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Europe and America hybrid grape variety Kyoho stress-resistant gene VlbZIP36 and application thereof

A grape and gene technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems affecting yield and quality

Active Publication Date: 2016-11-16
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because it is often exposed to various biotic and abiotic stresses, its yield and quality in the field are seriously affected.

Method used

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  • Europe and America hybrid grape variety Kyoho stress-resistant gene VlbZIP36 and application thereof
  • Europe and America hybrid grape variety Kyoho stress-resistant gene VlbZIP36 and application thereof
  • Europe and America hybrid grape variety Kyoho stress-resistant gene VlbZIP36 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Bioinformatics analysis of VlbZIP36

[0037] The phylogenetic tree analysis of the amino acid sequence of VlbZIP36 and the bZIP amino acid sequences of groups A and K of other studied plants and the multiple sequence alignment of highly conserved amino acid residues show that the amino acid sequence of VlbZIP36 is consistent with the amino acid sequence of Arabidopsis AtbZIP60 and rice OsbZIP74 The homology is the highest, and they all belong to the K group subfamily of the bZIP family ( figure 1 A). Analysis of the amino acid structures of VlbZIP36 and other bZIPs revealed that their amino acid sequences all contain a bZIP conserved domain consisting of a DNA binding domain and a leucine zipper domain ( figure 1 B).

Embodiment 2

[0038] Example 2: Analysis of the expression pattern of VlbZIP36

[0039] In order to analyze the expression pattern of VlbZIP36, the inventors analyzed the VlbZIP36: GUS activity of transgenic Arabidopsis with GUS expression vector. It was found that GUS was present in the cotyledon tips and roots of 5-day-old seedlings ( figure 2 C), there is expression in most tissues of 2-week-old seedlings ( figure 2 D), but no expression in newly germinated mature embryos ( figure 2 A). However, after mannitol stress treatment, GUS was expressed at the tips of newly germinated mature embryos ( figure 2 B), and the expression was significantly enhanced in all tissues in the 2-week-old seedlings ( figure 2 E). In adult seedlings, GUS was found in 3-week-old petioles ( figure 2 F), anthers, styles, siliques ( figure 2 Higher expression in L-O), weaker expression in guard cells and sepals ( figure 2 I), but no expression in petals. After drought stress treatment and ABA tre...

Embodiment 3

[0040] Example 3: Resistance of VlbZIP36 transgenic Arabidopsis lines to osmotic stress

[0041] The experimental results of osmotic stress treatment on the seeds of transgenic lines and wild-type plants showed that almost 100% of the transgenic lines and wild-type seeds could germinate after being cultured on MS medium for 3 days. After being cultured on the medium for 3 days, the transgenic lines showed a 21-28% higher seed germination rate than the wild type ( image 3 ). At the same time, after culturing on osmotic medium for 6 days, the cotyledon greening rate of the transgenic line was significantly higher than that of the wild type ( Figure 4 A, B). For example, when the concentration of mannitol was 300mM, the cotyledons of about 83% of the transgenic lines turned green, while only 66% of the wild-type plants turned green; when the concentration of mannitol was 350mM, the transgenic lines It showed a survival rate about 27% higher than that of the wild type. The i...

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Abstract

The invention discloses a Europe and America hybrid grape variety Kyoho stress-resistant gene VlbZIP36 and an application thereof. The disclosed gene complete open reading frame sequence is 969 bp, and 322 amino acids are encoded. Transgenic arabidopsis thaliana with overexpression of VlbZIP36 is found out to show enhanced resistance to drought stress in a seed germination stage, a seedling stage and a mature stage, and moreover, the resistance is related with changes of relative conductivity and malonaldehyde and expression enhancement of stress related genes. With use of the VlbZIP36 gene, water loss is restricted, the activity of antioxidase is regulated to remove ROS, the expression level of the related target gene is transcribed and regulated, and the VlbZIP36 gene plays a role in enhancing drought stress resistance in transgenic arabidopsis thaliana. The application disclosed by the invention is an application of the gene VlbZIP36 in improving the ability of plants to resist drought stress.

Description

technical field [0001] The invention relates to the technical field of plant stress resistance gene identification and genetic engineering, in particular to a European and American hybrid grape variety Kyoho stress resistance gene VlbZIP36. Background technique [0002] Basic leucine zipper bZIP transcription factor is the most widely distributed and conserved protein among eukaryotic transcription factors, and is found in humans, animals, plants, microorganisms and insects. At present, bZIP transcription factors have been discovered in various plants, for example, there are 75 in the Arabidopsis genome, 89 or 92 in the rice genome, 47 in soybean, 92 in sorghum, and 125 in maize. 47 or 55 were found in the grape genome and so on. According to the similarity and conservation of their structural domains, they can be divided into 10 subfamilies, namely A, B, C, D, E, F, G, H, I, S; or 13 subfamilies, namely A, B, C, D, E, F, G, H, I, J, K, L, S, etc. [0003] The structural ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 王西平涂明星
Owner NORTHWEST A & F UNIV
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