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Paddy rice leaf color regulation and control gene YL1 and use thereof

A technology for regulating genes and rice, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of rice leaf chlorosis and yellowing of rice leaves, etc.

Active Publication Date: 2017-02-15
HANGZHOU NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

OsCHL1 and OsCHL9 encode the magnesium ion chelatase subunit in the process of chlorophyll biosynthesis, and the inactivation of this gene will lead to light yellow-green leaves in the whole growth period of rice; FLG (OsPORB) encodes protochlorophyllin oxidoreductase B, the The enzyme can ensure the normal synthesis of chlorophyll in plants under strong light irradiation, and the mutation will cause the rice leaves to turn green and yellow; the OsDVR gene encodes diethylene chlorophyllate reductase, and diethylene chlorophyllin a is catalyzed by the enzyme to generate monochlorophyll. Ethylene chlorophyllin a, ethylene chlorophyllin a can generate chlorophyll b through a series of reactions under the action of chlorophyll a oxidase OsCAO1 / OsCAO2, and can also generate chlorophyll a under the catalysis of chlorophyll synthase YGL, so the process The function blockage or inactivation of any gene in will directly affect the synthesis of chlorophyll a and chlorophyll b, resulting in yellowing of rice leaves

Method used

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  • Paddy rice leaf color regulation and control gene YL1 and use thereof
  • Paddy rice leaf color regulation and control gene YL1 and use thereof
  • Paddy rice leaf color regulation and control gene YL1 and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Rice material

[0036] Rice (Oryza sativa L.) yl1-1 mutant plants were obtained from the indica rice variety Shuhui 527 (purchased from the Crop Variety Resource Bank of the Chinese Academy of Agricultural Sciences) through EMS mutagenesis treatment.

[0037] The yl1-1 mutant plants exhibited a distinct yellowish phenotype starting from the first fully expanded leaf (DAG6, true leaf expansion) of seedlings and continued throughout the growth period. Compared with the wild type (indica rice Shuhui 527), the yl1-1 mutant plants also showed delayed tillering period and reduced tillering number. In addition, there are also differences in plant height between the wild type and the mutant, and the difference is not obvious at the seedling stage (DAG6), and the plant height of the yl1-1 mutant is significantly lower than that of the wild-type plant at the mature stage, as shown in figure 1 shown.

[0038] 2. Analyze and target groups

[0039] f 2 F 1 Generation, F 1 F...

Embodiment 2

[0053] Plant Transformation:

[0054] Primers pCYL1-F (5'-tctagaAGTCCCATGTAAATAGGGTC-3') and pCYL1-R (5'-gtcgacGCTTCTCGGATAACGACTCT-3') were designed according to the target gene (LOC_Os02g05890). The full length of the target gene and 1382bp downstream of the 3'-end. Genomic DNA of japonica rice variety Nipponbare was used as a template, and pCYL1-F and pCYL1-R were used as primers, and PCR amplification was performed using the Q5 high-fidelity enzyme system of NEB Company. The reaction system was (20 μL): DNA template 2 μL, 2×Master Mix 10 μL, Each 1 μL of upstream and downstream primers was made up to 20 μL with ultrapure water. The PCR reaction conditions were pre-denaturation at 98°C for 30s; denaturation at 98°C for 10s, annealing at 65°C for 30s, extension at 72°C for 2min for 30s, 30 cycles; extension at 72°C for 10min. After the PCR product was subjected to electrophoresis, the target fragment was recovered and inserted into the T5-zero vector (purchased from Beijin...

Embodiment 3

[0056] 1. Chloroplast ultrastructure observation

[0057] In order to study the effect of YL1 gene mutation on rice chloroplast development, we observed and analyzed the chloroplast ultrastructure of 40-day-old wild-type and mutant heart leaves and fourth leaves by transmission electron microscopy. result Figure 6 shown. The heart leaf and the fourth leaf of the wild-type plants showed relatively complete chloroplast structure, and the grana lamellar accumulation was regular. However, the chloroplast morphology in the yl1-1 mutant exhibited an irregular phenotype, loosely packed thylakoid membranes, and significantly less grana lamellar packing than the wild type. This indicated that loss of YL1 function impaired chloroplast development.

[0058] 2. Subcellular localization of YL1 protein

[0059]We constructed the full-length subcellular localization vector of YL1 to explore the subcellular localization of YL1 in rice. The plasmid construction method is the same as that...

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Abstract

The invention discloses a paddy rice leaf color regulation and control gene YL1 and a use thereof. The paddy rice leaf color regulation and control gene YL1 has a gene nucleotide sequence shown in the formula of SEQ ID NO. 1. A coded protein sequence is shown in the formula of SEQ ID NO. 2. Through a map-based cloning technology, the novel paddy rice leaf color regulation and control gene YL1 is obtained. Through a transgenosis function complementation experiment, functions of the gene are verified. The YL1 and a chloroplast ATPase protein complex subunit AtpB produce synergism and YL1 gene mutation causes chloroplast ATPase activity reduction so that a photosynthesis-associated protein expression level is reduced, chloroplast development is influenced, chlorophyll content is reduced and a yellow leaf phenotype is produced. The protein and coding gene can further elucidate a paddy rice leaf color regulation and control molecule mechanism, can be used for crop genetic improvement and has an important meaning for high photosynthetic efficiency breeding.

Description

[0001] (1) Technical field [0002] The invention relates to a yellow leaf regulation gene, in particular to a rice yellow leaf regulation gene YL1 (YellowLeaf 1) and the application of the gene to regulate plant chloroplast development and improve photosynthesis efficiency. [0003] (2) Background technology [0004] Rice is one of the three major food crops in the world, and more than half of the population uses rice as a staple food. In my country, rice is the largest grain crop, its planting area accounts for about 28% of the total grain crop planting area, and its output accounts for about 35% of the total grain output (National data, 2012, http: / / www.stats.gov. cn / tjsj / ). Therefore, the high and stable yield of rice is of great significance to ensure the food security of our country. Leaf is the main site for photosynthesis of plants, which has an important impact on dry matter accumulation. Changes in rice leaf color will affect rice yield in most cases. Studies have s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/12
CPCC07K14/415C12N15/825
Inventor 于彦春陈飞武丽敏杨兴政
Owner HANGZHOU NORMAL UNIVERSITY
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