Oligogalacturonic acid regulation and control method for improving aluminum resistance of peas

Abstract

The invention discloses an oligogalacturonic acid regulation and control method for improving aluminum resistance of peas. The method comprises the following steps: soaking pea seeds in 7-8% sodium hypochlorite; washing with deionized water; the method comprises the following steps: soaking pea seeds in a 2mM CaCl2 solution, then laying the soaked pea seeds on a mesh screen of an air culture mist culture system, and growing the pea seeds at 20-25 DEG C for 48 hours; selecting seedlings with the root length of 2-3cm, transferring the seedlings into a 1 / 4 Hoagland solution, and culturing the seedlings in a day-night alternate culture mode; pre-adapting the seedlings in an environment in which the pH value of the seedlings is 4-5, and then treating the seedlings with an AlCl3 solution; the method comprises the following steps: respectively putting pea seedlings into sterile water, selecting pea seedlings with consistent growth vigor after water culture, and setting oligogalacturonic acid OGAs with the concentration of at least 50mg / L; the oligogalacturonic acid provided by the invention can be applied to enhancing the aluminum tolerance of peas, and can improve the aluminum tolerance of agricultural crops so as to promote agricultural income increase.

Description

Technical Field

[0001] The invention relates to the field of pea planting, and in particular to an oligogalacturonic acid regulation method for improving aluminum tolerance of pea. Background Art

[0002] Pea is one of the oldest domesticated species. It is widely cultivated because of its strong tolerance to abiotic stresses such as low temperature, drought, and salinity. It is the second largest cereal legume in the world after common beans. Pea is widely cultivated because of its strong cold resistance, short growth period, and strong adaptability. It is a multi-purpose crop that combines food, vegetables, feed, and fertilizer. Aluminum (Al) is the most abundant metal element in the earth's crust. It dissolves in soil water in the form of free ions under acidic conditions. 3+ 、Al(OH) 2+ The presence of aluminum in the form of plasma hinders the growth and development of plant roots. Acidic soils account for about 50% of the world's arable land, and aluminum toxicity is a serious agricultural problem. Therefore, regulating plant aluminum tolerance will help improve aluminum tolerance in pea crops and increase the food production potential of acidic soils. Summary of the invention

[0003] The purpose of the present invention is to propose the application of exogenous oligogalacturonic acid in enhancing the aluminum tolerance of pea.

[0004] The invention also provides a method for regulating oligogalacturonic acid for improving aluminum tolerance of pea.

[0005] To achieve this object, the present invention adopts the following technical solutions:

[0006] Application of exogenous oligogalacturonic acid in enhancing aluminum tolerance in pea.

[0007] A method for regulating oligogalacturonic acid for improving aluminum tolerance of peas, comprising the following steps:

[0008] (1) Soak pea seeds in 7-8% sodium hypochlorite; rinse with deionized water;

[0009] (2) pea seeds were soaked in a 2 mM CaCl2 solution and then spread on a mesh screen of an aeroponic mist culture system. The pea seeds were grown at 20-25°C for 48 h.

[0010] (3) Select seedlings with a root length of 2 to 3 cm, transfer them to 1 / 4 Hoagland solution for 4 days, and culture them in a growth chamber with alternating day and night culture at 26°C for 16 h and 24°C for 8 h;

[0011] (4) Pre-adapting the seedlings to an environment with a pH of 4 to 5 and then treating them with AlCl3 solution;

[0012] (5) Pea seedlings with a root length of 2 to 3 cm were placed in sterile water respectively, and pea seedlings with uniform growth were selected after hydroponics, and the concentration of oligogalacturonic acid (OGAs) was set to at least 50 mg / L.

[0013] Preferably, in step (5), the hydroponic conditions are: temperature 25±1°C, air humidity 75-85%, and light conditions of 16 hours of light, 14000-20000Lx of light intensity, and 8 hours of darkness.

[0014] Preferably, in step (5), the concentration of oligogalacturonic acid OGAs is 50-100 mg / L.

