CN-122012590-A - Transcription factor BnaMYB and application thereof

CN122012590ACN 122012590 ACN122012590 ACN 122012590ACN-122012590-A

Abstract

The invention discloses a transcription factor BnaMYB and application thereof, belonging to the technical field of genetic engineering. The invention provides an application of BnaMYB gene or protein in regulating drought resistance of rape and/or regulating anthocyanin content of rape under drought stress. BnaMYB30 through binding BnaUGT75C1 and BnaFLS1 promoters, participate in regulating anthocyanin glycosylation and flavonol synthesis, and significantly promote anthocyanin accumulation by activating key structural genes CHI and F3H, DFR in flavonoid synthesis pathways. BnaMYB30 over-expression strain lacks effective ABA and JA hormone signal regulation, has single stress response mode, leads to reduced antioxidant enzyme activity and reduced active oxygen scavenging capacity, and finally shows drought sensitive phenotype.

Inventors

CHEN WEIQI
ZHOU WEIJUN
HUANG QIAN
ZHOU YINGYING
QIN TONGJUN
ZHANG KANGNI
MA JIALI
SHEN YI
LAN WENQIANG
Hu Yiwa

Assignees

江苏省农业科学院

Dates

Publication Date: 20260512
Application Date: 20260226

Claims (10)

Application of BnaMYB30 gene or protein in regulating drought resistance of rape and/or regulating anthocyanin content of rape under drought stress.
2. The use according to claim 1, wherein BnaMYB is used for negative regulation of rape drought resistance and BnaMYB is used for positive regulation of rape anthocyanin content under drought stress.
3. The use according to claim 2, wherein BnaMYB regulates anthocyanin glycosylation and flavonol synthesis under drought stress by regulating BnaFLS and BnaUGT C1 genes, and wherein BnaMYB regulates BnaFLS and BnaUGT C1 expression by combining SEQ ID NO 13 and SEQ ID NO 14 sequences with BnaFLS and BnaUGT C1 promoters.
4. The use according to claim 1, wherein the BnaMYB gene is located at position 45820979 to 45822485 of the C07 chromosome of the brassica napus reference genome with version number zs11.v0.
5. The application of the recombinant vector of the over-expression BnaMYB in improving the anthocyanin content of the rape under drought stress.
6. The application of the recombinant vector according to claim 5, wherein the construction method of the recombinant vector comprises the steps of carrying out double enzyme digestion on the vector by utilizing restriction enzymes to obtain a linearization vector, obtaining a CDS target fragment of BnaMYB through PCR amplification, carrying out homologous recombination on the CDS target fragment of BnaMYB30 to the linearization vector, transferring a connection product into E.coli DH5 alpha competent cells through a heat shock method, and extracting plasmids after sequencing is correct.
7. The use according to claim 6, wherein the primer sequence for PCR amplification comprises SEQ ID NO. 1-SEQ ID NO. 2.
8. A method for regulating drought resistance of rape and/or regulating anthocyanin content of rape under drought stress is characterized by regulating and controlling drought resistance of rape and anthocyanin content of rape under drought stress by regulating and controlling expression level of BnaMYB in rape, improving expression level of BnaMYB in rape, reducing drought resistance of rape, improving anthocyanin content of rape under drought stress, reducing expression level of BnaMYB in rape, improving drought resistance of rape and reducing anthocyanin content of rape under drought stress.
9. The method of claim 8, wherein the means for increasing the expression level of BnaMYB in canola comprises any one or more of the following: (1) Introducing BnaMYB gene to make the gene over expressed; (2) Modifying endogenous BnaMYB gene regulatory region by gene editing to enhance transcription; (3) Administering BnaMYB a gene promoter activator to promote expression thereof; (4) A transcriptional regulator was introduced that up-regulated BnaMYB gene expression.
10. The method of claim 8, wherein the means for reducing the expression level of BnaMYB in canola comprises any one or more of the following: (1) Introducing BnaMYB gene silencing vector to inhibit the expression of the gene; (2) Gene editing knocks out or knocks down endogenous BnaMYB genes; (3) Administering BnaMYB gene promoter inhibitor to attenuate transcription; (4) An inhibitory regulatory factor was introduced which down-regulates BnaMYB gene expression.

