CN112715546B - Application of polygonatum polysaccharide in plant growth promotion and stress resistance - Google Patents
Application of polygonatum polysaccharide in plant growth promotion and stress resistance Download PDFInfo
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- 150000004676 glycans Chemical class 0.000 title claims abstract description 79
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 79
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 79
- 241000756042 Polygonatum Species 0.000 title claims abstract description 66
- 230000008635 plant growth Effects 0.000 title claims abstract description 13
- 241000196324 Embryophyta Species 0.000 claims abstract description 25
- 238000003973 irrigation Methods 0.000 claims abstract description 7
- 230000002262 irrigation Effects 0.000 claims abstract description 7
- 238000005507 spraying Methods 0.000 claims abstract description 5
- 235000013311 vegetables Nutrition 0.000 claims abstract description 3
- 230000035882 stress Effects 0.000 claims description 24
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical group ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 239000002244 precipitate Substances 0.000 claims description 6
- 238000009835 boiling Methods 0.000 claims description 5
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- 238000005238 degreasing Methods 0.000 claims description 3
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- 239000000126 substance Substances 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims 1
- 239000007921 spray Substances 0.000 claims 1
- 238000011282 treatment Methods 0.000 description 18
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 14
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 9
- 230000012010 growth Effects 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 229910002651 NO3 Inorganic materials 0.000 description 8
- 229930002875 chlorophyll Natural products 0.000 description 8
- 235000019804 chlorophyll Nutrition 0.000 description 8
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 8
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 7
- 241000209094 Oryza Species 0.000 description 7
- 235000007164 Oryza sativa Nutrition 0.000 description 7
- 229930003268 Vitamin C Natural products 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 239000003337 fertilizer Substances 0.000 description 7
- 235000009566 rice Nutrition 0.000 description 7
- 239000002689 soil Substances 0.000 description 7
- 235000019154 vitamin C Nutrition 0.000 description 7
- 239000011718 vitamin C Substances 0.000 description 7
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 240000001592 Amaranthus caudatus Species 0.000 description 5
- 235000009328 Amaranthus caudatus Nutrition 0.000 description 5
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 description 5
- 244000221633 Brassica rapa subsp chinensis Species 0.000 description 5
- 235000012735 amaranth Nutrition 0.000 description 5
- 239000004178 amaranth Substances 0.000 description 5
- 239000000706 filtrate Substances 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 241000037826 Polygonatum kingianum Species 0.000 description 2
- 241000037831 Polygonatum sibiricum Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000008641 drought stress Effects 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
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- 235000006408 oxalic acid Nutrition 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- CCBICDLNWJRFPO-UHFFFAOYSA-N 2,6-dichloroindophenol Chemical compound C1=CC(O)=CC=C1N=C1C=C(Cl)C(=O)C(Cl)=C1 CCBICDLNWJRFPO-UHFFFAOYSA-N 0.000 description 1
- 208000006820 Arthralgia Diseases 0.000 description 1
- 235000005340 Asparagus officinalis Nutrition 0.000 description 1
- 208000008035 Back Pain Diseases 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000234280 Liliaceae Species 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 108010020346 Polyglutamic Acid Proteins 0.000 description 1
- 241001468611 Polygonatum cyrtonema Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 244000193174 agave Species 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 206010003549 asthenia Diseases 0.000 description 1
- 208000019804 backache Diseases 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 235000021186 dishes Nutrition 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000013538 functional additive Substances 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- NCYVXEGFNDZQCU-UHFFFAOYSA-N nikethamide Chemical compound CCN(CC)C(=O)C1=CC=CN=C1 NCYVXEGFNDZQCU-UHFFFAOYSA-N 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000000361 pesticidal effect Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 229920002643 polyglutamic acid Polymers 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 208000012201 sexual and gender identity disease Diseases 0.000 description 1
- 208000015891 sexual disease Diseases 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/40—Liliopsida [monocotyledons]
- A01N65/42—Aloeaceae [Aloe family] or Liliaceae [Lily family], e.g. aloe, veratrum, onion, garlic or chives
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Pest Control & Pesticides (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses application of polygonatum polysaccharide in plant growth promotion and stress resistance. The polygonatum polysaccharide has a sugar content of more than 75%, an ash content of less than 5% and a protein content of less than 2%; the plants include grain crops and vegetables; the polygonatum polysaccharide can be used in a root irrigation or leaf surface spraying mode; the molecular weight of the polygonatum polysaccharide is 1-500kDa, and the concentration is 0.001% -5% (w/v). In the invention, the polygonatum polysaccharide can promote the growth of plants, enhance the stress resistance of crops and improve the quality of the crops, and is a novel plant biological stimulator which is plant-sourced, green and environment-friendly.
