CN110063161B - Method for promoting growth of andrographis paniculata and improving content of active ingredients of andrographis paniculata by utilizing sesbania stem nodule nitrogen-fixing rhizobia - Google Patents
Method for promoting growth of andrographis paniculata and improving content of active ingredients of andrographis paniculata by utilizing sesbania stem nodule nitrogen-fixing rhizobia Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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Abstract
The invention discloses a method for promoting the growth of common andrographis herb and improving the content of active ingredients of the common andrographis herb by utilizing sesbania stem tumor nitrogen-fixing rhizobium. The invention adopts the technique of inoculating nitrogen-fixing rhizobia by the non-root nodule plant to induce sesbania stem nodule nitrogen-fixing rhizobia of leguminous plant to infect andrographis paniculata plant, thereby not only promoting the growth of andrographis paniculata, but also increasing the biomass, especially the biomass of overground part; more importantly, the content of the effective components of the common andrographis herb is greatly increased, and each 667m2The total andrographolide yield can be improved by 112.32% to the maximum extent, which is beneficial to producing high-quality organic green andrographis paniculata, greatly improves the land utilization rate, has stronger practicability and popularization value, provides reference for the standardized planting sustainable development of andrographis paniculata, and also widens the application range of sesbania stem nodule nitrogen-fixing rhizobium.
Description
Technical Field
The invention belongs to the technical field of biological planting. More particularly, the method relates to a method for promoting the growth of andrographis paniculata and improving the content of active ingredients of andrographis paniculata by using sesbania stem tumor nitrogen-fixing rhizobium.
Background
Andrographis Paniculata is also named as Happy, is an Andrographis Paniculata Nees (Burm.f.) of Acanthaceae, and is cultivated in India, east China, south China and other areas. Bitter in property and cold in nature, it enters heart, lung, large intestine and bladder meridians. Has effects of clearing away heat and toxic materials, cooling blood, relieving swelling and pain, etc. It is mainly used for treating cold, fever, sore throat, aphtha, cough, diarrhea, dysentery, stranguria, carbuncle, sore, and snake bite. The main components of herba Andrographitis contain diterpene lactone and flavonoids. The diterpene lactones mainly comprise andrographolide, deoxyandrographolide, neoandrographolide and dehydroandrographolide, wherein the content of andrographolide is the highest and accounts for more than 1.5%.
The reality, safety, effectiveness, stability and controllability are the basic requirements of standardized planting of the common andrographis herb, and the production of the organic common andrographis herb is imperative. The traditional planting method for cultivating the common andrographis herb consumes a large amount of chemical fertilizer, especially nitrogen fertilizer. Excessive use of chemical fertilizer brings problems of environmental pollution, energy consumption, soil hardening, fertility reduction and the like, and the quality and sustainable development of the andrographis paniculata medicinal materials are seriously affected. How to improve the production capacity of the common andrographis herb in adverse circumstances, improve the planting benefit of the common andrographis herb in ecological vulnerable areas, reduce the use of chemical fertilizers, reduce the agricultural production cost and finally realize the organization of the common andrographis herb production is an important content of the common andrographis herb production, cultivation and planting technology and an important subject of the sustainable development of the common andrographis herb industry.
Biological nitrogen fixation means that some prokaryotes can convert molecular nitrogen into a form of ammonia nitrogen available for plants by using nitrogen fixation enzymes in the body, and the nitrogen fixation enzymes are required for plant growth. Biological nitrogen fixation can promote the growth of the interacting plants, enhance the resistance of the interacting plants to adverse stress, improve the immunity of the interacting plants to pathogenic bacteria, improve the soil fertility, improve the nitrogen content of the soil, reduce the input of nitrogen fertilizers, avoid the loss of the nitrogen fertilizers in the use process, save the production cost and be beneficial to maintaining ecological balance. The rhizobia can be symbiotic with most leguminous plants to form root nodules, fix free nitrogen and synthesize nitrogen compounds which can be utilized by the plants. Inducing rhizobium to carry out symbiotic nitrogen fixation with non-leguminous plants is a leading hot spot field of biological nitrogen fixation research, is successful in non-leguminous plants such as wheat, corn, barley, tobacco and the like at present, and opens up a new way for the research of high-yield and high-quality crop planting technology.
