B-type response adjustment gene ORR2 for regulating and controlling rice dwarfing and application of gene

A response regulator, rice technology, applied in the field of agricultural biology, can solve the problem of slow research in the field of cytokinin signal transduction

Active Publication Date: 2019-07-09
广东省科学院南繁种业研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, research in the field of cytokinin signal transduction is slow, especially in the study of rice plant type

Method used

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  • B-type response adjustment gene ORR2 for regulating and controlling rice dwarfing and application of gene
  • B-type response adjustment gene ORR2 for regulating and controlling rice dwarfing and application of gene
  • B-type response adjustment gene ORR2 for regulating and controlling rice dwarfing and application of gene

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preparation example Construction

[0033] Another embodiment of the present invention also provides a method for preparing the rice type B response regulator gene ORR2, comprising the following steps:

[0034] A) reverse transcribing the total RNA of true leaves of rice seedlings into cDNA;

[0035] B) Obtained by PCR amplification using the above cDNA as a template and using the primer pair shown in SEQ ID NO: 3 and SEQ ID NO: 4.

[0036] An example of the present invention also relates to the application of the rice B-type response regulator gene ORR2, one of which is the application in regulating the dwarfing of rice plants.

[0037] An example of the present invention also relates to an overexpression vector. An overexpression vector, which is transferred with the rice B-type response regulator gene ORR2; it can be realized by connecting the ORR2 gene to the vector pCAMBIA1390 plasmid through In-fusion technology.

[0038] An example of the present invention also relates to a method for preparing rice tra...

Embodiment 1

[0051] Embodiment 1 The acquisition of rice ORR2 gene and its encoded protein

[0052] (1) RNA extraction and cDNA cloning

[0053] The rice used was wild-type Nipponbare. Soak the seeds for 6-8 hours, then germinate them in an incubator at 30-32°C for 2-3 days, and sow when the seeds are white. After the seedlings grew to one leaf and one heart, the true leaves of the three seedlings were mixed and sampled. After adding liquid nitrogen, the tissues and cells were lysed with a mortar, and quickly transferred to a 2.0mL centrifuge tube. The plant total RNA extraction kit ( RNAprep pure Tissue Kit, TIANGEN); Use a spectrophotometer to detect the concentration of RNA, while agarose gel electrophoresis to detect the integrity of RNA. Next, use the reverse transcription kit (Prime Script Reverse Transcriptase kit, Takara) of Dalian Bao Biological Company to reverse transcribe into cDNA.

[0054] (2) Amplification of ORR2 gene

[0055] The rice ORR2 gene fragment was cloned usin...

Embodiment 2

[0057] Example 2 Study on the Expression of Rice ORR2 in Different Rice Tissues

[0058] According to the cDNA sequence of ORR2, design real-time quantitative PCR (qRT-PCR) primers:

[0059] ORR2-Q-F:AAGGTTCTTGAGACCCTCCT; SEQ ID NO.5

[0060] ORR2-Q-R: TGATGACTGGGAGATCCATTT SEQ ID NO. 6.

[0061] The internal reference primers used were rice Ubiqutin primers,

[0062] Ubiqutin-Q-F: GCTCCGTGGCGGTATCAT SEQ ID NO. 7;

[0063] Ubiqutin-Q-R: CGGCAGTTGACAGCCCTAG SEQ ID NO. 8.

[0064] Dilute the cDNA template 5 times and prepare a 20 μL reaction system: 10 μL SYBR Premix Ex Taq II (2×), 0.8 μL Forward Primer, 0.8 μL Reverse Primer, 2 μL cDNA, 0.4 μL Rox Reference Dye II, 6.0 μl ddH 2 O, and then carry out the amplification reaction on the fluorescent quantitative PCR instrument (ABI Prism7500HT, Applied Biosystems).

[0065]The reaction program is: 95°C, 30sec, 95°C, 5sec, 60°C, 34sec, a total of 40 cycles. Then carry out the amplification reaction on the fluorescent quantitat...

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Abstract

The invention relates to a rice B-type reaction regulation and control factor gene ORR2 and application of the gene. An ORR2 gene cDNA reading frame is in a nucleotide sequence shown in SEQ ID NO:1 ora nucleotide sequence completely in complementary pairing with SEQ ID NO:1 or a coding amino acid sequence, such as a nucleotide sequence shown in SEQ ID NO:2. An ORR2 gene overexpression vector is established, and the height of a rice overexpression plant with the established ORR2 gene is obviously lowered. The rice B-type reaction regulation and control factor gene ORR2 can be applied to regulation and control over rice plant-type dwarfing and dwarfing cultivation.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a rice type B response regulator gene ORR2 and an application thereof. Background technique [0002] Rice is one of the most important crops in the world. More than 50% of the world's population feeds on rice. The importance of rice is self-evident. With the reduction of arable land and the increase of population, the yield and quality of rice are also on the rise. The green revolution of rice marked by dwarf breeding has made a huge contribution to the increase of world food production. As an important agronomic trait, plant height directly affects rice plant type and yield. Rice plant type improvement plays an important role in increasing unit yield. Rice plant type traits are regulated by very complex regulatory mechanisms. Therefore, it is clear that crop type development Regulatory mechanism, excavation and utilization of new regulatory dwarf genes are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/10C12N15/82C12N1/21A01H5/04A01H6/46
CPCC07K14/415C12N15/1096C12N15/8261C12Q2531/113
Inventor 史发超安玉兴孙东磊卢颖林龚恒亮陈立君付建涛赵欢欢戴思行
Owner 广东省科学院南繁种业研究所
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