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Method for culturing transgenic plant with decreased wax

A plant, coding gene technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, plant peptides, etc.

Active Publication Date: 2013-03-20
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a lot of research has been made in this area, there are few reports on rice leaf waxy-deficient mutants related to drought tolerance traits. Therefore, using rice leaf waxy-deficient mutants to clone genes related to leaf waxy formation, And analyze its function, and further understand the correlation between rice leaf epidermis wax and drought tolerance, which has important theoretical and practical significance

Method used

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  • Method for culturing transgenic plant with decreased wax
  • Method for culturing transgenic plant with decreased wax
  • Method for culturing transgenic plant with decreased wax

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, discovery of gene

[0033] 1. Phenotype analysis of rice waxy mutant osgl1

[0034] The rice waxy deletion mutant osgl1 is a natural mutant of wild-type 3037. Under normal conditions, mutant osgl1 had no significant difference compared with wild-type 3037 ( figure 1 A), but in the case of rain or artificial shower, the leaves of mutant osgl1 showed a highly hydrophilic phenotype. In wild-type 3037, due to the presence of a large amount of wax in the leaf epidermis, water molecules tend to aggregate on the leaf surface to form spherical water droplets. As long as the leaves are shaken a little, these water droplets on the leaves will drip. In the mutant osgl1, due to the high hydrophilicity of the leaves, water molecules will be evenly distributed on the entire leaf surface without forming water droplets ( figure 1 B). figure 1 Middle, wild type on the left, mutant on the right.

[0035] 2. Electron microscope observation and analysis of mutant osgl1 ...

Embodiment 2

[0045] Example 2, the acquisition of OsGL1 and its coding gene OsGL1

[0046] 1. Map-based cloning of the genome gene of OsGL1

[0047] In order to clone the OsGL1 gene, the homozygous mutant leaf waxy deletion mutant was crossed with Zhonghua 11, and the obtained F 1 F 2 In the population, 180 F2 recessive individuals (F2 individuals with leaf wax loss phenotype) were initially mapped for the OsGL1 gene. Using STS molecular markers and using PCR method, it was found that the STS markers P1, P2, P3, P4, P5 and P6 on chromosome 9 had obvious linkage with the mutation site. Most of the exchanged plants between the mutation site and P3 were also exchanged between the mutation site and P1 and P2, and most of the exchanged plants between the mutation site and P4 were included in the mutation site In the exchange individual plants between P5 and P6. At the same time, the exchange individual plants between the mutation site and P3 were different from the exchange individual plant...

Embodiment 3

[0062] Embodiment 3, the application of gene OsGL1

[0063] 1. Complementation experiment of waxy deletion mutant osgl1 phenotype

[0064] 1. Construction of complementary vector pCGL and complementary control vector pCGLC

[0065] BAC OSJNBb0057D06 was digested with Xbal to obtain a full-length DNA fragment (8264bp) comprising 2899 bases upstream of the start codon ATG of OsGL1 and 1436 bases after the stop codon TGA, and cloned into pCAMBIA1300 (CAMBIA , Canberra, Australia) between the Xbal restriction sites to obtain the recombinant vector; digest the constructed complementary vector pCGL with HindIII, remove part of the coding region of OsGL1 and the 5' promoter region of the gene, and retain part of the coding region at the 3' end and the 3' regulatory region, that is, the complementary control vector pCGLC was constructed ( Figure 5 ).

[0066] pCGL, complementary expression vector, including 2899bp upstream and 1436bp downstream of OsGL1 gene;

[0067] pCGLC, the ...

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Abstract

The invention discloses a method for culturing a transgenic plant with decreased wax. The method for culturing the transgenic plant with the decreasing wax comprises the following steps of: losing the gene coding function of proteins shown in SEQ ID NO: 1 of an initial plant so as to obtain the transgenic plant with the decreased wax. In the method, the gene affects the formation of leaf wax and is the gene for controlling leaf wax to form, and particularly, the gene can positively regulate the formation of leaf wax. The gene has important application value on the understanding of the mechanism of forming the leaf wax of paddy rice, and the gene can be used for breeding paddy rice to improve cuticular properties of paddy rice leaves by the gene engineering method so as to improve the drought control capacity. In addition, the gene and the protein can control the content of cuticular wax which may affect glucuronidase (GUS) staining effect of paddy rice leaves, and the GUS staining effect of paddy rice leaves can be enhanced by lowering the content of cuticular wax, so that the gene can be used to culture transgenic test materials for detecting expression conditions of exogenous gene on leaves.

Description

technical field [0001] The present invention relates to a method for growing transgenic plants with reduced waxiness. Background technique [0002] Plant epidermis wax is the general term for a class of mixtures that cover the outermost layer of the plant surface and are insoluble in water but soluble in organic solvents (chloroform, etc.). They are synthesized and secreted by epidermal cells. It is generally believed that epidermal wax has the ability to prevent Non-porous loss of water in plant tissues, maintenance of plant surface cleanliness and plant surface waterproofing, protection of plants from harmful light damage, protection of plants from pathogens and insects, etc. In addition, the content of epidermal wax usually affects the change of epidermal structure, and the change of epidermal structure usually affects the adsorption and penetration of water molecules in the epidermis. For example, the content of epidermal wax affects the effect of GUS staining of rice le...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/63C12N15/82C07K14/415C12N15/29
Inventor 程祝宽覃宝祥李明唐丁王克剑
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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