Method and kit for promoting flowering time of rice and mutation genotypes

A technology of flowering time and mutated genes, applied in the field of molecular genetics, can solve problems such as unclear functions, achieve the effects of shortening the growth period, improving ecological adaptability, and earlier flowering time

Active Publication Date: 2019-11-08
JIANGHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the large number of F-box protein genes in rice, the functions of many members are still unclear

Method used

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  • Method and kit for promoting flowering time of rice and mutation genotypes
  • Method and kit for promoting flowering time of rice and mutation genotypes
  • Method and kit for promoting flowering time of rice and mutation genotypes

Examples

Experimental program
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specific Embodiment approach

[0042] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0043] Unless otherwise specified, the examples follow conventional experimental conditions, such as the Molecular Cloning Experiment Manual of Sambrook et al. (Sambrook J & Russell DW, Molecular cloning: a laboratory manual, 2001), or the conditions suggested by the manufacturer's instructions. Unless otherwise specified, the chemical reagents used in the examples are all conventional commercially available reagents, and the technical means used in the examples are conventional means well known to those skilled in the art.

Embodiment 1

[0044] Example 1 Rice OsFBO14 Gene Sequence and gRNA Sequence

[0045] The sequence of the coding region of the rice OsFBO14 (LOC_Os03g27250) gene is shown in SEQ ID NO.9, and the protein encoded by it contains the F-box domain ( figure 1 ), the sequence is shown in SEQ ID NO.10. There is no information about the function of this gene in rice and the specific traits it controls. In order to clarify the function of the gene in rice, the present invention intends to use the CRISPR / Cas9 gene editing method to mutate the OsFBO14 gene sequence and knock out the function of the gene in rice. The present invention selects the sequence shown in SEQ ID NO.1 as the target region for CRISPR / Cas9 gene editing, and synthesizes a gRNA sequence (see SEQ ID NO.2 for the sequence).

Embodiment 2

[0046] Example 2 The function of rice OsFBO14 gene and the use of CRISPR / Cas9 method to advance rice flowering time

[0047] 1. Construction of CRISPR / Cas9 vector.

[0048] The present invention selects the japonica rice variety Zhonghua 11 (ZH11, bred and provided by the Institute of Crop Science, Chinese Academy of Agricultural Sciences) as the receptor for gene editing. The Escherichia coli strain was from Tran1-T1 Phage Resistant Competent, purchased from Quanshijin Biotechnology Co., Ltd. Agrobacterium tumefaciens strain EHA105 was purchased from Shanghai Shifeng Biotechnology Co., Ltd. The sgRNA backbone vector pCXUN-U6 was obtained from Zhao Yunde's laboratory at the School of Cell and Developmental Biology at the University of California. The main reagents Taq enzyme was purchased from TIANGEN, endonuclease XbaI was purchased from NEB; KT, 6-BA, IAA, NAA, 2,4-D were purchased from Sigma; Cn, Hn, AS were purchased from GiBco; other inorganic reagents were from Sinopha...

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Abstract

The invention discloses a method and a kit for promoting flowering time of rice and mutation genotypes. The invention provides a method for achieving early flowering of rice by editing OsFBO14 gene sequences with CRISPR-Cas9 method and discloses genotype sequences having early flowering traits of rice after editing. The method of the invention mutates F-box protein-encoding genes OsFBO14 of rice with CRISPR / Cas9 method and discovers the regulation effect of the OsFBO14 genes on flowering time of rice. By CRISPR / Cas9 gene editing method, the rice can flower in advance, thereby shortening growthperiod of rice, reducing planting cost and improving ecological adaptability of rice. The invention also provides the OsFBO14 mutation genotypes of gene-edited plants with early flowering time, and these mutation genotype sequences can promote rice to flower in advance, and be used to improve flowering traits of rice.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and in particular relates to a method, a kit and a mutant genotype for advancing the flowering time of rice. Background technique [0002] In higher plants, flowering represents the transition process from vegetative growth to reproductive growth, which plays an important role in the whole growth and development stages of plants. The biological trait of when to flower is affected by the plant's own genetic factors and external environmental factors. Under the influence of this dual effect, a series of flowering induction processes have a general rule in higher plants, that is, the plant leaves produce flower-forming substances (or Called florigen), the flower-forming substance is transported from the leaf to the apex growth point through the transport tissue, stimulating the apical meristem to form a flower. [0003] Flowering period is an important trait in the evolution and adaptation proces...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/113C12N15/8213C12N15/827C12N2310/10C12N2310/20
Inventor 李甜甜花梦婷高利芬陈利红彭海周俊飞李论方治伟
Owner JIANGHAN UNIVERSITY
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