KR20170067440A

KR20170067440A - Plant growth accelerator and manufacturing methods therof

Info

Publication number: KR20170067440A
Application number: KR1020150174122A
Authority: KR
Inventors: 장윤석; 윤학원; 김재환; 황인환; 오영준
Original assignee: 포항공과대학교 산학협력단
Priority date: 2015-12-08
Filing date: 2015-12-08
Publication date: 2017-06-16

Abstract

The present invention relates to a method for increasing the carbon assimilation of a plant and thereby increasing biomass, and more particularly, to a method for increasing the carbon assimilation of a plant, thereby increasing the biomass And a method for producing the same.
The growth promoter according to the present invention comprises nanospheres. When the plants are exposed to nano-zero iron, the pores of the leaf become larger and the carbon assimilation becomes active, thus increasing the biomass.

Description

TECHNICAL FIELD The present invention relates to a plant growth promoter and a method for producing the plant growth promoter,

The present invention relates to a method for increasing the carbon assimilation of a plant and thereby increasing biomass, and more particularly, to a method for increasing the carbon assimilation of a plant, thereby increasing the biomass And a method for producing the same.

Since accelerating the increase in atmospheric carbon dioxide concentration has been identified as the main cause of climate change, there has been an active movement around the world to counteract climate change by reducing carbon dioxide. Carbon from the land occupies the largest portion of the global carbon flux, and land vegetation is the only organism that can remove carbon dioxide, except for marine plankton.

In particular, carbon dioxide abatement technology using plants not only has the advantage of reducing GHGs but also can be used as an energy source after conversion to biomass. The application is very limited because there is no data.

Most of the carbon dioxide removal studies using plants focus on genetically modified plants that can control the opening and closing of pores. This is because the only organ that undergoes gas exchange in plants is the pores surrounded by the guard cells.

There are various factors that control the opening and closing of pores. Among them, there is a correlation between the activity of the plasma membrane (PM) H ⁺ -ATPase called the proton pump and the pore opening phenomenon. It is a well-known fact that as the activity of PM H ⁺ -ATPAse in leaves increases, the protector cells become swollen and the pore becomes larger. Recently, it has been revealed that AHA2 (ARABIDOPSIS H + -ATPASE 2) among the isoforms of H ⁺ -ATPase is the main gene responsible for the opening and closing of pores of homologous proteins.

A problem to be solved by the present invention is to provide a new fertilizer capable of promoting plant growth and a method for producing the same.

Another problem to be solved by the present invention is to provide a new fertilizer capable of promoting the carbon monoxide function by controlling the opening and closing of the pore, thereby increasing the biomass, and a method for producing the same.

Another object of the present invention is to provide a method for promoting plant growth by using a fertilizer containing a substance capable of promoting carbon monoxide function by controlling the opening and closing of pores.

In order to solve the above-mentioned problems, the plant growth promoting agent according to the present invention is characterized by containing nano-zirconium iron (nZVI) as an active ingredient.

In the present invention, the term " growth " means an increase in the carbon percentage of the plant and / or the absolute amount of carbon (dry sample weight x carbon ratio).

In the present invention, the term " promoting " is intended to mean a plant growth promoter that is at least 10%, preferably at least 20%, more preferably at least 30%, and most preferably at least 50% Which means that growth is increased.

In the present invention, the term 'nano' is understood to mean a diameter of 1 to 1000 nm, preferably 5 to 500 nm, and more preferably 10 to 100 nm.

In the present invention, the 'soil' is understood to mean an environmental matrix supporting the roots of plants, and means soil and / or water containing soil.

In the present invention, the growth promoter enhances the H ⁺ -ATPase activity of a plasma membrane of a plant and promotes plant growth by increasing the expression of AHA2 gene among isoforms of H ⁺ -ATPase that regulates pore opening .

The nano-zirconium iron according to the present invention is nano-sized iron particles and is a conventional nanosized zero valent iron (hereinafter referred to as nZVI) which can be used as a donor.

nZVI can be used not only for commercial nZVI used for environmental purification but also for core-shell nano-iron including α-Fe and iron oxide on the surface. In addition, the nano-zirconium iron may have nanoparticles such as nickel, copper, and palladium on the surface thereof.

