A composition for alleviating waterlogging stress of chicken head and its application

Abstract

The present application belongs to the field of plant conservation, more particularly, relates to a composition for relieving waterlogging stress of Allium victorialis L. and application thereof. The composition of the present application is composed of matured cow dung and spermidine solution, the concentration of the spermidine solution is 0.1-1 mmol / L. Before planting Allium victorialis L., the matured cow dung is applied in the field, when the flood season comes and the plant height of Allium victorialis L. reaches about 60-80 cm, by using the conservation method of the composition, the harm of waterlogging stress on Allium victorialis L. can be continuously relieved, the content of proline in the leaf of Allium victorialis L. is increased, the content of malondialdehyde in the leaf of Allium victorialis L. is reduced, and the oxidative damage caused by waterlogging stress is reduced by increasing the activity of antioxidant enzyme in the leaf of the plant.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of plant maintenance, more particularly, relates to a composition for relieving waterlogging stress of Polygonatum sibiricum and application thereof. BACKGROUND

[0002] Polygonatum sibiricum is a shallow-rooted plant, and the main root system is distributed in the soil surface layer of 18-20 cm, and it grows naturally in the ecological environment with deep humus layer and good soil ventilation condition. However, in artificial cultivation environment, especially in the area where rainfall is concentrated in the rainy season, it is easy to suffer from waterlogging damage. Under waterlogging stress, Polygonatum sibiricum will show the damage performance of leaf yellowing, root damage, change of antioxidant enzyme system, change of endogenous hormone, damage of cell membrane system, increase of osmotic adjustment substances, decrease of photosynthesis and abnormal respiration.

[0003] Firstly, under the waterlogging stress of hypoxic environment, the leaves of Polygonatum sibiricum will show drooping, loss of green, yellowing, edge burning, wilting, shrinkage, patchy disease, and the root absorption capacity will decrease or even rot. Secondly, under waterlogging stress, due to the blocked normal oxidative phosphorylation level and respiration of Polygonatum sibiricum, the accumulation of active oxygen in the plant body will induce the up-regulation of the activity of various antioxidant enzymes, and the increase of antioxidant enzyme activity can quickly remove the accumulated active oxygen free radicals in the plant body, and reduce the toxic effect of active oxygen.

[0004] Therefore, it is very necessary to develop a product which can increase the antioxidant enzyme activity of Polygonatum sibiricum to relieve the physiological damage caused by waterlogging stress. SUMMARY

[0005] The present application aims to provide a composition for relieving waterlogging stress of Polygonatum sibiricum and application thereof.

[0006] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0007] The present application provides a composition for relieving waterlogging stress of Polygonatum sibiricum, which is composed of mature cow dung and spermidine solution, the concentration of the spermidine solution is 0.1-1 mmol / L, and the mass-volume ratio of mature cow dung to spermidine solution is 200-300 g:10-50 ml.

[0008] The mature cow dung in the composition can improve the soil structure, so that the soil becomes more loose, which is beneficial to the stretching and respiration of the plant root system in the adverse environment of waterlogging, and the spermidine can regulate the physiological process of the plant and enhance the antioxidant defense system. Through experimental screening of the two, scientific combination of the ratio can better relieve the waterlogging stress of Polygonatum sibiricum.

[0009] The application also provides application of the composition in relieving waterlogging stress of chicken head rhizoma polygonati.

[0010] Further, the composition is used for increasing the chlorophyll content in the leaves of chicken head rhizoma polygonati under waterlogging stress.

[0011] Further, the composition is used for reducing the malondialdehyde content in the leaves of chicken head rhizoma polygonati under waterlogging stress.

[0012] Further, the composition is used for increasing the proline content in the leaves of chicken head rhizoma polygonati under waterlogging stress.

[0013] Further, the composition is used for increasing the antioxidant enzyme activity in the leaves of plants to reduce the oxidative damage caused by waterlogging stress.

[0014] Further, the antioxidant enzyme includes peroxidase, superoxide dismutase and catalase.

[0015] The application also provides a method for relieving waterlogging stress of chicken head rhizoma polygonati by using the above composition, and the steps include: applying mature cow dung in a field before planting, transplanting chicken head rhizoma polygonati seedlings into the field, and when the chicken head rhizoma polygonati plants grow to a height of 60-80 cm, spraying spermidine in a 0.1-1 mmol / l water solution on the front and back of the plant leaves to prevent waterlogging stress.

[0016] Further, the application amount of the mature cow dung in the field is 10-13 m 3 / acre.

[0017] Further, the spraying frequency is once a day, 2-3 ml is sprayed per plant each time, and the total spraying times are 6-18 times.

[0018] The application has the following beneficial effects:

[0019] The application provides a composition of mature cow dung matrix and spermidine solution and application thereof, and by using the maintenance method of the composition, the harm of waterlogging stress on chicken head rhizoma polygonati can be continuously relieved, the proline content in the leaves of chicken head rhizoma polygonati is increased, the malondialdehyde content in the leaves of chicken head rhizoma polygonati is reduced, and the antioxidant enzyme activity in the leaves of plants is increased to reduce the oxidative damage caused by waterlogging stress before the end of July, i.e., before the coming of the rainy season in the vigorous growth season. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 Figure 2 is a graph of chlorophyll content changes under waterlogging for 1 day, wherein the vertical axis of the graph indicates that a represents chlorophyll a, b represents chlorophyll b, and T represents total chlorophyll in the treatment group.

[0021] Figure 2Figure 2 is a graph showing the change in chlorophyll content of waterlogged leaves for 2 days, wherein the vertical axis of the graph indicates chlorophyll a, chlorophyll b, and total chlorophyll in the treatment groups.

