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Application of camellia sinensis hexokinase CsHXK3 gene in regulating and controlling plant growth and development and enhancing cold resistance

A hexokinase and plant growth technology, applied in the field of biogenetic engineering, can solve the problems of complicated breeding procedures, immaturity, and low breeding efficiency

Active Publication Date: 2021-02-23
TEA RES INST CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented method allows researchers to verify that certain specific parts or organs are involved in controlling vegetative cell division during flowering by altering their expression levels within these organisms (Cs). By comparing this data from different plants grown on two types of soil they found out how much damage caused each type of root system when exposed to high temperatures compared to other roots systems tested previously. These results showed that Cs were highly expressed at higher rates than usual but also reduced green weights due to less water absorption. Overall, it suggests possible use of catheter directed DNA techniques to study various aspects related to stem cells such as leaf formation, stalk lengthening, and chlorophyll content.

Problems solved by technology

This patented technical problem addressed in this patent relates to improving the quality and quantity of green teas grown on high altitudes or at temperatures below zero degrees Celsius during periods called cool weather conditions. Current techniques involve selecting specific types of vegetables based upon their physical properties like water content level, root length, stem diameter, etc., without considering how these factors affect the longevity and productivity of certain crops.

Method used

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  • Application of camellia sinensis hexokinase CsHXK3 gene in regulating and controlling plant growth and development and enhancing cold resistance
  • Application of camellia sinensis hexokinase CsHXK3 gene in regulating and controlling plant growth and development and enhancing cold resistance
  • Application of camellia sinensis hexokinase CsHXK3 gene in regulating and controlling plant growth and development and enhancing cold resistance

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Cloning and overexpression vector construction of tea tree CsHXK3 gene

[0027] According to the CsHXK3 sequence published in the NCBI database, its nucleotide sequence is shown in SEQ ID NO. 1, and the amino acid sequence of its encoded protein is shown in SEQ ID NO. As a template, the CsHXK3 gene was amplified by PCR. Ligate the amplified sequence into pENTR TM / D-TOPO carrier, finally connected to the pH7FWG2 plant expression vector by LR reaction.

[0028] The upstream primer F used: 5'-CACCATGGGGAAGGTAGGGTTG-3' is shown in SEQ ID NO. 3; the downstream primer R: 5'-TAGCAACTGTACGGTATCCACATT-3' is shown in SEQ ID NO. 4.

Embodiment 2

[0029] Embodiment 2: Construction and screening of CsHXK3 gene overexpression plants

[0030] The vector successfully constructed in Example 1 was transformed into the Agrobacterium strain GV3101, and then transformed into wild-type Arabidopsis thaliana by the flower soaking method, and the seeds of the T0 generation of transgenic Arabidopsis thaliana were harvested. The selection resistance gene carried by the pH7FWG2 vector is hygromycin. Therefore, use hygromycin to positively select the transgenic seedlings of Arabidopsis thaliana, harvest the positive seedlings of the T1 generation with hygromycin resistance, and then test the T1 The seeds of the first generation were screened for hygromycin resistance, the segregation ratio was identified, and the line with resistant seedlings: non-resistant seedlings was about 3:1, indicating that in this line, the carrier sequence carrying the CsHXK3 gene was a single copy form into the plant genome. The T2 generation seedlings with h...

Embodiment 3

[0032] Example 3: Growth and development phenotypes of plants overexpressing CsHXK3

[0033] Seeds of wild-type, OE-1, OE-2 and OE-3 Arabidopsis were sown on 1 / 2 MS solid medium with 1.5% glucose and 6.0% glucose and 1.5% fructose and 6.0% fructose, and the seed germination growth state See figure 2 A, B, C and D. Wild-type seeds could germinate healthily on the medium containing 1.5% and 6.0% glucose, and there was no significant difference in survival rate ( figure 2 E), while the seed survival rate of the three overexpression lines on the medium containing 6.0% glucose was significantly lower than that of 1.5% glucose ( figure 2 e). Similarly, compared with the wild type, the seed survival rate of the three overexpression lines on the medium containing 6.0% fructose was significantly lower than that of 1.5% fructose ( figure 2 F). The results showed that CsHXK3 could sense the sugar concentration signal and participate in the regulation of plant seed growth and ger...

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Abstract

The invention discloses application of a camellia sinensis hexokinase CsHXK3 gene in regulating and controlling plant growth and development and enhancing cold resistance, and belongs to the technicalfield of biological genetic engineering. The invention comprises application of the camellia sinensis hexokinase CsHXK3 gene in regulating and controlling plant growth and development and enhancing cold resistance, and a method for regulating and controlling growth and development of arabidopsis thaliana and enhancing cold resistance by using the camellia sinensis hexokinase CsHXK3 gene. The biological function of the CsHXK3 gene is verified for the first time through an overexpression experiment, and a gene resource can be provided for breeding of a new camellia sinensis variety. The invention provides an important and possibly universal gene resource for regulating and controlling plant growth and development and enhancing cold resistance, and an excellent candidate gene is provided forbreeding of a cold-resistant plant variety.

Description

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Claims

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Application Information

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Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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