PyGSTf12 gene related to transport of anthocyanin in pear fruits, and recombinant expression vector and application of PyGSTf12 gene

An expression vector, anthocyanin technology, applied in application, genetic engineering, plant genetic improvement, etc., to reduce agricultural costs and achieve environmental friendliness

Inactive Publication Date: 2019-05-28
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether GSTs genes are involved in anthocyanin transport and which transcription factors are regulated in Hongpi pear fruit have not been reported yet.

Method used

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  • PyGSTf12 gene related to transport of anthocyanin in pear fruits, and recombinant expression vector and application of PyGSTf12 gene
  • PyGSTf12 gene related to transport of anthocyanin in pear fruits, and recombinant expression vector and application of PyGSTf12 gene
  • PyGSTf12 gene related to transport of anthocyanin in pear fruits, and recombinant expression vector and application of PyGSTf12 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Changes in the content of anthocyanins in different tissues and different development stages of 'Hongzaosu' and its green buds of the present invention

[0034] The total anthocyanin content of 'Hongzaosu' in different tissues was significantly different ( figure 1 B). At 20 days after flowering, the content of ‘Hongzaosu’ in young leaves was 3.26mg.g-1 FW, the young fruit was slightly lower than the leaves, and the petals and young stems were the lowest ( figure 1 B). During fruit development, the total anthocyanin content of 'Hongzaosu' fruit decreased to a relatively low level during the fruit swelling stage (60DAFB) until the fruit ripening stage (120DAFB). In 20DFAB, the highest value is 2.49mg.g-1FW, the lowest value is 0.86mg.g-1FW. The content of anthocyanins in the green mutants of ‘Hongzaosu’ was always low, with only slight changes in different tissues and developmental stages. The total anthocyanin content of the red tissue of 'Hongzaosu' was s...

Embodiment 2

[0035] Example 2 Screening of Differentially Expressed Genes in Four Tissues of Stems, Leaves, Flowers and Fruits of 'Red Zaosu' and Its Green Bud Variation of the Present Invention

[0036] A total of 7650 differentially expressed genes were identified by comparing the red and green tissues of 'Hongzaosu' and its green buds, and there were large differences in differentially expressed genes in different tissues. Among them, a total of 3,708 differentially expressed genes were identified, including 2,101 up-regulated genes and 1,607 down-regulated genes, compared with the green mutant of ‘Hongzaosu’ at 60 days after flowering. The stems of ‘Hongzaosu’ and its green mutant had the least differentially expressed genes (30 up-regulated genes and 10 down-regulated genes). These differentially expressed genes provide a large number of candidate genes related to anthocyanin biosynthesis and transport.

[0037] In addition, we further screened co-differentially non-expressed genes i...

Embodiment 3

[0038] Example 3 The flavonoid metabolic pathway map and the expression pattern heat map and correlation analysis of related genes in 'Hongzaosu' and its green mutants of the present invention.

[0039] The biosynthesis of flavonoids is catalyzed by a series of enzymes and regulated by transcription factors ( figure 2 A). Through RNA-Seq analysis, we identified 60 candidate genes involved in catalysis and regulation ( figure 2 B). Among these genes we found flavanone 3-hydroxylase (FLS, Pbr033501.1) and UDP-glucose:flavonoid-3-O-glucosyltransferase (UFGT, Pbr000426.1, Pbr000427.1, Pbr000428 .1, Pbr001279.1) had higher gene expression levels in red tissues than in green tissues. Notably, the expression level of the UFGT gene Pbr000427.1 was 10-fold higher than that of the gene Pbr000426.1. Since anthocyanin biosynthesis occurs in the cytoplasm, it requires transport to vacuolar storage. The gene Pbr012649.1 encodes glutathione S-transferase, which showed higher expressio...

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Abstract

The invention discloses a pear transcription factor PyGSTf12, and a recombinant expression vector and application thereof. The sequence difference between the PyGSTf12 and a promoter thereof is used as a molecular marker for identifying red and green peels, wherein the PyGSTf12 and the promoter are respectively separated from 'Red Zaosu' pears and yellow bud mutated fruits thereof, and the nucleotide sequences of the PyGSTf12 and the promoter are respectively as shown in SEQ ID No. 31 and SEQ ID No. 32. By a dual-luciferase report system, an agrobacterium-mediated instant genetic transformation method proves that the PyGSTf12 co-transformed by transcription factors PyHY5 and PyMYB114 promotes accumulation of anthocyanin in strawberry and pear fruits, and the PyGSTf12 promoter promotes thetransport function of the anthocyanin; through discovery of the PyGSTf12 gene, new genetic resources are provided for molecular breeding that promotes synthesis of the anthocyanin in pear peels, and new genetic resources are provided for the implementation of green agriculture; the development and utilization of the genetic resources is beneficial to agricultural cost reduction and environmental friendliness.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and relates to the recombinant expression vector and application of the PyGSTf12 gene related to anthocyanin transport in pear fruit, and specifically relates to the separation and cloning of a PyGSTf12 involved in the transport of anthocyanin in pear fruit peel from the 'Hongzaosu' pear Genes and their applications. Background technique [0002] Plant tissues with stems, leaves, flowers, and fruits are particularly attractive with red characteristics, and anthocyanins are the main pigments that make up the red color. This red color plays an important role in attracting insect pollinators and aiding in seed dispersal (Regan et al., 2001; Schaefer et al., 2004). In addition, anthocyanins also have important biological roles, such as resistance to disease and ultraviolet radiation, and reduction of reactive oxygen species in plants. As for human health, anthocyanins have potential effects...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/74C12N1/21C12N15/82A01H5/00A01H6/74C12N15/113C12Q1/6895
Inventor 吴俊姚改芳张绍铃杨广艳李甲明汪润泽齐开杰
Owner NANJING AGRICULTURAL UNIVERSITY
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