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Transcription factor EjMYB1 participating in control on synthesis of loquat fruit lignin and application thereof

A technology of transcription factor and lignin, which is applied in the field of transcription factor EjMYB1, can solve problems such as roughness and less juice, decline in commodity value, and light flavor, and achieve the effect of low economic benefit and heavy workload

Inactive Publication Date: 2014-09-10
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unlike other fruits that soften after harvesting, red meat loquat fruits will undergo lignification after harvesting, making the fruit firmer, rougher and less juicy, the flavor becomes weaker, and the commodity value decreases

Method used

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  • Transcription factor EjMYB1 participating in control on synthesis of loquat fruit lignin and application thereof
  • Transcription factor EjMYB1 participating in control on synthesis of loquat fruit lignin and application thereof
  • Transcription factor EjMYB1 participating in control on synthesis of loquat fruit lignin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 : ikB1 Acquisition of full-length sequences

[0027] (1) Experimental method

[0028] 1. Sequence search: In the Apple Genome V1.0 predicted peptides database of Rosaceae (http: / / www.rosaceae.org), apply the blastp algorithm to AtMYB58 Search for the amino acid sequence; select the top three sequences with the highest matching degree: MDP0000133542 (chr5:11008894..11011881), MDP0000435315 (chr3:7586759..7588715), MDP0000230141 (chr3:7611165..76131 21), according to these three sequences According to the location on the apple genome, the corresponding nucleotide sequence was obtained, primers were designed, and the loquat cDNA was used as a template to obtain the loquat sequence SEQ: NO. 1 with MDP0000230141 (chr3:7611165..7 613121) as the reference sequence

[0029] 2. ikB1 Sequence amplification: obtain a sequence SEQ: NO. 1 according to the found nucleotide sequence amplification, named as ikB1 . Loquat cDNA was used as template, and SEQ: NO. 9-10 w...

Embodiment 2

[0033] Example 2 : ikB1 Analysis of expression patterns in different tissues and fruits of different treatments of loquat

[0034] (1) Experimental method

[0035] 1. Sample Processing

[0036] Loquat fruits with consistent maturity, uniform size and no pests and diseases were treated as follows: 0°C, that is, the fruit was stored directly at 0°C; heat shock treatment, that is, after 4 hours of treatment at 40°C, and then stored at 0°C; program cooling For processing, after placing at 5°C for 6 days, store at 0°C. The pulp was cut into pieces and quickly placed in liquid nitrogen, and then stored at -80°C. Leaf and stem tissues were frozen in liquid nitrogen and stored at -80°C.

[0037] 2. Total RNA extraction

[0038] Weigh 0.2 g each of loquat leaves and stems and 1 g of fruit stored at -80°C, and extract total RNA according to the following steps: the samples are fully ground in liquid nitrogen, and added to a preheated 65°C container containing 4 ml CTAB / 80 Put μl...

Embodiment 3

[0043] Example 3 : ikB1 Analysis of Promoter Activity of Transcription Regulated Lignin Synthesis Genes

[0044] (1) Experimental method

[0045] 1. Construction of loquat lignin synthesis gene promoter vector

[0046] Design primers (SEQ: NO. 11-28), using the loquat DNA prepared by the GenomeWalker® Universal Kit (Clontech, USA) as a template, using Platinum ?Two rounds of PCR with Taq DNA Polymerase system to amplify loquat lignin synthesis gene JPAL , wxya s and EjCAD s promoter sequence (SEQ: NO. 29-37). The first round of PCR reaction program was denaturation at 94°C for 25 s, 72°C for 3 min, 7 cycles; denaturation at 94°C for 25 s, 67°C, 3 min, 32 cycles; extension at 67°C for 7 min. Using the first-round PCR product as a template, the second round of PCR amplification was performed. The reaction program was denaturation at 94°C for 25 s, 72°C for 3 min, 7 cycles; denaturation at 94°C for 25 s, 67°C, 3 min, 32 cycles Cycling; extension at 67°C for 7 min. Af...

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Abstract

The invention provides a transcription factor EjMYB1 participating in control on synthesis of loquat fruit lignin. By using AtMYB58 amino acid sequence for specifically controlling the lignin synthesis in Arabidopsis thaliana as the reference sequence, homologous cloning is performed according to the rose family database to obtain the transcription factor EjMYB1. The expression level of the EjMYB1 is positively related to the accumulation of the lignin in different tissues and different treatment processes of the loquat. The EjMYB1 can positively control the promoter activity of the lignin synthesis genes EjPAL1, Ej4CL1 and Ej4CL5. The EjMYB1 can obviously enhance the leaf lignin content when being instantaneously overexpressed in tobacco.

Description

technical field [0001] The invention belongs to the fields of plant molecular biotechnology and genetic engineering, and relates to a transcription factor involved in the regulation of loquat fruit lignin synthesis ikB1 . Background technique [0002] MYB is a class of functionally diverse transcription factors present in eukaryotes. MYB contains a highly conserved DNA domain, called the MYB domain, which is composed of 4 incompletely repeated amino acid sequences consisting of 52 amino acids. According to the number of MYB domains, it can be divided into several categories, among which R2R3-MYB is in plant There are 126 R2R3-MYB members in Arabidopsis, which are subdivided into 22 subfamilies. R2R3-MYB is involved in plant-specific growth processes, primary and secondary metabolism, cell building, developmental processes, and responses to biotic and abiotic stresses. [0003] Lignin is an aromatic heteropolymer and one of the main components of secondary cell walls. Lig...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/10C12N15/84A01H5/00
Inventor 殷学仁徐倩李鲜陈昆松
Owner ZHEJIANG UNIV
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