In the present invention, the nano-zirconium iron may be used in admixture with the soil, and the concentration in the soil is preferably 0.01 to 5.0 g / kg of soil, preferably 0.1 to 0.5 g / kg of soil.

In the present invention, the plant may include all plants as long as growth is promoted by nanospheres. More precisely, it can include plants whose pores are enlarged by the nano-zirconium iron contained in the soil, and in particular, plants that overexpress AHA2 and / or increase the activity of H + -ATPase in plants.

The plants are non-graminaceous plants such as rice, cucumber, tomato, and Arabidopsis, preferably herbaceous rice plants.

In one aspect, the present invention provides a method for promoting the growth of plants by adding nanosphereferrous iron to the rhizosphere of the plant, preferably to the soil where the plant is planted.

In one aspect, the present invention provides a method of overexpressing plant AHA2 by adding nano-zirconium iron to the rhizosphere of the plant, preferably to the planted vegetation.

In one aspect, the present invention provides a method for increasing the H ⁺ -ATPase activity of a plasma membrane in a plant by adding nanospheregulated iron to the rhizosphere of the plant, preferably to the planted vegetation.

In one aspect, the present invention provides a method of lowering the iron availability of plants by adding nano-zirconium iron to the rhizosphere of the plant, preferably to the soil where the plant is vegetated.

In one aspect, the present invention provides a method for increasing biomass by lowering the iron availability of plants by placing nanoparticles that emit electrons in the rhizosphere of the soil in which the plants are vegetated.

According to the present invention, it is possible to easily increase the carbon assimilation effect of plants, for example, Arabidopsis, by introducing nanospheres to an appropriate concentration in the soil, thereby increasing the biomass of the plant. Experiments have shown that biomass and leaf size increase about 1.5 and 1.6 times, respectively, compared to the control group grown in nZVI sprinkled soil.

FIG. 1 is a graph comparing the pore size and the size of the control group and the experimental group according to the present invention, the pore photographs of the control group (left) and the experimental group (right) (comparison group (growth in nZVI agitated soil)
Fig. 2 is a graph showing the H < + > -ATPase activity of the plant top (leaf) according to the present invention and the expression amount of AHA2 gene measured by qRT-
Figure 3 shows the phenotype and biomass of the control and experimental groups according to the invention
Figure 4 shows the leaf phenotype and size of the control and experimental groups according to the invention
Figure 5 shows the carbon ratio of the control and experimental groups
Figure 6 shows the ratio of iron content in the dry samples of the control and experimental groups

Hereinafter, the present invention will be described in more detail with reference to examples. It is to be understood by those skilled in the art that these embodiments are for further illustrating the present invention and that the scope of the present invention is not limited to these embodiments.

Example 1

Preparation of experimental material

RNIP (Toda, Japan), which is used for commercial use, was used for nZVI, soil for horticultural use was used, and tap water was used for plant culture.

Example 2

nZVI stirred soil preparation

0.1 g of nZVI was washed with ethanol and degased / deionized water, and then stirred with 85 g of dry soil and 115 ml of tap water. Arabidopsis thaliana seedlings were planted in the above soil and grown in a plant growth chamber under the following growth conditions for 3 weeks.

2-1. Growing conditions

Day / Night: 16 / 8h

Temperature: 23 ~ 24 ℃

Humidity: 30 to 40%

Example 3

Comparison of pore size, plasma membrane H + -ATPase activity and AHA2 gene expression

3-1. Pore size

The epidermal cells on the back of the plant leaf cultivated for 3 weeks were peeled off and their sizes were measured using a fluorescence microscope (Zeiss Axioplan). It can be seen from Fig. 1 that the pores of the Arabidopsis grown in the soil where the nZVI is stirred are larger than the control.

3-2. H ⁺ - ATPase Activity and AHA2 gene Expression level compare

The protoplast was prepared using plant leaves grown for 3 weeks and then the pumping out activity of H ⁺ -ATPase was measured (FIG. 2a). The expression level of AHA2 was also compared using qRT-PCT (Qunatitative real-time PCR) and RNeasy plant mini kit (Qiagen, USA) (Fig. 2B).

Example 4

Plant analysis

4-1. Biomass

Fresh weight of plants grown for 3 weeks was measured. Weights were measured on shoots except for plant roots. 10 modules per group The average weight was measured and repeated three times. As a result, as shown in Fig. 3, it can be seen that the weight of the plants grown in nZVI increased about 1.5 times as much as the weight of the control group.

