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Application of tobacco ntts3 gene in controlling tobacco leaf senescence

A leaf aging and genetic technology, applied in the field of genetic engineering, can solve the problems of unfavorable links and late maturity of flue-cured tobacco, and achieve the effect of delaying aging and efficiently controlling the aging process

Active Publication Date: 2020-07-24
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] As an important economic crop, tobacco leaves are the main harvesting organs; the state of tobacco leaves directly affects the increase of tobacco leaf production and the income of tobacco farmers; the untimely aging of tobacco leaves will have a significant impact on the quality of tobacco leaves; Tobacco leaves "turn green" caused by cloudy and rainy weather, and the tobacco leaves that should have matured under the influence of the environment cause late maturity; this brings disadvantages to the subsequent flue-cured tobacco link; therefore, in tobacco agricultural production, how to achieve tobacco leaf maturity through technical means The controllability of tobacco leaves has become an effective strategy to increase the yield and quality of tobacco leaves

Method used

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  • Application of tobacco ntts3 gene in controlling tobacco leaf senescence
  • Application of tobacco ntts3 gene in controlling tobacco leaf senescence
  • Application of tobacco ntts3 gene in controlling tobacco leaf senescence

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Cloning of NtTS3 gene

[0039] 1. Design NtTS3 gene amplification primers containing Gateway adapter

[0040] In this step, in order to connect the primers to the overexpression vector through the gateway recombination technique, the forward and reverse primers used here are both specific and can specifically clone the NtTS3 gene, and a gateway system is added to both ends of the primer The necessary part of the adapter can be amplified by nested PCR in the next step to obtain the NtTS3 gene containing the gateway recombination adapter. The specific method is as follows: the first forward primer (NtTS3-gatewayF) SEQ ID NO.1:

[0041] TACAA AAAAG CAGGC TTC AT GATGA TCGGA GAGAG AAACC;

[0042] The first reverse primer (NtTS3-gatewayR) SEQ ID NO.2:

[0043] GTACA AGAAA GCTGG GTC CT GGATC AGTTC AGAGA TCCA;

[0044] 2. Extract the RNA of Safflower Dajinyuan leaves (the main cultivated tobacco variety) and reverse transcribe it into cDNA, use this as a template, use th...

Embodiment 2

[0078] Acquisition of NtTS3 Gene RNAi Vector

[0079] 1. Design NtTS3-RNAi gene amplification primers containing Gateway adapter

[0080] In this step, in order to connect the primers to the overexpression vector through gateway recombination technology, the pair of primers can specifically amplify the RNAi fragment used for the NtTS3 gene, and part of the gateway linker is added to both ends of the pair of primers, which can pass through the nest The RNAi fragments directly used in the gateway system were obtained by formula PCR, as follows:

[0081] The third forward primer (NtTS3-RNAi-F) SEQ ID NO.6:

[0082] TACAA AAAAG CAGGC TTC GC TGAAC AAGGT GAAGT CCA;

[0083] The third reverse primer (NtTS3-RNAi-R) SEQ ID NO.7:

[0084] GTACA AGAAA GCTGG GTC TC ACTGG ATCAG TTCAG AG;

[0085] 2. Select the RNAi fragment directed at the NtTS3 gene as shown in SEQ ID NO.8, use the NtTS3-101 vector as a template, and use the third forward primer and the third reverse primer to car...

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Abstract

The invention discloses an application of a tobacco NtTS3 gene in tobacco leaf senescence control, belonging to the technical field of genetic engineering, which can promote or delay tobacco leaf senescence by regulating the NtTS3 gene, can carry out effective artificial intervention on tobacco leaf senescence, and can effectively control tobacco senescence process. A technical scheme comprise obtaining an NtTS3 gene contain a Zinc finger CCCH functional domain gene by using a gene cloning technique, Overexpression of NtTS3 gene promoted tobacco leaf senescence, and RNAi vector of NtTS3 gene is obtained by RNAi technology, which delayed tobacco leaf senescence by decreasing its expression. The invention can be applied to the regulation of tobacco leaf senescence.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to the application of tobacco NtTS3 gene in controlling the senescence of tobacco leaves. Background technique [0002] Senescence is the last stage of leaf development, and it is an orderly process affected by external factors (climate environment, biotic or abiotic stress, etc.) and internal factors (gene expression, hormone content, etc.). Leaf senescence is a manifestation of plant adaptation. In an adversity environment, plants recycle and reuse reusable substances through leaf senescence, and distribute them to seeds or young organs to increase plant resistance to external adverse environments. Early senescence or delayed senescence will have an impact on crops in agricultural production. Premature senescence of plant leaves and shorter photosynthesis time of leaves may lead to crop yield reduction, insufficient accumulation of organic matter, and reduc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/12A01H6/82
CPCC07K14/415C12N15/8261
Inventor 张增林郭永峰高晓明李伟徐萌萌
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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