IDL6 MATURE POLYPEPTIDE PLANT AGING PROMOTER, AND PREPARATION METHOD AND APPLICATION THEREOF

Abstract

The present application an IDL6 mature polypeptide plant aging promoter, and a preparation method and application thereof, belonging to the field of plant aging promoters. The IDL6 mature polypeptide plant aging promoter is with an IDL6 mature polypeptide as a main functional component, and the IDL6 mature polypeptide having the following amino acid sequence: F-G-S-L-V-L-N-A-L-P-K-G-S-V-P-A-S-G-P-S-K-R-I-N. The provided plant aging promoter can promote the leaf senescence of the plant, accelerate the senescence, and has no other additional bad performance, and has extremely strong field operability.

Claims

1. An IDL6 mature polypeptide plant aging promoter, wherein the plant aging promoter is with an IDL6 mature polypeptide as a main functional component, and the IDL6 mature polypeptide has the following amino acid sequence: TABLE-US-00002 F-G-S-L-V-L-N-A-L-P-K-G-S-V-P-A-S-G-P-S-K-R-I-N; the plant aging promoter includes the IDL6 mature polypeptide and a 2-(N-morpholino)ethanesulfonic acid solution, wherein the 2-(N-morpholino)ethanesulfonic acid solution is a solution obtained by dissolving 2-(N-morpholino)ethanesulfonic acid monohydrate in an MS liquid culture medium; a concentration of the IDL6 mature polypeptide in the plant aging promoter is 10-13 μmol/L, a concentration of 2-(N-morpholino)ethanesulfonic acid in the 2-(N-morpholino)ethanesulfonic acid solution is 2.8-3 mmol/L, and a pH of the 2-(N-morpholino)ethanesulfonic acid solution is 5.8-5.9.

2. The plant aging promoter according to claim 1, wherein the concentration of the IDL6 mature polypeptide in the plant aging promoter is 10 μmol/L, the concentration of 2-(N-morpholino)ethanesulfonic acid in the 2-(N-morpholino)ethanesulfonic acid solution is 2.8 mmol/L, and the pH of the 2-(N-morpholino)ethanesulfonic acid solution is 5.8.

3. The plant aging promoter according to claim 1, wherein the plant aging promoter further includes an auxiliary agent, the auxiliary agent is Tween-20, and an added amount of the auxiliary agent is 1-2 v/v ‰.

4. The plant aging promoter according to claim 2, wherein the plant aging promoter further includes an auxiliary agent, the auxiliary agent is Tween-20, and an added amount of the auxiliary agent is 1-2 v/v ‰.

5. A preparation method for the IDL6 mature polypeptide plant aging promoter according to claim 1, specifically including the following steps: Dissolving IDL6 mature polypeptide power in water to prepare an IDL6 mature polypeptide mother solution with a concentration of 10-13 mmol/L; Adding 2-(N-morpholino)ethanesulfonic acid monohydrate to a prepared MS liquid culture medium to prepare 2-(N-morpholino)ethanesulfonic acid solution, wherein a concentration of 2-(N-morpholino)ethanesulfonic acid in the 2-(N-morpholino)ethanesulfonic acid solution is 2.8-3 mmol/L; adjusting a pH value of the 2-(N-morpholino)ethanesulfonic acid solution to be 5.8-5.9, and mixing fully and dissolving evenly to obtain the 2-(N-morpholino)ethanesulfonic acid solution; Adding the obtained IDL6 mature polypeptide mother solution into the 2-(N-morpholino)ethanesulfonic acid solution and stirring evenly to obtain the IDL6 mature polypeptide plant aging promoter.

6. The preparation method according to claim 5, wherein after adding the obtained IDL6 mature polypeptide mother solution into the 2-(N-morpholino)ethanesulfonic acid solution, the preparation method further includes a step of adding 1-2 v/v ‰ of Tween-20.

7. An application of the IDL6 mature polypeptide plant aging promoter according to claim 1 in promoting plant aging, wherein during application, the IDL6 mature polypeptide plant aging promoter is directly sprayed on surfaces of plant leaves in full extension stage, or a cotton ball is immersed in the IDL6 mature polypeptide plant aging promoter and then applied onto surfaces of plant leaves in full extension stage.

8. The application according to claim 7, wherein a concentration of the IDL6 mature polypeptide plant aging promoter directly sprayed on surfaces of plant leaves in full extension stage is 10-12 μmol/L.

9. The application according to claim 7, wherein during application, the IDL6 mature polypeptide plant aging promoter is directly sprayed on surfaces of tobacco leaves in early mature stage, or a cotton ball is immersed in the IDL6 mature polypeptide plant aging promoter and then applied onto surfaces of tobacco leaves in early mature stage.