[0022] Figure 3 Figure 4 is a graph showing the change in chlorophyll content of waterlogged leaves for 4 days, wherein the vertical axis of the graph indicates chlorophyll a, chlorophyll b, and total chlorophyll in the treatment groups.

[0023] Figure 4 Figure 8 is a graph showing the change in chlorophyll content of waterlogged leaves for 8 days, wherein the vertical axis of the graph indicates chlorophyll a, chlorophyll b, and total chlorophyll in the treatment groups.

[0024] Figure 5 Figure 16 is a graph showing the change in chlorophyll content of waterlogged leaves for 16 days, wherein the vertical axis of the graph indicates chlorophyll a, chlorophyll b, and total chlorophyll in the treatment groups.

[0025] Figure 6 Figure 18 is a graph showing the change in malondialdehyde content of waterlogged leaves for different durations of waterlogging and treatment with urea oxidase.

[0026] Figure 7 Figure 20 is a graph showing the change in malondialdehyde content of waterlogged leaves for different durations of waterlogging and treatment with melatonin.

[0027] Figure 8 Figure 22 is a graph showing the change in malondialdehyde content of waterlogged leaves for different durations of waterlogging and treatment with salicylic acid.

[0028] Figure 9 Figure 24 is a graph showing the change in malondialdehyde content of waterlogged leaves for different durations of waterlogging and treatment with matured cow manure.

[0029] Figure 10 Figure 26 is a graph showing the change in malondialdehyde content of waterlogged leaves for different durations of waterlogging and treatment with spermidine.

[0030] Figure 11 Figure 28 is a graph showing the change in malondialdehyde content of waterlogged leaves for different durations of waterlogging and treatment with matured cow manure and spermidine.

[0031] Figure 12 Figure 30 is a graph showing the change in malondialdehyde content of waterlogged leaves for different durations of waterlogging and treatment with uniconazole.

[0032] Figure 13 Figure 32 is a graph showing the change in superoxide dismutase activity of waterlogged Liriope spicata treated with urea oxidase.

[0033] Figure 14 Figure 34 is a graph showing the change in superoxide dismutase activity of waterlogged Liriope spicata treated with melatonin.

[0034] Figure 15 Figure 36 is a graph showing the change in superoxide dismutase activity of waterlogged Liriope spicata treated with matured cow manure.

[0035] Figure 16Figure 1 is a graph showing the change in superoxide dismutase activity of waterlogged Rhizoma Polygonati Chinensis treated with spermidine.

[0036] Figure 17 Figure 2 is a graph showing the change in superoxide dismutase activity of waterlogged Rhizoma Polygonati Chinensis treated with mature cow manure and spermidine.

[0037] Figure 18 Figure 3 is a graph showing the change in superoxide dismutase activity of waterlogged Rhizoma Polygonati Chinensis treated with uniconazole.

[0038] Figure 19 Figure 4 is a graph showing the change in peroxidase activity of waterlogged Rhizoma Polygonati Chinensis treated with oxidized urea.

[0039] Figure 20 Figure 5 is a graph showing the change in peroxidase activity of waterlogged Rhizoma Polygonati Chinensis treated with melatonin.

[0040] Figure 21 Figure 6 is a graph showing the change in peroxidase activity of waterlogged Rhizoma Polygonati Chinensis treated with salicylic acid.

[0041] Figure 22 Figure 7 is a graph showing the change in peroxidase activity of waterlogged Rhizoma Polygonati Chinensis treated with mature cow manure.

[0042] Figure 23 Figure 8 is a graph showing the change in peroxidase activity of waterlogged Rhizoma Polygonati Chinensis treated with spermidine.

[0043] Figure 24 Figure 9 is a graph showing the change in peroxidase activity of waterlogged Rhizoma Polygonati Chinensis treated with mature cow manure and spermidine.

[0044] Figure 25 Figure 10 is a graph showing the change in peroxidase activity of waterlogged Rhizoma Polygonati Chinensis treated with uniconazole.

[0045] Figure 26 Figure 11 is a graph showing the change in catalase activity of waterlogged Rhizoma Polygonati Chinensis treated with oxidized urea.

[0046] Figure 27 Figure 12 is a graph showing the change in catalase activity of waterlogged Rhizoma Polygonati Chinensis treated with melatonin.

[0047] Figure 28 Figure 13 is a graph showing the change in catalase activity of waterlogged Rhizoma Polygonati Chinensis treated with salicylic acid.

[0048] Figure 29 Figure 14 is a graph showing the change in catalase activity of waterlogged Rhizoma Polygonati Chinensis treated with mature cow manure.

[0049] Figure 30 Figure 15 is a graph showing the change in catalase activity of waterlogged Rhizoma Polygonati Chinensis treated with spermidine.

[0050] Figure 31Figure of catalase activity change of water-flooded Jichouteng Rhizoma Polygonati after treatment of matured cow dung and spermidine.

[0051] Figure 32 Figure of catalase activity change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone.

[0052] Figure 33 Figure of proline content change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone.

[0053] Figure 34 Figure of proline content change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone.

[0054] Figure 35 Figure of proline content change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone.

[0055] Figure 36 Figure of proline content change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone.

[0056] Figure 37 Figure of proline content change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone.

[0057] Figure 38 Figure of proline content change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone.

[0058] Figure 39 Figure of proline content change of water-flooded Jichouteng Rhizoma Polygonati after treatment of dimethazone. DETAILED DESCRIPTION

[0059] The present application will be described in detail below with reference to the drawings and specific embodiments, but should not be understood as limiting the present application. If not specifically stated, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, if not specifically stated, can be obtained from commercial channels.

