Application of WRKY transcription factor gene CqWRKY1 in regulating drought resistance of benincasa hispida

A technology of transcription factors and drought resistance, applied in the fields of application, genetic engineering, plant genetic improvement, etc., to achieve the effect of improving drought resistance performance

Inactive Publication Date: 2019-04-12
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the effect of WRKY transcription factors on the drought resistance of Zucchini.

Method used

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  • Application of WRKY transcription factor gene CqWRKY1 in regulating drought resistance of benincasa hispida
  • Application of WRKY transcription factor gene CqWRKY1 in regulating drought resistance of benincasa hispida
  • Application of WRKY transcription factor gene CqWRKY1 in regulating drought resistance of benincasa hispida

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Acquisition of WRKY transcription factor gene CqWRKY1 and its encoded protein

[0022] (1) RNA extraction and cDNA cloning

[0023] The selected zucchini variety is a homozygous inbred line material selected from the natural pollination offspring of Xinnong gourd variety through pedigree method.

[0024] Soak the seeds in warm soup for 6-8 hours, then germinate 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 TissueKit, TIANGEN); the concentration of RNA was detected by a spectrophotometer, and the integrity of the RNA was detected by agarose gel electrophoresis. Next, use the reverse transcription kit (PrimeScript Reverse Transcriptase kit, T...

Embodiment 2

[0033] Construction of embodiment 2CqWRKY1 overexpression vector and detection of positive plants

[0034] (1) Construction of CqWRKY1 overexpression vector

[0035] Using the cDNA of the CqWRKY1 gene as a template, the 1746bp coding region was amplified by PCR, and the PCR amplification product was recovered and purified. The CqWRKY1 gene was connected to the overexpression vector pEarleyGate 101 plasmid using Gateway technology, and positive clones were selected for sequencing after transformation. The correct plasmid Agrobacterium GV3101 was transformed, and 50mg / L Rif and 50mg / L Kan were used as resistance screening, and the obtained single clone was identified as positive by PCR. Transform zucchini materials through the Agrobacterium-mediated method.

[0036] (2) Detection of overexpression plants

[0037] The specific primer CqWRKY1-F designed according to the overexpression vector sequence (pEarleyGate 101 vector is provided by Vegetable Research Institute of Guangdon...

Embodiment example 3

[0042] Implementation Case 3 Drought Resistance Identification of Overexpressed Plants

[0043] The T2 pure line Zygodon seeds of the overexpressed gene CqWRKY1 (T0 generation positive plants are self-pollinated to obtain T1 generation plants, and then T1 generation is self-pollinated to obtain T2 generation plants, and the pure line overexpression seeds are obtained through qPCR detection) and the control Seeds were primed under the same conditions and then sown on sterile substrate. When it grows to one leaf and one heart, carry out drought and water shortage treatment for 8 days, and observe the growth status of the plants. Then add water to recover for 3 days, the result is as follows: image 3 shown.

[0044] from image 3 It can be seen that under drought stress, the resistance of transgenic plants is significantly higher than that of the control, indicating that CqWRKY1 has a certain positive effect in regulating the drought resistance of zucchini. However, in terms...

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Abstract

The invention discloses the application of a WRKY transcription factor gene CqWRKY1 in regulating drought resistance of benincasa hispida. A nucleotide sequence of the WRKY transcription factor gene CqWRKY1 is shown in a SEQ ID NO:1. The invention obtains coding sequence information of the benincasa hispida CqWRKY1; and overexpressing of plant drought resistance of the constructed KRKY transcription factor gene CqWRKY1 is enhanced, so that the gene positively regulates the drought resistance of the benincasa hispida. Therefore, in the subsequent production practice, the drought resistance of the benincasa hispida can be improved by overexpressing the WRKY transcription factor gene CqWRKY1 in the benincasa hispida.

Description

technical field [0001] The invention belongs to the technical field of zucchini, and in particular relates to the application of WRKY transcription factor gene CqWRKY1 in regulating the drought resistance of zucchini. Background technique [0002] Drought is one of the most important abiotic factors affecting crop growth and yield. It seriously interferes with the normal physiological metabolism of plants, and the reduction in plant growth and crop yield caused by water deficit exceeds the sum of the effects of other adversities. [0003] Chieh-qua How (Benincasa hispida Cogn. var. Chieh-qua How) is an important vegetable crop of Cucurbitaceae Winter Melon. In recent years, zucchini production in southern China, especially in spring and summer, often faces drought caused by high temperature weather, which affects the yield and variety of zucchini. At present, the genome of Zucchini has not been published, and there are few studies on molecular genetics, beneficial genes rel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/34
CPCC07K14/415C12N15/8273
Inventor 王敏何晓明彭庆务林毓娥刘文睿江彪杜翔斐张金平梁肇均
Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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