Application of gene OsCOL16 to control of rice heading stage

A technology of heading stage and gene is applied in the application field of OsCOL16 gene in controlling the heading stage of rice, and can solve the problems of low seed maturity, reduced yield, shortened vegetative growth period, etc.

Active Publication Date: 2016-10-26
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the right growing season, late heading leads to a long vegetative growth period which promotes the accumulation of dry matter in the seeds, however too late heading may result in lower seed maturity at harvest
On the other hand, for crops with a short growing season, early heading is beneficial, however, premature heading shortens the vegetative period and leads to reduced yields

Method used

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  • Application of gene OsCOL16 to control of rice heading stage
  • Application of gene OsCOL16 to control of rice heading stage
  • Application of gene OsCOL16 to control of rice heading stage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Overexpression of OsCOL16 gene delayed heading

[0034] 1. Construction of overexpression vector

[0035] In order to obtain the phenotype of overexpressing the OsCOL16 gene in rice, an overexpression vector was constructed in this example. The specific construction method is as follows: plant the wild-type variety Nipponbare (English name Nipponbare, the whole genome sequencing has been completed) to about two weeks seedlings, use the plant RNA mini kit (purchased from Tiangen Biotechnology Co., Ltd.) to extract RNA, and then reverse transcribe into cDNA, the full-length cDNA sequence (SEQ ID NO: 1 ). The fragment was then recombined into the Xma I site of plasmid pCAMBIA2300 (purchased from Fermentas, Canada) by homologous recombination.

[0036] 2. Agrobacterium-mediated genetic transformation of rice

[0037] The obtained recombinant plasmid with correct sequencing was transferred into the callus of recessive genotype parent NIL (DTH2) through the rice...

Embodiment 2

[0043] The spatio-temporal expression pattern of embodiment 2 OsCOL16 gene

[0044] In order to study the rhythmic expression pattern of OsCOL16, we studied the expression level of OsCOL16 gene by real-time fluorescent quantitative PCR (qRT-PCR), and the primer sequences were qRT-OsCOL16-F (SEQ ID NO: 8) and qRT-OsCOL16-R (SEQ ID NO: 9). Leaves were collected every 4 hours during a 48-hour cycle under short-day (10 hours light, 14 hours dark) and long-day (14 hours light, 10 hours dark) conditions in a light incubator. Under short-day conditions, the expression level of OsCOL16 began to increase 4 hours after the light period, peaked at 10 hours after the dark period, then decreased rapidly and then gradually increased after the light period ( figure 2A). Similarly, under long-day conditions, the expression level of OsCOL16 began to increase after the light period, peaked at 6 hours after the dark period, then decreased rapidly and then gradually increased after the light p...

Embodiment 3

[0046] Example 3 Analysis of subcellular localization and transcriptional self-activation activity of OsCOL16 gene

[0047] 1. Subcellular localization of OsCOL16 gene

[0048] According to the Rice Genome Annotation Project database (http: / / rice.plantbiology.msu.edu), OsCOL16 encodes a predicted CCT / B-box zinc finger transcription factor. There is only one B-box domain (21-58aa) at the N-terminus, and one CCT domain (391-433aa) at the C-terminus. To determine whether OsCOL16 is a transcription factor with nuclear localization function, we performed subcellular localization experiments to verify its function. In this embodiment, a subcellular localization vector is constructed, and the vector construction process is as follows: use primers OsCOL16-GFP-BamHI-F (SEQ ID NO: 12) and OsCOL16-GFP-BamHI-R (SEQ ID NO: 13), to The full-length cDNA of OsCOL16, that is, the sequence shown in SEQ ID NO: 1 was amplified as a template, and then the fragment was recombined into the Bam HI ...

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Abstract

The invention provides application of a gene OsCOL16 to control of a rice heading stage. The gene OsCOL16 is derived from a rice variety Nipponbare, a cDNA sequence of the gene OsCOL16 is as shown in SEQ ID NO:1, and the gene OsCOL16 codes B-box / CCT zinc finger protein with 448 amino acids and belongs to a CONSTANS-like transcription factor gene family. According to genetic transformation experiments, overexpression of the gene OsCOL16 inhibits rice heading under both short-day and long-day conditions; according to further researches, the gene OsCOL16 is in day-night rhythm expression and located in cell nucleuses and has transcription self-activation activity, and expression of Ehd1, Hd3a and RFT1 is inhibited by up-regulation of Ghd7 to finally delay heading delay.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of OsCOL16 gene in controlling the heading stage of rice. Background technique [0002] Heading date (referred to as flowering time in crops) is an important agronomic trait that controls the regional adaptability of varieties, and precise control of flowering time is very important for reproductive transformation of varieties, thereby affecting crop yield (Cockram et al. (2007) J Exp Bot , 58(6):1231-1244). In the right growing season, late heading leads to a long vegetative growth period which promotes the accumulation of dry matter in seeds, whereas too late heading may result in lower seed maturity at harvest. On the other hand, for crops with a short growing season, early heading is beneficial, but premature heading will shorten the vegetative period and result in reduced yield. Therefore, the balance between dry matter accumulation and stress a...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/827
Inventor 曹立勇吴玮勋程式华郑晓明张迎信占小登沈希宏吴伟明陈代波于萍
Owner CHINA NAT RICE RES INST
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