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CN-122012529-A - CebHLH113 gene for regulating synthesis of taro anthocyanin and application thereof

CN122012529ACN 122012529 ACN122012529 ACN 122012529ACN-122012529-A

Abstract

The invention belongs to the technical field of plant molecular breeding, and particularly relates to CebHLH gene for regulating and controlling synthesis of taro anthocyanin and application thereof. The amino acid sequence of the protein coded by CebHLH113,113 genes is shown as SEQ ID NO. 1. The invention firstly separates a key gene CebHLH113,113 for positively regulating anthocyanin synthesis from Ganchun 3 red-meat taro. The CebHLH113,113 gene over-expression can obviously promote the accumulation of anthocyanin in the taro callus, and the silencing of the gene in the stem tip obviously reduces the accumulation level of anthocyanin. The invention fills the blank of the transcription regulation mechanism of the anthocyanin metabolism of the taro, is beneficial to providing important theoretical basis and technical support for improving the color quality of the taro and cultivating high-quality taro varieties with high anthocyanin content by a molecular breeding technology, and has wide application value.

Inventors

  • ZHU QIANGLONG
  • Jia Xinbi
  • WANG XINYU
  • Tong Ziyin
  • LIANG JIE
  • ZHOU QINGHONG
  • XIAO YAO

Assignees

  • 江西农业大学

Dates

Publication Date
20260512
Application Date
20260409

Claims (10)

  1. 1. A CebHLH gene for regulating and controlling synthesis of taro anthocyanin is characterized in that the amino acid sequence of protein coded by CebHLH gene is shown as SEQ ID NO. 1.
  2. 2. The CebHLH gene according to claim 1, wherein the nucleotide sequence of CebHLH gene is shown in SEQ ID No. 2.
  3. 3. A biological material for controlling the amount of CebHLH113,113 gene expression according to claim 1 or2, which is characterized by comprising a biological material that overexpresses CebHLH113,113 gene, a biological material that inhibits the CebHLH113,113 gene expression or knocks out the CebHLH113,113 gene expression; The method of inhibiting includes silencing.
  4. 4. The biological material according to claim 3, wherein the biological material over-expressing CebHLH gene comprises any one or more of amplification primers over-expressing the CebHLH gene, recombinant vector over-expressing the CebHLH gene, recombinant microorganism over-expressing the CebHLH gene, transgenic cell line over-expressing the CebHLH gene and transgenic plant tissue over-expressing the CebHLH gene; the basic skeleton of the recombinant vector comprises a pSuper1300 vector, and the basic microorganism of the recombinant microorganism comprises agrobacterium.
  5. 5. The biological material according to claim 4, wherein the amplification primer comprises a forward primer and a reverse primer, the nucleotide sequence of the forward primer is shown as SEQ ID NO. 3, and the nucleotide sequence of the reverse primer is shown as SEQ ID NO. 4.
  6. 6. The biological material according to claim 3, wherein the biological material for inhibiting the CebHLH gene expression or knocking out the CebHLH gene expression comprises any one or more of a silencing vector containing the CebHLH gene silencing target, a knocking-out vector containing the CebHLH gene knocking-out target, a transgenic cell line containing the silencing vector, a transgenic cell line containing the knocking-out vector, a recombinant microorganism containing the silencing vector, a recombinant microorganism containing the knocking-out vector, plant tissue for silencing CebHLH113 gene, and plant tissue for knocking out CebHLH113 gene; The basic backbone of the silencing vector comprises pTRV2.
  7. 7. Use of CebHLH gene according to claim 1 or 2 or biological material according to any one of claims 3-6 for regulating synthesis of taro anthocyanin and/or for breeding high anthocyanin taro varieties.
  8. 8. The use according to claim 7, wherein the regulation comprises over-expressing the CebHLH gene, promoting anthocyanin synthesis in taro, inhibiting the CebHLH gene expression or knocking out the CebHLH gene, and inhibiting anthocyanin accumulation in taro.
  9. 9. A method for promoting synthesis of anthocyanin in taro, which is characterized by comprising the following steps of overexpressing CebHLH gene as claimed in claim 1 or 2 in target taro and promoting synthesis of anthocyanin in taro.
  10. 10. A method for cultivating a high anthocyanin taro variety is characterized by comprising the following steps of overexpressing CebHLH gene according to claim 1 or 2 in target taro to obtain the high anthocyanin taro variety.

