Application of arabidopsis thaliana zinc finger protein gene ZFP6 to promoting growth of tobacco glandular hair

A technology of Arabidopsis thaliana zinc and gene, applied in the field of bioengineering, can solve problems such as shortage, and achieve the effects of increasing the number of glandular hairs, increasing the growth density of glandular hairs, and improving the quality of aroma

Inactive Publication Date: 2014-08-06
ZHEJIANG UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

Although a lot of research has been done on tobacco glandular trichomes at home and abroad, the regulation mechanism of glandular trichomes formation and development and the genetic law of glandular trichome density, especially the research on the relationship between glandular trichomes and tobacco aroma formation are still far away. Therefore, it is not only of great theoretical significance but also of potential application value to study the mechanism of controlling tobacco glandular hairs and aroma formation at the molecular level.

Method used

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  • Application of arabidopsis thaliana zinc finger protein gene ZFP6 to promoting growth of tobacco glandular hair
  • Application of arabidopsis thaliana zinc finger protein gene ZFP6 to promoting growth of tobacco glandular hair
  • Application of arabidopsis thaliana zinc finger protein gene ZFP6 to promoting growth of tobacco glandular hair

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Experimental program
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Effect test

Embodiment 1

[0021] Example 1 ZFP6 Gene Overexpression Vector Construction and Tobacco Genetic Transformation

[0022] (1) Take inflorescences of Arabidopsis thaliana, grind them with liquid nitrogen, add TRIzol reagent (Invitrogen), and extract RNA. 0.5 μg RNA was taken for reverse transcription reaction to obtain cDNA.

[0023] (2) Design specific primers according to the ZFP6 gene sequence on Genbank, and add SalI and NotI restriction enzyme sites to the 5‵ ends of the primers respectively. The sequences are as follows:

[0024] Primer 1: 5‵-GGTCGACGCTCACATATATGGCGACTGAA-3‵;

[0025] Primer 2: 5‵-AAGCGGCCGCTAATCATTCATGGCCCAAGG-3‵.

[0026] (3) Perform PCR amplification using the full-length cDNA as a template.

[0027] high fidelity enzyme HS DNA Polymerase (TaKaRa Code: DR010A) system:

[0028] 5×PrimeSTAR Buffer (Mg 2+ plus) 10 μl,

[0029] dNTP Mixture (2.5mM each) 4μl,

[0030] Primer 1 (10 μM) 1 μl,

[0031] Primer 2 (10 μM) 1 μl,

[0032] Template cDNA 1 μl,

[0033] ...

Embodiment 2

[0051] The character observation of embodiment 2 transgenic tobacco

[0052] The epidermal hairs of tobacco are divided into two categories according to the presence or absence of secretory glands. One is the glandular hairs that can produce secretions, and the other is the protective hairs that do not have the ability to secrete. The glandular hairs are divided into long-stalked glandular hairs and short-stalked glands according to their shape. hair.

[0053] The transgenic tobacco obtained in Example 1 is subjected to glandular trichome phenotype statistics, such as figure 1 As shown, overexpressing the Arabidopsis ZFP6 gene (sequence shown in SEQ ID NO.1) in tobacco can significantly promote the growth of the upper surface of the 3rd and 4th tobacco leaves at the age of 20 days compared with the wild type. The number of glandular hairs (long handle glandular hairs) and short glandular hairs (short glandular hairs) (P<0.01), but the number of epidermal hairs (protective hai...

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Abstract

The invention discloses an application of arabidopsis thaliana zinc finger protein gene ZFP6 to promoting the growth of tobacco glandular hair. The nucleotide sequence of the arabidopsis thaliana zinc finger protein gene ZFP6 is shown as SEQ ID NO. 1 (in the Specification). The application comprises the steps as follows: the arabidopsis thaliana zinc finger protein gene ZFP6 is connected to a plant expression carrier; a recombined expression carrier is obtained via construction; the recombined expression carrier is converted into tobacco. According to the application, the arabidopsis thaliana zinc finger protein gene ZFP6 is cloned, and is subjected to heredity conversion in tobacco; overly expressing outer source ZFP6 gene can remarkably improve the number of tobacco leaf glandular hair, and the number of protective hair not generating secondary metabolism product is not affected. The application disclosed by the invention conduces to the research of the regulation and control mechanism of glandular hair formation and development and the heredity law of glandular hair density, improvement on the quality of aroma of tobacco, and research and control of the formation mechanism of tobacco glandular hair and aroma formation.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to the application of an Arabidopsis zinc finger protein gene ZFP6 in promoting the growth of tobacco glandular hairs. Background technique [0002] Tobacco has become one of the most important economic crops in the global market in the 21st century. It is not only a source of income for tobacco farmers in more than 100 countries, but the entire tobacco industry involves different production departments and wholesale and retail links, and has become an important economic force in many countries. ; With the development of the tobacco industry, the local and local government taxes in many countries have been greatly increased. [0003] Tobacco is a high-efficiency crop. The planting area and output of tobacco in my country rank first in the world. Of all tobacco leaf types, flue-cured tobacco is the most widely grown. my country is the country that produces the mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/84C12N1/21C12R1/01
Inventor 甘银波颜安安丽君周忠静孟小芳刘伯涵张爱冬
Owner ZHEJIANG UNIV
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