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Application of citrus fcmyc2 gene and its encoded protein in regulation of citrus essential oil synthesis

A technology of citrus essential oil and gene coding, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem of low valencene content

Active Publication Date: 2021-06-11
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, some transcription factors that regulate the synthesis of sesquiterpenes and alkaloids have been found in other plants, such as cotton GaWRKY1, white woody AsMYC2, alfalfa bHLH transcriptional regulators TSAR1 and TSAR2, etc. Only AP2 / ERF transcriptional regulators have been found in citrus CitAP2.10 regulates the synthesis of the sesquiterpene valencene, but the content of valencene in essential oils is extremely low, while other transcription factors that regulate the synthesis of citrus monoterpenes and other sesquiterpene components have not been reported

Method used

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  • Application of citrus fcmyc2 gene and its encoded protein in regulation of citrus essential oil synthesis
  • Application of citrus fcmyc2 gene and its encoded protein in regulation of citrus essential oil synthesis
  • Application of citrus fcmyc2 gene and its encoded protein in regulation of citrus essential oil synthesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Titration of essential oil content in peel

[0037] In order to compare the difference of essential oil content in the pericarp of Huapi kumquat (HP) and Rongan kumquat (RA), the titration analysis of the essential oil content in the pericarp of the two varieties at the veraison stage was carried out by potassium bromide-potassium bromate titration. The result is as figure 1 As shown in middle B, the essential oil content of HP peel is 0.11 mg / g, and that of RA peel is 9.3 mg / g. The essential oil content of HP was extremely significantly lower than that of oleaginous normal variety RA (p<0.001), and also much lower than that of many other kumquat and citrus varieties.

Embodiment 2

[0039] Expression Analysis of FcMYC2 Gene and Limonene Synthase (LS) Gene in HP and RA Tissues

[0040] The FcMYC2 gene involved in this example is derived from kumquat (Fortunella crassifolia Swing.), and was screened from RNA-seq data of leaves and peels of HP and RA.

[0041] The limonene synthase gene involved in this example is derived from citrus, and limonene is the component with the highest content in the essential oil of citrus peel, which can reach more than 70% of the essential oil components.

[0042] The primer sequences used to detect the expression level of FcMYC2 gene are:

[0043] FcMYC2-qRT-F: 5'-TCCGCTCTGTGGTTCCAAAC-3' (SEQ ID NO.6)

[0044] FcMYC2-qRT-R: 5'-CTTGGCCCTGAGCTCTTTGA-3' (SEQ ID NO.7)

[0045] The primer sequences used to detect the LS gene are:

[0046] LS-qRT-F: 5'-ATGGAGATGGGCATGGTGTT-3' (SEQ ID NO.8)

[0047] LS-qRT-R: 5'-TGCCAGGAGATGCTGTGAAA-3' (SEQ ID NO.9)

[0048] Taking Actin (Cs1g05000) as an internal reference gene, its primer seq...

Embodiment 3

[0054] Correlation between the expression level of FcMYC2 gene and the essential oil content of citrus peel

[0055] Respectively in the present embodiment to Kumquat species HP, RA, Citrus species Citrus aurantium (Poncirus trifoliata (L.) Raf), Citrus species local early orange (Citrus reticulata Blanco), Yuanjiang sour orange (C.aurantium L.) , Newhall navel orange (C.sinensis(L.)Osbeck), Dongfengzaoyou (C.grandis(L.)Osbeck) and Eureka lemon (C.limon(L.)Burm) relative expression levels of FcMYC2 gene The essential oil content of the peel was analyzed. Among them, Dongfeng Zaoyou uses a blade to peel off the exocarp, and the other varieties peel off the whole peel. The result is as figure 2 As shown, the essential oil content in the peel of HP is only 0.09mg / g, and the essential oil content in the peel of several other citrus varieties is between 4.49 and 18.23mg / g, which is much higher than that in HP. Correlation analysis showed that the expression level of citrus FcMY...

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Abstract

The present invention provides an application of citrus FcMYC2 gene and its encoded protein in regulating the synthesis of citrus essential oil, and relates to the technical field of plant genetic engineering; the nucleotide sequence of the FcMYC2 gene is as SEQ ID NO.1 or SEQ ID NO.2 As shown, the amino acid sequence of the protein is shown in SEQ ID NO.3; the FcMYC2 gene has the function of regulating the synthesis of citrus essential oil, which is reflected in the synthesis of positively regulating the main components of citrus essential oil, monoterpene and sesquiterpene; The method of genetic engineering combined with physiological and biochemical tests determined the function of the FcMYC2 gene for the first time, providing valuable gene resources for the study of citrus essential oil synthesis regulation network and citrus breeding.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to the application of citrus FcMYC2 gene and its coded protein in regulating the synthesis of citrus essential oil. Background technique [0002] Citrus essential oils are stored in oil cells located in tissues such as peels, petals, and leaves. Its main components are monoterpenes and sesquiterpenes, accounting for more than 97% of the total essential oil content. There are nearly a hundred kinds of monoterpenes and sesquiterpenes in citrus essential oils, and their regulatory mechanisms and regulatory networks are still rarely reported. At present, some transcription factors that regulate the synthesis of sesquiterpenes and alkaloids have been found in other plants, such as cotton GaWRKY1, white woody AsMYC2, alfalfa bHLH transcriptional regulators TSAR1 and TSAR2, etc. Only AP2 / ERF transcriptional regulators have been found in citrus CitAP2.10 regul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/11C12N15/10C12N15/82A01H5/00A01H6/78
CPCC07K14/415C12N15/10C12N15/8247C12Q2531/113
Inventor 刘小丰江东赵晓春朱世平申晚霞余歆
Owner SOUTHWEST UNIV
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