[0060] Example 1

[0061] I. Experimental methods

[0062] 1. The test adopts a completely randomized block design, and is set as normal maintenance control, water-flooded control, water-flooded peroxide urea treatment, water-flooded melatonin treatment, water-flooded salicylic acid treatment, water-flooded cow dung treatment, water-flooded spermidine treatment, water-flooded cow dung + spermidine treatment, and water-flooded dimethazone treatment. Each treatment is set with 3 concentration levels, 10 pot-grown Jichouteng Rhizoma Polygonati seedlings, repeated three times, a total of 690, with a canopy density of 0.7, and the test is carried out under the shade of Populus canadensis. The outdoor water-flooded test lasts for 16 days, and the determination and data statistical analysis are carried out successively, and all the test work is completed at the end of August.

[0063] 2、First, in Beijing Forestry University test nursery, ready 690 high 26 cm, top end diameter 28 cm pottery basin, bottom water hole cover tile. Preparation of matrix: humus soil: vermiculite: river sand: grass carbon volume ratio is 5:2:2:1, cow dung treatment group respectively with rotten cow dung instead of grass carbon, river sand and vermiculite, cow dung accounts for 10%, 30%, 50% of matrix volume ratio. 0.5% potassium permanganate solution disinfection 4h. On April 3, from Huairou Yanshan mountain area to take two years old chicken head Jixiang, select the first stem section with terminal bud, head thick end diameter 4.0cm, length 8.0cm, plant 2 each pot. Dip 5% CE garlicin microemulsion and naphthalene acetic acid 500 times mixed solution. Fill the soil about to the height of the pot three quarters, parallel to the stem section with terminal bud, terminal bud thick end slightly high, tail thin end slightly low. Then cover the stem section with matrix, add matrix to 4cm from the upper edge of the pot, pour water thoroughly, pour water thoroughly again after 3 days. To May 30, chicken head Jixiang test plant growth to the height of 70cm, first in the treatment pot with urea peroxide, dig a circular ditch 5cm deep, bury 4 bags, 8 bags, 12 bags of bag control peroxide urea according to 1g per bag respectively in the set level, cover soil, start waterlogging treatment, irrigate with outdoor standing water for 1 day to 1cm above the pot, keep 3cm deep, to August 9, complete the whole test. The specific experimental design is shown in Table 1.

[0064] Table 1: chicken head Jixiang waterlogging test design.

[0065] Treatment (horizontal) Level 1 Level 2 Level 3 Normal maintenance control (CK) Waterlogging 0 Waterlogging 0 Waterlogging 0 Waterlogging control (WL) Waterlogging 3 cm Waterlogging 3 cm Waterlogging 3 cm Waterlogging urea peroxide treatment (UP) 4g 8g 12g Waterlogging melatonin treatment (MT) 100 μmol / L 500 μmol / L 1000 μmol / L Waterlogging salicylic acid treatment (SA) 80 mg / L 160 mg / L 240 mg / L Waterlogging cow manure treatment (CM) 10％ 30％ 50％ Waterlogging spermidine treatment (SP) 0.5 mmol / L 1.0 mmol / L 1.5 mmol / L Waterlogging cow manure + spermidine treatment (CS) 0.5 mmol / L 1.0 mmol / L 1.5 mmol / L Waterlogging uniconazole treatment (U) 20 mg / L 40 mg / L 80 mg / L

[0066] Note: in the three levels of waterlogging cow dung + spermidine treatment group, cow dung accounts for 30% of the volume ratio of matrix.

[0067] II. Detection and data statistics.

[0068] 1. Chlorophyll content: weigh 0.1g fresh leaves, cut into small pieces and put in centrifuge tube. Add 10mL of 95% ethanol and 80% acetone mixture with a volume ratio of 1:1, avoid light and stand for 48h until the green leaf tissue becomes colorless. Weigh 0.1g fresh leaves, cut into small pieces and put in centrifuge tube. Add 10mL of 95% ethanol and 80% acetone mixture with a volume ratio of 1:1, avoid light and stand for 48h until the green leaf tissue becomes colorless. Use the enzyme marker to measure the absorbance at 470nm, 645nm and 663nm respectively, and calculate the contents of Chl.a, Chl.b, Chl and Carotenoid.

[0069] 2. Malondialdehyde content: Fresh plant leaves were taken, 5% TCA 5 mL was added, and the obtained homogenate was grinded and centrifuged for 10 min. 2 mL of supernatant was taken, 0.67% TBA was added to 2 mL, mixed, and boiled in a 100°C water bath for 30 min, and centrifuged again. The absorbance values at 450 nm, 532 nm and 600 nm were measured, and the content of malondialdehyde (C, μmol / L) was calculated according to the following formula:

[0070] C = 6.45 (A 532 -A 600 )- 0.56A 450 .

[0071] A 532 is the absorbance at 532 nm, A 600 is the absorbance at 600 nm, and A 450 is the absorbance at 450 nm.

[0072] 3. Superoxide dismutase (SOD) activity determination: The leaves of the 5th-6th segments of the above-ground part and the underground part of the chicken head were collected, and the underground part was frozen in liquid nitrogen. 0.1 g of leaves was taken, grinded into homogenate with 6 mL of extraction solution and a little quartz sand, and centrifuged at 12000 g for 20 min at 4°C. 0.1 mL of enzyme solution supernatant was taken, 3 mL of reaction medium (containing 50 mmol·L -1 methionine, 750 μmol·L -1 NBT, 0.1 mmol·L -1 EDTA, 20 μmol·L -1 riboflavin) was added, mixed, irradiated for 20 min, darkened for 10 min, and the absorbance value was measured at 560 nm. The calculation formula of superoxide dismutase activity is as follows:

[0073] SOD = (Ack-As) × V × 60 (Ack × 0.5 × W × t).

