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Rice serine/threonine protein kinase gene OsSAPK8 coding sequence and applications thereof

A nucleotide sequence and rice technology, applied in the field of genetic engineering, can solve problems that need to be explored and the regulation network of low temperature stress is not clear

Inactive Publication Date: 2018-08-03
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been many studies on the downstream signaling mechanism of SnRK2.6 and the sensitivity to low temperature stress, the low temperature stress regulatory network in rice is still unclear. In addition, SAPK7 , 8 The mechanism of genes in plant drought and high-salt stress response remains to be explored

Method used

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  • Rice serine/threonine protein kinase gene OsSAPK8 coding sequence and applications thereof
  • Rice serine/threonine protein kinase gene OsSAPK8 coding sequence and applications thereof
  • Rice serine/threonine protein kinase gene OsSAPK8 coding sequence and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Rice genes OsSAPK8 Clone of

[0047] 1. Rice variety Zhonghua11 is cultivated in an incubator (SPX-250-GB, Shanghai, China): the growth conditions are 16h / 8h (L / D) photoperiod, 28℃;

[0048] 2. DNA extraction. Take about 500 mg of fresh rice plant tissue material, add 80 μl Lysis Buffer, grind the plant tissue with a grinding rod, and add 120 μl ddH 2 O. After centrifugation at 12000rpm for 15min, transfer the supernatant to a new centrifuge tube, measure the OD value, and electrophoresis;

[0049] 3. Gene cloning. Through the corresponding Arabidopsis AtSnRK2.6 Gene information analysis and comparison and search in gene library, target gene and Arabidopsis gene AtSnRK2.6 The homology is more than 78%. Using the extracted rice DNA as a template, PCR was performed using forward primers and reverse primers to obtain the full length of the gene. See SEQ ID NO. 1 for specific sequence information.

Embodiment 2

[0050] Example 2, Rice OsSAPK8 Analysis of gene expression patterns in organs

[0051] The total RNA from rice roots, stems, and leaves were extracted, and the total RNA was reverse-transcribed into cDNA using a reverse transcription kit. The primers SEQ ID NO.5 and SEQ ID NO.6 were used for Real-time PCR detection ( figure 1 ). The results showed that the gene was expressed constitutively, with the highest expression in leaves and the lowest expression in stems.

Embodiment 3

[0052] Example 3 Rice Gene OsSAPK8 Analysis of expression patterns under drought, high salt stress and plant hormone treatments

[0053] Two-week-old rice seedlings were treated with 150 mM NaCl, 20% PEG drought treatment and 6 ℃ low temperature treatment for 4 hours, and the total RNA from the roots was extracted. The total RNA was reverse transcribed into cDNA using the reverse transcription kit, and the primers SEQ ID NO. 5 and SEQ ID NO. 6 were used for Real-time PCR detection. It was found that this gene is extremely sensitive to abiotic stress, especially drought induced most obviously ( figure 2 A). Afterwards, the sequential induction expression patterns of the above three abiotic stresses were studied, namely 150mMNaCl, 20%PEG drought treatment for 12h, and total RNA in roots after 0, 2, 4, 8, and 12h treatments were extracted respectively; proceed to 6. ℃ low temperature treatment for 12h, extract the total RNA in the leaves after 0, 0.25, 0.5, 1, 1.5, 2, 4, 8, and 12...

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Abstract

The invention belongs to the technical field of genetic engineering, and specifically relates to a rice serine / threonine protein kinase gene OsSAPK8 coding sequence and applications thereof. The applications comprise cloning of gene OsSAPK8 gene OsSAPK8, the organ expression pattern of the OsSAPK8 gene, and the change of the expression level of the OsSAPK8 after treatment using different hormonesand different stresses. The invention also discloses sequencing identification of the Tilling mutant of the gene OsSAPK8. A result of phenotype identification of the mutant shows that the OsSAPK8 plays a great role in the resistance of the plant to cold stress. The gene OsSAPK8 can be used for improving the plant variety, for example, improving the resistance of rice to cold, drought and salt stress, improving the ability of rice to cope with abiotic stress and increasing the rice yield.

Description

Technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a kind of OsSAPK8 Gene coding sequence and its application. Background technique [0002] Rice is one of the most important food crops in the world. my country's rice planting area is about 1 / 4 of the world's total rice area, accounting for 1 / 3 of the country's grain farming area, and its output accounts for 37% of the world's total rice output and 45% of the country's total output. In recent years, with global climate change, low temperature and cold damage have occurred frequently, causing huge losses to agricultural production, especially some low temperature sensitive crops such as rice, wheat, soybeans and various tropical and subtropical fruits such as bananas, mangos, grapes, oranges, etc. Production will be reduced or even eliminated; the continuous low temperature in recent years has greatly affected the production of food crops, which may affect the c...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/12C12N15/11C12Q1/6895
CPCC12N9/12C12Q1/6895C12Q2600/13C12Q2600/156C12Y207/11
Inventor 明凤钟瑞龄王玉霞
Owner FUDAN UNIV
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