KR101128911B1 - Screening method for salinity tolerance in Panax ginseng C.A. Mayer - Google Patents

Abstract

The present invention relates to a method for assaying the resistance of ginseng to salts and a method for selecting salt resistant ginseng. The present invention is based on the fact that the reaction of the ginseng to the salts are different for each individual, by setting the test criteria and test indicators for selecting individuals resistant to salts, the criteria for selecting individuals with resistance to salts and Provide indicators to identify selected individuals.

Ginseng, Salt Resistance, Selection, Criteria, Criteria

Description

Screening method for salinity tolerance in Panax ginseng C.A. Mayer}

The present invention relates to a method for assaying the resistance of ginseng to salts and a method for selecting salt tolerant ginseng. Specifically, after treating ginseng by salt concentration, the chloroplast activity is analyzed according to the elapsed time to find an appropriate salt concentration. Based on this test, the protein expressed at that time was developed as a test index.

Salts limit the growth and production of plants, which is attributed to their effects on water and nutrient absorption (Munns & Tester 2008, Annu Rev Plant Biol 59: 651-681). Salt resistance studies are based on the analysis of related proteins or secondary metabolites (Kong-Negren et al. 2005, Sci Asia 31: 403-408) and the genetic map of salt resistance in the cellular machinery in which the gene is expressed. Enemy characteristics (Quesada et al. 2002, Plant Physiol 130: 951-963; Shinozaki & Yamaguchi Shinozaki 2007, J Exp Bot 58: 221-227). Recently, new techniques (microarrays and proteomics) have been used to investigate the cause more specifically. In the area of ginseng, the damage area of salt obstacles is increasing year by year (20-25% of total cultivation area, Ginseng special crops survey), and it is urgent to cultivate varieties resistant to salt. Development of test indicators should be a priority.

As a method for selecting ginseng that is resistant to salts, it is intended to develop assay criteria for selecting saline-resistant ginseng and indicators for identifying selected individuals.

In order to solve the above problems, the present invention provides a method for assaying the resistance of ginseng to salts.

The present invention also provides a method for selecting salt resistant ginseng.

It is possible to cultivate salt-resistant ginseng varieties by the method of assaying the resistance of ginseng to salts and selection methods of salt-resistant ginseng according to the present invention, thereby preventing the degradation of ginseng due to physiological disorders. In addition, it is possible to improve the first-class ratio of fresh ginseng yields from less than 5% to 10% by verifying the stability of selected individuals by the salt adaptation test criteria and the indicators.

In order to achieve the object of the present invention,

a) treating ginseng at a salt concentration of 4-6 ds / m for 13-17 days; And

b) providing a method for assaying the resistance of ginseng to salt comprising visually assessing susceptible and resistant individuals.

The old ginseng, geumpung, whirlwind, members, Yunpoong, Chunpoong, ground, vigilante, Jilin, three jusam, US Makita, hwagisam (Panax quinquefolius ), Panax ginseng ( Panax notoginseng ) or Panax ginseng japonicus ), but is not limited thereto.

In the step of treating the ginseng to salt, preferably ginseng may be treated for 15 days at a salt concentration of 5 ds / m.

Specifically, the salt composition may be composed of KNO ₃ , KH ₂ PO ₄ and MgSO _{4 ~} 7H ₂ O. Preferably, the concentration of the salt is KNO ₃ 2,000 ~ 2,200 mg / L, KH ₂ PO ₄ 800 ~ 900 ㎎ / ℓ and _{MgSO 4? 7H 2 O 1,400 ~} 1,700 ㎎ / ℓ may be, more preferably, the concentration of the salt is _{KNO 3 2,186.3 ㎎ / ℓ, KH} 2 PO 4 843.1 ㎎ / ℓ and MgSO _4? 7H may be ₂ O 1,529.5 ㎎ / ℓ.

Specifically, the visual evaluation that distinguishes the susceptible individual from the resistant individual may be based on the color of the plant, and more specifically, the susceptible individual may visually identify a red color due to salt damage.

The present invention also provides

a) treating ginseng at a salt concentration of 4-6 ds / m for 13-17 days;

b) visually assessing susceptible and resistant individuals to select resistant individuals; And

c) providing a method for selecting a salt-resistant ginseng comprising the step of identifying the selected resistant individuals by assay indicators.

The old ginseng, geumpung, whirlwind, members, Yunpoong, Chunpoong, ground, vigilante, Jilin, three jusam, US Makita, hwagisam (Panax quinquefolius ), Panax ginseng ( Panax notoginseng ) or Panax ginseng japonicus ), but is not limited thereto.

