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Grape hydrogen peroxide receptor gene and its encoded protein and application

A grape hydrogen peroxide, receptor gene technology, applied in the field of genetic engineering, can solve the problems of hydrogen peroxide receptor distribution and other information expression patterns and subcellular localization are not clear

Active Publication Date: 2022-07-15
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, until now, the distribution and other information (such as expression patterns and subcellular localization) of hydrogen peroxide receptors in the grape genome are still unclear.

Method used

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  • Grape hydrogen peroxide receptor gene and its encoded protein and application
  • Grape hydrogen peroxide receptor gene and its encoded protein and application
  • Grape hydrogen peroxide receptor gene and its encoded protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Identification and cloning of hydrogen peroxide receptor members in grapes

[0047] Based on the hydrogen peroxide receptor characteristics, we used the grape genome (12X.v2) and the corresponding annotation information (VCost.v3) to perform local BLAST search for HPCA in the grape whole genome (the E-value of the search result was less than 0.01), Screening for hydrogen peroxide receptors in grapes. We identified a total of four hydrogen peroxide receptors VvHPCA1, 2, 3 and 4 from the grapevine genome. The phylogenetic tree was constructed by MEGA software, and the characteristic sequences of HPCA in different species were analyzed by multiple sequence alignment ( figure 1 ). The above results indicate that the VvHPCAs identified by the present invention belong to the LRR-RK family and have a hydrogen peroxide characteristic domain.

[0048] Full-length cloning: high-fidelity DNA polymerase Max DNA Polymerase (Bao Bioengineering Co., Ltd., Dalian, China)...

Embodiment 2

[0054] Example 2 Motif and conserved domain analysis of proteins encoded by VvHPCAs

[0055] The above sequencing results (DNA sequences) were translated into amino acid sequences using Primer Premier v5 software. The amino acid sequence of VvHPCA1 protein is shown in SEQ ID NO.1, the amino acid sequence of VvHPCA2 protein is shown in SEQ ID NO.2, the amino acid sequence of VvHPCA3 protein is shown in SEQ ID NO.3, and the amino acid sequence of VvHPCA4 protein is shown in SEQ ID NO. .4 shown.

[0056] Using the Pfam database (http: / / pfam.xfam.org / search#tabview=tab1), NCBIConserved Domain Search (https: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi) and SMART ( Simple Modular Architecture Research Tool) (http: / / smart.embl-heidelberg.de) for LRR-RK and HP domain analysis.

[0057] The results showed that VvHPCAs contained a conserved motif at the C-terminus ( figure 2 A), both contain serine threonine kinase catalytic domain (Catalytic domain of serine threonine kinase, STK...

Embodiment 3

[0062] Example 3H 2 O 2 Treatment of 'Kyoho' grapes with ROS inhibitor (hypotaurine, HT)

[0063] 'Kyoho' grapes (Vitis vinifera cv. 'Kyoho') were collected during commercial harvesting, and grapes with the same ear size and fruit color were selected for processing. Refer to Guo et al. (Transcriptome analysis reveals mechanism of early ripening in Kyoho grape with hydrogen peroxide treatment. BMC Genomics. 21, 784. https: / / doi:10.1186 / s12864-020-07180-y .) method, configure 300mmol L - 1 H 2 O 2 , 150mmol L -1 HT solution (containing 0.01% Silwet_L-77) and double distilled water as the control, soak the selected ear in the above solution for 15min. Then put it into a fresh-keeping box, cover it with plastic wrap, and store it in a storage room at 25±1°C. Controls, H 2 O 2 and HT-treated fruit ( image 3 ), fully frozen in liquid nitrogen and stored at -80°C for subsequent analysis and experiments.

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Abstract

The present invention relates to grape hydrogen peroxide receptor gene and its encoded protein and application. The present invention identifies the grape hydrogen peroxide receptor VvHPCAs gene. The present invention uses hydrogen peroxide and hypotaurine to treat postharvest 'Kyoho' grapes, and measures the shedding rate, water loss rate, hardness, H 2 O 2 content and ROS metabolism-related enzyme activities, comprehensively monitor postharvest quality and ROS level changes, and analyze the expression of VvHPCAs in the above samples. 2 O 2 There was a positive correlation between the level of VvHPCA1 and VvHPCA3 expression and the degree of aging mediated 2 O 2 The content is negatively correlated with the degree of postharvest senescence; the invention also provides the application of VvHPCA1 and VvHPCA3 genes, and overexpressing VvHPCA1 gene or inhibiting the expression of VvHPCA3 can enhance the effect of grape fruit on H 2 O 2 Sensitivity of VvHPCA1 gene or overexpression of VvHPCA3 gene can reduce the sensitivity of grape fruit to H. 2 O 2 susceptibility to resist H 2 O 2 induced stress and aging.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a grape hydrogen peroxide receptor gene and its encoded protein and application. Background technique [0002] Reactive oxygen species (ROS) are a class of molecules / ions with active chemical properties and high oxidative activity, including hydrogen peroxide (H 2 O 2 ), hydroxyl radical (OH) . ) and so on. It can either be generated by cellular metabolic pathways (by-products) or actively synthesized by cells under specific physiological conditions (Mitler 2017). ROS can regulate the physiological activities of cells within a certain concentration range, otherwise it will lead to oxidative damage and even cell death. In recent years, the function of ROS in fruit ripening and senescence has attracted more and more attention. There are usually two accumulation peaks of ROS during fruit ripening and senescence. Generally, it rises sharply and reaches t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/11C12N15/82C12Q1/6851C07K14/415A01H5/00A01H5/08A01H6/88
CPCC07K14/415C12N15/8271C12N15/8266C12Q1/6851C12Q2531/113C12Q2521/107
Inventor 郭大龙刘海楠裴茂松余义和韦同路
Owner HENAN UNIV OF SCI & TECH
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