IDL7 MATURE POLYPEPTIDE PLANT SENESCENCE ACCELERATOR, PREPARATION METHOD AND APPLICATION THEREOF

Abstract

The present application discloses a plant senescence accelerator named IDL7 mature polypeptide, and its preparation method and an application thereof. The application belongs to the field of a plant senescence accelerator, and the IDL7 mature polypeptide serves as a major functional ingredient with the following amino acid sequence: F-G-S-L-V-L-N-A-L-P-K-G-S-R-P-G-S-G-P-S-K-K-T-N. The IDL7 mature polypeptide plant senescence accelerator may promote the leaf senescence of plants without other additional adverse manifestations, thus this application has strong field operability.

Claims

1. An IDL7 mature polypeptide plant senescence accelerator, wherein an IDL7 mature polypeptide serves as a major functional ingredient, and the IDL7 mature polypeptide has the following amino acid sequence SEQ ID No. 1:
F-G-S-L-V-L-N-A-L-P-K-G-S-R-P-G-S-G-P-S-K-K-T-N; the IDL7 mature polypeptide plant senescence accelerator comprises the IDL7 mature polypeptide and a 2-(N-morpholine) ethanesulfonic acid solution; wherein, the 2-(N-morpholine) ethanesulfonic acid solution is prepared by dissolving 2-(N-morpholine) ethanesulfonic acid monohydrate into an MS fluid medium; a concentration of the IDL7 mature polypeptide is 10-13 μmol/L, a concentration of the 2-(N-morpholine) ethanesulfonic acid solution is 2.8-3 mmol/L, and pH value of the 2-(N-morpholine) ethanesulfonic acid solution is 5.8-5.9.

2. The IDL7 mature polypeptide plant senescence accelerator according to claim 1, wherein a concentration of the IDL7 mature polypeptide is 10 μmol/L, a concentration of the 2-(N-morpholine) ethanesulfonic acid solution is 2.8 mmol/L, and the pH value of the 2-(N-morpholine) ethanesulfonic acid solution is 5.8.

3. The IDL7 mature polypeptide plant senescence accelerator according to claim 1, further comprising an assistant, the assistant comprises Tween-20, and an addition amount of the assistant is 1-2 v/v‰.

4. A preparation method of the IDL7 mature polypeptide plant senescence accelerator of claim 1, comprising following steps: dissolving IDL7 mature polypeptide powder into water to prepare an mother solution of IDL7 mature polypeptide with a concentration of 10 mmol/L for subsequent use; adding 2-(N-morpholine) ethanesulfonic acid monohydrate into a MS fluid medium to prepare a 2-(N-morpholine) ethanesulfonic acid solution of 2.8-3 mmol/L, adjusting pH of the solution to 5.8-5.9, and after fully mixing and dissolving, the 2-(N-morpholine) ethanesulfonic acid solution is obtained; adding the IDL7 mature polypeptide mother solution to the 2-(N-morpholine) ethanesulfonic acid solution, stirring uniformly and the IDL7 mature polypeptide plant senescence accelerator is obtained.

5. The preparation method of claim 4, wherein after adding the obtained IDL7 mature polypeptide mother solution to the 2-(N-morpholine) ethanesulfonic acid solution, the method further comprises a step of adding 1-2 v/v‰ Tween-20.

6. An application of the IDL7 mature polypeptide plant senescence accelerator according to claim 1 in promoting plant senescence, comprising the following steps: directly spraying the IDL7 mature polypeptide plant senescence accelerator on leaf surface of plant leaves staying at full extension period, or using cotton balls soaked with the IDL7 mature polypeptide plant senescence accelerator to apply the leaf surface of plant leaves staying at full extension stage.

7. An application of the IDL7 mature polypeptide plant senescence accelerator according to claim 2 in promoting plant senescence, comprising the following steps: directly spraying the IDL7 mature polypeptide plant senescence accelerator on leaf surface of plant leaves staying at full extension period, or using cotton balls soaked with the IDL7 mature polypeptide plant senescence accelerator to apply the leaf surface of plant leaves staying at full extension stage.

8. An application of the IDL7 mature polypeptide plant senescence accelerator according to claim 3 in promoting plant senescence, comprising the following steps: directly spraying the IDL7 mature polypeptide plant senescence accelerator on leaf surface of plant leaves staying at full extension period, or using cotton balls soaked with the IDL7 mature polypeptide plant senescence accelerator to apply the leaf surface of plant leaves staying at full extension stage.

9. An application of the IDL7 mature polypeptide plant senescence accelerator according to claim 1 in promoting plant senescence, comprising the following steps: directly spraying the IDL7 mature polypeptide plant aging accelerator is directly sprayed on leaf surface of tobacco leaves staying at early mature stage, or using cotton balls soaked with the IDL7 mature polypeptide plant senescence accelerator to apply the leaf surface of tobacco leaves staying at early mature stage.