In the step of treating the ginseng to salt, preferably ginseng may be treated for 15 days at a salt concentration of 5 ds / m.

Specifically, the salt composition may be composed of KNO ₃ , KH ₂ PO ₄ and MgSO _{4 ~} 7H ₂ O. Preferably, the concentration of the salt is KNO ₃ 2,000 ~ 2,200 mg / L, KH ₂ PO ₄ 800 ~ 900 ㎎ / ℓ and _{MgSO 4? 7H 2 O 1,400 ~} 1,700 ㎎ / ℓ may be, more preferably, the concentration of the salt is _{KNO 3 2,186.3 ㎎ / ℓ, KH} 2 PO 4 843.1 ㎎ / ℓ and MgSO _4? 7H may be ₂ O 1,529.5 ㎎ / ℓ.

Specifically, the visual evaluation that distinguishes the susceptible individual from the resistant individual may be based on the color of the plant, and more specifically, the susceptible individual may visually identify a red color due to salt damage.

Identifying the selected resistant individuals by assay indicators, specifically, beta-1,3-glucanase (β - 1,3-glucanase) or ribonuclease-like storage protein (Ribonuclease-like storage protein) It may be due to the expression level of, preferably, high expression of beta - 1,3-glucanase (β - 1,3-glucanase) or ribonuclease-like storage protein (Ribonuclease-like storage protein) Can be.

"Salt resistance" according to the present invention means the ability of any plant to grow under osmotic stress or stress caused by salts or ions in water and soil. For example, when moisturized with a liquid containing a mixture of water and ions that would be disadvantageous to the growth of other plants of the same species, or when cultivated with such a medium, increased compared to the same species and / or mutant plants. Plants with growth rates become salt resistant.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  1. Establishing Ginseng Salt Resistance Proper Test Criteria

In order to establish the ginseng salt resistance assay criteria, the chloroplast activity of each salt concentration after treatment was examined (Table 1). Chloroplast activity can be quantified as shown in Equation 1 as a result of chlorophyll fluorescence. That is, Rfd is a value obtained by subtracting the value of the second fluorescence emission threshold by the value of the second emission fluorescence duration threshold value minus the value of the second fluorescence emission threshold.

Chlorophyll Fluorescence The higher the Rfd value, the less the chloroplast performs the photosynthetic function and the less the damage caused by salts. The lower the Rfd value, the higher the chlorophyll can perform the photosynthetic function.

At 1 and 3 ds / m salt concentrations, the chloroplast activity did not change significantly after treatment, but at 5 ds / m salt concentrations began to decrease significantly after 5 days and gradually decreased until 15 days ( 2, 3). However, salt concentration of 7 ds / m necrosis 5 days after treatment. As a result, it was found that salt resistance evaluation was appropriate for 15 days treatment at 5 ds / m of the assay standard, and salt composition (mg / L) was KNO ₃ 2,186.3, KH ₂ PO ₄ 843.1, MgSO 4 -7H 2 O 1,529.5 (Table 2).

Table 1. Chloroplast activity according to elapsed time after treatment by salt concentration (chloroplast fluorescence Rfd value)

Salt concentration (ds / m) /
Days Since Processing  Chloroplast activity (chloroplast fluorescence Rfd value) One 5 10 15 One 1.013 1.109 1.018 1.018 3 1.098 1.102 0.912 0.842 5 0.845 0.561 0.433 0.367 7 0.217 Necrosis - -

Equation 1: Salt Damage (Rfd) = (Fp-Fs) / Fs

In the above formula, Fp represents the secondary fluorescence emission peak and Fs represents the secondary fluorescence emission sustained threshold.

Table 2. Content of Inorganic Components Used in Salt Composition by Salt Concentration

Salt concentration (ds / m) Ingredient Content (mg / l) KNO ₃ KH ₂ PO ₄ MgSO4? 7H2O One 437 168.6 305.9 3 1,311 505.9 917.7 5 2,186.3 843.1 1,529.5 7 3,060.9 1,180.4 2,141.3

Example  2. Selection of susceptible and resistant individuals using ginseng salt resistance test criteria

Salt Resistance Test Criteria (Salt Concentration 5ds / m, Salt Composition (mg / l) KNO ₃ 2,186.3, KH ₂ PO ₄ 843.1, MgSO4–7H2O 1,529.5, 15 days of treatment) were able to select susceptible and resistant individuals, and the susceptible individuals showed salt damage (red areas) after treatment, but resistant individuals did not show salt damage ( 4).