[0074] In the formula, Ack represents the absorbance value of the blank tube; As represents the absorbance value of the sample tube; V represents the total volume of the extraction solution; W represents the sample weight; and t represents the irradiation time.

[0075] 4. Peroxidase (POD) activity determination: The leaves of the 5th-6th segments of the above-ground part and the underground part of the chicken head were collected, and the underground part was frozen in liquid nitrogen. 0.1 g of leaves was taken, grinded into homogenate with 6 mL of extraction solution and a little quartz sand, and centrifuged at 12000 g for 20 min at 4°C. 50 μL of enzyme solution supernatant was taken, 3 mL of reaction solution (300 mL of phosphate buffer solution and 704 μL of H2O2 and 168 μL of guaiacol) was added, and colorimetry was performed at 470 nm. The calculation formula of peroxidase activity is as follows:

[0076] POD = ΔD470 x V1(0.1 x W x V2 x t).

[0077] Wherein, ΔD470 represents the change of absorbance within the reaction time; V1 represents the total volume of enzyme extract; W represents the sample weight; V2 represents the volume of enzyme solution when measured; t represents the reaction time.

[0078] 5. Catalase (CAT) activity determination: The leaves of the 5th-6th segments of the aerial parts of Allium victorialis-regel var. chinense and the underground parts were collected, packed, and then frozen in liquid nitrogen. 0.1 g of the leaves was ground into homogenate with 6 mL of extraction solution and a small amount of quartz sand. The homogenate was centrifuged at 12000 g at 4°C for 20 min. 50 μL of the enzyme solution supernatant was added to 3 mL of reaction solution (200 mL of phosphate buffer solution plus 227.2 μL of H2O2), and colorimetry was performed at 240 nm. The calculation formula of catalase activity is as follows:

[0079] CAT = (Ack-As) x V1(0.1 x W x V2 x t).

[0080] Wherein, Ack represents the absorbance of the blank tube; As represents the absorbance of the sample tube; V1 represents the total volume of enzyme extract; W represents the sample weight; V2 represents the volume of enzyme solution when measured; t represents the reaction time.

[0081] 6. Proline content: The proline (PRO) content was determined by the ninhydrin colorimetric method. 0.1 g of the middle leaves of Allium victorialis-regel var. chinense was ground after being added with 3% sulfosalicylic acid and quartz sand, and centrifuged at 10000 g at 4°C for 20 min. The supernatant was the proline extract. 2 mL of the extract was taken, 2 mL of glacial acetic acid and 2 mL of acidic ninhydrin solution were added, and the solution was heated in a boiling water bath for 30 min, and the solution turned red. After cooling, 5 mL of toluene was added, and the solution was shaken for 30 s, and then the upper liquid was taken and centrifuged at 3000 r / min for 5 min.

[0082] Colorimetry was performed on the supernatant at 520 nm, and the calculation formula of the proline content is as follows:

[0083] Proline = C x V(a x W).

[0084] Wherein, C is obtained from the standard curve; V represents the volume of the extract; a represents the volume taken when measured; and W represents the sample weight.

[0085] III. Results and analysis.

[0086] 1. Chlorophyll content.

[0087] From Figure 1It can be seen that the chlorophyll content of the leaves of the plants in each treatment changed little after 1 day of submergence, and only the chlorophyll a and total chlorophyll contents of the leaves of the plants in the water submergence control (WL) treatment decreased significantly.

[0088] From Figure 2 It can be seen that the chicken head rhizoma was sensitive to water submergence stress, and the root system had good oxygen permeability at the beginning of water submergence; in comparison, the effect of uniconazole was not obvious at the beginning of water submergence.

[0089] From Figure 3 It can be seen that the chlorophyll content decreased sharply on the fourth day of water submergence; the effect of urea peroxide was more prominent, and 8 g of bagged urea peroxide per pot could well alleviate the harm of water submergence stress; the stress resistance of melatonin was also manifested; uniconazole maintained cell viability by maintaining low-level metabolic activity, and ultimately protected the assimilation cells of the leaf mesophyll.

[0090] From Figure 4 It can be seen that after 8 days of water submergence, a large number of leaf edges were scorched and spots were obvious. The overall performance was that the chlorophyll content was already very low. Relatively speaking, the effect of cow dung plus spermidine was the best, and the effects of salicylic acid and uniconazole were also obvious.

[0091] From Figure 5 It can be seen that after 16 days of water submergence, a large number of leaves of the chicken head rhizoma died, and the chlorophyll content of the plants treated with high-dose spermidine

[0092] 1.5 mmol / L and cow dung and the chlorophyll content of the plants treated with 40 mg / L uniconazole at a medium dose remained relatively high.

[0093] 2. Changes in the content of malondialdehyde.

[0094] Water submergence has multiple effects on the physiology and biochemistry of plants, one of which is the effect on biological membrane systems and their functions. Water submergence stress can cause an increase in the content of malondialdehyde in plant leaves, leading to membrane lipid peroxidation and delipidization, and thus causing serious damage to cell membrane lipids and enzymes.

