Plant drought tolerance related protein oserf62 and its coding gene and application

A drought-tolerant, plant-based technology, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve the problems of large chance, unstable varieties, long cycle of drought-resistant rice varieties, etc.

Active Publication Date: 2017-11-10
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Using traditional breeding methods to cultivate drought-resistant rice varieties has a long cycle and high chance, the varieties cultivated are unstable, and the progress is relatively slow. However, the use of genetic engineering technology can directly modify the genetic background of plants at the genetic level, and the genetic traits of plants can be directional modified

Method used

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  • Plant drought tolerance related protein oserf62 and its coding gene and application
  • Plant drought tolerance related protein oserf62 and its coding gene and application
  • Plant drought tolerance related protein oserf62 and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Acquisition of Plant Drought Tolerance-Related Protein OsERF62 and Its Encoding Gene

[0047] The upland rice IRAT109 seedlings cultivated under normal conditions were taken, the total RNA was extracted by Trizol method, and after purification, the cDNA was obtained by reverse transcription with M-MLV reverse transcriptase. Using the cDNA as a template, primers F1 and R1 were designed for PCR amplification, the amplified product was subjected to agarose gel electrophoresis, and a DNA fragment of 1580 bp was recovered and purified for sequencing. The result is shown in sequence 2 of the sequence table.

[0048] The sequences of the above primers are as follows:

[0049] F1: 5′-CGG GGTACC AAAGGCATTCGCAACACACA-3′ (the underlined base is the restriction endonuclease KpnI restriction endonuclease recognition sequence);

[0050] R1: 5′-CC TTAATTAA CCAAAATACATTACGACTGGAC-3' (the underlined base is the recognition sequence for restriction endonuclease PacI).

...

Embodiment 2

[0052] Example 2, real-time fluorescent quantitative PCR analysis of the expression characteristics of the endogenous OsERF62 gene

[0053] 1. Coercion treatment

[0054] The rice variety Nipponbare and the upland rice variety IRAT109 seedlings with a seedling age of 4 weeks were subjected to the following treatments respectively:

[0055] 1. ABA treatment: Soak the roots of the seedlings in 100 μM ABA solution, culture in light for 1 hour, 2 hours, 4 hours, 6 hours, 9 hours, 12 hours, and 36 hours, then take the leaves, and freeze them with liquid nitrogen at -80°C Save for later.

[0056] 2. Dehydration treatment: put the seedlings in the air for 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, and 8 hours, then take the leaves, freeze them with liquid nitrogen, and store them at -80°C for later use.

[0057] 3. H 2 o 2 Treatment: Soak the roots of the seedlings in 1mM H 2 o 2 After being placed in the solution for 1 hour, 2 hours, 4 hours, 6 hours, 9 hours, 12 hou...

Embodiment 3

[0065] Embodiment 3, the construction of recombinant expression vector

[0066] After the 1580bp DNA fragment obtained in Example 1 was double-digested with KpnI and PacI, the digested product was connected to the backbone fragment of the vector pMDC32 that had been double-digested with KpnI and PacI, and confirmed by sequencing and digestion, the recombinant vector 35S was obtained: :OsERF62, the recombinant vector 35S::OsERF62 is inserted between the KpnI and PacI sites of the vector pMDC32 (that is, the downstream of the double tobacco mosaic virus promoter 35S) from the 10th to the 1570th nucleotide of Sequence Listing Sequence 2 DNA fragments shown in sequence.

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Abstract

The invention discloses a plant drought resistance related protein OsERF62, its coding gene and application thereof. The protein provided by the invention is derived from upland rice, is named as OsERF62, and is composed of the amino acid sequences shown as sequence 1 in the sequence table. Experiments prove that, the recombinant expression vector 35S::OsERF62 containing a DNA molecule (i.e., the coding gene of the protein) shown as the 102nd-1106th position of sequence 2 in the sequence table is employed to transform paddy rice Nipponbare to obtain OsERF62 gene overexpressed T2 generation transgenic lines, and under the following 3 drought stress treatment, i.e. 200g / L PEG6000, 200mmol / L mannitol and soil water cut-off, the survival rates or growth vigor of the plants are both significantly higher than those of the wild type paddy rice Nipponbare. The invention provides a new gene and method for improving plant drought tolerance, thus having important significance in plant genetic improvement and drought tolerance study.

Description

technical field [0001] The invention relates to a plant drought tolerance related protein OsERF62 and its coding gene and application. Background technique [0002] Abiotic stress is the main stress factor that affects the normal growth and development of plants and limits the increase of crop yield. At present, the impact of drought on crop production in the world ranks first among all stress factors, and its damage to crop growth is equivalent to the sum of all natural disasters, and has become a bottleneck in agricultural development in many places. Rice is one of the main food crops in the world and is the staple food for more than 60% of the population in my country. According to statistics, rice water accounts for about 70% of agricultural water use. Since 1997, my country's rice cultivation area has continued to shrink, and more than 20 million mu of rice suffers from drought every year, and even tens of millions of mu of rice cannot be planted. Therefore, overcomin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10C12N1/15C12N1/19C12N1/21C12N7/01A01H5/00
CPCC07K14/415C12N15/8273
Inventor 李自超熊海燕刘鹏丽李金杰张洪亮
Owner CHINA AGRI UNIV
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