10. An application of the IDL7 mature polypeptide plant senescence accelerator according to claim 2 in promoting plant senescence, comprising the following steps: directly spraying the IDL7 mature polypeptide plant aging accelerator is directly sprayed on leaf surface of tobacco leaves staying at early mature stage, or using cotton balls soaked with the IDL7 mature polypeptide plant senescence accelerator to apply the leaf surface of tobacco leaves staying at early mature stage.

11. An application of the IDL7 mature polypeptide plant senescence accelerator according to claim 3 in promoting plant senescence, comprising the following steps: directly spraying the IDL7 mature polypeptide plant aging accelerator is directly sprayed on leaf surface of tobacco leaves staying at early mature stage, or using cotton balls soaked with the IDL7 mature polypeptide plant senescence accelerator to apply the leaf surface of tobacco leaves staying at early mature stage.

12. The application according to claim 6, wherein a concentration of the plant senescence accelerator directly sprayed on the leaf surface is 10-12 μmol/L.

13. The application according to claim 9, wherein a concentration of the plant senescence accelerator directly sprayed on the leaf surface is 10-12 μmol/L.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] FIGS. 1A-1B are schematic diagrams showing a result of Arabidopsis thaliana detached leaves treated by an artificially synthesized IDL7 mature polypeptide plant senescence accelerator;

[0028] FIGS. 2A-2B are schematic diagrams showing a result of in vivo tobacco leaves treated by an IDL7 mature polypeptide plant senescence accelerator.

DETAILED DESCRIPTION OF THE INVENTION

[0029] To more clearly and specifically introduce the IDL7 mature polypeptide plant senescence accelerator and its preparation method and application provided by the embodiments of the present application the technical solution in the embodiments of the present application will be further described clearly and integrally. Apparently, the examples described herein are merely a portion of embodiments of the present application, rather than all the embodiments. Based on the embodiments of the present application, all the other embodiments obtained by a person skilled in the art without inventive effort shall fall within the protection scope of the present application.

[0030] It is to be noted that the term “mother solution” used in the embodiments refers to a solution with a higher concentration, and the solution needs to be diluted in subsequent use to serve as a working solution with a lower concentration.

Embodiment 1

[0031] An IDL7 mature polypeptide plant senescence accelerator comprising 10 μmol/L IDL7 mature polypeptide, a 2.8 mmol/L 2-(N-morpholine) ethanesulfonic acid solution; the 2-(N-morpholine) ethanesulfonic acid solution was prepared by dissolving 2-(N-morpholine) ethanesulfonic acid monohydrate into an MS fluid medium, and the 2-(N-morpholine) ethanesulfonic acid solution has a pH value of 5.8.

[0032] The preparation method is as follows:

[0033] (1) IDL7 mature polypeptide powder was weighed and dissolved into water, and an IDL7 mature polypeptide mother solution having a concentration of 10 mmol/L was prepared for further use;

[0034] (2) 2-(N-morpholine) ethanesulfonic acid monohydrate was weighed quantitatively and added to the MS fluid medium, and the 2.8 mmol/L 2-(N-morpholine) ethanesulfonic acid solution is prepared, pH of the solution was adjusted to 5.8, and materials were fully mixed and dissolved uniformly to obtain the 2-(N-morpholine) ethanesulfonic acid solution;

[0035] (3) 40 μL of the IDL7 mature polypeptide mother solution obtained in the step (1) were taken and added to 40 mL of the 2-(N-morpholine) ethanesulfonic acid solution prepared in the step (2), and stirred uniformly with a glass rod to obtain 40 mL of the IDL7 mature polypeptide plant senescence accelerator.

Embodiment 2

[0036] An IDL7 mature polypeptide plant senescence accelerator comprising 12 μmol/L IDL7 mature polypeptide, a 2.9 mmol/L 2-(N-morpholine) ethanesulfonic acid solution; the 2-(N-morpholine) ethanesulfonic acid solution was prepared by dissolving 2-(N-morpholine) ethanesulfonic acid monohydrate into an MS fluid medium, and the 2-(N-morpholine) ethanesulfonic acid solution has a pH value of 5.9.

[0037] The preparation method was as follows:

[0038] (1) IDL7 mature polypeptide powder was weighed and dissolved into water, and an IDL7 mature polypeptide mother solution having a concentration of 10 mmol/L was prepared for further use;

[0039] (2) 2-(N-morpholine) ethanesulfonic acid monohydrate was weighed quantitatively and added to the MS fluid medium, and the 2.9 mmol/L 2-(N-morpholine) ethanesulfonic acid solution is prepared, pH of the solution was adjusted to 5.9, and materials were fully mixed and dissolved uniformly to obtain the 2-(N-morpholine) ethanesulfonic acid solution;

[0040] (3) 120 μL of the IDL7 mature polypeptide mother solution obtained in the step (1) were taken and added to 100 mL of the 2-(N-morpholine) ethanesulfonic acid solution prepared in the step (2), and stirred uniformly with a glass rod to obtain 100 mL of the IDL7 mature polypeptide plant senescence accelerator.