[0015] Compared with the prior art, one of the above technical solutions has the following beneficial effects:

[0016] This study analyzed the biomass, endogenous antioxidant enzyme activity, and gene expression of peas, indicating that exogenous addition of oligogalacturonic acid (OGAs) can enhance aluminum tolerance in peas and act as a signal substance to participate in the regulation of aluminum tolerance in peas. Thus, the oligogalacturonic acid in this study can be used to enhance aluminum tolerance in peas, improve aluminum tolerance in agricultural crops, and promote agricultural income. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This paper compares and analyzes the effects of different concentrations of OGAS on pea plant height (A), root length (B), root activity (C), aboveground fresh weight (D), and root fresh weight (E).

[0018] Figure 2 Oligogalacturonic acid (OGAs) are involved in the response of pea to Al toxicity. Root scanning analysis of pea seedlings (A, B), Al content determination by ICP-OES (C), and hematoxylin and morin staining (D, E, F). Bar graphs represent mean ± SD (n = 4).

[0019] Figure 3 Oligogalacturonic acid (OGAs) are involved in the response of pea root tips to aluminum stress. The content of endogenous oligogalacturonic acid (OGAs) was determined by ESI-MS mass spectrometry (A).

[0020] Figure 4 The effects of oligogalacturonic acid OGAs on total reactive oxygen species (A), SOD, CAT and POD enzyme activities (B, C, D) in pea were detected by fluorescence staining: root apex total reactive oxygen species staining and fluorescence intensity (E, I); root apex superoxide anion staining and fluorescence intensity (F, J); root apex hydrogen peroxide staining and fluorescence intensity (G, K); root apex hydroxyl radical staining and fluorescence intensity (H, L).

[0021] Figure 5 This is the expression profile of RBOH, SOD, CAT and POD under oligogalacturonic acid OGAs treatment.

[0022] in:

[0023] Figure 1 , Figure 2 , Figure 3 , Figure 5 Asterisks indicate statistically significant differences according to Duncan's test (ns, not significant; *p<0.05; **p<0.01; ***p<0.001). DETAILED DESCRIPTION

[0024] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0025] For ease of understanding of the present invention, the present invention is described more fully below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive. If no specific technology or conditions are indicated in the embodiments, the technology or conditions described in the documents in this area or the product instructions are carried out. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained by commercial purchase.

[0026] Application of exogenous oligogalacturonic acid in enhancing aluminum tolerance in pea.

[0027] A method for regulating oligogalacturonic acid for improving aluminum tolerance of peas, comprising the following steps:

[0028] (1) Soak pea seeds in 7-8% sodium hypochlorite; rinse with deionized water;

[0029] (2) pea seeds were soaked in a 2 mM CaCl2 solution and then spread on a mesh screen of an aeroponic mist culture system. The pea seeds were grown at 20-25°C for 48 h.

[0030] (3) Select seedlings with a root length of 2 to 3 cm, transfer them to 1 / 4 Hoagland solution for 4 days, and culture them in a growth chamber with alternating day and night culture at 26°C for 16 h and 24°C for 8 h;

[0031] (4) Pre-adapting the seedlings to an environment with a pH of 4 to 5 and then treating them with AlCl3 solution;

[0032] (5) Pea seedlings with a root length of 2 to 3 cm were placed in sterile water respectively, and pea seedlings with uniform growth were selected after hydroponics, and the concentration of oligogalacturonic acid (OGAs) was set to at least 50 mg / L.

[0033] Preferably, in step (5), the hydroponic conditions are: temperature 25±1°C, air humidity 75-85%, and light conditions of 16 hours of light, 14000-20000Lx of light intensity, and 8 hours of darkness.

[0034] Preferably, in step (5), the concentration of oligogalacturonic acid OGAs is 50-100 mg / L.

[0035] Plant material and growth conditions:

[0036] Seeds were soaked in 7.5% sodium hypochlorite for 30 minutes and then rinsed 6 times with deionized water. Seeds were soaked in 2mM CaCl2 solution for 8h and then evenly spread on the mesh screen of the aeroponic mist culture system. The mist was atomized for 60s every 8min, and the plants were grown at 24℃ for 48h. Uniform seedlings with a root length of 2-3cm were selected and transferred to 1 / 4 Hoagland solution (25μM H3BO3) for 4 days in a growth chamber with a 16h (26℃) / 8h (24℃) day and night alternation. After pre-adapting to an environment with a pH of 4.5 (containing 0.5mM CaCl2, 25μM H3BO3) for 3h or 12h, the seedlings were treated with 15μM AlCl3, 30μM AlCl3 (containing 0.5mM CaCl2, 25μM H3BO3, pH 4.5) solution.

