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Application of gene OsLUX in promoting rice heading and improving plant disease resistance

A technology of transgenic rice and disease resistance, applied in the field of plant genetic engineering

Pending Publication Date: 2022-04-15
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

CCA1 overexpression plants showed a more susceptible phenotype, but GRP7 overexpression plants did not have the corresponding phenotype, indicating that CCA1 may also participate in the resistance response of Arabidopsis through other pathways

Method used

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  • Application of gene OsLUX in promoting rice heading and improving plant disease resistance
  • Application of gene OsLUX in promoting rice heading and improving plant disease resistance
  • Application of gene OsLUX in promoting rice heading and improving plant disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Phenotypes of 3 late heading mutants

[0056] In order to identify new genes regulating heading date of rice, EMS mutagenesis was carried out on Nipponbare variety (Nipponbare), and three late-drawing individual plants elh1, elh2 and elh3 were found in the mutant library. Under long-day and natural short-day conditions, they showed stable extreme late-drawing and late-drawing phenotypes ( figure 1 ).

Embodiment 2

[0057] Embodiment 2Mutmap method identifies the mutant of 3 kinds of OsLUX genes

[0058] To identify the genes controlling the extreme late heading, the mutants were backcrossed with the wild type, and in the backcross F 2 Mutmap identification using extremely late heading individual plants in segregated populations ( figure 2 ), according to the candidate gene, referring to the LOC_Os01g74020 site information of the rice genome sequence database [GenBank (http: / / www.ncbi.nlm.nih.gov) registration number: NC_029256], a pair of expansions were designed on both sides of the OsLUX sequence The primers OsLUX-F and OsLUX-R were amplified by PCR to obtain the complete coding region sequence of OsLUX for sequence comparison with the wild-type CDS sequence.

Embodiment 3

[0059] Functional identification of embodiment 3 OsLUX gene

[0060] To confirm that the phenotypes of the extreme late-twitch mutants elh1 / oslux-1, elh2 / oslux-2, and elh3 / oslux-3 were caused by loss of OsLUX function, the inventors performed genetic complementation of elh1 / oslux-1 and OsLUX Knockout experiments. The specific implementation is as follows: using Nipponbare genomic DNA as a template, PCR amplification is performed with primers OsLUX-EcoRI-F (SEQ ID NO: 7) and OsLUX-EcoRI-R (SEQ ID NO: 8) to obtain the full-length sequence of the OsLUX gene, Subsequently, homologous recombination was carried out with the pCAMBIA1305.1 vector digested with EcoRI, and the positive strain obtained by using elh1 / oslux-1 as the transformed recipient had its heading stage phenotype restored to the wild-type phenotype ( image 3 Middle A,C,E). The target sequence of OsLUX (SEQ ID NO: 9) was designed and constructed on the vector BGK032 (purchased from Baige Gene Technology Co., Ltd.),...

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Abstract

The invention discloses application of a gene OsLUX in promoting rice heading and improving plant disease resistance. The OsLUX gene is a core component of a rice biological clock, three mutants of the OsLUX are identified by a Mutmap method, and the three mutants are represented as an extreme late extraction phenotype and a late extraction phenotype. Further research finds that the OsLUX gene shows circadian rhythm expression, is positioned in a cell nucleus and a plasma membrane and has transcriptional self-activation activity, expression of Ehd1, Hd3a and OsMADS14 is promoted by inhibiting OsGI, Hd1 and Ghd7, and heading is finally promoted. Meanwhile, the elh1 / oslx-1 inoculated with the xanthomonas oryzae is more susceptible to diseases compared with the wild type, which indicates that the OsLUX has a positive regulation effect on the resistance of the xanthomonas oryzae. The invention further enriches and perfects the theoretical basis of the rice biological clock gene for regulating and controlling the photoperiod flowering.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to the application of gene OsLUX in promoting rice heading and improving plant disease resistance. Background technique [0002] The circadian clock is endowed with the function of the "central processing unit" of plants, which is involved in the regulation of almost all growth and development, material metabolism and stress response processes of rice. The dynamic changes of external environmental factors are transmitted to the circadian clock system through the input pathway, so that plants can perceive the signal changes at a specific time, and then regulate the corresponding growth and development process of plants to achieve synchronization with external environmental changes. In the field environment, the entire life cycle of crops is in changing natural conditions (mainly light and temperature), and the biological clock coordinates the internal time mechani...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/46
CPCC07K14/415C12N15/8281C12N15/8261
Inventor 吴玮勋徐鹏曹立勇程式华张迎信占小登
Owner CHINA NAT RICE RES INST
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