CN-120648736-B - Application of HD-ZIP transcription factor in regulation and control of tomato fruit quality

CN120648736BCN 120648736 BCN120648736 BCN 120648736BCN-120648736-B

Abstract

The application discloses an application of an HD-ZIP transcription factor in regulating tomato quality, wherein the protein has functional redundancy, and the function of the protein is regulated by two genes, wherein the CDS sequence of a gene Solyc03g034130.2 is shown as SEQ ID NO.1, and the CDS sequence of a gene Solyc03g034110.2 is shown as SEQ ID NO. 2. According to the application, a tomato transcription factor mutant library is constructed by CRISPR technology, and a mutant Tom6-155 which can inhibit the expression of two homologous genes of the HD-ZIP transcription factor is screened. Based on observations of this mutant phenotype, the inventors determined that HD-ZIP transcription factors are associated with tomato fruit quality. The mutant has an increased brix content of tomato fruits compared to the wild type. It is explained that Solyc03g034130.2 and Solyc03g034110.2 exert important negative regulation effects in the process of tomato fruit quality formation. The application lays a good genetic resource foundation for the cultivation of new varieties of high-quality tomatoes.

Inventors

ZHANG YUQIN
Elon Chagny
LIU JIA
SU NING
Qin Yuntai

Assignees

中国科学院大学
特拉维夫大学拉莫特有限公司

Dates

Publication Date: 20260508
Application Date: 20250618

Claims (3)

The application of the HD-ZIP transcription factor in regulating tomato quality is characterized in that the amino acid sequences of the HD-ZIP transcription factor are respectively shown as SEQ ID NO.3 and SEQ ID NO.4, the coding genes are Solyc03g034130.2 genes and Solyc03g034110.2 genes, the CDS sequence of the Solyc03g034130.2 genes is shown as SEQ ID NO.1, the CDS sequence of the Solyc03g034110.2 genes is shown as SEQ ID NO.2, and the regulating tomato quality is shown in that the content of soluble solids in tomatoes of a knockout strain is improved after the Solyc03g034130.2 genes and the Solyc03g034110.2 genes are knocked out simultaneously.
2. A breeding method for improving tomato fruit quality is characterized in that Solyc03g034130.2 genes and Solyc03g034110.2 genes in tomato plants are knocked out simultaneously to obtain plants with improved tomato fruit quality, the CDS sequence of the Solyc03g034130.2 genes is shown as SEQ ID NO.1, the CDS sequence of the Solyc03g034110.2 genes is shown as SEQ ID NO.2, and the improvement of tomato fruit quality is shown as improvement of soluble solid content.
3. The breeding method for improving tomato fruit quality according to claim 2, wherein Solyc03g034130.2 gene and Solyc03g034110.2 gene in tomato genome are knocked out to obtain Solyc03g034130.2 gene and Solyc03g034110.2 gene knocked out plants, and homozygous Solyc03g03g034130.2 gene and Solyc03g034110.2 gene knocked out plants are obtained from selfed offspring of Solyc03g03g034130.2 gene and Solyc03g034110.2 gene knocked out plants.

Description

Application of HD-ZIP transcription factor in regulation and control of tomato fruit quality Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to application of an HD-ZIP transcription factor in regulating and controlling tomato fruit quality. Background In recent years, with the inclination of breeding targets to high yield, stress resistance and other characters, the modern cultivated tomato (Solanum lycopersicum) variety has the prominent problems of reduced flavor quality while remarkably improving yield and resistance. A large number of researches show that the phenomenon is mainly represented by systematic deterioration of key quality indexes such as unbalanced sugar-acid ratio of fruits, obviously reduced content of characteristic aroma substances, substantial change of fruits and the like. In-depth analysis shows that the quality degradation is closely related to disturbance of the transcription factor regulation network in the process of fruit development and ripening. Specifically, MYB, NAC, WRKY and MADS-box members form a complex molecular network affecting fruit quality formation through fine control of sugar metabolism, organic acid synthesis, volatile matter accumulation and other key ways. However, the key transcriptional regulatory mechanism for tomato quality formation still lacks systematic knowledge, and especially the development of novel transcription factors and the regulatory network analysis thereof are in need. In order to systematically improve the quality traits of tomatoes, the research identifies new key transcription factors through modern molecular experiment technology, and analyzes the regulation and control mechanisms of the key transcription factors on important quality traits such as accumulation of tomato sugar, aroma synthesis and the like. The method provides a new theoretical basis and technical support for realizing the precise improvement of tomato quality through molecular design breeding. Disclosure of Invention The invention aims to provide an application of an HD-ZIP transcription factor in regulating and controlling tomato fruit quality. In order to achieve the above object, the technical scheme of the present invention is as follows: According to the application, a tomato transcription factor mutant library is constructed by CRISPR technology, and a mutant Tom6-155 which can inhibit the expression of two homologous genes of the HD-ZIP transcription factor is screened. Based on observation of the mutant phenotype, the inventor determines that the HD-ZIP transcription factor is related to tomato fruit quality, the protein has functional redundancy, and the functions of the protein are respectively regulated and controlled by two genes, wherein the CDS sequence of the gene Solyc03g034130.2 is shown as SEQ ID NO.1, and the CDS sequence of the gene Solyc03g034110.2 is shown as SEQ ID NO. 2. The corresponding amino acid sequences are shown as SEQ ID NO.3 and SEQ ID NO. 4. Wherein, the Solyc03g034130.2 gene has a gene sequence ID (SequenceID) of LOC101251158 and a CDS sequence length of 675bp in NCBI, comprising 224 amino acids. The Solyc03g034110.2 gene has a gene sequence ID (SequenceID) in NCBI of LOC101254766 and a CDS sequence length of 642bp, including 213 amino acids. The two genes and the corresponding amino acids are subjected to sequence alignment by DNAMAN software, and the result shows that the CDS sequence similarity is 91.26 percent and the amino acid sequence similarity is 87.5 percent. Through preliminary researches on Solyc03g034130.2 and Solyc03g034110.2 genes of tomato, the inventor finds that Solyc03g034130.2 and Solyc03g034110.2 have a certain influence on tomato fruit quality, and the Solyc03g034130.2 and Solyc03g034110.2 genes of tomato are inversely related to tomato fruit quality, namely, after gene knockout or knockout, the content of soluble solids of tomato fruits is increased, and the fruit quality is improved. Lays a certain genetic resource foundation for the cultivation of new species of high-quality tomatoes. The invention also constructs a series of plant expression vectors, expression vectors containing the genes, transgenic plant lines and host cells containing the vectors, and the functions of regulating and controlling the quality of tomato fruits also fall into the protection scope of the invention. Wherein the recombinant vector comprises a gene editing recombinant vector and an over-expression recombinant vector. The Solyc03g034130.2 and Solyc03g034110.2 genes according to the present invention can be easily mutated by a person skilled in the art using known methods, such as directed evolution and point mutation. Those artificially modified, which have 75% or more identity to the nucleotide sequences of the genes Solyc03g034130.2 and Solyc03g034110.2, are all nucleotide sequences derived from the present invention and are equivalent to the sequences of the present in