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Method for increasing salicylic acid content in plant, and its special carrier

A technology of pyruvate and chloroplast, applied in the field of increasing the content of salicylic acid in plants, can solve the problems of difficult to meet medical and health care, difficult to use value of willow, high cost of extraction and purification

Active Publication Date: 2008-08-20
SINOBIOWAY BIO AGRI GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, long-term use of chemically synthesized acetylsalicylic acid will have adverse effects on the stomach, while aspirin-like components in natural plants and fruits have no related side effects, but the content is too low to meet the needs of medical and health care, and extraction and The cost of purification is higher
Now people mostly use white willow bark as raw material to extract natural aspirin analogues, but the resources of white willow are limited, and the stripped willow is difficult to have other utilization value

Method used

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  • Method for increasing salicylic acid content in plant, and its special carrier
  • Method for increasing salicylic acid content in plant, and its special carrier
  • Method for increasing salicylic acid content in plant, and its special carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, construction is used to improve the carrier of salicylic acid content in the plant

[0039] Now refer to figure 1 (represent corresponding gene with ICS and IPL according to the function of gene, IPL comprises pmsB, pchB; ICS comprises entC, icsl, pchA. ss, chloroplast localization signal; 35S, CaMV 35S promoter; Ter, 35S terminator.), Construction is used to improve the plant transformation carrier of salicylic acid content in the plant, and specific method is as follows:

[0040] 1. Amplification of the isochoristate synthase gene (ICS) and the isochoristate-pyruvate lyase gene (IPL)

[0041] 1. Amplification of the heterochoristate synthase gene (entC) derived from Escherichia coli

[0042] Using the genomic DNA of Escherichia coli DH5α as a template, under the guidance of primers p1: 5'-GGCAGAACGGCGCAGGACAT-3' and p2: 5'-CGATAGCGACGGGCAAACTCTTC-3', the heterochorismate synthase gene was amplified by PCR. After the reaction, the PCR amplified produc...

Embodiment 2

[0060] Embodiment 2, the acquisition of transgenic rice and the mensuration of SA content thereof

[0061] Taking rice as an example, the impact of the carrier for increasing the salicylic acid content in plants of the present invention on the SA content in plants is detected, and the specific methods are as follows:

[0062] 1. Obtaining genetically modified rice

[0063] Through the Agrobacterium-mediated method, pmBE / pHOS, pcBE / pHOS, pmBA / pHOS, pcBI / pHOS and pmBI / pHOS were respectively transformed into the calli of rice Zhonghua No. 11, and the transgenes transformed with the above-mentioned different vectors were obtained after cultivation. Plants, after identification, each transgenic plant obtained more than 10 independent transgenic lines, the transgenic rice transformed with pmBE / pHOS was named pmBE (containing pmsB and entC), and the transgenic rice transformed with pmBI / pHOS was named pmBI (containing pmsB and entC), the transgenic rice transformed with pmBA / pHOS wa...

Embodiment 3

[0089] Embodiment 3, semi-quantitative RT-PCR method detects the relationship between gene expression and SA content

[0090] Detect the expression level of exogenous gene (containing pchB and entC) in the transgenic rice pcBE that embodiment 2 obtains with semi-quantitative RT-PCR method, and analyze the correlation that gene expression level and salicylic acid content improve, to determine salicylic acid content Whether the increase is caused by the overexpression of the introduced exogenous gene. The specific method is as follows: select 13 pcBE transgenic rice plants with different free SA contents, extract total RNA, synthesize cDNA through reverse transcription, and use the cDNA as a template to use the primers of the chloroplast localization signal peptide coding sequence linked to the 5' of the exogenous gene (p13: 5'-ATGGCTTCCTCTGTCATTTC-3' and p14: 5'-GCTAACTTCTTCCACCATTGC-3') and primers for the endogenous reference gene Actin (p15: 5'-TCCGTGACATCAAGGAAAAG-3' and p1...

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Abstract

The invention discloses the method of improving the content of the salicin in the plant and the appropriative carrier. The carrier is the plant expressing carrier containing the separating offset acid composing enzyme gene and the separating offset acid-the pyruvic splitting dividing enzyme gene. The method uses the salicin composing approach and guides the different ICS gene and the IPL gene into the plant. The invention guides the said carrier into the plant group or the cell to gain the turning gene plant of the content of the salicin improved. The transferring gene test indicates that the dissociating state SA content in the transferring gene rice is mostly nine times than the wildness type. The method of the invention and the appropriative carrier can improve the content of the salicin in the plant greatly and provides the new method of amending the plant breed and improving the crop.

Description

technical field [0001] The invention relates to a method for increasing salicylic acid content in plants and a special carrier thereof. Background technique [0002] Salicylic acid (Salicylic acid, SA) is a kind of phenolic compound, and its structural formula is shown in Formula I. The pure product is powdery crystal, soluble in water, easily soluble in polar organic solvents, melting point is 157-159°C, easy to sublime after drying (Yongbing Yuan, Zongxun Cao. 1994. The role of salicylic acid in plants, Plant Bulletin of Science 11(3):1-9). [0003] [0004] Salicylic acid is ubiquitous in plants, including free and bound salicylic acid. According to research, exogenous application of salicylic acid can affect the opening and closing of plant stomata, seed germination, fruit production, ion absorption, heat production, and flowering. , sexual differentiation and inhibition of ethylene biosynthesis and other physiological activities, so SA and its salts are considered ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/52A01H4/00A01H5/00
Inventor 王喜萍于鲲陈丽娜谢莹夏勉张朝晖
Owner SINOBIOWAY BIO AGRI GRP CO LTD
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