[0095] Malondialdehyde is the end product of membrane lipid peroxidation under water submergence stress conditions, and can reflect the integrity of cell membrane structure to some extent, and is closely related to plant resistance. From Figure 6It can be seen that, simple waterlogging without mitigation stress measures (WL) and normal management of the control CK, the leaf malondialdehyde content of Allium victorialis var. platyspermum reached a peak of 61.21 nmol / g after 4 days of waterlogging; then the malondialdehyde content decreased rapidly after 8 days of waterlogging; the leaves wilted on the 16th day of waterlogging. The application of urea peroxide (UP1, UP2, UP3 are level 1, level 2, level 3 respectively) can alleviate the problem of root hypoxia after waterlogging, and the malondialdehyde content increases slightly in the first two days of waterlogging, especially the high concentration of oxygen supply, which can well protect the leaf cell membrane system and prevent serious membrane lipid peroxidation due to waterlogging. After adding urea peroxide, urea peroxide slowly releases oxygen in the water solution. The oxygen content in the soil solution is increased, the aeration of the substrate is improved, the stress resistance of the plant is enhanced, the damage of waterlogging stress is effectively alleviated, and the damage degree of stress is reduced.

[0096] From Figure 7 It can be seen that, with the increase of waterlogging time, waterlogging stress significantly increases the malondialdehyde content of Allium victorialis var. platyspermum. Under the condition of waterlogging alone, the malondialdehyde content of leaves increases significantly after 1d, 2d and 4d of waterlogging. Compared with the control, different concentrations of melatonin treatment (MT1, MT2, MT3, level 1, level 2, level 3 respectively) reduces the malondialdehyde content of Allium victorialis var. platyspermum under waterlogging stress.

[0097] From Figure 8 It can be seen that, the plants treated with salicylic acid show slow rising speed and low peak value, and the malondialdehyde content is reduced by 40% compared with the waterlogging control on the 4th day of waterlogging, indicating that salicylic acid can inhibit the increase of malondialdehyde content in the early stage of waterlogging, thereby improving the waterlogging stress adaptation ability of Allium victorialis var. platyspermum; then the malondialdehyde content decreases rapidly with the leaf apoptosis of the waterlogging control WL, while the malondialdehyde of the plants treated with different concentrations of salicylic acid (SA1, SA2, SA3, level 1, level 2, level 3 respectively) decreases slowly, showing strong anti-flooding ability, indicating that the Allium victorialis var. platyspermum treated with salicylic acid has strong adaptability to membrane system peroxidation after waterlogging, so that the cell membrane has certain stability, thereby reducing the damage of plasma membrane under water stress.

[0098] From Figure 9 It can be seen that, the substrate mixed with high proportion of composted cow manure (CM1, CM2, CM3, level 1, level 2, level 3 respectively) can better maintain the aeration of the substrate in the early stage of waterlogging, which is conducive to maintaining the normal state of cell metabolism, so the degree of membrane lipid peroxidation is obviously weaker than that of WL treatment; and in the peak period of malondialdehyde on the 4th day, the peak value of the cow manure treatment is relatively low, indicating that the damage to the leaf cells is lighter.

[0099] Polyamines are a class of low-molecular-weight aliphatic nitrogen-containing bases with biological activity. Under abiotic stress, polyamines can scavenge free radicals in plant bodies and are closely related to the regulation of plant stress tolerance. They have strong functions against abiotic stress and are widely used in stress resistance research. Figure 10 It can be seen that spermidine (SP1, SP2, SP3, respectively, level 1, level 2, level 3) can reduce the content of malondialdehyde during flooding, thereby protecting the cell membrane system from or reducing the damage of membrane lipid peroxidation.

[0100] From Figure 11 It can be seen that in this test, 30% of the rotten cow dung as a potting substrate combined with spermidine (CS1, CS2, CS3, respectively, level 1, level 2, level 3) showed the most ideal protective function of mesophyll cell membrane system. Not only reduced the content of malondialdehyde, but also showed good stress resistance in terms of peak height and appearance time. After applying spermidine to the potting chicken head of the rhizoma of Polygonatum sibiricum in the cow dung substrate, the malondialdehyde slowly rose after flooding stress, and the peak appeared on the 8th day of flooding, while the WL treatment of the control group appeared on the 4th day after flooding, and the maximum content of malondialdehyde was 37% lower than that of the control group. It shows that CS treatment can effectively protect the chicken head of the rhizoma of Polygonatum sibiricum, and the CS-treated plants show less damage symptoms such as loss of green, yellowing, and scorched leaves after suffering from flooding stress, and the symptoms appear later, which is of great significance in actual production.

[0101] Uniconazole belongs to gibberellin inhibitors and is a broad-spectrum azole plant growth regulator. It has a strong inhibitory effect on the growth of herbaceous or woody monocotyledonous or dicotyledonous crops. As a commonly used economic crop growth regulator, uniconazole and paclobutrazol can control the overgrowth of stems and leaves, promote root growth, and improve the root crown ratio. The medicinal part of the chicken head of the rhizoma of Polygonatum sibiricum is in the rhizome, and there is a problem of overgrowth and lodging during cultivation, so proper chemical control and regulation can achieve a win-win effect.

[0102] The mechanism of action of uniconazole is similar to that of paclobutrazol, which can regulate vegetative growth, shorten internodes, dwarf plants, and promote the growth and differentiation of lateral buds and flower buds, and enhance the stress resistance of plants. Uniconazole has the advantages of small dosage, strong activity, no plant deformation, long effective period, and safety to humans and animals, and has been widely used in rice, wheat, corn, fruit trees, flowers, and other crops.

[0103] From Figure 12It can be seen that although uniconazole (U1, U2, U3, respectively, level 1, level 2, level 3) has a certain role in stress at the beginning of the flooding test, but the role is not very significant; in the later stage of the flooding test, the stress resistance of uniconazole gradually comes into play, not only the peak of malondialdehyde is reduced, but also the peak is delayed, therefore, the application of uniconazole can significantly inhibit the accumulation of malondialdehyde content in the leaves of Allium victorialis var. platyspermum under flooding stress. Related research has also found that uniconazole can increase the content of osmotic regulation substances, reduce the generation and accumulation of malondialdehyde, improve the cell membrane permeability, maintain the stability of the cell membrane, and enhance the ability of the cell membrane to resist adversity.

