CN-120944941-B - Application of pineapple calcium-dependent protein kinase gene in pineapple water heart disease prevention and treatment

Abstract

The invention relates to the technical field of molecular biology, in particular to application of pineapple calcium-dependent protein kinase genes in pineapple aquatic heart disease prevention and treatment, wherein the pineapple calcium-dependent protein kinase genes comprise AcCPK, acCPK and/or AcCPK genes, CDS sequences of the pineapple calcium-dependent protein kinase genes are shown as SEQ ID NO.1-3, and the pineapple calcium-dependent protein kinase genes are expressed in pineapple fruits instantly, so that pineapple aquatic heart disease can be obviously inhibited, and important theoretical support is provided for analysis of pineapple aquatic heart disease mechanism and cultivation of high-quality water-resistant heart disease pineapple varieties.

Inventors

• LI CHUANLING
• FU QIONG
• ZHU ZHUYING
• ZHANG MIAOLIN
• Chen Linpan
• ZHANG XIUMEI
• YAO YANLI
• HE JUNJUN
• WU QINGSONG
• LIN WENQIU
• GAO YUYAO

Assignees

• 中国热带农业科学院南亚热带作物研究所

Dates

Publication Date

20260508

Application Date

20250807

Claims (6)

1. 1. The application of pineapple calcium-dependent protein kinase genes in pineapple water heart disease prevention and treatment or preparation of pineapple water heart disease prevention and treatment reagents is characterized in that the pineapple calcium-dependent protein kinase genes comprise AcCPK4, acCPK8 and/or AcCPK genes, a CDS sequence of the AcCPK4 genes is shown as SEQ ID NO.1, a CDS sequence of the AcCPK8 genes is shown as SEQ ID NO.2, and a CDS sequence of the AcCPK15 genes is shown as SEQ ID NO. 3.
2. 2. The application of pineapple calcium-dependent protein kinase genes in breeding of pineapple varieties with water-resistant heart disease is characterized in that the pineapple calcium-dependent protein kinase genes comprise AcCPK, acCPK and/or AcCPK genes, a CDS sequence of the AcCPK gene is shown as SEQ ID NO.1, a CDS sequence of the AcCPK gene is shown as SEQ ID NO.2, and a CDS sequence of the AcCPK gene is shown as SEQ ID NO. 3.
3. 3. A method for inhibiting pineapple aquatic heart disease is characterized by comprising the step of transforming an expression vector for inhibiting pineapple aquatic heart disease into pineapple fruits by using a method for infecting pineapple fruits by agrobacterium, wherein the expression vector for inhibiting pineapple aquatic heart disease comprises a pineapple calcium-dependent protein kinase gene as set forth in claim 1.
4. 4. The method for inhibiting pineapple heart disease according to claim 3, wherein the AcCPK gene amplification primer is shown in SEQ ID NO.7-SEQ ID NO.8, the AcCPK gene amplification primer is shown in SEQ ID NO.9-SEQ ID NO.10, and the AcCPK gene amplification primer is shown in SEQ ID NO.11-SEQ ID NO. 12.
5. 5. The application of the expression vector for inhibiting pineapple aquatic heart disease or the engineering bacteria for inhibiting pineapple aquatic heart disease in pineapple aquatic heart disease prevention and treatment or preparation of pineapple aquatic heart disease prevention and treatment reagent, wherein the expression vector for inhibiting pineapple aquatic heart disease or the engineering bacteria for inhibiting pineapple aquatic heart disease comprises the pineapple calcium-dependent protein kinase gene in claim 1.
6. 6. The application of the expression vector for inhibiting pineapple aquatic heart disease or the engineering bacteria for inhibiting pineapple aquatic heart disease in breeding pineapple varieties with water-resistant heart disease, wherein the expression vector for inhibiting pineapple aquatic heart disease or the engineering bacteria for inhibiting pineapple aquatic heart disease comprises the pineapple calcium-dependent protein kinase gene in claim 1.

