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Cloning and Application of Rice Heading Time Gene haf1

A technology of heading stage and rice, applied in the field of functional verification and application, the isolation and cloning of the gene HAF1, can solve the problems of delayed heading, prolonged growth period, decreased yield, etc.

Active Publication Date: 2019-07-16
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the "south-north shift" in the introduction of rice is not suitable. The main reason is that the growth period of southern varieties in the north is prolonged, and the heading is delayed, which leads to the failure of normal fruiting in the later period and the decline in yield.

Method used

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  • Cloning and Application of Rice Heading Time Gene haf1
  • Cloning and Application of Rice Heading Time Gene haf1
  • Cloning and Application of Rice Heading Time Gene haf1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: the isolated clone of HAF1 gene

[0054] 1. Obtaining late heading mutants

[0055]The mutants identified in the present invention are from rice T-DNA insertion mutants (for the creation method of the mutant library, please refer to the paper: Wu et al., Development of enhancer trap lines for functional analysis of therice genome, Plant J (2003): 418-427; Zhang et al., Non-random distribution of T-DNA insertions at various levels of the genome hierarchy as revealed by analyzing13,804 T-DNA flanking sequences from an enhancer-trap mutant library, Plant J(2007):947-959). The screening and identification method of mutants is as follows: from the above-mentioned rice T-DNA insertion mutant library (see http: / / rmd.ncpgr.cn / ), the T0 generation transgenic rice seeds are planted in the field after conventional seed soaking and germination procedures After getting each T 1 A total of 20 plants were planted in the generation family. The plants were planted in the...

Embodiment 2

[0065] Example 2: Experimental verification of functional complementarity

[0066] 1. Construction of complementary vectors

[0067] The complementary vector constructed in Example 1 is pC2301-HAF1 (see image 3 Figure A in and Figure 8), the constructed vector was electrotransformed into Agrobacterium tumefaciens EHA105 (purchased from CAMBIA, http: / / www.cambia.org / daisy / cambia / materials / overview.html) strain.

[0068] 2. Genetic transformation

[0069] The Agrobacterium-mediated genetic transformation method (Wu et al., Development of enhancer trap lines for functional analysis of the rice genome.2003, Plant J.35:418-427) was used to introduce the complementary strain into the callus of the haf1 mutant. Cultivation, infection, co-cultivation, screening of calli with resistance to G418 (purchased from Beijing Yuanpinghao Biotechnology Co., Ltd.), differentiation, rooting and seedling hardening and transplanting in the field to obtain transgenic plants (Agrobacterium-mediat...

Embodiment 3

[0071] Example 3: HAF1 regulates known heading date related regulatory genes

[0072] In order to analyze the molecular mechanism of HAF1 gene regulating rice flowering, the present invention uses quantitative RT-PCR technology to detect the expression of related heading date regulation genes in wild-type (Zhonghua 11) and haf1 mutant plants. The result is as Figure 4 As shown, the expression rhythm of HAF1 gene gradually increased at night and peaked at night, and gradually increased during the day under long-day light (16 hours of light / 8 hours of darkness) or short-day (8 hours of light / 16 hours of darkness) conditions. decreased and reached a trough in the evening, but in the haf1 mutant, the expression of the HAF1 gene was significantly reduced (such as Figure 4 panels A and B in). By detecting the expression of the circadian clock gene OsGI (LOC_Os01g08700), it was found that there was no difference in the expression of OsGI in the wild-type and haf1 mutants, indicat...

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Abstract

The invention relates to to the technical field of plant genetic engineering, in particular to isolation cloning, functional verification and application of gene HAF1 (heading date associated factor 1) mediating paddy rice heading date. Gene functional complementation tests prove that the HAF1 gene cloned by the invention has the function of regulating the paddy rice heading date. HAF1 encodes a ubiquitin ligase, and geographical distribution of paddy rice varieties can be controlled through ubiquitination regulation of the paddy rice heading date gene HAF1, thus deciding the growth habits and yield of paddy rice. The cloned gene and its encoded protein provided by the invention can be widely used for regulating the flowering characteristics of plants, and improving the growth period and regional adaptation range of plant varieties, thus having important application value.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. Specifically, it is the isolation, cloning, functional verification and application of a gene HAF1 that mediates heading date in rice. Transforming the gene into rice and other plants through genetic engineering technology can regulate the heading date of rice and other crops, thereby determining the growth habit and yield of rice. Background technique [0002] The transition from vegetative growth to reproductive growth (shown as heading in rice) is one of the most important growth and development processes in its life cycle. Because people use different organs for different plants, plants are required to have a suitable flowering time. For example, delaying or avoiding flowering of Moso bamboo can increase the yield of useful vegetative organs and improve economic benefits. Proper flowering time in rice is essential for successful sexual reproduction to obtain more seeds. ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00C12N9/00C12N15/52A01H6/46
Inventor 吴昌银杨莹符德保
Owner HUAZHONG AGRI UNIV