[0037] Related tests:

[0038] ① Determine the effects of different OGS concentrations on pea growth and root activity:

[0039] Pea seedlings with a root length of 2-3 cm were placed in polyethylene plastic pots filled with 1L of sterile water. The hydroponic box was placed at a temperature of 25±1°C, an air humidity of 75%-85%, and a light condition of 16h of light, 8h of darkness, and a light intensity of 14000-20000Lx. After three days of hydroponic culture, pea seedlings with consistent growth were selected and treated with hydroponic nutrient solutions with oligogalacturonic acid OGAs concentrations of 0 (CK), 10, 20, 50, 100, and 200 mg / L. The hydroponic nutrient solution was replaced every 3 days, and the growth and phenotypic changes of each pea plant were regularly observed and recorded. Samples were taken for determination after 7 days of treatment. Root activity was determined by triphenyltetrazolium chloride method (TTC method).

[0040] ② Determination of the effect of OGs on aluminum tolerance of pea root tips:

[0041] After seedlings were pretreated with OGs and co-treated with aluminum toxicity, root length was scanned and analyzed using root image analysis software Win-RHIZO Pro (Regent Instruments, QC, Canada). Each replicate had at least 10 plants, and all experiments were repeated 3 times. To quantitatively determine the Al concentration, lateral root tips (0-10 mm) treated with 15 μM AlCl3 for 24 h were collected after rinsing three times in ultrapure water. Al was extracted by soaking in 2 mL of HCl for 48 h, and the Al concentration in the extract was determined by ICP-OES.

[0042] To determine the hematoxylin and Morin staining of pea under different treatments, 5-day-old seedlings were pretreated with OGs for 12 hours and then treated with 0.15 μmol / L AlCl3 solution for 24 hours. The roots were then immersed in deionized water for 5 minutes to remove the aluminum treatment solution remaining on the root tip surface. The roots were then stained in hematoxylin staining solution (containing 0.1% hematoxylin, 0.01% KIO3 and 0.2 mmol / L NaOH) for 15 minutes. After staining, the root tips were again immersed in deionized water for 15 minutes. Finally, the root tips were photographed and observed using a stereo microscope (Olympus SZX16).

[0043] The 0-10 mm section samples after aluminum treatment were stained in 0.01% Morin staining solution for 30 min, rinsed with pure water for 30 s, and then directly observed and photographed under a fluorescence microscope (Olympus BX53);

[0044] ③Analysis of endogenous OGAS content in pea under different treatments:

[0045] Peas with root length of about 3-4 cm were selected and transferred to Hoagland nutrient solution for culture with a pH of 5.5. Different treatments were added after 12 hours. After 24 hours, 50 lateral roots with a root tip of 1 cm were taken from each treatment. The final result was the average of three results. MALDI-TOF-MS was used for detection to determine the endogenous OGAS content in the root tip of peas under different treatments.

[0046] ④Analysis of ROS staining in pea root tips under different treatments:

[0047] Take about 30 aluminum-treated root tips of 0-10 mm in length, rinse with deionized water for 5 minutes, place the root in a 1.5 ml centrifuge tube (be careful not to pinch the root tip), and then take an appropriate amount of 1 mMol / L CM-H2DCFDAY and Dihydrorhodamine 123 (DHR 123), 10 mMol / L Dihydroethidium and 5 mMol / L Hydroxyphenyl Fluorescein dyes that have been fully shaken under light-proof conditions to ensure that the root is completely immersed in the dye. The light-proof dyeing lasts for 30 minutes, and the roots are rinsed three times with deionized water. The root tips are photographed and observed with a stereo microscope (Olympus SZX16). Image J is used to process the fluorescence images and grayscale value analysis is performed. The experiment is repeated 3 times.

