Application of oscol16 gene in controlling rice heading stage

A technology of transgenic rice and heading date, applied in the field of genetic engineering, can solve the problems of reduced yield, low seed maturity, shortened vegetative growth period, etc.

Active Publication Date: 2019-09-20
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 oscol16 gene in controlling rice heading stage
  • Application of oscol16 gene in controlling rice heading stage
  • Application of oscol16 gene in controlling 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

[0042] Example 2 Spatio-temporal expression pattern of OsCOL16 gene

[0043] 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 2 A). 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 period ...

Embodiment 3

[0045] Example 3 Analysis of subcellular localization and transcriptional self-activation activity of OsCOL16 gene

[0046] 1. Subcellular localization of OsCOL16 gene

[0047] 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 the application of the OsCOL16 gene in controlling the heading stage of rice. The gene OsCOL16 is from the rice variety Nipponbare, and its cDNA sequence is shown in SEQ ID NO: 1. The gene encodes a B-box / CCT zinc finger protein with 448 amino acids, belonging to CONSTANS‑like transcription factor gene family, genetic transformation experiments showed that overexpression of OsCOL16 gene inhibited rice heading under both short-day and long-day conditions. Further studies found that the OsCOL16 gene was expressed in a circadian rhythm, localized in the nucleus and had transcriptional self-activation activity. It inhibited the expression of Ehd1, Hd3a and RFT1 by up-regulating Ghd7, and finally delayed heading.

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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/46
Inventor 吴玮勋曹立勇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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