Promoter used for corn low phosphor stress inducible expression, and applications thereof

A technology for inducing expression and promoters, applied in the fields of molecular biology and genetic engineering, can solve problems such as phosphorus poisoning, affecting plant growth and production, and achieve the effect of avoiding energy waste

Active Publication Date: 2018-08-21
CROP SCI RES INST SHANXI ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the excessive consumption of plant nutrients and energy in transgenic plants due to the use of constitutive promoters for high-intensity expression under any phosphorus conditions in the curr

Method used

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  • Promoter used for corn low phosphor stress inducible expression, and applications thereof
  • Promoter used for corn low phosphor stress inducible expression, and applications thereof
  • Promoter used for corn low phosphor stress inducible expression, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: ZmPHR1 Gene 5' Upstream Sequence P 2205 -ZmPHR1 and the acquisition of missing fragments

[0020] first step: ZmPHR1 Gene 5' Upstream Sequence P 2205 -ZmPHR1 the acquisition

[0021] by ZmPHR1 The sequence of the coding region was used as a probe, and BLAST searched the maize B73 genome sequence database (http: / / maizesequence.com), and the sequence about 2.0 kb upstream of the homologous gene was used as the target region, and the DNA of the maize inbred line 478 was used as a template, and the primer SF1 ( Italics are Sal I restriction sites, SEQ ID NO:6) and SR (italics are Bgl II restriction sites, SEQ ID NO:7) amplified fragments P 2205 -ZmPHR1 Ligated to pMD18-T vector.

[0022] (1) Prepare the PCR reaction solution according to the following components:

[0023] DNA 1 μL

[0024] dNTP Mixture (2.5 mΜ each) 8 μL

[0025] 10×LA PCR Buffer II (Mg 2+ plus) 5 μL

[0026] TaKaRa LA Taq enzyme (5 U / μl) 0.5 μL

[0027] Primer SF1 (10 pmol...

Embodiment 2

[0037] Embodiment 2: Construction of recombinant plant expression vector

[0038] All recombinant plasmids and plant expression vector pCAMBIA1305 were used Sal I and Bgl II double enzyme digestion and ligation, the correct recombinant plant expression vector was obtained after enzyme digestion detection and sequencing verification, and the recombinant plant expression vector contained a GUS reporter gene ( figure 2 , image 3 ), named as P 2205 -ZmPHR1::GUS , P 1763 -ZmPHR1::GUS , P 1502 -ZmPHR1::GUS , P 972 - ZmPHR1::GUS and P 482 -ZmPHR1::GUS . Transform the recombinant vector into Agrobacterium GV3101 middle.

Embodiment 3

[0039] Example 3: Transforming Arabidopsis thaliana with Agrobacterium to obtain pure transgenic lines

[0040] recombinant plant expression vector P 2205 -ZmPHR1::GUS , P 1763 -ZmPHR1::GUS , P 1502 - ZmPHR1::GUS , P 972 -ZmPHR1::GUS and P 482 -ZmPHR1::GUS and pCAMBIA1305 vector (35S::GUS) were transformed into wild-type Arabidopsis Col-0, and the seeds were harvested in the greenhouse. got a T 1 Substitute seeds contained 25 μg mL -1 MS medium screening and PCR detection (SEQ ID NO:12, SEQ ID NO:13) ( Figure 4 ) after transplanting and growing in the greenhouse, self-crossing to get T 2 generation, and then through selfing and hygromycin selection to obtain the transgenic pure line T 3 generation, respectively named as T P 2205 -ZmPHR1 , T P 1763 -ZmPHR1 , T P 1502 - ZmPHR1 , T P 972 -ZmPHR1 , T P 482 -ZmPHR1 and TpCAMBIA1305, for subsequent functional analysis.

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Abstract

The invention belongs to the technical field of molecular biology and gene engineering, and provides a promoter used for corn low phosphor stress inducible expression, and applications thereof. The promoter is used for solving problems in the prior art that transgenic plant nutrient and energy are over consumed, plant growth and yield are influenced, and phosphorus poisoning is caused because of high expression of a conventional combined promoter under any phosphor conditions. The promoter comprises a ZmPHR1 gene initiation codon ATG upstream 2205bp promoter sequence represented by SEQ ID NO:1, and 5'-terminal deleted promoter deletion fragment sequences represented by SEQ ID NO:2, 3, 4, and 5. The promoter can be used for replacing the combined promoter in plant engineering, ensuring thefunction of phosphor high efficiency absorption and utilization genes under low phosphor stress, avoiding energy waste caused by unnecessary expression at normal phosphor conditions, and culturing ideal high phosphor crop varieties.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and genetic engineering, and specifically relates to a corn low-phosphorus stress-induced expression promoter and application thereof. Background technique [0002] Phosphorus is an indispensable macronutrient for plant growth and development. It is not only a component of many important molecules in plants, but also plays an irreplaceable role in energy transfer and many metabolic regulation. Phosphorus is absorbed by plants mainly through the root system in the form of phosphate. Although the total phosphorus content in the soil is high, the phosphate radical is easily adsorbed and fixed by various metal cations and minerals in the soil or converted into organic phosphorus without being absorbed. The lack of available phosphorus in the soil and inefficient utilization are two important factors that limit plant growth. The application of inorganic phosphorus fertilizers is currently a...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82C12N15/66A01H5/00A01H6/20
CPCC12N15/66C12N15/8222C12N15/8271
Inventor 白建荣闫蕾杨瑞娟苏亮王秀红李锐关超张丛卓
Owner CROP SCI RES INST SHANXI ACADEMY OF AGRI SCI
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