[0048] ⑤Analysis of antioxidant enzyme activity and total reactive oxygen species in peas under different treatments:

[0049] Reactive oxygen species (ROS) mainly include hydrogen peroxide (H2O2), singlet oxygen ( 1 O2), superoxide anion (O2 - ) and hydroxyl radicals (OH·). ROS are inevitable byproducts of aerobic metabolism in plant cells. Plants have evolved complex mechanisms, including enzymatic and non-enzymatic mechanisms, to remove excess ROS and keep the intracellular ROS level in a dynamic balance. However, under stress conditions, the production and removal of intracellular ROS are unbalanced, resulting in excessive accumulation of ROS and oxidative damage to cells.

[0050] SOD, CAT, POD and total reactive oxygen species were detected using kits purchased from Suzhou Keming Biotechnology Co., Ltd. and Kanglang Biotechnology Co., Ltd., respectively. For each treatment, about 0.1 g of pea lateral root was weighed, 1 mL of extract was added, and ice bath homogenization was performed. Centrifuge at 8000g for 10 min at 4℃, and the supernatant was taken. The experiment was repeated three times.

[0051] ⑥RNA extraction and qRT-PCR analysis:

[0052] use Total RNA was isolated from pea root tips using the Super Total RNA Extraction Kit (Promega, Shanghai). TM First-strand cDNA was synthesized from 1 μg of total RNA using RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara). Gene expression levels were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR) using TB Premix Ex Taq TM II (Tli RNaseH Plus) (Takara) was detected on a LightCycler 480II machine (Roche Diagnostics, Switzerland). Relative expression was normalized to the expression level of actin. Primers were designed using the NCBI primer design tool. Relative expression was calculated by the double ΔΔCt method. Experiments were performed with three biological and technical replicates.

[0053] result:

[0054] ① Effects of different oligosaccharide concentrations on peas:

[0055] In order to explore the effect of oligogalacturonic acid (OGAs) on the growth of pea, this study measured the plant height, root length, aboveground fresh weight and root fresh weight of pea under the treatment of different concentrations of oligogalacturonic acid (OGAs) from 0 to 200 mg / L. Figure 1 , the results showed that oligogalacturonic acid OGAs in the low concentration range (0-50 mg / L) had no significant effect on pea root growth. However, as the concentration of OGAs increased, it showed a significant inhibitory effect on pea growth, specifically manifested as shortened root length, and the fresh weight of the roots was significantly reduced at a concentration of 200 mg / L. Similarly, the effect of OGAs on the aboveground growth of peas also followed this trend: low concentration treatment had no significant restriction on aboveground growth, while high concentration inhibited its growth. In summary, when the concentration of OGAS in the solution was lower than 50 mg / L, it had no significant effect on pea growth.

[0056] ②Effects of OGs on aluminum tolerance of pea root tips:

[0057] This study further explored the effect of exogenous oligogalacturonic acid (OGAs) on aluminum tolerance in pea. Figure 2 The experimental results showed that aluminum treatment significantly inhibited the elongation of pea roots, and this inhibitory effect was reflected at different aluminum concentrations. However, exogenous addition of oligogalacturonic acid (OGAs) can effectively alleviate the inhibitory effect of aluminum toxicity on root elongation (see Figure 2 A and 2B). In particular, under the condition of high aluminum concentration, the aluminum content of the 0-1 cm section of the root tip of different varieties of pea was determined by ICP-OES technology, and it was found that oligogalacturonic acid OGAs pretreatment could significantly reduce the accumulation of aluminum in the root tip. In addition, the results of hematoxylin staining and Morin staining also showed that oligogalacturonic acid OGAs could reduce aluminum toxicity to the root tip and reduce aluminum accumulation, further confirming that oligogalacturonic acid OGAs pretreatment could alleviate aluminum toxicity to the root tip of pea.