Description

Application of pineapple calcium-dependent protein kinase gene in pineapple water heart disease prevention and treatment Technical Field The invention relates to the technical field of molecular biology, in particular to application of pineapple calcium-dependent protein kinase gene in pineapple water-borne heart disease prevention and treatment. Background Pineapple [ Ananascomosus (l.) Merr ] is the third largest tropical fruit worldwide next to bananas and citrus, with extremely high production applications and economic value. The global climate abnormal fluctuation greatly increases the incidence rate of pineapple water-borne heart disease, and the edible quality and commodity value of pineapple fruits are greatly reduced. Pineapple water-borne heart disease is a complex physiological and biochemical process, the cause of which is not completely resolved, and researches show that the incidence of the pineapple water-borne heart disease is closely related to external environmental factors, plant growth and development states, endogenous hormone levels, mineral nutrition in fruits, sugar content and the like. The study shows that the pineapple water-borne heart disease can be obviously induced by carrying out the treatment at the high temperature of 38 ℃ and the treatment at the low temperature of 10 ℃ 50 days after the pineapple flowers are scored. In addition, the size of pineapple coronary buds is inversely related to the incidence of the water-borne heart disease, the incidence of the pineapple coronary buds is lower, the size of plant leaves is positively related to the incidence of the water-borne heart disease, and the incidence of the pineapple coronary buds is higher when the leaves are more exuberant. Calcium is a very important nutrient element in plants, and calcium in plants exists mainly in the form of free calcium and bound calcium, and is widely distributed in cell nuclei, cytoplasm, vacuoles, cell membranes and cell walls, wherein vacuoles are calcium stores in cells and are important for regulating osmotic pressure in cells. The calcium in the plant body is mainly obtained from the soil by the root, and the calcium ions in the soil pass through apoplast and symplastomeric paths under the action of transpiration tension and are transported from the xylem to plant tissues with strong transpiration, wherein the transpiration of the leaves is greater than that of the fruits, so that more calcium can be unloaded into the leaves. This is why the more vigorous the plant grows, the more likely the fruit is to be calcium deficient. Calcium is not only a very important nutrient element in plants, but also plays an important role in stabilizing plant cell membranes and cell walls, regulating and controlling enzymes, regulating permeation and the like. For example, ca 2+ can be used as a second messenger to participate in the response of plants to biotic and abiotic stress, plays an important role in the plants, can be combined with exposed carboxyl of methyl galacturonate to form calcium pectate, so that the cell wall structure is more stable, and can be coupled with phospholipid and protein on the cell membrane to stabilize the cell membrane structure. In the later stage of cell mitosis, the cell plate separating two daughter cells mainly consists of calcium pectate, and calcium deficiency influences the formation of the cell plate and spindle filaments, so that the daughter cells cannot be separated in the cell division process, a binuclear phenomenon occurs, and finally the cells die. Calcium-dependent protein kinases (CPKs) are a typical class of serine/threonine protein phosphatases with four characteristic domains, the Calcium-binding domain (Calcium-BindingDomain, CBD), the serine/threonine protein kinase domain (ProteinKinaseDomain, PKD), the self-inhibiting junction region (AutoinhibitoryJunction, AJ), and the N-terminal variable domain (NTD) (N-TerminalVariableDomain). In the process of plant response to environmental stress, the concentration of intracellular Ca 2+ is increased, so that Ca 2+ is combined with CBD, further the conformational change of the protein is induced, the inhibition of a kinase domain by a self-inhibition domain is relieved, and a CPKs signal transduction function is activated. Studies have shown that CPKs is ubiquitous in plants, protists, oomycetes, and green algae, but has not been found in animals and fungi. CPKs constitute a large family of polygenic kinases in plants involved in regulating growth and development and stress response. Currently, 34, 31 and 17 CPKs family members have been identified in Arabidopsis, rice and pineapple, respectively, with some members having deeply resolved in function. For example, qinetal (2020) identified 17 AcoCPKs family members by systematic analysis of the pineapple (Ananascomosus) genome and found that expression of AcoCPK, acoCPK3 and AcoCPK6 was significantly inhibited by abiotic stresses (e.g., drought, salt stress, et