Description

Transcription factor BnaMYB and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a transcription factor BnaMYB and application thereof. Background Anthocyanin is an important secondary metabolite of plants, and synthesis and accumulation of anthocyanin are closely related to response of plants to adversity stress. However, the dynamic regulation and control mechanism of anthocyanin in drought resistance response of rape is not completely clear, and particularly, the synthesis route, key genes and the action difference in different rape varieties are still in urgent need of systematic research. Rape is not only an important oil crop, but also has higher ornamental value. Therefore, research on the synthesis regulation mechanism of anthocyanin in rape is particularly important for development of drought-resistant breeding of colored rape. Disclosure of Invention In view of the above, the invention aims to provide an application of BnaMYB gene or protein in regulating drought resistance of rape and/or regulating anthocyanin content of rape under drought stress, wherein BnaMYB30 overexpression obviously increases anthocyanin content under drought stress and improves drought sensitivity. The invention further aims to provide an application of the recombinant vector overexpressing BnaMYB in improving the anthocyanin content of rape under drought stress. The invention also aims to provide a method for regulating and controlling the drought resistance of the rape and/or regulating and controlling the anthocyanin content of the rape under drought stress. In order to achieve the above object, the present invention provides the following technical solutions: the invention provides an application of BnaMYB gene or protein in regulating drought resistance of rape and/or regulating anthocyanin content of rape under drought stress. Preferably, bnaMYB is used for negatively regulating and controlling the drought resistance of the rape, and BnaMYB is used for positively regulating and controlling the anthocyanin content of the rape under drought stress. Preferably, bnaMYB regulates anthocyanin glycosylation and flavonol synthesis under drought stress by regulating BnaFLS and BnaUGT C1 genes. Preferably, bnaMYB controls the expression of BnaFLS and BnaUGT C1 by the sequence of SEQ ID NO. 13 and SEQ ID NO. 14 in combination with the BnaFLS and BnaUGT C1 promoters. Preferably, the BnaMYB gene is located at position 45820979 to 45822485 of the C07 chromosome of the brassica napus reference genome with a version number of ZS11.v0. The invention also provides application of the recombinant vector of the over-expression BnaMYB to improving the anthocyanin content of the rape under drought stress. Preferably, the construction method of the recombinant vector comprises the steps of carrying out double enzyme digestion on the vector by utilizing restriction enzymes to obtain a linearization vector, obtaining a CDS target fragment of BnaMYB through PCR amplification, carrying out homologous recombination on the CDS target fragment of BnaMYB to the linearization vector, transferring a connection product into a DH5 alpha competent cell of escherichia coli by a heat shock method, and extracting plasmids after sequencing correctly. Preferably, the primer sequence for PCR amplification comprises SEQ ID NO 1-SEQ ID NO 2. The invention also provides a method for regulating drought resistance of rape and/or regulating anthocyanin content of rape under drought stress, which comprises regulating drought resistance of rape and anthocyanin content of rape under drought stress by regulating and controlling expression level of BnaMYB in rape, improving expression level of BnaMYB in rape, reducing drought resistance of rape, improving anthocyanin content of rape under drought stress, reducing expression level of BnaMYB in rape, improving drought resistance of rape and reducing anthocyanin content of rape under drought stress. Preferably, the means for increasing the expression level of BnaMYB in canola comprises any one or more of the following: (1) Introducing BnaMYB gene to make the gene over expressed; (2) Modifying endogenous BnaMYB gene regulatory region by gene editing to enhance transcription; (3) Administering BnaMYB a gene promoter activator to promote expression thereof; (4) A transcriptional regulator was introduced that up-regulated BnaMYB gene expression. Preferably, the means for reducing the expression level of BnaMYB in canola comprises any one or more of the following: (1) Introducing BnaMYB gene silencing vector to inhibit the expression of the gene; (2) Gene editing knocks out or knocks down endogenous BnaMYB genes; (3) Administering BnaMYB gene promoter inhibitor to attenuate transcription; (4) An inhibitory regulatory factor was introduced which down-regulates BnaMYB gene expression. The invention has the beneficial effects that: The invention sc