[0058] ③Analysis of endogenous OGAS content in pea under different treatments:

[0059] To investigate whether oligogalacturonic acid (OGAs) are involved in the response of pea root tips to aluminum stress, Figure 3 In this study, mass spectrometry was used to analyze the effects of aluminum treatment on the content and composition of endogenous oligogalacturonic acid (OGAs) in pea root tips. The results showed that even under control conditions, pea root tips contained endogenous oligogalacturonic acid (OGAs). Figure 3 A). However, under aluminum treatment, the content of oligogalacturonic acid OGAs in the root tip increased significantly, with an increase of about 100%. Further analysis of the composition of oligogalacturonic acid OGAs revealed that endogenous oligogalacturonic acid OGAs were mainly products with a degree of polymerization of 2, accounting for about 60%-70%. However, after aluminum treatment, the content of oligogalacturonic acid OGAs with a degree of polymerization of 6 and 7 increased. The above results clearly show that aluminum stress can induce the production of oligogalacturonic acid OGAs and increase the proportion of high-polymerization oligogalacturonic acid OGAs.

[0060] ④Differences in antioxidant enzyme activity and reactive oxygen metabolism in peas under different treatments:

[0061] In order to investigate the effects of oligogalacturonic acid (OGAs) on reactive oxygen metabolism in pea root tips, Figure 4 The results showed that compared with the normal treatment group, under aluminum treatment, exogenous addition of oligogalacturonic acid OGAs reduced the total active content of pea root tips ( Figure 4 A); while the activities of SOD, CAT and POD increased ( Figure 4 BD); further detection by fluorescence staining showed that, under the conditions of Al0 and Al15, the total ROS, O2·- and H2O2 in the root tip were significantly reduced by exogenous addition of oligogalacturonic acid OGAs, while there was no significant difference in ·OH ( Figure 4 EL).

[0062] ⑤ Expression profiles of RBOH, SOD, CAT and POD under oligogalacturonic acid OGAs treatment:

[0063] In order to further clarify the relationship between the regulation of oligogalacturonic acid OGAs on aluminum tolerance in pea and the expression of RRBOH, SOD, CAT, and POD, such as Figure 5 , this protocol selected 8 genes and verified them by real-time quantitative reverse transcription PCR (qRT-PCR). qRT-PCR results showed that there was no significant difference in the expression of LOC127101393 / RBOH gene among different treatments ( Figure 5 A), under Al treatment, the expression levels of all genes were much higher than those under control conditions ( Figure 5AH), and exogenous addition of oligogalacturonic acid (OGAs) treatment, the expression levels of three genes in the SOD family and three genes in the POD family were upregulated. These results indicate that in pea, there are significant differences in transcriptional regulation among different genotypes.

[0064] illustrate:

[0065] The main site of aluminum poisoning is the root tip, which is the most serious site of aluminum poisoning in plants. In crops, oligogalacturonic acid OGAs were initially identified as having physiological activity because they can induce crops to synthesize antitoxins. This study aims to explore the effect of oligogalacturonic acid OGAs on pea growth and analyze the pea phenotype. The results show that when the OGA concentration in the solution is lower than 50 mg / L, it has no significant effect on pea growth ( Figure 1 ).

[0066] Oligogalacturonic acid OGAs have a positive effect on promoting plant growth and development, and can promote the thickening of stems and the elongation of plants. This study further explored the effect of exogenous oligogalacturonic acid OGAs on aluminum tolerance in pea. Under Al treatment conditions, root elongation was significantly inhibited compared with normal conditions, and exogenous addition of oligogalacturonic acid OGAs treatment could alleviate aluminum toxicity ( Figure 2 AB); and the aluminum content in the root tip was different under different treatments. Oligogalacturonic acid OGAs pretreatment can significantly reduce the accumulation of aluminum in the root tip. The results of hematoxylin and morin staining also showed that oligogalacturonic acid OGAs can reduce the enrichment of active aluminum in the root tip ( Figure 2 DF), further verifying that oligogalacturonic acid (OGAs) pretreatment could alleviate aluminum toxicity.

[0067] Oligogalacturonates (OGs) are oligomers of α-1,4-linked galacturonic acid residues released from plant cell walls after partial degradation of homogalacturonic acid. OGs can trigger defense responses, including the accumulation of reactive oxygen species and pathogenesis-related proteins, and protect plants from pathogen infection. The test results of this scheme show that under aluminum treatment, the content of oligogalacturonic acid OGAs in the root tip increased, and there were also differences in the content of oligogalacturonic acid OGAs with different polymerization degrees, indicating that aluminum can induce the production of oligogalacturonic acid OGAs ( Figure 3 A).

