A group of pear peel qrt-PCR reference genes and their primers and applications

A technology of peel and internal reference, applied in the field of plant genetic engineering, can solve the problems of poor stability of internal reference, lack of combined application of multiple internal reference, unstable expression of traditional internal reference gene, etc.

Active Publication Date: 2022-07-15
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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  • A group of pear peel qrt-PCR reference genes and their primers and applications
  • A group of pear peel qrt-PCR reference genes and their primers and applications
  • A group of pear peel qrt-PCR reference genes and their primers and applications

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Embodiment 1

[0077] The present embodiment provides the internal reference gene sequence of pear peel development process, bagging and postharvest low-temperature storage, including:

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

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

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

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

[0082] (5) The ACT6 / 7 / 8 / 9 gene transcript is shown in SEQ ID NO: 5 in the sequence listing.

Embodiment 2

[0084] The present embodiment provides a group of methods for screening pear pericarp development process, bagging and postharvest low-temperature storage internal reference genes, including the following steps:

[0085] From a botanical point of view, pear fruit is composed of seeds, receptacle (flesh), 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 the internal reference gene screening of pear peel development process, bagging and postharvest low temperature storage.

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

[0087] 1.1) At present, in pear peel qRT-PCR researc...

Embodiment 3

[0105] The present embodiment provides a group of pear pericarp development process, bagging and postharvest low temperature storage processing in the application of the internal reference gene, including the following steps:

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

[0107] 1.1) Expandin (Expansins, EXP) plays an important role in prolonging cell growth and degrading cell wall in pear peel. Combined with pear genome and RNA-seq data, the present invention identified an EXP gene (PbrEXPA8a) expression profile that is closely related to the pericarp development and postharvest low temperature storage of different pear varieties. The detailed investigation found that the expression of PbrEXPA8a increased in the early stage of pear fruit division, increased sharply and reached a peak in the later stage of division, and gradually decrease...

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Abstract

The present invention provides a set of new internal controls that can be used for gene expression analysis in the development process of pericarp of a wide range of pear varieties, bagging and postharvest low-temperature storage, among which ACT6 / 7 / 8 / 9 and NAP1 can be used as optimal internal reference gene combinations for pear pericarp genes Expression analysis. The expression stability of the five new internal reference genes screened based on the genome and transcriptome of the present invention is better than that of traditional internal reference genes (Actin, EF1α, TUB, UBI, GAPDH and 26 ‑18S rRNA); developed a pair of ACT6 / 7 / 8 / 9 and NAP1 reference combinations from 5 new reference controls for gene expression homogenization during pericarp development, cryopreservation and bagging of a wide range of pear cultivars The best effect; the five new internal reference and the developed ACT6 / 7 / 8 / 9 and NAP1 internal reference gene combination can be applied to the research of gene expression in the process of pericarp development, bagging and postharvest low temperature storage treatment of a wide range of pear varieties.

Description

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

Claims

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Application Information

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Patent Type & AuthorityPatents(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