Description
Technical Field
The invention belongs to the field of agricultural functional additives, and relates to application of polygonatum polysaccharide in plant growth promotion and stress resistance
Background
The biological stimulator is naturally available in nature, is rich in active substances, and can directly act on plants without artificial chemical synthesis. Such as polysaccharides of plant origin, polysaccharides of microbial origin, polyglutamic acid, etc., which we are familiar with. The biological stimulin is neither a pesticide nor a traditional fertilizer; the target of the bio-stimulin is the crop itself, which can improve the physiological and biochemical state of the plant, improve the pesticide effect and the utilization rate of the fertilizer, improve the level of the crop for resisting the adverse environment, and also improve the final yield of the crop and the quality of agricultural products.
The plant of Polygonatum belongs to Asparagus, and is widely distributed in northern hemisphere temperate zone. Rhizoma Polygonati can be used for treating fatigue, asthenia, dyspepsia, anorexia, sexual disorder, backache, gonalgia, and premature gray hair. Researches show that the polysaccharide in the sealwort plays a vital role in the processes. However, no research on polygonatum polysaccharide in the field of plant growth promotion and stress resistance exists at present.
Disclosure of Invention
The invention aims to provide application of polygonatum polysaccharide in plant growth promotion and stress resistance; aims to provide a brand-new high-efficiency plant source polysaccharide for regulating and enhancing the stress resistance of plants.
In order to achieve the purpose, the invention is realized by the following technical scheme:
the invention provides application of polygonatum polysaccharide in plant growth promotion and stress resistance.
The growth and stress resistance promotion includes the growth promotion of plants and the stress resistance improvement of the plants. The stress resistance mainly comprises abiotic stresses such as salt stress, drought stress and low temperature stress.
The molecular weight of the polygonatum polysaccharide is 1-500 kDa.
Preferably, the large molecular weight polygonatum polysaccharide with the molecular weight of 400-500kDa has better effect in promoting the growth of plants, and can obviously improve the yield of the plants, the quality of crops and the like.
Wherein, the high molecular weight polygonatum polysaccharide has better effect of being sprayed on the leaves.
Preferably, the polygonatum polysaccharide with a small molecular weight of 10-50kDa, more preferably the polygonatum polysaccharide with a small molecular weight of 10-20 kDa has a better effect in improving the stress resistance of plants, and can significantly improve the abiotic stress resistance of crops, such as salt stress, drought stress and the like. Wherein, the small molecular weight polygonatum polysaccharide has better root irrigation effect.
Preferably, the polygonatum polysaccharide can promote the growth of plants and enhance the stress resistance of the plants, and the use concentration of the polygonatum polysaccharide is 0.001-5% (w/v);
dissolving 1g of rhizoma Polygonati polysaccharide in 1L of pure water to obtain 0.1% (w/v) rhizoma Polygonati polysaccharide solution, and preparing rhizoma Polygonati polysaccharide with different concentrations by the method.
The application mode comprises root irrigation and foliage spraying.
The plants comprise grain crops, vegetables, flowers, trees and the like.
The polygonatum polysaccharide is extracted from polygonatum kingianum, which is rhizome of Polygonatum kingianum Coll et Hemsl, Polygonatum sibiricum Red or Polygonatum cyrtonema Hua of Liliaceae.
The extraction method of the polygonatum polysaccharide comprises the following steps:
step (1): steaming a fresh rhizoma Polygonati sample at 100 deg.C for 4-8h, drying in a vacuum oven at 40-55 deg.C, and grinding the sample into powder;
step (2): refluxing and degreasing the sample powder obtained in the step (1) by using petroleum ether, filtering, taking a precipitate, extracting for 1-2h by using boiling water, and repeating for 3 times;
and (3): the extract was concentrated by rotary evaporation and deproteinized with Sevag reagent (chloroform: n-butanol ═ 4:1, v/v). And then, using ethanol with the final concentration of 50-90% for alcohol precipitation, removing pigments and other small molecular substances, simultaneously centrifuging and collecting precipitates, and removing water and ethanol through vacuum drying to obtain the polygonatum polysaccharide. Preferably, the final concentration of ethanol is 75%.
