Application of rice PS1 protein and rice PS1 protein coding gene in adjustment of plant senescence

A technique for encoding genes and transgenic plants, which is applied in the application field of rice PS1 protein and its encoding genes in regulating plant senescence, and can solve problems such as the lack of unified understanding of leaf senescence ideas and methodologies, the cloning and research lag of key genes of leaf senescence, etc.

Active Publication Date: 2013-12-25
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

However, at present, people's understanding of the basic scientific issues in leaf senescence is still very superficial, and there is still a lack of unified understanding of the thinking and methodology of leaf senescence research, and the cloning and research of key genes of leaf senescence are still very lagging behind

Method used

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  • Application of rice PS1 protein and rice PS1 protein coding gene in adjustment of plant senescence
  • Application of rice PS1 protein and rice PS1 protein coding gene in adjustment of plant senescence
  • Application of rice PS1 protein and rice PS1 protein coding gene in adjustment of plant senescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1. Obtaining PS1 protein related to plant senescence and its coding gene

[0066] 1. Mutant acquisition and phenotype

[0067] A premature senescence mutant was found from the rice mutant library (from the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences). Compared with the wild type, the mutant phenotype has obvious premature aging phenotype, and the phenotype of the mutant is as follows figure 1 Shown in A. Wherein, WT is wild-type rice, and ps1-D is the mutant rice obtained from the above screening. There is almost no difference between the mutant rice seedling stage and the wild type. With the progress of development, the aging of the old leaves accelerated after entering the tillering stage. Compared with the wild type, the older leaves were more prematurely senescent, and the chlorophyll content was relatively lower. After entering the filling stage, the aging phenotype was further intensified. Take the sword leaf as an exa...

Embodiment 2

[0082] Example 2, PS1 gene positively regulates rice senescence

[0083] 1. Expression of PS1 gene in different tissues

[0084] Different organs or tissues of the wild-type rice (Oryza.sativa L.) variety Nipponbare, as well as samples of the same organ at different stages, were extracted for RNA analysis of PS1 gene tissue expression characteristics, specific detection methods, data processing, internal reference primer sequences and PS1 gene The detection primer sequence was the same as in Example 1, and the experiment was repeated three times. The expression of PS1 gene in different organs or tissues, and in the same organ in different periods is as follows: Figure 4 As shown, the vertical axis represents the ratio of the expression levels of PS1 gene and ACTIN1 gene in various organs and tissues. in, Figure 4 Middle A is the expression of PS1 gene in root, stem, leaf, leaf sheath, young ear and endosperm, Figure 4 Middle B is the expression of PS1 gene in different ...

Embodiment 3

[0094] Example 3, Obtaining and functional identification of PS1 overexpressed rice

[0095] The gene involved in this example is the PS1 gene obtained in Example 1 from rice (Oryza. (named PS1 protein). Sequence 1 consists of 1179 nucleotides, and Sequence 2 consists of 392 amino acids.

[0096] 1. Obtaining of PS1 overexpressed rice

[0097] 1. Construction of recombinant expression vector pCAMBIA2300-Actin-PS1

[0098] (1) Extraction of rice total RNA and acquisition of cDNA

[0099] Rice total RNA was extracted using the Trizol reagent method (Invitrogen). The specific process is as follows: put 1-2g leaves of rice (Oryza.sativa L.) variety Nipponbare in a pre-cooled mortar, quickly grind them into powder with liquid nitrogen, and weigh 0.2g to 2ml of liquid nitrogen pre-cooled In a centrifuge tube; add 1ml Trizol reagent to each tube, mix well, add 200μl chloroform, shake on a vortex shaker for 1min, let stand at room temperature for 10min, then centrifuge at 12,000r...

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Abstract

The invention discloses an application of a rice PS1 protein and a rice PS1 protein coding gene in the adjustment of plant senescence. The application disclosed by the invention particularly relates to the application of the protein formed by amino acid sequences shown in a sequence 2 in a sequence table or the protein coding gene in the adjustment of plant senescence. A rice premature senescence mutant is cloned into the PS1 gene, so that the PS1 gene is adjusted and controlled by development ages and subjected to the peculiar induction of abscisic acids. The transgenosis experiments prove that the over-expression of the PS1 gene in wild-type rice can result in premature senescence; and the senescence can be delayed to a certain degree by inhibiting the expression of the PS1 gene in rice, so that the thousand seed weight of grains is increased, and the PS1 gene has good application potentiality in the increase of rice yields. The rice PS1 protein and the rice PS1 protein coding gene have potential application value in the terms of adjusting and controlling the plant senescence by using a genetic engineering means so as to carry out high-yield molecular breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of rice PS1 protein and its coding gene in regulating plant aging. Background technique [0002] Senescence is an inevitable life phenomenon in the process of plant development, which is subject to fine genetic regulation. It is the adaptability formed by plants in the long-term evolution process, and has important biological significance for plants themselves. Plant senescence is accompanied by the transfer of nutrients from aging organs to new organs and reproductive organs, so it is also considered to be a process of plant nutrient recycling. Although senescence is a normal physiological phenomenon actively regulated by plants, in agricultural production, due to some unfavorable factors, it is extremely easy to cause senescence to occur in advance, resulting in premature aging. Premature senescence leads to the premature loss of photosynthetic function and assimilat...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N15/84A01H5/10
Inventor 储成才梁成真王义琴唐九友
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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