Group of pyrus peel qRT-PCR reference genes and primers and application of reference genes

An internal reference gene and gene technology, which is applied in the field of plant genetic engineering, can solve the problems of no comparative verification of expression stability, lack of combined application of multiple internal references, and poor stability of internal references.

Active Publication Date: 2020-05-08
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 1) The application of these traditional internal reference genes in the study of pear peel is only a reference to other species, without systematic comparison and verification of expression stability within the genome, and many studies have shown that the expression of these traditional internal reference genes under different experimental conditions does not vary. Stablize;
[0009] 2) At present, only a single internal reference is used for homogenization in the study of gene expression in pear peel, and there is a lack of more accurate combination of multiple internal references;
[0010] 3) There is a lack of a suitable internal reference gene system for the study of gene expression in the pericarp of a wide range of pear varieties
[0011] Therefore, it is necessary to establish a suitable internal reference gene system on a genome-wide scale based on extensive transcriptome data as soon as possible to solve the problems of poor stability of traditional internal references and unsatisfactory gene expression equalization effects in pear peels

Method used

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  • Group of pyrus peel qRT-PCR reference genes and primers and application of reference genes
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  • Group of pyrus peel qRT-PCR reference genes and primers and application of reference genes

Examples

Experimental program
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Effect test

Embodiment 1

[0077] This example provides internal reference gene sequences for pear peel development, bagging and postharvest low temperature storage, including:

[0078] (1) The NAP1 gene transcript is shown in the sequence table as SEQ ID NO: 1;

[0079] (2) The BPS1 gene transcript is shown in the sequence table as SEQ ID NO: 2;

[0080] (3) The Alba-like1 gene transcript is shown in the sequence table as SEQ ID NO: 3;

[0081] (4) The UBC32 gene transcript is shown in the sequence table as SEQ ID NO: 4;

[0082] (5) ACT6 / 7 / 8 / 9 gene transcripts are shown in SEQ ID NO: 5 in the sequence table.

Embodiment 2

[0084] This embodiment provides a set of pear peel development process, bagging and postharvest low temperature storage internal reference gene screening methods, including the following steps:

[0085] From a botanical point of view, pear fruit is composed of seeds, receptacle (pulp), endocarp, mesocarp and exocarp, and is a false fruit developed from the receptacle. Pear peel is an important part of fruit quality and commerciality, and is the most noticeable fruit trait ( figure 1 ), the present invention relates to screening of internal reference genes for pear peel development process, bagging and post-harvest low temperature storage.

[0086] 1. The candidate internal reference genes provided by the present invention are obtained based on the transcriptome analysis of the Dangshansu pear (P.bretschneideri) genome and pear peel development process, bagging and postharvest low-temperature storage

[0087] 1.1) At present, in the pear peel qRT-PCR research, the most commonl...

Embodiment 3

[0105] This embodiment provides a set of application of internal reference genes in pear peel development process, bagging and postharvest low temperature storage, including the following steps:

[0106] The present invention utilizes the expression pattern of PbrEXPA8a in the pear peel to verify the practicability of the internal reference genes (RGs) recommended by the above analysis

[0107] 1.1) Expansins (EXP) play an important role in prolonging cell growth and degrading cell walls in pear peel. Combining pear genome and RNA-seq data, the present invention identified an expression profile of an EXP gene (PbrEXPA8a) that is closely related to pericarp development and postharvest low temperature storage of different pear varieties. Detailed investigation found that the expression level of PbrEXPA8a increased in the early division of pear fruit, sharply increased and reached the peak in the late division, and gradually decreased after the ripening period. During postharves...

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Abstract

The invention provides a group of new reference genes which can be used for gene expression analysis in peel development process, bagging and postharvest low-temperature storage of wide pyrus varieties, wherein ACT6 / 7 / 8 / 9 and NAP1 can be used as an optimal reference gene combination for pyrus peel gene expression analysis. The expression stability of five new reference genes screened on the basisof genomes and transcriptomes in the pyrus peel development process, the bagging and the postharvest cryopreservation treatment is superior to that of traditional reference genes (Actin, EF1[alpha], TUB, UBI, GAPDH and 26-18S rRNA); a pair of the reference gene combination of the ACT6 / 7 / 8 / 9 and the NAP1 is developed from the five new reference genes, and has the best homogenization effect when used for gene expression in the peel development process and the low-temperature storage and bagging treatment of the wide pyrus varieties; and the five new reference genes and the developed reference gene combination of the ACT6 / 7 / 8 / 9 and the NAP1 can be applied to gene expression research of the peel development process, the bagging and the postharvest low-temperature storage treatment of the widepyrus varieties.

Description

technical field [0001] The present invention relates to the field of plant genetic engineering, in particular to 5 new internal reference genes that can be used for gene expression analysis in the pericarp development process, bagging and post-harvest low temperature storage of a wide range of pear varieties, among which ACT6 / 7 / 8 / 9 and NAP1 are recommended as the most The combination of excellent internal reference genes was used for the analysis of gene expression in the pericarp of a wide range of pear varieties. Background technique [0002] Pear (Pyrus L.), one of the most economically valuable fruits in the world, can be roughly divided into two categories: Western pear and Oriental pear. Since the genome sequence (~512Mb) of P. bretschneideri has been completed, it has become a model plant for the study of Pyrus. The development process of pear fruit is divided into fruit setting period, division period, expansion period, maturity period and senescence period. From a...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/166
Inventor陈建清陈昌妹谢桂烽夏忠恒袁梦程翔宇陈清西
OwnerFUJIAN AGRI & FORESTRY UNIV