Nicotiana benthamiana secreting type glandular hair regulation gene NbJAZ3 as well as expression vector and application thereof

A technology for regulating genes and expression vectors, applied in the field of molecular biology, can solve problems such as the unknown mechanism of tobacco secretory glandular hairs and the lack of regulatory means

Pending Publication Date: 2021-09-14
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to provide a kind of Nicotiana benthamiana secretory glandular hair regulation gene wxya , and use it to construct corresponding expression vectors and transform plants, in order to solve the current technical prob...

Method used

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  • Nicotiana benthamiana secreting type glandular hair regulation gene NbJAZ3 as well as expression vector and application thereof
  • Nicotiana benthamiana secreting type glandular hair regulation gene NbJAZ3 as well as expression vector and application thereof
  • Nicotiana benthamiana secreting type glandular hair regulation gene NbJAZ3 as well as expression vector and application thereof

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Effect test

Embodiment 1

[0027] Example 1: Regulatory genes of Nicotiana benthamiana secretory glandular hairs wxya Cloning and functional verification route

[0028] The epidermal hairs of Nicotiana benthamiana are all secretory type, which can be divided into long-stalked and short-stalked secretory glandular hairs according to the length of the epidermal hair gland stalk. Therefore, using Nicotiana benthamiana as a material to explore the molecular mechanism of JA regulating the development of secretory glandular hairs It has important research value.

[0029] The research of the present invention adopts homologous cloning technology, utilizes Solanaceae genome website Nicotiana benthamiana ( Nicotiana benthamiana ) genome sequence for reverse transcription PCR (Reverse Transcription-Polymerase Chain Reaction, RT-PCR), using the first strand of cDNA synthesized by reverse transcription of the total RNA of Nicotiana benthamiana leaves as a template, the negative regulation of glandular hair grow...

Embodiment 2

[0043] Embodiment two: wxyaTissue expression of genes and expression pattern analysis in response to MeJA treatment

[0044] The previous studies of the present invention showed that after 2 weeks of treatment with 1 and 5 μM MeJA, the plants of Nicotiana benthamiana gradually dwarfed, and after 2 weeks of treatment with 10 μM MeJA, the plants of Nicotiana benthamiana died. Therefore, the present invention collected Nicotiana benthamiana leaves treated with 1 μM MeJA for 2 weeks as materials to observe the morphology of glandular hairs. The results showed that the number of long-stalked secretory glandular hairs increased significantly, while the number of short-stalked secreted glandular hairs did not change significantly; in addition, the length of glandular hairs did not change significantly. These results suggest that JA signaling has an important role in the development of tobacco secretory glandular hairs.

[0045] As the core repressor of JA signaling, JAZ protein par...

Embodiment 3

[0047] Embodiment three: wxya Location Validation Study

[0048] Constructed pCXDG-GFP- wxya The subcellular localization vector was transformed into Nicotiana benthamiana leaves by transient transformation. It was observed by laser confocal microscopy that the green fluorescence was concentrated in the nucleus and cell membrane (see Figure 4 ),This indicates wxya Localized in the nucleus and cell membrane. In order to further verify the test results, the pCXDG-GFP- wxya The co-expression of the vector and the nuclear localization vector transformed tobacco leaves, and it was found that the green fluorescence and the red fluorescence could overlap and localize in the nucleus, which indicated that wxya indeed localized to the nucleus.

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Abstract

The invention discloses a nicotiana benthamiana secreting type glandular hair regulation gene NbJAZ3 as well as an expression vector and application thereof, and aims to solve the technical problems that the generation mechanism of tobacco secreting type glandular hair is unknown and an effective regulation means is lacked at present. The gene NbJAZ3 capable of regulating secreting type glandular hair generation is identified, an overexpression vector and a silence vector of the gene are constructed, and a corresponding overexpression strain and a corresponding gene silence strain are respectively obtained through transformation; and phenotypic and physiological feature identification and detection show that the NbJAZ3 gene can negatively regulate the generation of tobacco secreting type glandular hair, and has important application value in the aspects of glandular hair generation density regulation, glandular hair type directional improvement and the like.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a regulatory gene of Nicotiana benthamiana secretory glandular hairs wxya , its expression vector and application. Background technique [0002] Cuticle hairs are specialized structures on the surface of plants. According to whether the epidermis has branches, it can be divided into branched epidermal hairs and non-branched epidermal hairs; according to whether there are glands, it can be divided into glandular epidermal hairs and non-glandular epidermal hairs; according to the presence or absence of secretory ability, it can be divided into Divided into secretory and non-secretory; according to the number of cells in the epidermis, divided into unicellular and multicellular epidermal trichomes. Plant epidermis plays an extremely important role. Plants are subjected to many different types of biotic and abiotic stresses during their growth and development. Plant epide...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/84C07K14/415A01H5/00A01H6/82
CPCC07K14/415C12N15/8218C12N15/8261
Inventor 闫筱筱崔丽鹏王召军张洪映崔红陈亮
Owner HENAN AGRICULTURAL UNIVERSITY
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