Promoter for abiotic stress specificity expression of sweet potato IbCBF3 gene and application thereof

A kind of technology of abiotic stress and promoter, applied in the field of abiotic stress-specific expression promoter of sweet potato IbCBF3 gene, and abiotic stress-specific expression promoter field, can solve the low temperature intolerance of sweet potato, restrict the development of sweet potato industry, etc. question

Active Publication Date: 2018-11-23
XUZHOU INST OF AGRI SCI IN JIANGSU XUHUAI DISTRICT (JIANGSU XUZHOU SWEETPOTATO CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The intolerance of low temperature of sweet potato seri

Method used

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  • Promoter for abiotic stress specificity expression of sweet potato IbCBF3 gene and application thereof
  • Promoter for abiotic stress specificity expression of sweet potato IbCBF3 gene and application thereof
  • Promoter for abiotic stress specificity expression of sweet potato IbCBF3 gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Cloning of embodiment 1 sweet potato IbCBF3 gene promoter

[0028] According to the cDNA sequence of the important transcription factor IbCBF3 in the sweet potato low temperature stress signal transduction pathway, according to the requirement of the genome walker kit specification sheet of Clontech company, 3 specific primers (SEQ ID NO.2: SP1:5'-GAAGTTGAGACAGGCGGAGCAG-3'; SEQ ID NO.3SP2:5'-GTCTCCCGAAACTTCTTCCTCCC-3'; SEQ ID NO.4SP3:5'-CCAACAACACCTCTTCATCCGACA-3', the design direction is the direction of the unknown region to be amplified, SP2 The position is located on the inside of SP1, and SP3 is located on the inside of SP2) and four uniquely designed degenerate primers AP1, AP2, AP3, and AP4 with lower annealing temperatures provided in the kit are used as primers for nested PCR amplification. The amplification results were sequenced after gel recovery. Design SEQ ID NO.5~6 (SEQ ID NO.5:5'-TGATAGTGAGTTGGGTTAGC-3; SEQ ID NO.6:5'-TCCATATACGGCGGTACTT-3'), using swee...

Embodiment 2

[0029] Example 2 Sequence Analysis of the IbCBF3 Gene Promoter in Sweet Potato

[0030] The plant care software in PLACE analyzed the sequence, and the results showed that in the IbCBF3 promoter, in addition to the conserved elements such as TATA-box, GATA-box and CAAT-box, which are necessary for transcription, there are also some elements that can sense environmental stress, light, Elements of water, hormone response and growth and development regulation, such as ACE, ATCT-motif, GATT-motif, G-box and I-box involved in light regulation; abscisic acid response factor ABRE; ethylene response factor ERE and jasmonate A Ester response factor CGTCA-motif et al. In addition, the promoter also contains multiple CANNTG cis-acting elements that can combine with the upstream regulatory factor ICE gene and MYCRE.

Embodiment 3

[0031] Example 3 Construction of dual fluorescent expression vectors

[0032]Using the cDNA of Xushu 29 as a template, the promoters of IbCBF3 were amplified with primers SEQ ID NO.12-13 (SEQ ID NO.125'-GATTGGGAAGAAAGTAT-3'; SEQ ID NO.135'-ATACGGCGGTACTTAGTAAAC-3') The full-length sequence and primers SEQ ID NO.15~16 (SEQ ID NO.155'-ATGTTATCAAGAGTGACAAGC'; SEQ ID NO.165'-GATCATCCCGTTGAAGCTAG') amplify the IbICE1 gene, connect to the T-blunt vector, and transform it into coli DH5α competent cells. Kanamycin was added to the LB medium, and a single colony was obtained by plate screening. The genome of the single colony was extracted for PCR verification, and the verified single colony was shaken to extract the plasmid for sequencing. The IbCBF3 promoter (IbCBF3pro for short) and the IbICE1 gene were respectively constructed into the vectors pBI101 and pCAMBIA1200 according to the instructions of the In-fusion kit of TAKARA Company to obtain the recombinant vectors IbCBF3pro-GUS...

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Abstract

The invention relates to a promoter for abiotic stress specificity expression, in particular to a promoter for abiotic stress specificity expression of a sweet potato IbCBF3 gene and an application thereof, which belongs to the field of plant genetic engineering. The invention provides a promoter for abiotic stress specificity expression for the sweet potato IbCBF3 abiotic stress specificity expression. A nucleotide sequence of the promoter is as shown by SEQ ID NO.1; and a DNA fragment of the invention is the promoter of a key transcription factor in the sweet potato low-temperature stress way, and the promoter contains a plurality of adversity and hormone response elements. By virtue of dual-luciferase systematic analysis in tobacco leaves, the promoter is rapidly combined with an upstream gene IbICE1 under the low-temperature stress to induce the expression situation of a downstream gene and can be used for researching the low temperature resistance of the sweet potatoes and for thetransgenic breeding of the sweet potatoes.

Description

technical field [0001] The invention relates to a promoter specifically expressed by abiotic stress, in particular to a promoter specifically expressed by abiotic stress of sweet potato IbCBF3 gene and application thereof, belonging to the field of plant genetic engineering. Background technique [0002] Temperature is one of the most important environmental factors in the process of plant growth and development. Low temperature not only affects the growth and yield of crops, but also restricts the cultivation time and geographical distribution of crops. According to the latest statistics from the Food and Agriculture Organization of the United Nations (FAO, Food and Agriculture Organization of the United Nations), my country's sweet potato planting area is 3.37 million hectares, accounting for 40% of the world's planting area, and it is the largest sweet potato planting country; the total output is 7.13 million tons , accounting for 73% of the world's sweet potato productio...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00A01H6/00
CPCC12N15/8205C12N15/8222
Inventor 靳容刘明李洪民张爱君陈晓光唐忠厚
Owner XUZHOU INST OF AGRI SCI IN JIANGSU XUHUAI DISTRICT (JIANGSU XUZHOU SWEETPOTATO CENT)
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