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Nicotiana tabacum slowly activating anion channel homologue (NtSLAH1) and application thereof

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

Active Publication Date: 2018-06-22
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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  • Nicotiana tabacum slowly activating anion channel homologue (NtSLAH1) and application thereof
  • Nicotiana tabacum slowly activating anion channel homologue (NtSLAH1) and application thereof
  • Nicotiana tabacum slowly activating anion channel homologue (NtSLAH1) and application thereof

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Experimental program
Comparison scheme
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 SEQID 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 sequ...

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Abstract

The invention belongs to the technical field of tobacco genetic engineering, and in particular relates to a nicotiana tabacum slowly activating anion channel homologue (NtSLAH1) and application thereof. The CDS sequence of the gene comprises 1107 bp basic group, and the sequence of the basic group is shown in SEQ ID NO. 1, wherein the 270th-615th nucleotides are specific nucleic acid fragments. The NtSLAH1 is a key protein of tobacco chlorine ion metabolism; after the expression of the NtSLAH1 gene is inhibited by using a virus-induced gene silencing (VIGS) technology, the chloride ion contentin transgenic silencing plants is significantly reduced. Therefore, a good foundation is laid for cultivating new varieties of low-chlorine tobacco, and a fundamental technical support is also provided for the stability of tobacco 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 Applications(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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