformation solution on LB solid medium containing 50mg / L hygromycin positive clones were screened. After sequencing verification, the plasmid was extracted to obtain the pMDC83-GmMYB181 plant overexpression vector ( figure 2 ), pMDC83-GmMYB181 was transformed into Agrobacterium tumefaciens strain EHA105 (Biovector Co., LTD) by freeze-thaw method. pMDC83-GmMYB181 was transformed into Arabidopsis thaliana through the mediation of Agrobacterium strain EHA105, cultured on MS medium containing 50mg / L hygromycin, and the transgenic plants with hygromycin resistance were obtained through preliminary screening.

[0044] Genomic DNA of transgenic Arabidopsis thaliana with hy...

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Abstract

The invention belongs to the field of plant genetic engineering and relates to application of a soybean MYB transcription factor GmMYB181. A gene GmMYB181 playing important regulation and control effects in a plant type and reproductive organ development control process is cloned from soybeans and has a sequence as shown in SEQ ID NO.1; the mRNA (messenger ribonucleic acid) expression analysis shows that the gene is a floral organ specifically expressed gene, an expression model of the gene is related with the reproductive development of a plant, and further, the phenotypic change of arabidopsis thaliana of GmMYB181 over-expression shows that the gene GmMYB181 plays the important regulation and control effects in the plant type and reproductive organ development control process. The invention discloses a genetic engineering improvement method for modifying the plant type and the reproductive organ of the plant by the gene; the method has a certain effect on cultivation of plant varieties with changed plant types and reproductive organs, especially soybean varieties, and the plant types and the reproductive organs of crops can be directionally modified to improve the seed production efficiency of the crops and increase the yield of the crops, especially the soybeans.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and relates to the application of a soybean MYB transcription factor GmMYB181, in particular to the application of soybean-derived MYB transcription factor GmMYB181 specifically expressed in floral organs and related to the development and growth of floral organs in plants. Applications in plant type and plant reproductive organ modification. Background technique [0002] Reproductive growth is the most important stage in the life history of higher plants. It is the means and basis for plant reproduction and population dispersal, and it is also a critical period for agricultural production to obtain products. The flower organ is an important functional organ in the reproductive process. The development of the flower is an important symbol of the beginning of the reproductive growth period of the plant. Its development directly affects the formation and quality of fruits and seeds. Flower...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00
Inventor 喻德跃薛茜黄方阚贵珍王慧
Owner NANJING AGRICULTURAL UNIVERSITY
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