Embodiment 3

[0041] An IDL7 mature polypeptide plant senescence accelerator comprising 13 μmol/L IDL7 mature polypeptide, a 3 mmol/L 2-(N-morpholine) ethanesulfonic acid solution; the 2-(N-morpholine) ethanesulfonic acid solution was prepared by dissolving 2-(N-morpholine) ethanesulfonic acid monohydrate into an MS fluid medium, and the 2-(N-morpholine) ethanesulfonic acid solution has a pH value of 5.8.

[0042] The preparation method was as follows:

[0043] (1) IDL7 mature polypeptide powder was weighed and dissolved into water, and an IDL7 mature polypeptide mother solution having a concentration of 10 mmol/L was prepared for further use;

[0044] (2) 2-(N-morpholine) ethanesulfonic acid monohydrate was weighed quantitatively and added to the MS fluid medium, and the 3 mmol/L 2-(N-morpholine) ethanesulfonic acid solution is prepared, pH of the solution was adjusted to 5.8, and materials were fully mixed and dissolved uniformly to obtain the 2-(N-morpholine) ethanesulfonic acid solution;

[0045] (3) 78 μL of the IDL7 mature polypeptide mother solution obtained in the step (1) were taken and added to 60 mL of the 2-(N-morpholine) ethanesulfonic acid solution prepared in the step (2), and stirred uniformly with a glass rod to obtain 60 mL of the IDL7 mature polypeptide plant senescence accelerator.

[0046] Performance Test

[0047] Laboratory Test

[0048] Embodiment 1 was set as an example; the IDL7 mature polypeptide plant senescence accelerator prepared in Embodiment 1 was used to treat detached leaves of Arabidopsis thaliana and in vivo leaves of tobacco respectively according to method of applying.

[0049] Treatment of Arabidopsis thaliana leaves: Arabidopsis thaliana grew for 30 days around under continuous illumination to obtain detached leaves from the same leaf position (the 6th leaf position); the detached leaves were treated by a puncher having a diameter of 0.5 cm, then placed flat on a petri dish; treated with 2.8 mmol/L 2-(N-morpholine) ethanesulfonic acid solution (hereafter refer as MES) as a control group; meanwhile, 10 μmol/L plant senescence accelerator was evenly and directly sprayed on the detached leaf surface of Arabidopsis thaliana as an experimental group, and placed the leaves of the control group and the experimental group for 4 days, then phenotype was observed.

[0050] As shown in FIG. 1A, Arabidopsis thaliana leaves with the same growth vigor in the same leaf position (the 6th leaf position) were selected, and treated by a puncher with a diameter of 0.5 cm. Then the Arabidopsis thaliana leaves were put on a Petri dish regularly, and treated by spraying with the IDL7 mature polypeptide plant senescence accelerator to obtain the experimental group, meanwhile, the control group was added and treated by the same method except that the plant senescence accelerator was replaced by 2.8 mmol/L EMS. The leaves of the control group and the experimental group were put for 4 days under continuous illumination.

[0051] It can be seen from the results that the IDL7 mature polypeptide plant senescence accelerator may promote the leaf senescence of Arabidopsis thaliana. 4 days later, the Arabidopsis thaliana detached leaves sprayed with the IDL7 mature polypeptide plant senescence accelerator had obvious premature phenotype. Meanwhile, chlorophyll content was measured by ethanol method, as shown in FIG. 1B; compared with the control group, the chlorophyll content in the leaves treated by the IDL7 mature polypeptide plant senescence accelerator decreased significantly, and the chlorophyll content in the control group decreased to 0.517 μg/mg, and the chlorophyll content in the treatment group decreased to 0.311 μg/mg.

[0052] Field Test

[0053] As shown in FIG. 2A, the tobacco variety K326 cultivated for about 60 days in field conditions served as a material, and was sprayed with 10 μmol/L IDL7 mature polypeptide plant senescence accelerator. Meanwhile, a control group was sprayed with a solution free of IDL7 mature polypeptide; 14 days later, it can be obviously observed that the tobacco sprayed with the IDL7 mature polypeptide plant senescence accelerator showed premature phenotype. Meanwhile, ethanol method was used to measure the chlorophyll content of tobacco leaves being sprayed with the IDL7 mature polypeptide plant senescence accelerator and the tobacco leaves of the control group after 14 days; and the results showed that compared with the control group (the chlorophyll content of the control group is 0.598 μg/mg), as shown in FIG. 2B, the chlorophyll content of the tobacco leaves treated by the IDL7 mature polypeptide plant senescence accelerator is 0.225 μg/mg, which is decreased significantly. The results showed that the IDL7 mature polypeptide plant senescence accelerator had obvious promotion effect on the leaf senescence of in vivo plants indeed.