CN-119709837-B - Application of watermelon KNOX transcription factor ClSPR in creation of reticulate pericarp watermelon

Abstract

The invention discloses an application of a watermelon KNOX transcription factor ClSPR in creating a reticulate pattern pericarp watermelon, and belongs to the technical field of plant genetic engineering. The application of the watermelon KNOX transcription factor ClSPR in creating reticulate peel watermelons disclosed by the invention adopts a CRISPR/Cas9 gene editing technology to carry out fixed-point editing on the watermelon KNOX transcription factor ClSPR so as to obtain novel germplasm of reticulate peel watermelons. Compared with wild type, the reticulate peel watermelon germplasm provided by the invention has no visible phenotype change in other tissues such as female flowers, leaves, tendrils, stems, roots and the like except that the peel fringe phenotype is changed, and can realize directional genetic improvement of excellent watermelon germplasm peel fringe phenotype, so that the reticulate peel watermelon germplasm has important production application potential for breeding and accumulating new materials of new varieties.

Inventors

• WEI CHUNHUA
• LI HAO
• ZHANG YONG
• SHEN JUNJUN
• ZHANG XIAN
• YUE ZHEN
• YU YUHONG
• LI JUNHUA
• WANG ZHONGYUAN
• YUAN LI
• MA JIANXIANG
• YANG JIANQIANG

Assignees

• 西北农林科技大学
• 开封市农林科学研究院

Dates

Publication Date

20260508

Application Date

20241226

Claims (3)

1. 1. The application of the knocked-out watermelon KNOX transcription factor ClSPR in creating reticulate pericarp watermelons is characterized in that the CDS sequence of the watermelon KNOX transcription factor ClSPR is shown in SEQ ID NO. 1.
2. 2. The application of the biological material for knocking out the watermelon KNOX transcription factor ClSPR in creating reticulate pattern pericarp watermelons is characterized in that the CDS sequence of the watermelon KNOX transcription factor ClSPR is shown as SEQ ID NO. 1; The biological material is any one of the following: An expression cassette capable of silencing watermelon KNOX transcription factor ClSPR; b, a recombinant vector containing the expression cassette A; And C, a recombinant microorganism containing the expression cassette A or the recombinant vector B.
3. 3. A method for creating new germplasm of a reticulate pattern peel watermelon is characterized in that a CRISPR/Cas9 gene editing technology is adopted to knock out a watermelon KNOX transcription factor ClSPR at a fixed point, and a CDS sequence of a watermelon KNOX transcription factor ClSPR is shown as SEQ ID NO. 1.

Description

Application of watermelon KNOX transcription factor ClSPR in creation of reticulate pericarp watermelon Technical Field The invention relates to the technical field of plant genetic engineering, in particular to application of watermelon KNOX transcription factor ClSPR in creating reticulate pericarp watermelons. Background Watermelon (Citrullus lanatus) is a cucurbitaceae (Cucurbitaceae) genus watermelon (Citrullus), which is an important cucurbitaceae crop widely planted in the world, and is a fruit type cash crop. According to statistics, the seeding area of the watermelons in 2022 is 139.19 ten thousand hectares, the total yield reaches 6054.22 ten thousand tons, and the total cultivation area and the yield respectively account for 47.73% and 60.57% of the total world cultivation area and the yield (FAO of grain and agriculture organization of China, http:// www.fao.org /). Therefore, the watermelon is taken as one of important melon crops cultivated and planted in China, and plays an important role in agricultural structure adjustment and rural economic development. The peel strips are important appearance quality of watermelons, and can directly influence the selection preference of consumers in the market, for example Crimson types of watermelons, the peel of which is covered with green and wide strips and the edges of the strips are dispersed, the watermelon strips are popular in the United states, brazil and Europe, and the peel of which is covered with dark green, medium-wide strips and the edges of the strips of Jubille types of watermelons are clear, so that the watermelon strips have large market sales in China and Korea. Therefore, the peel stripe is taken as a typical agronomic character of the watermelon, has important significance for improving the commodity of the watermelon, and can influence the purchase selection of consumers to a certain extent, thereby influencing the market sales of the watermelon. At present, researches on watermelon streak traits are mainly focused on genetic analysis and candidate gene positioning cloning levels, molecular mechanism researches on related regulatory genes are not reported, for example, kim and the like position major QTL loci of watermelon peel streaks to a range of 2.29Mb of chromosome 6, heeBum and Park and the like position peel streak regulatory loci S to a range of 3.99Mb of chromosome 6, maragal and the like show that the watermelon peel streaks and inter-streak peel color candidate genes are Cla97C09G175170 and Cla97C09G175150 on chromosome 9 respectively, and Wang and the like find that the intermittent streak formation of watermelon peel is regulated by ClIS (Cla 019202). Therefore, providing the use of watermelon KNOX transcription factor ClSPR in creating a textured peel watermelon is a problem that one skilled in the art would need to solve. Disclosure of Invention In view of this, the present invention provides the use of watermelon KNOX transcription factor ClSPR in creating a reticulate peel watermelon. The invention completes the positioning of watermelon peel stripe regulation gene ClSPR by the traditional gene positioning technology in the early stage, and preliminarily determines the best candidate gene by sequence and expression analysis. Based on the gene, the CRISPR/Cas9 technology can directionally change the stripe phenotype of the watermelon peel, further enrich the appearance characters of excellent watermelon materials and provide material resources for the enrichment of new varieties of special watermelons. In order to achieve the above purpose, the present invention adopts the following technical scheme: the CDS sequence of the watermelon KNOX transcription factor ClSPR in creating reticulate pattern pericarp watermelon is shown in SEQ ID NO. 1. Further, the application of the biological material for knocking out the watermelon KNOX transcription factor ClSPR in creating reticulate pattern pericarp watermelons, wherein the CDS sequence of the watermelon KNOX transcription factor ClSPR is shown as SEQ ID NO. 1; The biological material is any one of the following: An expression cassette capable of silencing watermelon KNOX transcription factor ClSPR; b, a recombinant vector containing the expression cassette A; And C, a recombinant microorganism containing the expression cassette A or the recombinant vector B. Furthermore, a novel germplasm creating method of the reticulate pericarp watermelon adopts CRISPR/Cas9 or other gene editing technology to carry out fixed-point editing or knockout on the watermelon KNOX transcription factor ClSPR or the conserved domain sequence thereof, so that the gene protein is in a loss or mutation state to generate a pericarp covered reticulate phenotype. The CDS sequence of the watermelon KNOX transcription factor ClSPR is shown in SEQ ID NO. 1. The invention firstly utilizes the positioned watermelon KNOX transcription factor ClSPR and combines the CRISPR/Cas9 technology to create a new ge