[0104] 3. The role of antioxidant enzymes.

[0105] The antioxidant enzyme system of Allium victorialis var. platyspermum is an important system for the plant to adapt to the environment, maintain normal growth and development, and defend against stress. Among them, glutathione peroxidase, catalase and peroxidase mainly participate in the metabolic activity of the leaves of Allium victorialis var. platyspermum.

[0106] 3.1, Superoxide dismutase SOD.

[0107] Superoxide dismutase (SOD) is an antioxidant enzyme that has been studied more in recent years. SOD is a metal-containing antioxidant enzyme that exists universally in the plant kingdom. It has the function of catalyzing O·2- to disintegrate, catalyzing O·2- to form H2O2 and O2, and H2O2 is converted into H2O and O2 under the action of CAT, POD, etc., thereby reducing the toxic effect of active oxygen. Therefore, SOD is considered to be a key enzyme for preventing oxidative stress. After flooding stress, too much water will cause poor soil aeration and lack of oxygen around the rhizosphere. This poor rhizosphere environment will affect the absorption and transport of water and minerals by the roots, and will produce redundant active oxygen, including superoxide anion free radicals (Οˉ2·), hydrogen peroxide (H2O2), hydroxyl radicals (·OH) and singlet oxygen (O2), which will cause imbalance of the redox state of plant cells, have toxic effects on cells, and may cause cell death. 1

[0108] Figure 13 It is shown that the flooding stress is effectively improved by the application of peroxide urea, especially in the early stage of flooding, the treatments of UP1, UP2 and UP3 all show the ability to enhance the activity of SOD, among which the 8g dose of UP2 is the best, on the fourth day of flooding, the SOD activity of the leaves of Allium victorialis var. platyspermum in this treatment is significantly higher than that of CK and WL, which is 53% and 25% respectively, thereby effectively removing ROS. It can be seen that the removal of redundant ROS through the antioxidant enzyme system is the main mechanism for Allium victorialis var. platyspermum to adapt to the stress of flooding.

[0109] From the above, it can be seen that the application of uniconazole and peroxide urea can effectively improve the activity of antioxidant enzymes in the leaves of Allium victorialis var. platyspermum under flooding stress, and can effectively reduce the accumulation of malondialdehyde in the leaves of Allium victorialis var. platyspermum under flooding stress. Figure 14 ​It can be seen that under waterlogging stress, the activity of SOD of chicken head lily significantly increased. The treatment of 500 μmol / L melatonin can increase the activity of SOD by 55%.

[0110] From Figure 15 It can be seen that the mixing of rotten cow manure in the mechanism can improve the health status of the leaves under waterlogging stress. The internal mechanism is that the addition of 50% can significantly improve the activity of SOD.

[0111] Figure 16 It shows that exogenous spermidine can protect chicken head lily leaf cell membrane system from free radical attack under waterlogging stress.

[0112] Figure 17 It shows that the addition of 30% rotten cow manure in the medium combined with the spraying of 1.0 mmol / L or more spermidine on the leaves can keep the SOD in the leaves of chicken head lily at a high activity level during waterlogging, and its activity is significantly higher than that of the normal management CK control and waterlogging control WL.

[0113] Figure 18 It shows that under waterlogging stress conditions, spraying 20-40 mg / L of uniconazole can promote the increase of antioxidant enzyme SOD, thereby inhibiting the accumulation of free radicals and reducing the degree of membrane lipid peroxidation. In this test, uniconzole works slowly, and its effect on increasing the activity of SOD in leaves is particularly obvious from the 8th to the 16th day after waterlogging stress. Therefore, in the summer cultivation of chicken head lily, low-dose uniconzole should be used early to prevent waterlogging damage to chicken head lily.

[0114] 3.2, Peroxidase POD.

[0115] From Figure 19 It can be seen that peroxide urea can quickly start the stress response in the first 4 days of waterlogging stress, and quickly improve the activity of POD, which is particularly suitable for reducing the damage of short-term waterlogging to chicken head lily. The use of 8-12 g of higher dose of bagged controlled release can still increase the activity of POD by 40% from the 8th to the 16th day of waterlogging compared with the waterlogging control WL, that is, from 1067.34 U·g -1 ·min -1 to 1494.57 U·g -1 ·min -1 .

[0116] Figure 20It is shown that melatonin treatment makes the chicken head lily leaves maintain a higher activity during the waterlogging stress process. After 2 days of stress, the POD activity of leaves treated with 100 μmol / L, 500 μmol / L and 1000 μmol / L melatonin is significantly higher than that of the control WL. Until the 16th day of waterlogging, the POD activity of chicken head lily leaves treated with 500 μmol / L melatonin is 1.5 times that of the WL treatment.

[0117] Figure 21 It is shown that salicylic acid (SA) or salicylic acid (SA) is a plant phenolic compound, which is automatically synthesized by plants under normal conditions or under biological and non-biological stress. It plays a prominent role in regulating plant growth and response to environmental stress. For chicken head lily under waterlogging stress, the application of 160 mg / L salicylic acid can significantly improve the activity of leaf POD; when the concentration increases to 240 mg / L, the positive effect will decrease.

[0118] Figure 22 It is shown that adding 50% of the rotten cow dung to the cultivation substrate is beneficial to enhance the POD activity of chicken head lily leaves under waterlogging stress.

[0119] From Figure 23 It can be seen that after 4 days of waterlogging stress, the POD activity of WL waterlogging control plants decreases rapidly, while the POD activity of plants treated with 1.0 mmol / L SP2 reaches the highest, which is 34% higher than that of WL treatment plants. At the end of waterlogging stress, the POD activity of plants treated with high concentration of spermidine 1.5 mmol / L, i.e. SP3, is significantly higher than that of other treatments.

