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Cucumber CsERF004 gene as well as encoding protein and application thereof

A technology that encodes proteins and cucumbers, and is used in applications, genetic engineering, plant genetic improvement, etc.

Active Publication Date: 2017-06-20
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research of ERF on cucumber disease resistance has not been reported

Method used

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  • Cucumber CsERF004 gene as well as encoding protein and application thereof
  • Cucumber CsERF004 gene as well as encoding protein and application thereof
  • Cucumber CsERF004 gene as well as encoding protein and application thereof

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Experimental program
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Effect test

specific Embodiment approach 1

[0024] Embodiment 1: The nucleotide sequence of the cDNA of the cucumber CsERF004 gene in this embodiment is shown in SEQ ID NO: 1 in the sequence listing.

specific Embodiment approach 2

[0025] Specific embodiment 2: The amino acid sequence of the encoded protein of the cucumber CsERF004 gene in this embodiment is shown in SEQ ID NO: 2.

[0026] ERF is an important transcriptional regulator in the SAR signaling pathway, and plays a very important role in the induced plant disease resistance. The present invention reports the Arabidopsis ERF-related gene CsERF004 in cucumber for the first time, and clones the full-length cDNA of cucumber CsERF004 by PCR technology. CsERF004 was introduced into cucumber and overexpressed to obtain transgenic offspring cucumber. The first functional verification showed that the overexpression of CsERF004 gene significantly increased the content of free salicylic acid in cucumber, and it had obvious resistance to downy mildew and Corynespora leaf spot at the seedling stage. In order to reduce cucumber diseases and insect pests and improve resistance to cucumber downy mildew and Corynespora leaf spot, a new solution is provided. ...

Embodiment 1

[0028] Example 1, Trizol method to extract the RNA of cucumber susceptible variety 'D0401'

[0029] Total RNA was extracted from cucumber leaves by Trizol method.

[0030] (1) Take 100 mg of fresh cucumber leaf tissue, add liquid nitrogen to grind thoroughly, and transfer the powder into a 1.5 mL sterile centrifuge tube;

[0031] (2) Add 1mL Trizol, shake fully, and let stand at 15-30°C for 5min;

[0032] (3) Add 300 μL of chloroform, shake vigorously for 15 seconds, and centrifuge at 12000 rpm for 15 minutes at 4°C;

[0033] (4) Transfer the supernatant into a new sterile centrifuge tube, add an equal volume of pre-cooled isopropanol, place at -20°C for 20min, and centrifuge at 12000rpm at 4°C for 15min;

[0034] (5) Discard the supernatant, add 1 mL of 75% DEPC-ethanol, vortex wash for 10 s, and centrifuge at 7500 rpm at 4°C for 5 min (repeat the operation twice);

[0035] (6) Abandon the ethanol, and place the centrifuge tube on a clean filter paper to dry;

[0036] (7)...

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Abstract

The invention relates to a cucumber CsERF004 gene as well as an encoding protein and application thereof. The invention provides a cucumber disease-resistance related gene CsERF004 gene as well as an encoding protein and application thereof. A cDNA (complementary Deoxyribonucleic Acid) nucleotide sequence of the cucumber CsERF004 gene is shown as SEQ ID NO: 1 in a sequence table. An amino acid sequence of the encoding protein is shown as SEQ ID NO: 2. The CsERF004 gene overexpression obviously improves the content of free-state salicylic acid in cucumbers, and the cucumber CsERF004 gene has obvious resistance on downy mildew and target leaf spot at a seedling period. The invention provides a new solution way for reducing diseases and insect pests of the cucumbers and improving the disease resistance of the downy mildew and target leaf spot of the cucumbers. The cucumber CsERF004 gene provided by the invention is used for improving the resistance on the downy mildew and target leaf spot of the cucumbers.

Description

technical field [0001] The present invention relates to CsERF004 gene and its coded protein and application. Background technique [0002] Cucumber is a staple vegetable that is eaten as a tender fruit. In production, cucumbers are susceptible to infection by a variety of pathogenic bacteria, which seriously affects their yield and quality. Cucumber downy mildew and Corynespora leaf spot are important diseases in cucumber production. The disease spreads rapidly and poses a great threat to cucumber production. The long-term use of chemical agents for disease control has resulted in resistance to downy mildew and Corynesporium leaf spot. At the same time, a large amount of pesticide residues will cause environmental pollution and endanger human health and other problems. The green, economical and effective control method is still breeding and utilizing disease-resistant varieties. Breeding single disease-resistant varieties can no longer solve the serious impact of diseas...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415A01H5/00
CPCC07K14/415C12N15/8282
Inventor 辛明秦智伟刘东周秀艳张艳菊
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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