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Application of os07g0503900 protein in regulating plant resistance to weeds

A protein and plant technology, applied in the application field of Os07g0503900 protein in regulating plant resistance to weeds, can solve problems such as genetic mechanism and weed suppression mechanism are not very clear

Active Publication Date: 2022-07-19
ZHONGKENONGFU (BEIJING) BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]The flavonoids found in rice that can inhibit the growth of weeds (allelopathic effect) are mainly triflavones, and whether other flavonoids have allelopathy However, the genetic mechanism of rice allelopathy and the mechanism of weed suppression are not very clear

Method used

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  • Application of os07g0503900 protein in regulating plant resistance to weeds
  • Application of os07g0503900 protein in regulating plant resistance to weeds
  • Application of os07g0503900 protein in regulating plant resistance to weeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1. Compound purification

[0072] (1) Collect the fresh leaves of rice (indica) about 60 days after sowing, and grind them into powder after drying with a freeze dryer. 8 times the volume of 75% ethanol was added, and ultrasonic extraction was performed three times for 20 min each time.

[0073] (2) Filter the extract, and use a rotary evaporator to concentrate it until there is no alcohol smell, and sequentially add equal volumes of petroleum ether, dichloromethane, and n-butanol to extract 3 times respectively.

[0074] (3) After concentrating the n-butanol phase obtained with a rotary evaporator, a medium and low pressure rapid preparation liquid chromatograph (Biotage Isolera TM Prime) using wet loading for coarse fractionation. The silica gel column packing is YMC*GEL C18 spherical packing with a particle size of 50 μm and a pore size of 12 nm. The weight of the filler used in this example is 100 g. The mobile phase is water (containing 0.1% formic aci...

Embodiment 2

[0080] Example 2, Os07g0503900 gene cloning

[0081] (1) The genomic DNA of the leaves of the japonica rice Zhonghua 11 (Oryza sativa L.ssp.japonica cv.Zhonghua11, ZH11) at the seedling stage was extracted.

[0082] (2) According to the existing Os07g0503900 sequence information on the Rice Genome Annotation Project, primers F1 / R1 containing BamHI and HindШ were used to amplify the CDS sequence of the Os07g0503900 gene containing the restriction site with the ZH11 genomic DNA as the template.

[0083] Primer F1: 5'-ggatccATGGCTCCAGCGATGGCGAG-3';

[0084] Primer R1: 5'-aagcttCTATATGGATGACATGTGGGC-3'.

[0085] (2) ligating the obtained gene fragment to pEASY-T 3 Vector (TransGen Biotech, -T3Cloning Kit), transformed Escherichia coli DH5α competent cells, and screened positive clones using blue-white spots.

[0086] (3) The positive clones were identified by PCR using primers F2 / R2, and the amplified fragment size was 310bp as positive clones. The primers are as follows:

...

Embodiment 3

[0091] Embodiment 3, the application of Os07g0503900 gene

[0092] 1. Construction of prokaryotic expression vector

[0093] (1) The recombinant vector pEASY-T obtained in Example 2 was transformed with restriction enzymes BamHI and HindШ 3 -Os07g0503900 was completely digested, and the expression vector pMAL-c2X (Hua Yue Yang, VECT-570) was digested simultaneously. The digestion system was: 5 μg plasmid, 2.5 μL 10× digestion buffer, 2 μL BamHI, 2 μL HindШ, plus ddH 2 O replenish the reaction system to 50 μL. The restriction enzyme digestion reaction conditions were: 37°C for 4 hours.

[0094] (2) Separate the digested products by agarose electrophoresis, and recover a fragment of about 1.5Kb containing Os07g0503900 and a pMAL-c2X vector fragment of about 6.6Kb, respectively, and dissolved in 30 μL of ddH 2 O in.

[0095] (3) ligating the gene fragment obtained in step (2) with the vector backbone fragment. The ligation reaction system is: 1 μL 10× ligase buffer, 0.5 μL ...

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Abstract

The invention discloses the application of Os07g0503900 protein in regulating the resistance of plants to weeds. The invention provides applications of Os07g0503900 protein or related biological materials in any of the following: regulating plant allelopathy; regulating plant resistance to weeds; regulating tricin-5-O-glucoside content in plants; Glycosylation generates tricin-5-O-glucoside; as or prepares glycosyltransferase; inhibits weed growth. Experiments show that Os07g0503900 can catalyze the glycosylation of tricin to generate tricin‑5‑O‑glucopyranoside, and tricin‑5‑O‑glucopyranoside can regulate plant allelopathy and inhibit the growth of weeds. It can be seen that Os07g0503900 has important application value in regulating the allelopathy of plants. The invention has important guiding significance for cultivating rice varieties with high allelopathic ability and regulating the biosynthesis of tricin-5-O-glucoside.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of Os07g0503900 protein in regulating the resistance of plants to weeds. Background technique [0002] Flavonoids are a class of natural metabolites that are widely distributed in plants and contain a C6-C3-C6 basic skeleton. The main natural flavonoids can be divided into flavonoids, flavonols, and dihydroflavonoids according to the degree of oxidation of the central three-carbon chain, whether it is cyclic or not, and the position of B-ring connection (2- or 3-position). Classes, dihydroflavonols, anthocyanins, isoflavones, etc. Most flavonoids are combined with sugar to form glycosides in plants, and some of them exist in free state (in the form of aglycone). Flavonoids have strong anti-inflammatory and anticancer effects, and have great application prospects in the treatment of cardiovascular disease, coronary heart disease, tumors and other diseases. In additi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12N15/82A01H5/00A01H6/46
CPCC12N9/1048C12N15/8218C12N15/8245
Inventor 漆小泉马爱民宋波
Owner ZHONGKENONGFU (BEIJING) BIOTECHNOLOGY CO LTD
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