Preferably, the polygonatum polysaccharide obtained in the extraction process has a sugar content of more than 75%, an ash content of less than 5% and a protein content of less than 2%.
Has the advantages that: the invention provides application of polygonatum polysaccharide in plant growth promotion and stress resistance. In the invention, the polygonatum polysaccharide is derived from polygonatum, is environment-friendly, green and pollution-free, and can promote the growth of crops and enhance the stress resistance of the crops only with extremely low amount.
Detailed Description
In order to make the objects, features and advantages of the present invention more obvious and understandable, the technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the embodiments described below are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments that can be derived by one skilled in the art from the embodiments given herein are intended to be within the scope of the invention.
Example 1 extraction method of Polygonatum polysaccharide
A fresh sample of rhizoma Polygonati (purchased from Tan Bao-Jiuhua house flagship shop, dried rhizoma Polygonati, production place: Anhui Chinese) was taken at 1 kg, steamed at 100 deg.C for 6h, then dried in a vacuum oven at 60 deg.C to constant weight, and after drying, the sample was ground into powder with a mortar. Refluxing and degreasing the sample powder for 1h by using petroleum ether, filtering by suction filtration, taking a precipitate, extracting for 2h by using 1.5L of boiling water, and repeating for 3 times; the extract was rotary-evaporated, concentrated to 500mL, and 125 mL of Sevag reagent (chloroform: n-butanol 4:1, v/v) was added thereto, and the mixture was repeatedly shaken, centrifuged to remove precipitated protein, and the aqueous phase was retained and repeated three times. Adding 1500mL of ethanol into the water phase liquid after removing the protein to precipitate extracellular polysaccharide, centrifuging to collect precipitate, and vacuum drying to remove water and ethanol to obtain rhizoma Polygonati polysaccharide, named as HJPS.
Through determination, the polygonatum polysaccharide extracted by the method has the sugar content of more than 75 percent, the ash content of less than 5 percent, the protein content of less than 2 percent and the molecular weight of 496 kDa.
The sugar content was measured by the phenol-sulfuric acid method, the ash content was measured with reference to GB 5009.4-2016, the protein content was measured by the Brad-ford method, and the molecular weight was measured by the GPC method.
Example 2 preparation of Low molecular weight Polygonatum polysaccharide
The polygonatum polysaccharide in example 1 was dissolved to 10g/L, pH was adjusted to 3 using HCl, and then heated in a boiling water bath for 2 h. Regulating pH to neutral with KOH, and precipitating with ethanol to obtain small molecular rhizoma Polygonati polysaccharide. The molecular weight of the micromolecule polygonatum polysaccharide is 35.4kDa through determination, and the micromolecule polygonatum polysaccharide is named as HJPS-L1.
Example 3 preparation of Low molecular weight Polygonatum polysaccharide
The polygonatum polysaccharide in example 1 was dissolved to 10g/L, pH was adjusted to 2 using HCl, and then heated in a boiling water bath for 2 h. Regulating pH to neutral with KOH, and precipitating with ethanol to obtain small molecular rhizoma Polygonati polysaccharide. The molecular weight of the micromolecule polygonatum polysaccharide is 13.6kDa through determination, and the micromolecule polygonatum polysaccharide is named as HJPS-L2.
Example 4 enhancement of salt stress resistance of rice seedlings by root irrigation treatment of polygonatum polysaccharides rice seeds were soaked in 1% (w/v) NaClO for 10min and washed clean with distilled water repeatedly. Soaking in distilled water for 6h, placing in dark at 25 deg.C for accelerating germination for 24h, selecting full and uniform seeds when wheat seeds are just white, and sowing into water culture dishes with 100 granules per dish. Placing the culture medium in an illumination incubator for culture under the following culture conditions: photoperiod 14/l0h (day/night), temperature 27/20 deg.C (day/night), light intensity 800. mu. mol-2s-1Relative humidity 65. + -. 5%.
After one week of growth and culture of rice seedlings, the rice seedlings were placed in 1/2MS nutrient solution containing 100mM, and 0.001% (w/v) of HJPS, HJPS-L1 and HJPS-L2 were additionally added, while a blank Control (CK) was designed. And after 14d, measuring related indexes of salt stress on fresh weight, plant height, root length, chlorophyll content and the like of the rice seedlings.
