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Tobacco cadmium transporter gene NtPLA1 and application

A technology of transgenic plants and genes, applied in the field of genetic engineering, can solve the problems of affecting the biological yield and quality, affecting the growth of tobacco roots and the formation of chloroplasts, etc.

Active Publication Date: 2022-05-27
GUIZHOU TOBACCO SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, cadmium is one of the non-essential elements for plant growth. A higher concentration of cadmium in the soil will affect the growth of tobacco roots and chloroplast formation, thereby affecting its biological yield and quality.

Method used

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  • Tobacco cadmium transporter gene NtPLA1 and application

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Experimental program
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Embodiment Construction

[0018] 1. Cloning of tobacco NtPLA1 gene

[0019] After sowing of tobacco K326, when 5-6 true leaves were cultivated in the greenhouse, the fresh leaves were taken to extract total RNA using the plant total RNA extraction kit of Axygen Company, and the reverse transcription kit of TaKaRa Company was used for reverse transcription to synthesize cDNA.

[0020] The PLA gene in Genbank database (accession number: XM_016598298.1) was used as the reference sequence, and Primer 5 was used to design forward and reverse primers. The nucleotide sequence of the forward primer is SEQ NO: 3: 5'-ATGGGAAGGTTATTATTCTT-3', and the nucleotide sequence of the reverse primer is SEQ NO: 4: 5'-TCAGTTGGTGTATCGAAGCT-3'.

[0021] Tobacco PLA1 gene was amplified using KOD Fx NEO high-fidelity enzyme from NEB Company using the cDNA template of tobacco ‘K326’. The reaction volume is 50 μL: 2×PCR Buffer 25 μL, dNTPs 10 μL, KOD Fx NEO 1 μL, forward primer 2 μL, reverse primer 2 μL, tobacco cDNA 2 μL, dd H...

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Abstract

The invention relates to a tobacco cadmium transport related NtPLA1 gene and application thereof. The nucleotide sequence of the tobacco NtPAL1 gene and the coded protein sequence of the tobacco NtPAL1 gene are respectively shown as SEQ ID NO: 1 and SEQ ID NO: 2. The NtPAL1 gene is cloned from tobacco for the first time, and the tobacco NtPAL1 gene is proved to have a cadmium transport function by transferring a cadmium-sensitive yeast mutant strain delta ycf1 and gene editing. The tobacco NtPAL1 gene has an important application value in creation of low-cadmium-accumulation tobacco germplasm.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the NtPLA1 gene related to tobacco cadmium transport and its application. Background technique [0002] Tobacco has a strong cadmium accumulation ability, and tobacco plants absorb too much cadmium, which will increase the cadmium concentration in the smoke and endanger the health of smokers. In addition, cadmium is one of the non-essential elements for plant growth. Higher concentrations of cadmium in soil will affect the growth of tobacco roots and the formation of chloroplasts, thereby affecting its biological yield and quality. The accumulation of cadmium in plants involves many processes such as cadmium absorption, transport, and distribution, and its regulatory network and molecular mechanism still need to be further analyzed. [0003] At present, the cadmium transporter genes found in tobacco are relatively scarce, and only a few have been reported, such as NtHMA2 / 4, Nt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/55C12N9/18C12N15/84C12N15/81A01H5/00A01H6/82
CPCC12N9/18C12N15/8271C12N15/81C12Y301/01032Y02A40/146
Inventor 张吉顺张孝廉王仁刚王志红林世锋孔德钧林英超
Owner GUIZHOU TOBACCO SCI RES INST