[0120] Figure 24 It is shown that the substrate mixed with 30% of the rotten cow dung combined with the spraying of 1.0 mmol / L spermidine shows a strong effect of improving POD activity, and it remains until the end of waterlogging.

[0121] Figure 25 It is shown that in the early stage of waterlogging stress, uniconazole does not significantly improve the POD activity, and even at the 1st day of waterlogging stress, the POD activity of WL waterlogging control plants is the strongest. With the continuation of waterlogging stress, the stress resistance of uniconazole gradually comes out. On the 8th day of waterlogging, the POD activity treated with 40 mg / L U2 reaches the highest, and until the 16th day, the POD activity of plants treated with uniconazole is significantly higher than that of waterlogging control.

[0122] 3.3, Catalase CAT.

[0123] Catalase (CAT) is an antioxidant enzyme mainly existing in plant and microbial cells, and is a physiological index of plant adaptation to adversity. It is a major component of the antioxidant defense system formed by plants during long-term evolution. CAT and POD are the main enzymes for removing active oxygen in plants.

[0124] From Figure 26 It can be seen that from the second day of stress, the CAT activity of plants treated with urea peroxide was significantly higher than that of other treatments.

[0125] Figure 27 It is shown that melatonin at a moderate dose of 500 μmol / L can rapidly increase CAT activity, and significantly higher than the WL control as the waterlogging stress continues.

[0126] Figure 28 It is shown that the combination of exogenous SA and CAT can enhance the antioxidant capacity of Allium victorialis var. platyspermum under water stress, while reducing the degree of destruction of related photosynthetic pigments, thereby alleviating the degradation of enzymes related to the dark reaction in the photosystem and reducing the degree of destruction of the photosynthetic system, thereby improving the resistance of the plant to water stress. From the fourth day of waterlogging treatment, the CAT activity of plants treated with 160 mg / L salicylic acid was significantly higher than that of the WL waterlogging control plants.

[0127] Figure 29 It is shown that in this test, the highest mixing ratio of 50% of the rotten cow manure substrate can effectively improve the CAT activity of Allium victorialis var. platyspermum under waterlogging stress.

[0128] Figure 30 It is shown that under waterlogging stress, the CAT activity of Allium victorialis var. platyspermum plants treated with spermidine peaked on the fourth day, and there was no significant difference between the three concentrations, all of which were significantly higher than the waterlogging control. However, in the late waterlogging period, the SP3 high concentration, i.e. 1.5 mmol / L treatment, performed best, with the highest CAT activity of the waterlogged plants.

[0129] Figure 31 It is shown that the combination of 30% rotten cow manure mixed with the substrate and foliar spraying of spermidine results in the best performance of Allium victorialis var. platyspermum under waterlogging stress. During the entire waterlogging stress period, the CAT activity of the plants under this treatment was higher than that of the control, and on the 2nd, 4th, 8th and 16th days, it reached a significant level. Moreover, the peak value appeared on the 8th day of waterlogging, and from the 4th to the 8th day, the CAT activity was at a very high level, effectively protecting the cell membrane system of the stressed plants.

[0130] Figure 32It shows that, in the initial stage of flooding stress, the three concentrations of uniconazole treatment have no significant difference with the control, from the second day, the CAT activity of uniconazole treated plants is significantly higher than that of the control, and the difference reaches a very significant level at 8, 16 days of flooding. Therefore, uniconazole is an effective cultivation option for Allium chinense to deal with rain season floods. In this test, the U2 treatment concentration of 40 mg / L works very well, and there is no obvious difference with the U3 treatment concentration of 80 mg / L, and it remains stable and effective in the later stage of stress.

[0131] 4. Proline.

[0132] Proline is an important amino acid, known to participate in the biosynthesis of primary metabolism during growth and development, in addition, it is a kind of osmotic adjustment substance to reduce osmotic damage, and a large amount of proline will be produced under stress, and its accumulation is considered as a kind of osmotic adaptation mechanism. The accumulation of proline can improve the waterlogging tolerance and eliminate the damage of flooding stress.

[0133] Figure 33 It shows that under flooding stress, the proline content in leaves rises rapidly. After peroxide urea treatment, the proline content rises more slowly, the peak is higher, and it will decrease in the later stage. The reason is that the oxygen supply in the root is sufficient due to peroxide urea in the early stage of stress, and the stress signal transmission is not strong, and the accumulation of proline is not triggered quickly.

[0134] From Figure 34 It can be seen that after the treatment of melatonin, the proline content rises rapidly, and the high content is maintained for a long time to maintain the osmotic balance of cells under stress, and protect the normal function of cells.

[0135] Under flooding stress, salicylic acid is used to link the effect of stress damage. Figure 35 In this test, the proline of salicylic acid treated plants rises quickly, and after the proline of the control treated plants begins to decrease rapidly on the 4th day, the salicylic acid treatment still maintains the rising trend, and begins to decrease on the 8th day, but the proline content is significantly higher than that of the control at this time, which provides protection for the stressed plants.

[0136] Figure 36 It shows that the matrix mixed with rotten cow dung delays the increase speed of proline content, but maintains a high content of proline for a long time, and delays the time of damage to mesophyll cells.

[0137] Figure 37The results show that spermidine can promote the accumulation of proline in the mesophyll under waterlogging stress, not only increasing the absolute content of proline from 784 μg / g to 837 μg / g, but also maintaining a high proline accumulation state during waterlogging stress. Waterlogging stress causes an increase in proline content in the body of Rhizoma Polygonati Chinensis, thereby playing an important role in stabilizing cell protein structure, reducing cell acidity, and removing active oxygen, reducing the damage caused by stress to the leaves and maintaining the normal growth and development of Rhizoma Polygonati Chinensis.