10. The application according to claim 9, wherein a concentration of the IDL6 mature polypeptide plant aging promoter directly sprayed on surfaces of tobacco leaves in early mature stage is 10-12 μmol/L.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0026] FIG. 1 is a schematic diagram of the results of treating Arabidopsis thaliana in-vitro leaves with the IDL6 mature polypeptide plant aging promoter;

[0027] FIG. 2 is a schematic diagram of the results of measuring the chlorophyll content in the Arabidopsis thaliana in-vitro leaves treated with the IDL6 mature polypeptide plant aging promoter;

[0028] FIG. 3 is a schematic diagram of the results of treating tobacco in-vivo leaves with the IDL6 mature polypeptide plant aging promoter; and

[0029] FIG. 4 is a schematic diagram of the results of measuring the chlorophyll content in the tobacco in-vivo leaves treated with the IDL6 mature polypeptide plant aging promoter.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] The technical solutions of the present application will be described in detail below in combination with specific embodiments. However, it should be understood that elements, structures and features in one embodiment may also be advantageously incorporated into other embodiments without further description.

[0031] The embodiments are only described as preferred embodiments of the present application, and are not intended to limit the scope of the present application. Various modifications and improvements made on the technical solutions of the present application by ordinary skill in the art without departing from the design spirit of the present application shall fall within the protective scope confirmed by the claims of the present application.

[0032] The technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present application.

[0033] 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 an operating solution with a lower concentration.

Embodiment 1

[0034] An IDL6 mature polypeptide plant aging promoter was provided. The plant aging promoter included an IDL6 mature polypeptide and a 2-(N-morpholino)ethanesulfonic acid solution, wherein the concentration of IDL6 in the IDL6 mature polypeptide was 10 μmol/L, and the concentration of 2-(N-morpholino)ethanesulfonic acid in the 2-(N-morpholino)ethanesulfonic acid solution was 2.8 mmol/L. The 2-(N-morpholino)ethanesulfonic acid solution was a solution obtained by dissolving 2-(N-morpholino)ethanesulfonic acid monohydrate in an MS liquid culture medium, and the pH of the 2-(N-morpholino)ethanesulfonic acid solution was 5.8.

[0035] The preparation method included the following steps. (1) IDL6 mature polypeptide power was weighed and dissolved in water to prepare an IDL6 mature polypeptide mother solution, wherein the concentration of the IDL6 mature polypeptide in the mother solution was 10 mmol/L.

[0036] (2) A certain amount of 2-(N-morpholino)ethanesulfonic acid monohydrate was weighed and added to an MS liquid culture medium to prepare a 2-(N-morpholino)ethanesulfonic acid solution, wherein the concentration of the 2-(N-morpholino)ethanesulfonic acid was 2.8 mmol/L. The pH value of the solution was adjusted to be 5.8, fully mixed and dissolved evenly to obtain the 2-(N-morpholino)ethanesulfonic acid solution.

[0037] (3) 40 μL of the IDL6 mature polypeptide mother liquid obtained in step (1) was added to 40 mL of the 2-(N-morpholino)ethanesulfonic acid solution prepared in step (2) and stirred evenly by a glass bar to obtain 40 mL of the IDL6 mature polypeptide plant aging promoter.

Embodiment 2

[0038] An IDL6 mature polypeptide plant aging promoter was provided. The plant aging promoter included an IDL6 mature polypeptide and a 2-(N-morpholino)ethanesulfonic acid solution, wherein the concentration of IDL6 in the IDL6 mature polypeptide was 12 μmol/L, and the concentration of 2-(N-morpholino)ethanesulfonic acid in the 2-(N-morpholino)ethanesulfonic acid solution was 2.9 mmol/L. The 2-(N-morpholino)ethanesulfonic acid solution was a solution obtained by dissolving 2-(N-morpholino)ethanesulfonic acid monohydrate in an MS liquid culture medium, and the pH of the 2-(N-morpholino)ethanesulfonic acid solution was 5.9.

[0039] The preparation method included the following steps.

[0040] (1) IDL6 mature polypeptide power was weighed and dissolved in water to obtain an IDL6 mature polypeptide mother solution, wherein the concentration of the IDL6 mature polypeptide in the mother solution was 10 mmol/L.

[0041] (2) A certain amount of 2-(N-morpholino)ethanesulfonic acid monohydrate was weighed and added to an MS liquid culture medium to prepare a 2-(N-morpholino)ethanesulfonic acid solution, wherein the concentration of the 2-(N-morpholino)ethanesulfonic acid was 2.5 mmol/L. The pH value of the solution was adjusted to be 5.9, fully mixed and dissolved evenly to obtain the 2-(N-morpholino)ethanesulfonic acid solution.

[0042] (3) 120 μL of the IDL6 mature polypeptide mother liquid obtained in step (1) was added to 100 mL of the 2-(N-morpholino)ethanesulfonic acid solution prepared in step (2) and stirred evenly by a glass bar to obtain 100 mL of the IDL6 mature polypeptide plant aging promoter.

