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PSY2 gene sequence related to carrot carotenoid synthesis and application thereof

A carrot and red carrot technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of organic matter accumulation affecting growth and development, physiological and metabolic disorders, and unbalanced carrot nutrition supply.

Inactive Publication Date: 2020-10-13
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Abiotic stress can also lead to unbalanced nutrient supply of carrots and disorder of physiological metabolism, which in turn affects its growth and development and the accumulation of organic matter

Method used

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  • PSY2 gene sequence related to carrot carotenoid synthesis and application thereof
  • PSY2 gene sequence related to carrot carotenoid synthesis and application thereof
  • PSY2 gene sequence related to carrot carotenoid synthesis and application thereof

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Embodiment Construction

[0011] 1. Extraction of carrot total RNA and synthesis of cDNA: total RNA was extracted from carrot leaves according to the procedure described in the kit (Beijing Tiangen Biochemical Technology Co., Ltd.). The concentration of RNA was determined by Nano-Drop, a micro-ultraviolet detector. Subsequently, the RNA reverse transcription operation was completed with reference to the HiScript II Q RT SuperMix for qPCR (+gDNA wiper) kit (Nanjing Novizan Biological Co., Ltd.).

[0012] 2. Cloning of carrot DcPSY2 gene: the carrot DcPSY2 gene sequence was retrieved from the carrot transcriptome and genome database in our laboratory, and the sequence was cloned and analyzed. Using Primer Premier 5.0 software to design a pair of cloning primers (upstream DcPSY2-F: 5'-TGGTGTGAATGGTTGAGGCCTCT-3'; downstream DcPSY2-R: 5'-GAATTTTTACATGGCCAATTGTG-3'), using the high-fidelity enzyme Prime STAR Max, with carrot 'Jun Chuan Red' cDNA was used as template, and 30μL system was used for amplificati...

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Abstract

A PSY2 gene is a key gene participating in carotenoid synthesis. A new PSY gene DcPSY2 is cloned from carrots. The open reading frame length of the gene is 1317bp, and 438 amino acids are encoded. Theprotein encoded by DcPSY2 belongs to hydrophilic protein. The protein is closest to the evolutionary relationship of leguminous rosewood PSY and has high conservative property. Fluorescent quantitative expression analysis shows that the relative expression level of the DcPSY2 gene reaches a peak value after treatment for 2h under high temperature and high salt stress, and the relative expressionlevel is significantly different from that of a control group (P is less than 0.05). Under low temperature and drought stress, the expression level of the DcPSY2 gene of the carrots is increased mostobviously after treatment for 1h. The DcPSY2 gene of the carrots is cloned from the carrots by using a polymerase amplification technology, and the DcPSY2 gene of the carrots responds to the adversitystress of carrot plants.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering and relates to a plant PSY2 gene and its application. The present invention obtains a gene DcPSY2 related to carotenoid synthesis from carrot variety 'Junchuanhong', and the gene can be applied to the research and application of carotenoid synthesis pathway in carrot. Background technique [0002] Carrot (Daucus carota L.) is a biennial root vegetable crop belonging to the genus Carrot in the family Apiaceae, and is a variety of wild carrot. Carrots are rich in nutritional value and medicinal value, and are very common in our country. Carrots have fleshy taproots as their main edible organs. Carrots can be divided into different types according to the length, shape, and color of the fleshy roots. Carrots are rich in nutrients, such as carotenoids, vitamins, starch, sugar and lipids, which are mainly derived from the secondary phloem. [0003] In plant production, abiotic stresses such a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/10
CPCC12N9/1085C12N15/10C12Y205/01
Inventor 熊爱生丁旭徐志胜
Owner NANJING AGRICULTURAL UNIVERSITY
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