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Application of rice OsUBC27 gene or protein coded by rice OsUBC27 gene in increasing rice yield

A rice and gene technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of unreported application, lack of understanding of UBCs function, limited understanding of biological function, etc., and increase the grain length of rice , the effect of increasing the thousand-grain weight

Active Publication Date: 2022-07-05
武汉艾迪晶生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the roles of several plant UBCs have been reported, our understanding of the biological functions of most of these enzymes is still very limited, especially for UBCs in food crops such as rice.
In addition, the application of rice OsUBC27 gene or its encoded OsUBC27 protein in improving rice yield has not been reported.

Method used

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  • Application of rice OsUBC27 gene or protein coded by rice OsUBC27 gene in increasing rice yield
  • Application of rice OsUBC27 gene or protein coded by rice OsUBC27 gene in increasing rice yield
  • Application of rice OsUBC27 gene or protein coded by rice OsUBC27 gene in increasing rice yield

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Cloning and transformation of the rice OsUBC27 gene.

[0030] RNA was extracted from young leaves of rice, and reverse-transcribed into cDNA using MLV reverse transcription reagent (Takara).

[0031] Total RNA was extracted using NucleoZOL kit (MACHEREY-NAGEL, Germany), the specific steps are as follows:

[0032] (1) Put 100 mg of leaves into a 2 mL sterilized centrifuge tube, add a small steel ball to each tube, quickly put the sample into liquid nitrogen, and use a proofing machine to break the leaf tissue;

[0033] (2) Add 1 mL of NucleoZOL extract, and after mixing, add 400 μL RNase-free water to each 1 mL of NucleoZOL extract to lyse the leaves, shake vigorously for 15s, and incubate at room temperature for 5-15 minutes;

[0034] (3) Centrifuge the sample for 15 minutes at 12000 rpm at room temperature;

[0035] (4) Transfer 1 mL of supernatant to a new 2 mL centrifuge tube;

[0036] (5) Add 1 mL of isopropanol to each 1 mL of supernatant to precipitate RNA, inc...

Embodiment 2

[0054] Identification of OsUBC27 gene expression in overexpression lines.

[0055] The seeds of hygromycin-positive transgenic T0 generation rice plants were collected, and the T2 generation lines were planted in the field from May to October 2021. When the rice tillering stage, young leaves were selected to extract total RNA, and reverse transcribed into cDNA for qRT-PCR ( The steps are the same as in Example 1). Using the rice constitutively expressed gene Actin as the internal reference, the primer sequences are as follows:

[0056] ActinF: GAAATGGAGACTGCCAAGACC;

[0057] ActinR: TTGGCAATCCACATCTGCTG;

[0058] UBC27-CDS-F and UBC27-CDS-R primers were used to detect the expression of OsUBC27 gene in different T2 rice lines. The results showed that the expression levels of OsUBC27 gene in the transgenic lines numbered OE78 and OE79 in the T2 generation transgenic lines were significantly higher than those in the wild-type lines, and the lines OE78 and OE79 were OsUBC27 gen...

Embodiment 3

[0060] Expression characteristics of OsUBC27 in different organs of rice.

[0061] Extract the RNA of rice roots (35 days), stems (35 days), leaves (35 days), ears (5-8em), seeds (12 days), reverse transcribed into cDNA for qRT-PCR analysis, the steps are the same as the embodiment 1. Using the constitutively expressed gene Actin in rice as an internal reference, cDNA templates from different rice tissues were amplified with primers UBC27-CDS-F and UBC27-CDS-R, and the expression of OsUBC27 gene in different tissues was detected. The results showed that the expression level of OsUBC27 gene was comparable in roots and ears, and higher than that in stems, leaves and seeds (e.g. image 3 shown).

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Abstract

The invention discloses an application of a rice OsUBC27 gene or a protein coded by the rice OsUBC27 gene in increasing the rice yield, and relates to the technical field of bioengineering. The base sequence of the OsUBC27 gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the protein coded by the OsUBC27 gene is as shown in SEQ ID NO. 2. The constructed plant overexpression vector pCUbi1390-OsUBC27 is expressed in wild Nipponbare, the tiller number of a transgenic plant is obviously increased compared with that of the wild plant, and compared with the wild plant, the grain length and thousand grain weight of rice grains are increased.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the application of rice OsUBC27 gene or its encoded protein in improving rice yield. Background technique [0002] Rice is an important food crop in the world. With the continuous growth of population, how to increase rice production and ensure food security has become an important scientific issue. There are many factors affecting rice yield, such as internal genetic factors such as tiller number, grain number per panicle and grain weight; and environmental factors including abiotic stress such as light, temperature, water and salinity, and biotic stress from diseases and insect pests; In addition, artificial cultivation management, fertilization patterns, and pest control methods also play an important role in rice yield. [0003] The plant ubiquitin / protease system is the main pathway for protein degradation in cells, and plays an important role in plant growth and dev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C12N9/10C12N15/54A01H5/10A01H5/00A01H6/46
CPCC12N15/8261C12N15/8205C12N9/104Y02A40/146
Inventor 鄂志国吴丽娟唐立群韩聪王惠梅王磊
Owner 武汉艾迪晶生物科技有限公司
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