Description

CebHLH113 gene for regulating synthesis of taro anthocyanin and application thereof Technical Field The invention belongs to the technical field of plant molecular breeding, and particularly relates to CebHLH gene for regulating and controlling synthesis of taro anthocyanin and application thereof. Background The tuber of taro (Colocasia esculenta (L.) Schott) is also called as a root of Odonkey, and is a plant of the genus Odonkey of the family Araceae, namely a perennial monocotyledonous plant She Caoben, which is used as a traditional vegetable and grain crop in China, and the cultivation history of the tuber of taro is over 9000 years. The taro not only bears abundant historical culture background, but also is rich in nutritional ingredients such as starch, high-quality protein, polysaccharide, vitamins, dietary fibers and the like, and has the effects of nourishing, strengthening spleen, nourishing stomach, detoxifying and the like in the theory of traditional Chinese medicine, and has great value for human health. In recent years, taros are increasingly favored by consumers and have a wide range of researchers due to their unique flavor and rich nutrition. Anthocyanin is an important water-soluble secondary metabolite in plants, is mainly stored in cell vacuoles, not only endows flowers, fruits, tubers and other organs with various colors, but also can enhance the resistance of plants to drought, saline-alkali, diseases and insect pests and other stresses. In addition, the strong antioxidant activity makes the composition widely applied in the fields of food and medical treatment, and has important development value. The white, yellow, red, purple and other color cores of taro bulbs are determined by anthocyanin, and the detection of components such as pelargonidin-3-glucoside, cyanidin-3-rhamnoside and the like in the bulbs has been studied, and anthocyanin-3-rutinoside is also identified in the bulbs, so that anthocyanin is proved to be a key substance for forming the color of taros. Basic/helix-loop-helix (bHLH) transcription factors are the second largest family of transcription factors in plants, and are widely involved in numerous biological processes such as growth, signal transduction, metabolic regulation, and stress response. Members of this family have been identified in a variety of plants and their function has been intensively studied. Research shows that bHLH transcription factors play a role in diverse regulation in different plants. For example, about 10% of the bHLH members in Arabidopsis are involved in maintaining iron homeostasis, rice OsbHLH107,107 affects grain size by regulating the proliferation of ear shell cells, and in cotton, bHLH transcription factors are also closely related to the accumulation of secondary metabolites such as cottonseed oil, gossypol, anthocyanin, etc. Although there is much knowledge of the function of bHLH transcription factors in plant growth and development, the mechanism of this family to regulate anthocyanin biosynthesis may have species specificity due to species-to-species variability, and related studies are still not comprehensive enough to be explored further. The bHLH gene family has the characteristics of huge number of members and obvious functional differentiation, the roles of different members in the biological process are obviously different, and whether the bHLH gene family participates in and regulates the synthesis process of the taro anthocyanin is not clear. Therefore, the key genes for regulating anthocyanin biosynthesis in the taros are excavated, which is helpful for providing important theoretical basis and technical support for improving the color quality of the taros and cultivating high-quality taro varieties with high anthocyanin content by a molecular breeding technology. Disclosure of Invention The invention aims to provide CebHLH gene for regulating and controlling synthesis of taro anthocyanin and application thereof. The CebHLH113,113 gene over-expression can obviously promote the accumulation of anthocyanin in the taro callus, and the silencing of the gene in the stem tip obviously reduces the accumulation level of anthocyanin. The discovery is helpful for providing important theoretical basis and technical support for improving the color quality of taros and cultivating high-quality taro varieties with high anthocyanin content by a molecular breeding technology, and has wide application value. The invention provides CebHLH gene for regulating synthesis of taro anthocyanin, wherein the amino acid sequence of protein encoded by CebHLH gene is shown as SEQ ID NO. 1. As a preferable scheme, the nucleotide sequence of CebHLH113,113 gene is shown as SEQ ID NO. 2. The invention also provides a biological material for regulating CebHLH113,113 gene expression quantity, which comprises a biological material for over-expressing CebHLH113,113 gene, a biological material for inhibiting CebHLH113,113 gene expression or knocking o