[0138] Figure 38 The results show that in this test, Rhizoma Polygonati Chinensis accumulates a large amount of proline in the body when subjected to waterlogging stress, thereby improving the adaptability of Rhizoma Polygonati Chinensis to adversity stress. The combination of 30% composted cow manure and arginine treatment effectively increases the proline accumulation rate at the beginning of stress, and can continuously maintain a high concentration of proline, with a maximum value of 934 μg / g, which is higher than that of spermidine alone at 12%. In this test, 1.0 mmol / L and 1.5 mmol / L spermidine doses are significantly higher than 0.5 mmol / L SP1 treatment, so the water stress relief dose can be determined as 1.0 mmol / L to cope with the expected waterlogging damage to Rhizoma Polygonati Chinensis.

[0139] Figure 39 The results show that S3307, a plant growth retardant that has been proven to be quite effective in practice, is commonly used to regulate crop growth and development, enhance stress resistance, and improve crop resistance to environmental stress, such as waterlogging, weak light, heavy metals, and saline-alkali stress. Waterlogging stress is one of the factors that restrict crop growth, and an increase in soil moisture can cause a lack of oxygen in a short period of time, and anaerobic respiration and its metabolites can cause damage to cells. S3307 can regulate proline content, soluble protein content, and soluble sugar content, thereby regulating cell osmotic regulation and improving plant waterlogging tolerance. In this test, proline accumulation increased rapidly under S3307 treatment, with a peak value of 883 μg / g on the 4th day of waterlogging, and in subsequent waterlogging tests, S3307 treatment was significantly higher than waterlogging control WL. From this test, S3307 has the best sustained effect in the later stages of waterlogging stress.

[0140] It is worth noting that uniconazole can also increase the root crown ratio of plants, increase root dry weight, promote the transportation of nutrients from the aboveground part to the underground part, and make the plant root system develop. For the ecological cultivation of chicken head lily under the forest, there is a vigorous growth season of stem and leaf, and after 10-13 nodes, there is a climbing growth, and the hooked leaf tips of the alternate leaves are gathered together with the fallen and remote branches and leaves, which creates conditions for the occurrence of spider mites, brown spot disease, mildew and the like. Therefore, before the end of July, that is, before the rainy season, during the vigorous growth season, spraying uniconazole with a concentration of 40mg / L or less in advance can not only cope with waterlogging in advance, but also improve the health of chicken head lily and increase the economic yield of rhizome.

[0141] It should be noted that when the present application claims involving numerical ranges, it should be understood that each numerical range of two endpoints and any numerical value between the two endpoints can be selected. In order to prevent repetition, the present application describes the preferred embodiments.

[0142] Although the preferred embodiments of the present application have been described, those skilled in the art can make further changes and modifications to these embodiments once they know the basic creative concept. Therefore, the appended claims are intended to include the preferred embodiments and all changes and modifications falling within the scope of the present application.

[0143] Obviously, those skilled in the art can make various modifications and variations to the present application without departing from the spirit and scope of the present application. Thus, if these modifications and variations of the present application fall within the scope of the claims of the present application and their equivalents, the present application also intends to include these modifications and variations.

Claims

1. A composition for alleviating waterlogging stress in chicken head of ginseng, characterized in that, The composition is composed of matured cow dung and spermidine solution, the concentration of the spermidine solution is 0.1-1mmol / L, and the mass-volume ratio of the matured cow dung and the spermidine solution is 200-300g:10-50ml.

2. The composition of claim 1 is used for relieving the waterlogging stress of chicken head lily.

3. Use of the composition according to claim 2 for alleviating waterlogging stress in chicken head of ginger, characterized in that, The composition is used for increasing the chlorophyll content in the leaves of chicken head lily under waterlogging stress.

4. The use of a composition according to claim 2 for alleviating waterlogging stress in chicken head of ginger, characterized in that, The composition is used for reducing the malondialdehyde content in the leaves of chicken head lily under waterlogging stress.

5. The use of a composition according to claim 2 for alleviating waterlogging stress in chicken head of ginger, characterized in that, The composition is used for increasing the proline content in the leaves of chicken head lily under waterlogging stress.

6. Use of the composition according to claim 2 for alleviating waterlogged stress in chicken head of ginger, characterized in that, The composition is used for increasing the antioxidant enzyme activity in the leaves of plants to reduce the oxidative damage caused by waterlogging stress.

7. Use of the composition according to claim 6 for alleviating waterlogging stress in chicken head of ginger, characterized in that, The antioxidant enzymes include peroxidase, superoxide dismutase and catalase.

8. A method for alleviating waterlogging stress in chicken head using the composition as claimed in claim 1, wherein the steps of The composition comprises: Before planting, the matured cow dung is applied in the field, and then the chicken head lily seedlings are transplanted into the field, when the chicken head lily plants grow to a height of 60-80cm, the spermidine is configured into a 0.1-1mmol / l aqueous solution, and the solution is uniformly sprayed on the front and back of the plant leaves to prevent waterlogging stress.

9. The composition according to claim 8, for use in a method for alleviating waterlogging stress in chicken head shorthorn, characterized in that, The application amount of the matured cow dung in the field is 10-13 m 3 / acre.

10. The composition according to claim 8, for use in a method for alleviating waterlogging stress in chicken head shorthorn, characterized in that, The spraying frequency is once a day, 2-3ml is sprayed per plant each time, and the total spraying times are 6-18 times.

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