[0068] When plants encounter adversity, they will produce reactive oxygen species, causing oxidative stress reactions, disrupting the normal metabolism of the plant, and thus affecting the growth and development of the plant. In order to reduce the damage caused by adversity to plants, plants have evolved a defense system for removing reactive oxygen species in the body during the long-term adaptation process. The main forms of ROS are singlet oxygen ( 1 O2), superoxide anion (O 2·-), hydrogen peroxide (H2O2) and hydroxyl radicals (OH · ). Through fluorescence staining, it was found that under the treatment of exogenous addition of oligogalacturonic acid OGAs, compared with the normal treatment group, total ROS, O2·- and H2O2 had significant differences, while ·OH had no significant difference ( Figure 4 EL). In plants, SOD catalyzes stress-induced O2·- and converts it into H2O2, while POD and CAT are mainly involved in the removal of H2O2, thereby maintaining the steady-state balance of reactive oxygen species in cells. According to the antioxidant enzyme activity analysis of this experiment, oligogalacturonic acid OGAs treatment significantly affected the antioxidant enzyme system of plants, including the enzyme activities of SOD, POD and CAT, which increased with oligogalacturonic acid OGAs pretreatment and the increase of aluminum concentration, and there were significant differences in total reactive oxygen activity; this indicates that the antioxidant enzyme system plays an important role in plant response to aluminum stress. Exogenous addition of oligogalacturonic acid OGAs treatment showed higher activity. In order to further explore whether these genes play a role in the regulation of oligogalacturonic acid OGAs and aluminum tolerance in pea, the expression levels of these eight genes in pea were further determined by real-time fluorescence quantitative PCR. It was found that the expression of genes in the SOD, CAT, and POD families increased after oligogalacturonic acid OGAs treatment, and under aluminum stress conditions, the expression of all genes increased. After oligogalacturonic acid OGAs induction, under Al stress conditions, SOD and POD genes were also upregulated ( Figure 5 AH). The increase in expression may enhance aluminum tolerance in pea.

[0069] in conclusion:

[0070] This study analyzed the biomass, endogenous antioxidant enzyme activity, and gene expression of peas, indicating that exogenous addition of oligogalacturonic acid (OGAs) can enhance aluminum tolerance in peas and act as a signal substance to participate in the regulation of aluminum tolerance in peas. Thus, the oligogalacturonic acid in this study can be used to enhance aluminum tolerance in peas, improve aluminum tolerance in agricultural crops, and promote agricultural income.

[0071] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A method for regulating oligogalacturonic acid to improve aluminum tolerance in peas, characterized in that: The following steps are involved: (1) Soak pea seeds in 7-8% sodium hypochlorite; rinse with deionized water; (2) pea seeds were soaked in a 2 mM CaCl2 solution and then spread on a mesh screen of an aeroponic mist culture system. The pea seeds were grown at 20-25°C for 48 h. (3) Select seedlings with a root length of 2 to 3 cm, transfer them to 1 / 4 Hoagland solution for 4 days, and culture them in a growth chamber with alternating day and night culture at 26°C for 16 h and 24°C for 8 h; (4) Pre-adapting the seedlings to an environment with a pH of 4 to 5 and then treating them with AlCl3 solution; (5) Pea seedlings with a root length of 2 to 3 cm were placed in sterile water respectively, and pea seedlings with uniform growth were selected after hydroponics, and the concentration of oligogalacturonic acid (OGAs) was set to at least 50 mg / L.

2. The method for regulating oligogalacturonic acid for improving aluminum tolerance of pea according to claim 3, characterized in that: In the step (5), the hydroponic conditions are: temperature 25±1°C, air humidity 75-85%, and light conditions of 16 hours of light, 14000-20000Lx of light intensity, and 8 hours of darkness.

3. The method for regulating oligogalacturonic acid for improving aluminum tolerance of pea according to claim 1, characterized in that: In the step (5), the concentration of oligogalacturonic acid OGAs is 50-100 mg / L.

4. Application of exogenous oligogalacturonic acid in enhancing aluminum tolerance of pea.

Images