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A tobacco slow anion channel protein ntslah1 and its application

A channel protein and anion technology, applied in the field of tobacco slow anion channel protein NtSLAH1 and its application patent application, can solve the problems of large functional differences

Active Publication Date: 2021-05-07
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

but due to SLAC There are many genes in the gene family, and their functions vary greatly in different plants. Therefore, for different plants and different SLAC The function of the gene still needs to be further studied and differentiated, so that it can be targeted for future plant improvement

Method used

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  • A tobacco slow anion channel protein ntslah1 and its application
  • A tobacco slow anion channel protein ntslah1 and its application
  • A tobacco slow anion channel protein ntslah1 and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Present embodiment mainly with regard to tobacco slow anion channel protein NtSLAH1 The process of gene acquisition is briefly introduced as follows.

[0048] Using the cultivar tobacco leaf as a sample, the total RNA of the tobacco leaf was extracted with an RNA extraction kit, and reverse-transcribed into cDNA for future use;

[0049] Through the method of homologous comparison, refer to Arabidopsis AtSLAH1 The sequence of the gene and the known partial gene sequence of tobacco, the designed amplification primer sequence is as follows:

[0050] F: 5'-CGCGAGCTCGGTACCATGGGGGAAGAAGTTTTTG-3',

[0051] R: 5'-GCTCACCATGGATCCCTAATTACGTTTAGTGAAGT-3';

[0052] Using the above prepared cDNA as a template, PCR amplification was performed using the above primers.

[0053] During PCR amplification, the reference design of the 50 μL reaction system is as follows:

[0054] Upstream primer F, 1 μL;

[0055] Downstream primer R, 1 μL;

[0056] cDNA template, 1 μL;

[0057] 1...

Embodiment 2

[0069] to be sure NtSLAH1 Gene function in tobacco, selection NtSLAH1 The specific nucleic acid fragment in the gene (the 270th-615th nucleotide sequence of SEQ ID NO.1 in the sequence table) is used as a guide sequence to construct the silencer NtSLAH1 The VIGS vector was used for the transient silencing of the gene, and the tobacco plants were further transformed to construct transgenic plants. The relevant experimental procedures are briefly introduced as follows.

[0070] (1) Construction of VIGS vector for transient silencing

[0071] First, the primer sequences for PCR amplification were designed as follows:

[0072] NtSLAH1 -F: 5'- GACGACAAGACCCTGCAGCCTTTCATCCTCCTCTTTTC-3',

[0073] NtSLAH1 -R: 5'-TGAGGAGAAGAGCCCTGCAGGCACTCTCTTTCCATTCCT-3';

[0074] Perform PCR amplification (amplification length: 346bp) with the above primer sequences to obtain the guide sequence of VIGS;

[0075] Secondly, using the In-Fusion method, the above-mentioned amplified guide seq...

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Abstract

The invention belongs to the technical field of tobacco genetic engineering, and specifically relates to a tobacco slow anion channel protein NtSLAH1 and a patent application for its application. The gene CDS sequence includes 1107bp bases, and its base sequence is shown in SEQ ID NO.1; wherein the 270th-615th nucleotides are specific nucleic acid fragments. Tobacco slow anion channel protein NtSLAH1 is a key protein in tobacco chloride ion metabolism, which can be inhibited by virus-induced gene silencing (VIGS) NtSLAH1 After gene expression, the chloride ion content in the transgenic silenced plants was significantly reduced. Based on this, it can lay a good foundation for the cultivation of new tobacco varieties with low chlorine content, and also provide fundamental technical support for the stability of tobacco leaf quality and the improvement of cigarette quality.

Description

technical field [0001] The invention belongs to the technical field of tobacco genetic engineering, and specifically relates to a tobacco slow anion channel protein NtSLAH1 and a patent application for its application. Background technique [0002] Existing studies on tobacco generally believe that the appropriate content of chloride ions in tobacco leaves is 0.3-0.8, and when the content reaches 1%, it will affect the smoldering fire retention, and when it is further higher than 1%, there will be black ash flameout phenomenon; On the one hand, when the content of chloride ions in tobacco leaves is too high, starch will accumulate more, the leaves will be thick and brittle, and have high hygroscopicity, which will make the color darken easily during storage and produce bad smell. In a word, the chloride ion content in tobacco leaves has a relatively important and direct impact on the quality of tobacco leaves. [0003] Based on the direct impact and importance of chloride i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/82
CPCC07K14/415C12N15/8243
Inventor 张慧徐国云金立锋周会娜翟妞刘萍萍郑庆霞陈霞陈千思王燃
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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