Embodiment 3

[0043] An IDL6 mature polypeptide plant aging promoter was provided. The plant aging promoter included an IDL6 mature polypeptide and a 2-(N-morpholino)ethanesulfonic acid solution, wherein the concentration of IDL6 in the IDL6 mature polypeptide was 13 μmol/L, and the concentration of 2-(N-morpholino)ethanesulfonic acid in the 2-(N-morpholino)ethanesulfonic acid solution was 3 mmol/L. The 2-(N-morpholino)ethanesulfonic acid solution was a solution obtained by dissolving 2-(N-morpholino)ethanesulfonic acid monohydrate in an MS liquid culture medium, and the pH of the 2-(N-morpholino)ethanesulfonic acid solution was 5.8.

[0044] The preparation method included the following steps.

[0045] (1) IDL6 mature polypeptide power was weighed and dissolved in water to obtain an IDL6 mature polypeptide mother solution, wherein the concentration of the IDL6 mature polypeptide in the mother solution was 10 mmol/L.

[0046] (2) A certain amount of 2-(N-morpholino)ethanesulfonic acid monohydrate was is weighed and added to an MS liquid culture medium to prepare a 2-(N-morpholino)ethanesulfonic acid solution, wherein the concentration of the 2-(N-morpholino)ethanesulfonic acid was 3 mmol/L. The pH value of the solution was adjusted to be 5.8, fully mixed and dissolved evenly to obtain the 2-(N-morpholino)ethanesulfonic acid solution.

[0047] (3) 78 μL of the IDL6 mature polypeptide mother liquid obtained in step (1) was added to 60 mL of the 2-(N-morpholino)ethanesulfonic acid solution prepared in step (2) and stirred evenly by a glass bar to obtain 60 mL of the IDL6 mature polypeptide plant aging promoter.

Performance Test

Laboratory Test

[0048] By taking Embodiment 1 as an example, Arabidopsis thaliana in-vitro leaves and tobacco in-vivo leaves were treated with the IDL6 mature polypeptide plant aging promoter prepared in Embodiment 1, respectively.

[0049] The Arabidopsis thaliana leaves were treated in the following way.

[0050] Control group 1: Arabidopsis thaliana grew under continuous illumination for about 30 days. In-vitro leaves with the same growth potential at the same leaf site (the sixth leaf site) were selected by a pair of forceps and placed flatly on a culture dish. 2.8 mmol/L 2-(N-morpholino)ethanesulfonic acid solution (MES) was directly spayed on the surfaces of the Arabidopsis thaliana in-vitro leaves, and the leaves were kept under continuous illumination for 4 days to observe the phenotype.

[0051] Test group 1: Arabidopsis thaliana grew under continuous illumination for about 30 days. In-vitro leaves with the same growth potential at the same leaf site (the sixth leaf site) were selected by a pair of forceps and placed flatly on a culture dish. The IDL6 mature polypeptide plant aging promoter at 10 μmol/L was directly spayed on the surfaces of the Arabidopsis thaliana in-vitro leaves, and the leaves were kept under continuous illumination for 4 days to observe the phenotype.

[0052] It could be known from the results that, as shown in FIG. 1, the IDL6 mature polypeptide plant aging promoter could promote the aging of Arabidopsis thaliana is leaves. Four days later, the Arabidopsis thaliana in-vitro leaves sprayed with the IDL6 mature polypeptide plant aging promoter in the test group 1 showed obvious premature senescence phenotype. Meanwhile, the chlorophyll contents of the leaves in the control group 1 and the test group 1 were measured by ethanol. As shown in FIG. 2, compared with the control group 1, the chlorophyll content of the leaves treated with the IDL6 mature polypeptide plant aging promoter in the test group 1 decreased after the leaves were kept under continuous illumination for 3 days. The chlorophyll content in the control group 1 decreased to 0.503 μg/mg, while the chlorophyll content in the test group 1 decreased to 0.276 μg/mg.

Field Test

[0053] Tobacco K326 cultivated for about 60 days in the field was used as raw material. The tobacco was sprayed with the IDL6 mature polypeptide plant aging promoter at 10 μmol/L and used as a test group 2, and the tobacco was sprayed with a solution not containing the IDL6 mature polypeptide and used as a control group 2. As shown in FIG. 3, fourteen days later, it could be observed that the tobacco sprayed with the IDL6 mature polypeptide plant aging promoter in the test group 2 showed premature senescence phenotype. Meanwhile, the chlorophyll contents of tobacco leaves in the test group 2 sprayed with the IDL6 mature polypeptide plant aging promoter and in the control group 2 after fourteen days were measured by ethanol. As shown in FIG. 4, the results showed that, compared with the control group 2 (0.598 μg/mg), the chlorophyll content (0.372 μg/mg) of tobacco leaves treated with the IDL6 mature polypeptide plant aging promoter in the test group 2 was reduced obviously. The results showed that the IDL6 mature polypeptide plant aging promoter really had an obvious effect of promoting the aging of in-vivo plant leaves.