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Clone and application of a gene improving rice drought tolerance and relative with ABA synthesis

A technology of drought tolerance and genetics, applied in the field of cultivating plants with improved drought tolerance, can solve the problems of limited systematic research and less research, and achieve the effects of broad application prospects, improved drought tolerance, and improved tolerance

Active Publication Date: 2008-05-07
未名三农生物农业技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, water stress and its signal transduction mechanism have been extensively studied in Arabidopsis (Shinozaki K, Yamaguchi-Shinozaki K. Gene expression and signal transduction in water-stress response. Plant Physiology, 1997, 115: 327-334) , but systematic studies on monocotyledonous plants are still very limited, especially on crops, especially the relationship between the response of crops to early-drying signals and their NCED gene expression and ABA levels and the mechanism remains to be elucidated

Method used

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  • Clone and application of a gene improving rice drought tolerance and relative with ABA synthesis
  • Clone and application of a gene improving rice drought tolerance and relative with ABA synthesis
  • Clone and application of a gene improving rice drought tolerance and relative with ABA synthesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1. Obtaining drought tolerance genes OsNCEDs

[0070] 1. Obtaining the gene sequence

[0071] We used the cDNA sequence of Arabidopsis AtNCED3 as the basis for blast analysis in the rice genome database. Three OsNCEDs genes with high homology to AtNCED3 sequence were found, named OsNCED3 (AY838899), OsNCED4 (AY838900), OsNCED5 (AY838901).

[0072] 2. PCR amplification and sequence analysis of OsNCED3, OsNCED4 and OsNCED5

[0073] Using the total DNA of Oryza sativa “Zhonghua NO.11” as a template, the PCR method was used to amplify OsNCED3, OsNCED4, and OsNCED5. The primer sequences are as follows:

[0074] (1) Design primers according to the CDS sequence of OsNCED3:

[0075] OsNCED3-F 5’-CACCACAAAATGGCGACGATCACGACG-3’

[0076] OsNCED3-R 5’-GGCCTGGGTGGTGAGCTC-3’

[0077] (2) Design primers according to the CDS sequence of OsNCED4:

[0078] OsNCED4-F 5’-CACCACAAAATGGCGTCCTCCGCGCCTT-3’

[0079] OsNCED4-R 5’-AGCTTGTCGCTGCAGCTCGTT-3’

[0080] (3) Design primers according...

Embodiment 2

[0086] Example 2. Obtaining of transgenic rice with OsNCEDs genes related to drought tolerance in rice

[0087] The drought tolerance-related genes OsNCED3, OsNCED4 and OsNCED5 obtained in Example 1 were respectively transformed into rice by Agrobacterium-mediated method, and the specific methods are as follows:

[0088] 1) Construction of pHORD vector:

[0089] The pH2GW7 vector (Invitrogen's Gateway TM vector vector containing the CaMV 35S constitutive promoter) The CaMV 35S constitutive promoter in the Arabidopsis thaliana rd29A inducible promoter ( Narusaka Y, Nakashima K , Shinwari ZK , Sakuma Y , Furihata T , Abe H, Narusaka M , Shinozaki K , Yamaguchi-Shinozaki K .Interaction between two cis-acting elements, ABRE and DRE, inABA-dependent expression of Arabidopsis rd29A gene in response to dehydration and high-salinity stresses. Plant Journal, 2003, 34(2): 137-48.). Specific method: Using Arabidopsis genomic DNA as template, use rd29A-F1 (5'CACCGAGCTCATTTTCGTTCTTGACA...

Embodiment 3

[0096] Example 3. Genetic analysis and molecular identification of transgenic T1 rice plants

[0097] Take the OsNCED3-pH2GW7, OsNCED3-pHORD, OsNCED4-pH2GW7 and OsNCED5-pHORD T1 transgenic rice seeds harvested in Example 2, and soak the seeds in water for 3 days to germinate, and then use 50mg / L hygromycin for resistance screening. The non-transgenic rice Zhonghua 11 was used as a control. After 5 days of selection, all the control plants died. The number of resistant and dead seedlings of the transgenic plants was counted, the resistance segregation ratio of the transgenic plants was analyzed, and the separation of resistant and non-resistant seedlings was selected. For strains with a ratio of about 3:1, the results showed that OsNCED3-pH2GW7, OsNCED3-pHORD, OsNCED4-pH2GW7 and OsNCED5-pHORD transgenic strains with single point insertion were obtained.

[0098] Select the plants of the transgenic line inserted at the unit point with a growth period of 15 days, extract the total RN...

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Abstract

The invention discloses an OsNCEDs gene relative to drought resistance originating from rice. Coded protein has one of the following amino acid sequences: 1). SEQ ID NO: 1, 3 or 5 in sequence list; 2). The amino acid residue sequence for the SEQ ID NO: 1, 3 or 5 in the sequence list pass through replacing, lacking or adding of one to ten amino acid residue and derived protein has the function of controlling drought resistance of plants. The experiment proves that: the gene conversion rice can obviously improve drought resistance of rice. The invention has the advantages of having an important theory and actual meaning upon research of drought resistance mechanism for plants and improving drought resistance and relative characteristics, playing an important role in drought resistance gene engineering improvement for plants (especially for rice crops) and having wide application prospect.

Description

Technical field [0001] The present invention relates to transcription factors related to ABA synthesis, and particularly uses genetic engineering methods to manipulate the expression of OsNCEDs genes of source rice, thereby regulating the level of ABA in plants, and is applied to cultivating plants with improved drought tolerance. Background technique [0002] Phytohormones are very small amounts of non-nutritive substances that are synthesized by some organs (or tissues) of plants and transferred to another organ (or tissues), and specifically affect their physiological processes. Plant growth and development are regulated by nutrition, endogenous hormones and growth environment. [0003] There are five main types of plant hormones, namely auxin, gibberellin, cytokinin, abscisic acid and ethylene. Phytohormones, as trace signal molecules in plants, participate in almost every process of regulating plant growth and development, including regulating the growth and development of p...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H1/00
Inventor 王喜萍孟秀萍黄晓翠刘春霞辛莉谢莹夏勉
Owner 未名三农生物农业技术有限公司
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