Example  3. Establishment of Test Indicators from Resistant Subjects by Salt Resistance Test Criteria

Twenty-two upper and lower proteins responding to salt stress were found in sensitive and resistant individuals selected by the salt resistance assay criteria and classified as shown in FIG. 5. Among these proteins, β- 1,3-glucanase and Ribonuclease-like storage proteins in resistant individuals were strongly expressed at the mRNA level, which was related to salt resistance and was set as an indicator.

1 shows the fluorescence response of chloroplasts using the Kautsky Kinetics parameter. Fo represents the primary fluorescence emission start point and Fm represents the primary fluorescence emission peak. Fp represents the secondary fluorescence peak and Fs represents the secondary fluorescence sustained threshold.

Figure 2 shows the chloroplast activity according to the post-treatment period in salt concentration 5 ds / m conditions. It was shown that the treatment condition for 15 days at the salt concentration of 5 ds / m is suitable for the resistance test criteria.

Figure 3 shows the leaf damage symptoms according to the elapsed time after treatment in salt concentration 5 ds / m conditions.

4 shows susceptible and resistant individuals selection by salt resistance assay criteria (5 ds / m). Sensitive subjects: Chlorophyll fluorescence Rfd value 0.5 or less, Resistant subjects: Chlorophyll fluorescence Rfd value 1.0 or higher

Figure 5 shows the difference in the indicators of susceptibility and resistant individuals at salt concentration 5 ds / m. The figure below shows the β- 1,3-glucanase and Ribonuclease-like storage proteins, assay markers expressed from salt-resistant individuals at salt concentrations of 5 ds / m.

Claims (14)

a) treating ginseng at a salt concentration of 4-6 ds / m for 13-17 days; And b) A method for assaying the resistance of ginseng to salt comprising visually assessing susceptible and resistant individuals. The method of claim 1, wherein step a) is performed for 15 days at a salt concentration of 5 ds / m. The method of claim 1, wherein the salt composition comprises KNO ₃ , KH ₂ PO _4, and MgSO ₄ ˜7H ₂ O. The method of claim 3, wherein the salt according to claim wherein the salt composition is _{KNO 3 2,000 ~ 2,200 ㎎ / ℓ} , KH 2 PO 4 800 ~ 900 ㎎ / ℓ and MgSO _4? 7H ₂ O 1,400 ~ that consists of 1,700 ㎎ / ℓ Ginseng tolerance test method. The method of claim 1, wherein the visual evaluation is based on the color of the plant. The ginseng of the salt according to claim 1, wherein the ginseng is an old wind, a golden wind, a whirlwind, a seongwon, a breeze, a thunderstorm, a plaque, a vigilant, a jilin, a recommendation I, a recommendation II, a mito or a panax quinquefolius . Tolerance Assay Method. a) treating ginseng at a salt concentration of 4-6 ds / m for 13-17 days; b) visually assessing susceptible and resistant individuals to select resistant individuals; And c) A method for selecting salt-resistant ginseng, comprising the step of identifying the selected resistant individuals by assay indicators. 8. The method of claim 7, wherein step a) is performed at a salt concentration of 5 ds / m for 15 days. The method of claim 7, wherein the salt composition is selected from salt resistant ginseng, characterized in that consisting of KNO ₃ , KH ₂ PO ₄ and MgSO _{4 ~} 7H ₂ O. The method of claim 9 wherein the salt composition is _{KNO 3 2,000 ~ 2,200 ㎎ / ℓ} , KH 2 PO 4 800 ~ 900 ㎎ / ℓ and MgSO _4? 7H salt tolerance according to claim ₂ O 1,400 ~ that consists of 1,700 ㎎ / ℓ Selection method of ginseng. 8. The method of claim 7, wherein the visual evaluation is based on the color of the plant. The method of claim 7, wherein the assay indicator is characterized in that the expression level of beta - 1,3-glucanase (β - 1,3-glucanase) or ribonuclease-like storage protein (Ribonuclease-like storage protein) Selection method of salt resistant ginseng to be. According to claim 7, wherein the ginseng is the old wind, gold wind, whirlwind, seongwon, lotus, hurricane, plaque, vigilante, Jilin, recommendation I, recommendation II, mima or ginseng ( Panax quinquefolius ) Selection method. delete

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