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Regulatory gene NtMADS1 during tobacco flowering period as well as cloning method and application thereof

A technology of regulating genes and cloning methods, applied in the field of genetic engineering, can solve the problems of tobacco farmers' economic losses, manpower consumption, and affecting farmers' income in remote mountainous areas

Active Publication Date: 2018-03-20
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, early flowering of tobacco will bring significant economic losses to tobacco farmers and affect the income of farmers in remote mountainous areas
[0004] At present, agronomic measures (such as topping and branching) are mainly used in production to solve the problem of early flower bud differentiation induced by low temperature stress, but this is only a very labor-intensive remedial measure

Method used

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  • Regulatory gene NtMADS1 during tobacco flowering period as well as cloning method and application thereof
  • Regulatory gene NtMADS1 during tobacco flowering period as well as cloning method and application thereof
  • Regulatory gene NtMADS1 during tobacco flowering period as well as cloning method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Cloning NtMADS1 Gene

[0038] Tobacco leaf cDNA was used as a template, and primers were designed according to the tobacco genome database information, and the NtMADS1 The PCR amplification of the full-length fragment of the gene, the PCR reaction system and the amplification conditions are shown in Table 1; the PCR amplification product was obtained.

[0039] Primers are:

[0040] NtMADS1 -F:5'-ATGGTGAGAGGAAAAACTCAGATG-3';

[0041] NtMADS1 -R: 5'-TCAATTCTGTAGGGCGCGCTTCGTTC-3'.

[0042] The amplified PCR product was electrophoresed on a 0.8% agarose gel to obtain a fragment of 650 bp size, and the gel electrophoresis results were as follows: figure 1 shown. After electrophoresis, use a clean blade to cut off the gel with the target fragment and put it into a centrifuge tube. Do not cut the gel too large to avoid a large amount of impurities in the DNA fragment solution during recovery; add 3 to the centrifuge tube After doubling the volume of QG solu...

Embodiment 2

[0043] Example 2: Obtaining homologous genes

[0044] Using the NtMADS1 polypeptide sequence as the alignment sequence on NCBI, BlastP was used for homologous sequence alignment, and the N. benthamiana polypeptide sequence NibenMADS1 with the highest homology was selected. obtained in cigarettes benthamiana Niben MADS1 The gene sequence is shown in SEQ ID No: 3, and the amino acid sequence of the encoded polypeptide is shown in SEQ ID No: 4.

Embodiment 3

[0045] Embodiment 3: the cultivation of Nicotiana benthamiana

[0046] Sow plump and healthy Nicotiana benthamiana seeds in square plastic culture pots with a diameter of 10 cm containing a substrate (nutrient soil: vermiculite 1:4 volume ratio configuration), and sow 1-2 seeds in each pot. Put the sown pots into an incubator with a temperature of 22°C, light for 16 hours during the day / 8 hours at night, and a humidity of 60% to cultivate until the 2-3 leaf stage for later use. Water every 2 weeks during the cultivation process.

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Abstract

The invention discloses a regulatory gene NtMADS1 during a tobacco flowering period as well as a cloning method and application thereof. A sequence of nucleotide contained in the regulatory gene NtMADS1 during the tobacco flowering period is as shown in SEQ ID NO:1; a sequence of amino acid contained in a polypeptide coded by the regulatory gene NtMADS1 during the tobacco flowering period is as shown in SEQ ID NO:2. In tobacco, by reducing the expression of the regulatory gene NtMADS1, the tobacco flowering period can be ahead of time; in addition, by comparing a polypeptide sequence of the regulatory gene NtMADS1, homologous genes having the same functions can be searched, and the functions of the homologous genes can be verified by utilizing a VIGS (Virus-induced Gene Silencing) technology, so that the regulatory gene NtMADS1 has a wide application range; through the regulatory gene NtMADS1 during the tobacco flowering period, genetic and technical supports are provided for optimizing breeding of the tobacco during the flowering period.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a gene for regulating the flowering period of tobacco NtMADS1 And its cloning method and application. Background technique [0002] Low temperature is one of the most important environmental factors limiting crop production. Low temperature stress can change the flowering period of plants, thereby affecting crop yield and the economic value of some flower plants. In southern China, the phenomenon of tobacco early flowering induced by low temperature stress seriously affects tobacco production and farmers' income. [0003] Tobacco originated in South America and is a typical temperature-loving short-day crop. The optimum temperature for growth and development is 25-28°C. When the temperature is minus 2-3°C, the tobacco plant will die. The most suitable growth temperature for the underground is 31°C. When tobacco is at the age of 6-7 leaves, if it is subje...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/10C12N15/82C07K14/415A01H5/00A01H6/82
CPCC07K14/415C12N15/827
Inventor 谢贺白戈邹保红杨大海姚恒李永平肖炳光李军营张谊寒
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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