Ornithogalum thyrsoides QtHIPP37 gene and application thereof

A technology of evergreen and gene, applied in the field of evergreen QtHIPP37 gene and its application

Active Publication Date: 2019-04-05
四川天艺优境环境科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Ornithogalum thyrsoides is a perennial bulbous flower of the genus Ornithogalum thyrsoides in the family Liliaceae. It has the advantages of strong vitality, wide adaptability, strong reproductive ability, and easy cultivation, especially for drought, saline-alkali and other adversity stresses. Strong stress resistance, but its molecular mechanism needs to be further studied
[0006] Molecular chaperones are key proteins that regulate the safe transport of metal ions in cells, and are closely related to plant resistance; heavy metal-associated isoprenylated plant protein (HIPP) is a plant-specific Molecular chaperones containing two conserved sites of metal binding site (HMA) and C-terminal prenylation site (Ca

Method used

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  • Ornithogalum thyrsoides QtHIPP37 gene and application thereof
  • Ornithogalum thyrsoides QtHIPP37 gene and application thereof
  • Ornithogalum thyrsoides QtHIPP37 gene and application thereof

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Experimental program
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Embodiment 1

[0034] Example 1 Extraction of the QtHIPP37 gene of Ornithogalum leucocephala:

[0035] Cut the leaves of the aseptic tissue cultured seedlings of Ornithogalum ornithogalum on a sterilized ultra-clean workbench and transfer them to a medium containing cytokinin. After 20-30 days, the surface of the leaves gradually grows multiple greens. The beads on the leaves began to form. Using the beads on the leaves of Ornithogalum vulgaris as the material, RNA was extracted and reverse transcribed into cDNA. The corresponding primers were designed for PCR, agarose gel electrophoresis, and the target bands were recovered. The pMD19-T vector was ligated, transformed into E. coli, sequenced and analyzed. Pick positive clones for plasmid extraction, design two seamlessly fused primers based on the sequence information of the PHBYFP plant expression vector, and use high-fidelity enzymes for PCR amplification of the positive plasmid, and perform the linearization process with the PHBYFP plant ex...

Embodiment 2

[0050] Example 2 Gene function verification

[0051] Firstly, the plant expression vector of QtHIPP37 gene of Ornithogalum sibiricum was constructed and transformed into wild-type tobacco for eukaryotic expression and phenotypic observation, identification and analysis of transgenic tobacco plants.

[0052] (1) Construction of a constitutive plant expression vector of QtHIPP37 gene

[0053] 1. The ORF of the QtHIPP37 gene was cloned into the PHBYFP vector and placed under the control of the ubiquitin promoter. The specific method is:

[0054] 2. Design seamless connection primers based on the above-mentioned cDNA sequence of Dieffenbachia QtHIPP37 gene and the 15-20 bases beside each restriction site of HindIII and BamHI of PHBYFP vector. The sequence is as follows:

[0055] PHBY-F-HindIIIQtHIPP37F:

[0056] ACCAGTCTCTCTCTCAAGCTTatgactaaagatgaagagttc;

[0057] PHBY-R-BamHIQtHIPP37R:

[0058] gctcaccatactagtggatccCATCACAGAACAACTGCCAGTG;

[0059] 3. Use HindIIIQtHIPP37F and BamHIQtHIPP37R pri...

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Abstract

The invention discloses an ornithogalum thyrsoides QtHIPP37 gene and application thereof. The nucleotide sequence of the gene is as shown in SEQ ID NO.1, and the amino acid sequence coded by the geneis as shown in SEQ ID NO.2. According to the ornithogalum thyrsoides QtHIPP37 gene and application thereof, on the basis of establishing a high-efficiency regeneration system and transcriptome analysis of a bulbil on leaves of ornithogalum thyrsoides, the QtHIPP37 gene is cloned and an over-expression vector is constructed, the QtHIPP37 gene and the over-expression vector are introduced into modelplant tobacco, the stress resistance of the transgenic tobacco to the drought stress is remarkably improved, and therefore it is proved that the QtHIPP37 gene can improve the stress resistance performance such as the drought resistance and the saline-alkali resistance of plants from the molecular level and has a good application prospect in plant biotechnology application.

Description

Technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to a white flower ornithogalum QtHIPP37 gene and its application. Background technique [0002] Drought and salinity are currently global problems that severely restrict agricultural production. China itself is a country with severe water shortages and very scarce freshwater resources, as well as being affected by climate warming and excessive use of chemical fertilizers, drought and water shortage and soil compaction. Problems such as salinization are becoming more and more serious, posing a great threat to agriculture, forestry and horticulture. In particular, many flowers and horticulture plants are cultivated annually in protected areas and greenhouses. Watering and irrigation consumes a lot of resources, manpower and material resources, and water resources are in urgent need of high efficiency. At the same time, the problem of salinization of cultivated ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/82
CPCC12N15/8273C07K14/415
Inventor 姜福星黄远祥周鹏李西何伟王丽娜
Owner 四川天艺优境环境科技有限公司
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