Soybean photosynthesis related gene GmDeg1 and applications thereof

A photosynthesis and soybean technology, applied in the field of genetic engineering, can solve problems such as unclear, unreported Deg gene family, etc.

Inactive Publication Date: 2013-03-20
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the mechanism of how the reduction of the activity of the PSII reaction center converts the excitation energy to heat dissipation is still unclear.
[0005] So far, in soybean, the Deg gene family has not been reported

Method used

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  • Soybean photosynthesis related gene GmDeg1 and applications thereof
  • Soybean photosynthesis related gene GmDeg1 and applications thereof
  • Soybean photosynthesis related gene GmDeg1 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the cloning of soybean GmDeg1 gene

[0038] The total RNA of the leaves of soybean variety Kefeng 1 was extracted with a plant total RNA extraction kit (purchased from Tiangen Company), and the integrity of the RNA was detected by 1% agarose gel electrophoresis. The cDNA was synthesized according to the instruction of SYBR PrimeScript RT-PCR Kit II kit from TaKaRa Company. Design primers:

[0039] GmDeg1-F: 5'-GGGGACAAGTTTGTACAAAAAAAGCAGGCTTCCCAACCAAAAACAATGGCCAC-3' (SEQ ID NO. 3)

[0040] GmDeg1-R: 5'-GGGGACCACTTTGTACAAGAAAGCTGGGTCTGATTCATCAGGCTTTGGCTCC-3' (SEQ ID NO. 4)

[0041]Prepare PCR reaction solution (50μl system) according to the following component order

[0042]

[0043] Carry out the PCR reaction according to the following procedure

[0044]

[0045] PCR products were detected by 1% agarose gel electrophoresis ( figure 1 ) and recovery and purification, the recovered fragment was TA-cloned into pMD19-TSimple Vector (purchased from T...

Embodiment 2

[0046] Example 2, Fluorescent Quantitative Analysis of Soybean GmDeg1 Gene

[0047] Using MV (methyl viologen) as simulated photooxidation treatment, 600 μmol L containing 0.01% Tween-20 -1 MV was sprayed on the leaves, and the light was set at 500 μmol m -2 ·s -1 , the leaves were quickly taken at 0h, 1h, 3h, 6h and 12h respectively, quickly frozen in liquid nitrogen, and stored in a -80°C refrigerator. Using 18S rRNA as an internal reference gene, Primer Premier (V5.0) software was used to design primers. The primer sequences are as follows:

[0048] Specific primers for amplifying GmDeg1:

[0049] Upstream primer: 5'-TGTCTACATAACCAACCTTGC-3' (SEQ ID NO.5)

[0050] Downstream primer: 5'-GACCTTGAGATCAGAGGC-3' (SEQ ID NO.6)

[0051] Specific primers for amplifying 18S rRNA:

[0052] Upstream primer: 5'-CGGCTACCACATCCAAGGAA-3' (SEQ ID NO.7)

[0053] Downstream primer: 5'-GCTGGAATTACCGCGGCT-3' (SEQ ID NO.8)

[0054] Prepare the PCR reaction system (20μl system) according...

Embodiment 3

[0059] Embodiment 3, expression analysis of soybean GmDeg1 gene in Escherichia coli

[0060] Design specific primers, add HindIII (AAGCTT), XhoI (CTCGAG) restriction sites and protective bases, use the cDNA prepared in Example 1 as a template, amplify the truncated GmDeg1 fragment, and construct the pET28a-GmDeg1 vector. Sex primers are:

[0061] Upstream primer: 5'-CCCAAGCTTTTGTTGTCACCCTCCCCAAGGAA-3' (SEQ ID NO.9)

[0062] Downstream primer: 5'-CCGCTCGAGTGATTCATCAGGCTTTGGCTCC-3' (SEQ ID NO.10)

[0063] After the PCR reaction procedure, the PCR product was recovered and TA cloned into pMD19-T Simple Vector, the bacterial solution with the correct sequencing result was selected to extract the plasmid, and then pMD19T-GmDeg1 and pET28a were double digested to recover the digested product. Transform Escherichia coli, extract the plasmid and detect pET28a-GmDeg1 by double enzyme digestion. Transform pET28a-GmDeg1 into BL21(DE3) competent cells.

[0064] Prokaryotic expression ...

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Abstract

The invention belongs to the field of genetic engineering, and discloses a soybean photosynthesis related gene GmDeg1 and applications thereof. The nucleotide sequence of the soybean photosynthesis related gene GmDeg1 is shown as SEQ ID NO. 1, and the amino acid sequence of coded proteins is shown as SEQ ID NO. 2. By using a carrier which can guide an exogenous gene to be expressed in a plant, the gene GmDeg1 disclosed by the invention is guided into plant cells, so that a transgenic plant with an improved photoinhibition resistance capacity can be obtained. The gene disclosed by the invention has an anti-photoinhibition effect on soybeans, and especially has a great significance on cultivating high-photosynthetic-efficiency soybean varieties.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and relates to a soybean photosynthesis-related gene GmDeg1 and an application thereof. Background technique [0002] Photosynthesis is the source and basis for the survival of the biological world. However, under strong light, the photosynthetic function of plants is easily inhibited, and the photosynthetic mechanism is destroyed, thereby reducing the photosynthetic efficiency. This process is called photoinhibition. During the long-term evolution process, plants have formed a variety of biophysical and biochemical mechanisms to protect them from strong light damage, flexibly resist complex and changeable light stress, and minimize photoinhibition damage. In addition to some morphological adaptations, several protective mechanisms exist in the photosynthetic apparatus, such as heat dissipation, photorespiration, and turnover of the D1 protein. The main target of light damage is the D1 protein...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/57C12N9/50C12N15/63C12N15/82A01H5/00
Inventor 杨守萍孔星张璟曜
Owner NANJING AGRICULTURAL UNIVERSITY
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