4-2. Leaf size

The size of the leaves was measured using Image J program and the number of leaves used was 50. Figure 4 shows the phenotype and measurement values of the leaves.

4 -3. Increase carbon assimilation

Carbon assimilation was analyzed using a stable isotope consumption mass spectrometer (IsoPrime-EA, UK), and it was confirmed that the carbon isotope ratio (δ) of the Arabidopsis grown in the nZVI stirred soil was further reduced. This is consistent with the fact that higher photosynthetic activity results in more lighter 12C than 13C and lower δ values. The carbon content of the dried samples was measured using an elemental analyzer (Flash EA 1112, USA) and compared. Referring to FIG. 5, it can be seen that the carbon ratio is increased as compared with the control group, which means that the carbon assimilation is increased.

4-4. Iron element analysis in plants

Fig. 6 shows the analysis results of the iron component of the dried sample. Plants that have been grown for 3 weeks are washed in flowing water except for roots and dried in an oven at 80 ° C for more than 10 hours. After finely crushing the sample using a grinder, add 2 ml of nitric acid (60%) and dissolve completely at 105 ° C using a heat block. The iron content was measured using ICP-AES (Thermo, USA). The results showed that the iron concentration in the plants grown in the nZVI agar soil was low as shown in Fig. This confirms that nZVI lowers the iron availability of plants.

Although it is not theoretically possible, the main role of H + -ATPase in plants is to emit hydrogen ions (H +) from the roots to acidify the rhizosphere and help in the absorption of iron, which is invisible. When nZVI is in the rhizosphere, which is not readily accessible by plants, electrons from nZVI break down water to produce OH-, which lowers the availability of iron ions in the soil. As a result, the activity of the plasma membrane H + -ATPase is increased in plants to overcome the lowered iron availability due to nZVI.

In the present invention, as shown in FIG. 2, the increase in the activity of the plasma membrane H + -ATPase appears not only in the roots but also in the upper part, more precisely in the leaf part, and thus the activity of H + -ATPase AH2, which is a related homologue gene, was also overexpressed (Santi et al., New Phytol . 2009, 183, 1072). In a recent academic study AHA2 gene it was proven to be an important factor leading to development of the open pores of the plasma membrane H + -ATPase (Wang et al. , Proc. Natl. Acad. Sci. USA 2014, 111, 533). This was confirmed in FIG. 1 that the experimental group had larger pores than the control group.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims

A plant growth promoter characterized by containing nano-spirit ferrite (nZVI) as an active ingredient.

The plant growth promoter according to claim 1, wherein the nano-zirconium iron is in the form of a core-shell.

The plant growth promoter according to claim 1, wherein the nano-zirconium iron has a concentration in the soil of 0.01 to 5.0 g / kg of soil.

The plant growth promoter according to claim 1, wherein the growth promoter increases H ⁺ -ATPase activity of the plasma membrane of the plant.

2. The plant growth promoter according to claim 1, wherein the growth promoter enhances the expression of AHA2 gene.

The plant growth promoter according to claim 1, wherein the plant is non-graminaceous plants.

The plant growth promoter according to claim 6, wherein the plant is a Arabidopsis thaliana.

The plant growth promoter according to claim 1, wherein the plant growth promoter is located in the rhizosphere.

Wherein plant growth is promoted by adding nano-valent iron as an effective ingredient to a plant-grown soil.

10. The method of claim 9, wherein the nano-zirconium iron has a concentration in the soil of 0.01 to 5.0 g / kg of soil.

10. The method of claim 9, wherein the nano-zirconium iron is introduced near the root.

10. The method of claim 9, wherein the plant is non-graminaceous plants.

10. The method of claim 9, wherein the concentration of iron ions in the plant is lowered.

10. The method of claim 9, wherein said plant overexpresses AHA2.

10. The method of claim 9, wherein the H ⁺ -ATPase activity of the plasma membrane is increased in the plant.

16. The method according to claim 14 or 15, wherein the plant is over-expressed in the aerial part and the H ⁺ -ATPase activity of the plasma membrane is increased.

Characterized in that the nanoparticles which emit electrons are placed in the rhizosphere of the soil in which the plant is vegetated, thereby lowering the iron availability of the plant and increasing the biomass.