The results of the experiment are shown in table 1. Under the simulated stress treatment of 100mM, compared with a blank control CK group, exogenous addition of polygonatum polysaccharides with different molecular weights can remarkably relieve salt stress damage to rice seedlings. Specifically, the plant height, fresh weight and chlorophyll of the HJPS group added with the macromolecular polygonatum polysaccharide are respectively increased by 17.6%, 51.8% and 53.2% compared with those of the CK group; the plant height, fresh weight and chlorophyll of the HJPS-L1 group added with the micromolecule polygonatum polysaccharide are respectively improved by 27.0%, 78.1% and 108.5% compared with those of the CK group; the plant height, fresh weight and chlorophyll of the HJPS-L2 group added with the micromolecule polygonatum polysaccharide are respectively improved by 36.8%, 69.8% and 140.4% compared with the plant height, fresh weight and chlorophyll of the CK group; therefore, the micromolecule polygonatum polysaccharide has better capability of enhancing the salt stress resistance of crops in the root irrigation treatment. It is worth noting that the addition amount of the polygonatum polysaccharides with different molecular weights is only 0.001%.
Wherein, the content of chlorophyll is measured according to the following method: 0.5g leaf (fresh weight) was minced, dark bleached with 50mL of 80% (v/v) aqueous acetone at room temperature for 48h, then centrifuged at 10000g for 5min, and the absorbance of the supernatant was measured at 645nm and 663 nm. The chlorophyll content (C) was then calculated using the following formulaChl)：CChl(mg/g)＝8.05A663+20.29A645。
TABLE 1 Effect on plant height, fresh weight and chlorophyll of Rice seedlings under different treatments
Example 5 leaf surface spraying treatment of Polygonatum sibiricum polysaccharide to promote growth of amaranth
The experiment is carried out by adopting polygonatum polysaccharide solutions with different molecular weights and the final concentration of 0.01 percent (w/v) and blank contrast, amaranth is taken as a research object, and the experiment is carried out in Puhao ecological park in Putongku city of Nanjing province of Jiangsu province. The specific treatment was as follows, and 4 treatments were designed for the experiment, each sprayed with 5L of 0.01% (w/v) HJPS, HJPS-L1, HJPS-L2 polysaccharide solution and an equivalent amount of water as a blank Control (CK). Sampling is carried out after one week of treatment, and the fresh weight, the vitamin C content and the nitrate content of the amaranth are determined. Wherein each is processed for 4m20.25m of sampling area2And the rest is processed according to normal management.
The results of the experiment are shown in table 2. The treatment of the polygonatum polysaccharides with different molecular weights obviously improves the yield of the amaranth, the vitamin content and reduces the nitrate content. Specifically, the HJPS group added with the macromolecular polygonatum polysaccharide has the fresh weight and the vitamin C respectively increased by 45.0 percent and 60.0 percent compared with the CK group, and the nitrate content is reduced by 46.6 percent; the HJPS-L1 group added with the micromolecular polygonatum polysaccharide has the fresh weight and the vitamin C respectively increased by 19.3 percent and 33.4 percent compared with the CK group, and the nitrate content is reduced by 19.8 percent; the HJPS-L2 group added with the micromolecular polygonatum polysaccharide has the fresh weight and the vitamin C respectively increased by 21.0 percent and 40.4 percent compared with the CK group, and the nitrate content is reduced by 30 percent. Therefore, the macromolecular polygonatum polysaccharide has the capability of better promoting the growth of crops and improving the quality of the crops in the spraying treatment of the leaves.
Wherein, the content of the vitamin C is determined according to the following method: weighing about 2.0g of sample, and leaching the sample with 30mL of 1% (W/V) oxalic acid in a triangular flask; transferring into a 100mL volumetric flask, and fixing the volume with 1% oxalic acid; fully shaking up, and leaching for 10-15 min; filtering and reserving for later use. The sample filtrate, 10.0mL, was pipetted into a 250mL Erlenmeyer flask and titrated with the 2, 6-dichloroindophenol dye solution until the solution was pink and persists for 15 seconds. Each sample was run in 3 replicates and the dye was calibrated before each run. A standard curve was made using vitamin C as a standard.
