Sweet potato leaf development and flavonoid enhancement related protein IbBBX29 as well as coding gene and application thereof

A technology that encodes genes and flavonoids, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of lack of genetic resources, incompatibility of intraspecific hybridization, narrow genetic basis, etc.

Active Publication Date: 2022-05-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sweet potato is a vegetatively propagated crop, the incompatibility of interspecific and intraspecific hybridization, the lack of genetic resources, and the narrow genetic basis all severely limit the application of conventional breeding

Method used

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  • Sweet potato leaf development and flavonoid enhancement related protein IbBBX29 as well as coding gene and application thereof
  • Sweet potato leaf development and flavonoid enhancement related protein IbBBX29 as well as coding gene and application thereof
  • Sweet potato leaf development and flavonoid enhancement related protein IbBBX29 as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Example 1, the acquisition of IbBBX29 gene

[0104] The steps to obtain the IbBBX29 gene are as follows:

[0105] 1. According to the transcriptome analysis, find a highly expressed gene IbBBX29 in sweet potato leaves, obtain the diploid CDS sequence of the gene by comparing with the sweet potato diploid genome database, and synthesize the primer IbBBX29-F:5'- ATGGCGAAAGAAACGAAGAG-3' and IbBBX29-R: 5'-TCAATCAAGCAGTGACGACG-3'. Total RNA from young leaf organs of flavonoid-rich Sushu 33 was extracted with a plant total RNA extraction kit, and the total RNA was extracted with PrimeScript TM 1stStrand cDNA Synthesis Kit reverse-transcribes the first-strand cDNA.

[0106] 2. Using the cDNA obtained in step 1 as a template, using IbBBX29-F and IbBBX29-R as primers to carry out PCR amplification, and ligating the amplified product with the cloning vector pMD19-T to obtain the recombinant vector pMD19-T-IbBBX29. The recombinant vector pMD19-T-IbBBX29 was sequenced to obtain...

Embodiment 2

[0108] Example 2. Application of IbBBX29 protein and its encoding gene in regulating leaf development and / or flavonoid content in leaves

[0109] 1. Construction of recombinant plasmids

[0110] A. Construction of recombinant plasmid pCambia1300-IbBBX29-GFP

[0111] 1. The double-stranded DNA molecule shown in SEQ ID No. 2 of the artificially synthesized sequence listing. Using this double-stranded DNA molecule as a template, IbBBX29-OE-F: 5'-GG GGTACC ATGGCGAAAGAAACGAAGAG-3' (KpnI restriction site is underlined), and IbBBX29-OE-R: 5'-ACGC GTCGAC ATCAAGCAGTGACGACG' (the underlined part is the Sal I restriction site) was used as a primer for PCR amplification to obtain a double-stranded DNA molecule containing the restriction endonuclease KpnI at the N-terminal and the restriction endonuclease SalI at the C-terminal.

[0112] 2. The vector pCambia1300-GFP was double-digested with restriction enzymes KpnI and SalI to recover the large fragment, while the double-stranded DNA...

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Abstract

The invention discloses a sweet potato leaf development and flavonoid enhancement related protein IbBBX29 as well as a coding gene and application of the sweet potato leaf development and flavonoid enhancement related protein IbBBX29. The invention specifically discloses a protein with an amino acid sequence of SEQ ID No.1, a coding gene, a related biological material and application of the protein to regulation and control of plant leaf development and / or flavonoid content in leaves. According to the invention, the IbBBX29 gene is introduced into a wild sweet potato, so that a transgenic sweet potato plant over-expressing the IbBBX29 gene is obtained, and meanwhile, an IbBBX29 gene knockout plant is constructed. Compared with a wild plant, the fresh weights of the overground part, the leaves and the stems of the overexpressed plant are obviously increased, and the flavonoid content in the leaves is also obviously increased. Experiments show that the IbBBX29 gene plays a key role in regulation and control of sweet potato leaf development and flavonoid synthesis, and important functional genes and breeding materials are provided for genetic improvement of sweet potato leaf development and / or regulation and control of the flavonoid content in leaves.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to sweet potato leaf development and flavonoid enhancement related protein IbBBX29 and its encoding gene and application. Background technique [0002] Sweet potato (Ipomoea batatas (L.) Lam.) is an autohexaploid plant belonging to the Convolvulaceae, Ipomoea, and Section Batatas groups, and is an important food, feed, industrial raw material and new Root crops for energy use. Sweet potatoes are widely distributed in more than 100 countries and regions around the world, and my country is the largest producer of sweet potatoes in the world. Sweet potato has a well-developed aerial part, stems and leaves reproduce and grow quickly, and can be harvested multiple times during the growth cycle. Sweet potato stems and leaves are edible, and are rich in nutrients such as protein, polyphenols, vitamins, and minerals. It is a high-quality vegetable resource. According to research ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/12A01H5/00A01H6/20A01H6/00
CPCC07K14/415C12N15/8205C12N15/8243C12N15/8241C12N15/8218
Inventor 何绍贞张欢刘庆昌翟红高少培赵宁高晓茹
Owner CHINA AGRI UNIV
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