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Loquat ejpfpb and method for improving tobacco sugar level by its coding gene

A gene, loquat technology, applied in the field of plant transgenics, to achieve the effect of improving flavor quality and accelerating growth

Active Publication Date: 2015-08-05
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

Sugar is an important component of fruit flavor, and fructose plays a more critical role in flavor composition. Therefore, if the regulation of sugar composition and sugar level can be realized through PFP gene, it can provide a technical basis for the improvement of fruit flavor; sugar It is closely related to the quality of tobacco and is one of the most important chemical components that affect the taste of tobacco. It has a direct impact on the acid-base balance and tar content of smoke. Undoubtedly, the use of PFP to realize the regulation of tobacco sugar is very important for improving tobacco quality. Beneficial and feasible, but so far no relevant data reports and new plants have come out

Method used

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  • Loquat ejpfpb and method for improving tobacco sugar level by its coding gene
  • Loquat ejpfpb and method for improving tobacco sugar level by its coding gene
  • Loquat ejpfpb and method for improving tobacco sugar level by its coding gene

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Embodiment Construction

[0017] The present invention will be further described in detail below in combination with embodiments and with reference to the accompanying drawings. The following experimental methods, unless otherwise specified, are conventional methods. The reagents used, unless otherwise specified, are conventional reagents purchased from Biochemical Reagent Company. The control tobacco or wild-type tobacco are all non-transgenic tobaccos.

[0018] (1) Isolate and clone the cDNA fragment sequence of loquat EJPFPb gene

[0019] The improved CTAB method was used to extract total RNA from loquat leaves. The specific steps were as follows: Preheat 500 μl of RNA extraction solution (2% CTAB, 2% PVP, 100mM Tris, 25mM EDTA, 2M Nacl, pH8.0) in a 65°C water bath; Add 0.2g of loquat leaves into liquid nitrogen and grind into powder, quickly transfer the powder into the preheated extract solution, then quickly add 40μl of mercaptoethanol and a small amount of spermidine and vortex to mix; c...

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Abstract

The invention relates to a method for improving the sugar level of nicotiana tabacum by using eriobotrya japonica EJPFPb and a coding gene thereof. The method comprises the following five steps: (1) separating and cloning cDNA (complementary Deoxyribonucleic Acid) fragment sequences of an eriobotrya japonica EJPFPb gene; (2) separating and cloning a cDNA full-length sequence of the eriobotrya japonica EJPFPb gene; (3) constructing a binary expression vector of the eriobotrya japonica EJPFPb gene; (4) carrying out nicotiana tabacum genetic transformation; (5) testing and assaying the growth speed and sugar composition level of transgenic nicotiana tabacum. According to transgenic nicotiana tabacum plants obtained by the method disclosed by the invention, the growth speed is high, the content of saccharose in laminae is obviously lowered, the content of glucose is basically unchanged, and the content of fructose is increased by 3-10%, so that the aims of regulating and controlling the composition and level of sugar and improving the flavor and quality of nicotiana tabacum by using the EJPFPb gene are achieved; moreover, by broadening the newly-obtained eriobotrya japonica EJPFPb gene to other vegetable, fruit tree and foodstuff crops through transferring, the quality and flavor of receptor crops are expected to be improved.

Description

technical field [0001] The invention relates to the field of plant transgenic technology, in particular to a loquat ( Eriobotrya japonica Lindl.) EJPFPb and its coding gene improve tobacco ( Nicotiana tabacum ) sugar level approach. Background technique [0002] Glycolysis is the most important way to regulate sugar metabolism in plants. There are three rate-limiting sites in the glycolytic pathway, which are acted on by three enzymes, hexokinase, phosphofructokinase, and pyruvate kinase. The reactions catalyzed by these three enzymes are all irreversible reactions, so these three enzymes are the rate-limiting enzymes of glycolysis. Among them, phosphofructokinase is a type of kinase that can act on fructose-6-phosphate, and the reaction product is fructose-1,6-diphosphate or fructose-2,6-diphosphate. Phosphofructokinase in plants is divided into two types, one is ATP-dependent ATP: fructose-6-phosphate 1-phosphotransferase (ATP: Fru-6-P 1-phosphotransferase, EC 2.7.1....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00C12N15/54C12N15/10
Inventor 秦巧平崔永一林飞凡
Owner ZHEJIANG FORESTRY UNIVERSITY
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