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Application of rice blast resistance related gene oscol9

A resistance-related, rice blast technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as unreported, and achieve the effect of great application value

Active Publication Date: 2019-07-09
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of transcription factor plays an important role in regulating plant perception of circadian rhythm changes and flowering. Studies have shown that banana MaCOL1 is involved in regulating plant responses to biotic and abiotic stress responses, but whether CO and COL proteins are involved in regulating rice blast resistance? The relevant research has not been reported

Method used

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  • Application of rice blast resistance related gene oscol9
  • Application of rice blast resistance related gene oscol9
  • Application of rice blast resistance related gene oscol9

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Cloning and sequence analysis of rice OsCOL9 gene

[0028] 1) Extraction of rice total RNA

[0029] Zhongerruanzhan (Zhongerruanzhan) (disclosed in the document "High-quality and high-yielding rice variety Zhongerruanzhan [J]. Chinese Agricultural Sciences, 2001, (06): 49.") and its rice strain with high blast resistance H4 (near isogenic line H4) (in the literature "rice blast resistance protein Pik 2 Cloning of -H4 gene and screening of interacting proteins[J].Guangdong Agricultural Sciences, 2014, (04):156-160.") Four-leaf stage rice seedlings were frozen and ground with liquid nitrogen in a ceramic mortar into powder, transferred to a 1.5mL centrifuge tube, and added Trizol reagent (Invitrogen) according to the ratio of 1mL extraction reagent per 100mg of material, and mixed uniformly; Centrifuge at ℃ for 15 min, discard the middle layer and the lower organic phase, collect the upper aqueous phase and transfer it to a new centrifuge tube; add 600 μL isop...

Embodiment 2

[0035] Example 2 Analysis of the expression trend of OsCOL9 in different rice plants after inoculation with Magnaporthe grisea

[0036] The expression pattern of OsCOL9 gene after inoculation with Magnaporthe grisea was analyzed by quantitative RT-PCR. The rice blast fungus (Magnaporthe oryzae) GD0193 was inoculated in the disease-resistant variety H4 (in the literature "Evaluation of rice blast resistance and resistance genetic analysis of three space-induced mutagenesis rice lines [J]. Journal of South China Agricultural University, 2012, 33 (03 ): 273-276." Disclosed) after different time points (0h, 6h, 12h, 24h, 36h, 48h, 72h) collect leaf and extract its total RNA, utilize reverse transcription kit ReverTraAce to carry out reverse transcription cDNA Synthesis of the first strand. Then, according to the instructions of the SYBR Premix ExTaq kit, the expression of OsCOL9 was analyzed using the AB StepOne Plus fluorescent quantitative PCR detector, and Actin was used as an...

Embodiment 3

[0043] Example 3 Construction of rice OsCOL9 overexpression vector and acquisition of transgenic seedlings

[0044] The OsCOL9 cDNA fragment amplified by OsCOL9-F and OsCOL9-R was digested with KpnI (NEB) and BamHI (NEB), and connected to the overexpression vector pOX (in the literature "The cloning and functional analysis of rice SDG711 and SDG723 [D ]. South China Agricultural University, 2012 "published in the corresponding site, the ligation product was transformed into E. coli (E.coli) DH5α competent cells, and the product after the overexpression vector pOX digestion and recovery was also transformed into E. coli DH5α competent cells state cells (as a control); positive clones were selected to extract plasmid DNA for enzyme digestion identification ( Figure 4 , 1, 3, and 4 are correct positive pOX-OsCOL9 plasmids), and finally the OsCOL9 overexpression vector pOX-OsCOL9 driven by the rice Ubi promoter was obtained. Transform pOX-OsCOL9 into Agrobacterium by electric sh...

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Abstract

The invention discloses application of a rice blast resistance related gene OsCOL9and belongs to the technical field of plant gene engineering. The expression level of the gene OsCOL9 is increased after the gene OsCOL9 is induced by rice blast pathogenic bacteria. The expression of the gene is up-regulated after the gene is induced by Magnaporthe oryzae of Oryza Sativa, and the rice blast resistance of the Oryza Sativa can be obviously improved through overexpression. A plant overexpression vector is constructed by OsCOL9 and a Ubi promoter so as to convert the Oryza Sativa, and a result shows that the rice blast resistance can be remarkably improved through the overexpression of the gene. An expression vector can be constructed by using the gene disclosed by the invention and is applied to the rice blast resistant breeding of the Oryza Sativa. According to the application, better understanding of an action mechanism of OsCOL9 is facilitated, a foundation for further understanding the interaction of Oryza Sativa-Magnaporthe oryzae and a disease resisting signal conduction path is laid through OsCOL9cloning, and the application value in breeding is relatively high.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering and relates to the application of a rice disease resistance gene, in particular to the application of a rice blast resistance related gene OsCOL9. Background technique [0002] Rice (Oryza Sativa) is one of the most important food crops in the world, feeding more than half of the world's population (Khush, 2005), and it is also the main source of food for nearly half of my country's population (Liu Guoquan et al., 2004). Rice blast, caused by Magnaporthe oryzae infection, is an important limiting factor in rice production (Liu et al, 2013). It has the characteristics of fast transmission, wide occurrence range, and serious damage. In epidemic years, rice blast severe disease areas generally cause rice production to decrease by 10% to 20%, sometimes 40% to 50%, and some fields even fail to harvest, which seriously threatens rice production (Pennisi, 2010). Aggregating multiple ma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8282
Inventor 王加峰刘浩董双玉陈志强王慧
Owner SOUTH CHINA AGRI UNIV
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