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Rice lesion aging regulation gene and application thereof

A technology for regulating genes and rice, applied in the field of agricultural biology, can solve problems such as the decline of cell proliferation and differentiation ability and physiological functions, the decrease of cell membrane permeability, and the impact cannot be restored.

Active Publication Date: 2021-06-11
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) Irreversible aging stage. At this time, the organelles in the cell are cleaved, the permeability of the cell membrane is reduced, the cell proliferation and differentiation ability and physiological function gradually decline, and at the same time, the chromatin begins to degrade, and PCD occurs, that is, the impact cannot be recovered.
Only a few rice senescence genes have been identified so far, and there are still a large number of plant senescence-related genes to be identified

Method used

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  • Rice lesion aging regulation gene and application thereof
  • Rice lesion aging regulation gene and application thereof
  • Rice lesion aging regulation gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Obtaining of Lesion-Like Progeria Mutant Materials

[0022] Through chemical mutagenesis of japonica rice variety Yundao 32 (Y32) by EMS, a lesion-like premature senescence mutant lmes3 (lesion mimic and early senescence 3) was screened. The traits of the mutant have been stably inherited after multiple generations of selfing, and the aboveground part of the seedling stage is normal, but the development of the root system is hindered; the leaves of the tillering stage appear rusty spots, and the number of spots increases with the growth of the plant; At the booting stage, the tip of the leaf begins to wither and turn yellow, showing a premature senescence phenotype; at the heading stage, the withered yellow starts from the tip of the leaf and continues along the edge of the leaf to the bottom of the leaf; at the mature stage, spots exist in succession, aging is accelerated, the leaves are all withered and yellow, and the whole plant is yellowish-brown until p...

Embodiment 2

[0024] Example 2: Population Construction and Genetic Analysis

[0025] The mutant lmes3 was used as the female parent to cross with japonica rice variety Nipponbare (NIP), indica rice variety Taichung Local No. 1 (TN1) and Nanjing No. 6 (NJ06), respectively, and the F 1The plants showed normal wild-type phenotype, indicating that lmes3 is controlled by recessive nuclear genes. Statistics F 2 Segregation population segregation ratio (Table 1), the results show that the segregation ratio of normal phenotype plants and mutant phenotype plants is close to 3:1 segregation by Chi-square test, which shows that the lesion-like premature senescence phenotype of lmes3 is caused by single hidden Sex nuclear gene control.

[0026] Table 1 Genetic analysis of lesion-like progeria mutant lmes3

[0027]

[0028]

Embodiment 3

[0029] Example 3: Fine mapping of the mutant gene LMES3

[0030] Using 163 pairs of SSR primers and STS primers evenly distributed on 12 rice chromosomes, the mutant and NJ06 were screened for polymorphism. Mutant pool was established by BSA method (F 2 About 20 single plants with mutant phenotypes were selected from the population, the leaves were weighed and mixed, and DNA was extracted). The screened molecular markers with better polymorphisms amplify the DNA of parents, F1 and mutant pools, and find the markers linked to the target gene; then use the markers that express linkage to analyze the genes of individual plants with mutant phenotypes in the F2 population Type, and initially confirm the chromosomal location of the target gene. Plant genomic DNA was extracted by CTAB method.

[0031] Use 10 μL system for PCR reaction: 1 μL DNA template, 5 μL 2×PCR MasterMix, 0.5 μL each for forward and reverse primers (10 μmol / L), add ddH 2 O to make up 10 μL. The PCR amplifica...

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Abstract

The invention discloses a rice lesion aging regulation gene and application thereof. The rice lesion aging regulation gene is named as rice lesion aging regulation gene LEMS3, and the nucleotide sequence of the rice lesion and senescence regulation gene LEMS3 is as shown in SEQ ID NO.1. The gene LMES3 for controlling rice lesion senescence is cloned from a rice lesion premature senescence mutant lmes3 through a map-based cloning technology, and a functional complementation experiment proves that the LMES3 is a related gene for controlling rice senescence. Through observation of a transmission electron microscope, a photosynthetic rate, a photoreaction detection technology and a dark reaction detection technology, the premature senility gene cloned by the invention is proved to have an obvious regulation effect on growth and development of rice, completeness of thylakoid in chloroplast and the like. The invention also provides application of the gene. Some specific marker characters of the senescence gene can be applied to rice variety improvement, yield increase and resistance screening by utilizing a gene engineering technology.

Description

technical field [0001] The invention relates to the field of agricultural biotechnology, in particular to a rice lesion senescence regulation gene and its application. Background technique [0002] Rice is an important food crop in the world. The total output of rice accounts for the third place in the world's food crop production, only lower than corn and wheat. However, it is easy to process and convenient to eat, and can maintain the lives of a large number of people. There are about 3.5 billion people in the world, and half of the population eats rice, mainly distributed in Asia, southern Europe, tropical America and parts of Africa. In my country, more than 60% of the population uses rice as a staple food. [0003] Plant leaf senescence is the final stage of leaf development and an adaptive mechanism formed during the long-term evolution of plants. As the final stage of leaf growth and development, leaf senescence is induced by external and internal signals, and the pr...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415A01H5/00A01H5/12A01H6/46
CPCC07K14/415C12N15/8281C12N15/8266C12N15/8218
Inventor 方云霞薛大伟饶玉春王小虎张晓勤张弦
Owner HANGZHOU NORMAL UNIVERSITY
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