Wherein, the content of nitrate is measured according to the following method: weighing about 5g of sample, adding 10mL of distilled water, fully grinding, transferring the residue and the juice into a 100mL volumetric flask, and fixing the volume. Mixing thoroughly, filtering to obtain filtrate, adding 1.5g of active carbon into the filtrate, stirring in warm water at 60-70 deg.C while heating for 5min, and filtering to obtain filtrate. Taking 4.0mL of filtrate, adding 0.2mL of concentrated hydrochloric acid into a 50mL small-volume bottle, oscillating for 1-2 min, adding 0.5mL of 1% phenol solution and 5mL of 0.1 mol.L-1 sulfuric acid solution, adding distilled water to fix the volume, and standing for 15 min. And (3) taking the reagent blank as a reference solution, measuring an absorbance value at the wavelength of 390nm, and calculating the concentration of the nitrate through a nitrate standard curve.
TABLE 2 Effect of different treatments on the fresh weight, vitamin C content and nitrate content of amaranth
Example 6 comparison of growth promoting effects of Macro-molecular Polygonatum polysaccharide of different concentrations
Macromolecular polygonatum polysaccharide was prepared according to the method of example 1. The potted pakchoi is taken as an experimental object, 3 treatments are set in the experiment, each treatment is repeated for 3 times, wherein the treatment 1(T1) is an experimental control common fertilizer (150 mg per kg of soil), the treatment 2(T2) is a common fertilizer (150 mg per kg of soil) + macromolecular polygonatum polysaccharide (0.1 mg per kg of soil), the treatment 3(T3) is a common fertilizer (150 mg per kg of soil) + macromolecular polygonatum polysaccharide (1 mg per kg of soil), the treatment 3(T4) is a common fertilizer (150 mg per kg of soil) + macromolecular polygonatum polysaccharide (10 mg per kg of soil), and the experimental results are shown in Table 3.
TABLE 3 influence of macromolecular Polygonatum polysaccharide on growth characteristics and yield of potted pakchoi
As can be seen from Table 3, the macromolecular polygonatum polysaccharide mixed with the fertilizer can promote the growth of the roots of the pakchoi, thicken and enlarge the roots, has obvious promotion effects on the plant height, the fresh weight of the underground part and the fresh weight of the overground part of the pakchoi, and can improve the yield of the pakchoi by 31 percent, 46.5 percent and 49.1 percent by the treatment of T2, T3 and T4 respectively. This shows that, in a certain concentration range, the increase of the usage amount of the polygonatum polysaccharide can further improve the yield of crops.
Claims (10)
1. The application of polygonatum polysaccharide in promoting plant growth and enhancing plant stress resistance; the plant stress resistance is abiotic stress.
2. The use of claim 1, wherein said polygonatum polysaccharide has a sugar content of greater than 75%, an ash content of less than 5%, and a protein content of less than 2%.
3. Use according to claim 1, wherein the plants comprise cereals and vegetables.
4. The use of claim 1, wherein the polygonatum polysaccharide is applied by root irrigation or foliar spray.
5. The use of claim 1, wherein the polygonatum polysaccharide has a molecular weight of 1-500 kDa.
6. The use of claim 1, wherein the polygonatum polysaccharide is used at a concentration of 0.001-5% (w/v).
7. The application as claimed in claim 5, wherein the rhizoma Polygonati polysaccharide with molecular weight of 400-500kDa is large molecular weight rhizoma Polygonati polysaccharide, and is applied by spraying on leaves to promote plant growth.
8. The use of claim 5, wherein the molecular weight of the polygonatum polysaccharide is 1-50kDa and the plant stress resistance is improved by root irrigation.
9. The use of claim 1, wherein the extraction process of the polygonatum polysaccharides comprises the following steps:
(1) steaming fresh rhizoma Polygonati sample at 100 deg.C for 4-8 hr, drying in vacuum oven at 40-60 deg.C, and pulverizing the sample;
(2) refluxing and degreasing the sample powder obtained in the step (1) by using petroleum ether, filtering, taking a precipitate, extracting for 1-2 hours by using boiling water, and repeating for 3 times;
(3) concentrating the extract by rotary evaporation, deproteinizing with Sevag reagent; and then, using ethanol with the final concentration of 50-90% for alcohol precipitation, removing pigments and other small molecular substances, simultaneously centrifuging and collecting precipitates, and removing water and ethanol through vacuum drying to obtain the polygonatum polysaccharide.
10. The use according to claim 9, wherein the Sevag reagent is chloroform mixed with n-butanol at a volume ratio of 4: 1.
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