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Application of gene Loc_Os01g12810

A gene and gene silencing technology, applied in the field of genes, can solve the problem of little research on chloroplast development in rice

Inactive Publication Date: 2018-08-07
RICE RES INST GUANGDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is little research on rice chloroplast development in the existing scientific research, so it is urgent to find a gene to study the development mechanism of rice chloroplast

Method used

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  • Application of gene Loc_Os01g12810
  • Application of gene Loc_Os01g12810
  • Application of gene Loc_Os01g12810

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The construction of embodiment 1 recombinant plasmid

[0033] This embodiment constructs the recombinant plasmid, and the specific steps are as follows:

[0034] The seedling leaf parts of rice variety Lijiang Xintuan Heigu (LTH) were taken, and the total RNA of the leaves was extracted with TriZol Reagent (Invitrogen Company, its article number: 15596026), and the purity of the total RNA was detected by agarose gel electrophoresis and ultraviolet spectrophotometer And the amount, take 1 μg of total RNA to do the initial reverse transcription reaction, the reverse transcriptase used is PrimeScript (TAKARA company), the steps of the reverse transcription reaction refer to the instruction of the reverse transcriptase. Using the reverse transcription product as a template, primer 1 was used for PCR amplification, and the forward and reverse nucleotide sequences of the primer 1 are respectively shown in SEQ ID NO: 4 and SEQ ID NO: 5 in the sequence listing. The polymerase ...

Embodiment 2

[0037] The genetic transformation method mediated by Agrobacterium EHA105 was used to transform the recombinant plasmid obtained in Example 1 into the seeds of normal japonica rice. Transformed primary (T 0 Generation) Through PCR and fluorescent quantitative PCR detection, the expression level of the target gene Loc_Os01g12810 was detected at the RNA level, and the result of the wild line was used as a control. Real-time quantitative PCR was used to identify the expression effect of the Loc_Os01g12810 gene in transgenic plants, and the steps were as follows:

[0038] Take the leaves of three-leaf stage seedlings of different transgenic lines that are positive in PCR detection and carry out total RNA extraction. The reagent used is TriZol Reagent (Invitrogen Company, its article number is: 15596026). Gel electrophoresis and UV spectrophotometer were used to detect the purity and quantity of total RNA, and 1 μg of total RNA was used for initial reverse transcription reaction. ...

Embodiment 3

[0040] Put the 8 T in embodiment 2 0 The seeds of the generation transgenic line and the seeds of the wild line ZH11 were placed in an incubator at 49°C for 72 hours to break the dormancy of the seeds, soaked in water for 24 hours at room temperature, placed at 32°C for 48 hours, and sowed on a plastic container filled with soil after the seeds germinated. In the dish (3 biological repetitions were designed), grown under the cover of plastic film, the transgenic plants interfered with by Loc_Os01g12810 showed albinism in the aerial parts (see image 3 ). Growing under normal conditions with the same temperature conditions, Loc_Os01g12810 interfering transgenic plants showed partial albinism of leaves, and when transferred to a normal growth environment, the degree of albinism would gradually weaken and gradually return to normal green (see Figure 4 ), when growing to the heading stage, the ears also lack chlorophyll, showing albinism, but compared with the wild line, it does...

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Abstract

The invention belongs to the technical field of genes and especially relates to application of a gene Loc_Os01g12810. The gene Loc_Os01g12810 can be applied to rice chloroplast development regulationmechanism researches; a gene method is utilized to regulate an expression level of the gene Loc_Os01g12810 to research rice chloroplast development regulation mechanisms; a nucleotide sequence of thegene Loc_Os01g12810 is shown in SEQ ID NO:1 in a sequence table; an amino acid sequence of encoding protein of the gene Loc_Os01g12810 is shown in SEQ ID NO:2 in the sequence table. According to the application of the gene Loc_Os01g12810 disclosed by the invention, that the rice gene Loc_Os01g12810 is a gene related with rice chloroplast development is proved for the first time. The Loc_Os01g12810is a PPR gene. Cloning and biological function verification of the gene have important reference significance in rice chloroplast development molecule mechanism researches and researches on effects of PPR family gene in rice development.

Description

technical field [0001] The invention belongs to the field of gene technology, and in particular relates to the application of a gene Loc_Os01g12810. Background technique [0002] Rice is one of the most important food crops in the world. More than half of the world's population depends on rice as a staple food. Therefore, rice production plays a decisive role in world food security. However, in the past 10 years, with the increase of world population and the decrease of arable land, there has been a food crisis all over the world. [0003] 90% to 95% of the dry matter accumulated in rice comes directly or indirectly from photosynthesis. As an important photosynthetic organ of plants, the development of chloroplast directly affects the carbon fixation efficiency of plants, which in turn affects various growth and development processes of plants. Understanding the development mechanism of chloroplasts has very important theoretical significance for improving photosynthetic ef...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/10A01H6/46A01H1/04
CPCA01H1/04C07K14/415C12N15/8218C12N15/8269
Inventor 董景芳刘斌
Owner RICE RES INST GUANGDONG ACADEMY OF AGRI SCI
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