Chinese patent document (publication No. CN 106134728A) discloses a method for promoting the growth of non-leguminous plants by using rhizobia, which is a non-leguminous plant selected from rice, corn, sorghum, wheat and millet, by diluting inoculated cells obtained by inoculating and culturing and centrifuging a liquid of rhizobia, and then inoculating the diluted bacterial liquid to the roots of seedlings of the non-leguminous plants, wherein the rhizobia is selected from alfalfa rhizobia, pea rhizobia, bean rhizobia, soybean rhizobia and lupin rhizobia, and the interacting plants of the rhizobia are mainly concentrated in gramineae plants which are researched at present. It is known that there is selectivity between rhizobia and host plants, rhizobia has the characteristics of species diversity, habitat diversity and host diversity, and the mutual assistance between rhizobia and plants varies depending on the species of rhizobia, the physiological and biochemical characteristics of host plants, the growth substrate (pH, redox conditions, texture, organic matter content, root secretions, rhizosphere microorganisms, rhizosphere minerals, etc.), external environmental conditions, and other factors. The effect of different rhizobia on plant treatment is different, and the mechanism is different. The Cao Yanzhen et al (Cao Yanzhen, Li Kuen. Amelan. non-leguminous crops and nitrogen-fixing microorganisms form nodule-like [ J ] under the influence of helper bacteria, proceedings of university of agriculture in Huazhong, 1994(4): 313-; in the root nodules of cabbage, not only the presence in the cells was observed, but also the nitrogenase activity was measured by the acetylene reduction method. How to explore and construct a nitrogen fixation system of non-leguminous crops and expand the nitrogen fixation function to non-leguminous plants except the radiorhizobium plants, particularly Chinese herbal medicine plants, is still extremely challenging.
The formation of functional neoplasia is a complex process involving different phenotypic stages, each stage being dependent on both bacterial genes and host plant genes. At present, related researches on influences of biological nitrogen fixation and nodulation nitrogen fixation of non-leguminous medicinal materials on the quality of medicinal materials are few, the influence researches on the andrographis paniculata are rare, and the development and utilization of rhizobium resources and the improvement of the quality of andrographis paniculata raw medicinal materials are urgent market requests.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects and shortcomings of the prior art and provide a method for promoting the growth of andrographis paniculata and improving the content of active ingredients of andrographis paniculata by using sesbania stem nodule nitrogen-fixing rhizobium. By adopting the technology of inoculating nitrogen-fixing rhizobia to the non-rhizomatous plant, inducing sesbania stem tumour nitrogen-fixing rhizobia of leguminous plants to infect andrographis paniculata plants, not only can the growth of andrographis paniculata be promoted, but also the biomass, particularly the biomass of overground parts, can be increased; more importantly, the content of the active ingredients of the common andrographis herb is greatly increased, and the quality of the medicinal materials is improved.
The above purpose of the invention is realized by the following technical scheme:
the invention researches for the first time to find that after the sesbania stem tumor nitrogen-fixing rhizobium is used for treating, the growth of common andrographis herb can be promoted, and the biomass, particularly the biomass of the overground part, can be increased; more importantly, the content of the effective components of the common andrographis herb is greatly increased, and each 667m2The yield of the andrographolide can be improved by 112.32% to the maximum, which is beneficial to producing high-quality organic green medicinal materials of andrographis paniculata.
Firstly, the invention discloses the application of sesbania stem tumor nitrogen-fixing rhizobium in promoting the growth of andrographis paniculata plants and/or improving the content of effective components of andrographis paniculata, and the application is within the protection scope of the invention.
Further, in the preferred embodiment of the present invention, the promoting of the growth of andrographis paniculata plants refers to the application of sino-nodus azorhizobium in promoting the growth of aerial parts of andrographis paniculata plants.
Further, in a preferred embodiment of the present invention, the content of the effective components of andrographis paniculata is the content of andrographolide.
Further, in a preferred embodiment of the present invention, the andrographolide comprises andrographolide and dehydroandrographolide.
Further, in the preferred embodiment of the present invention, the use of Sinorhizobium sessiliflorum for increasing the activity of total superoxide dismutase (T-SOD) of Andrographis paniculata Nees is also within the scope of the present invention. Researches show that the irrigation treatment of sesbania stem tumor nitrogen-fixing rhizobium can promote the physiological action of an antioxidant enzyme system of plant cells of andrographis paniculata plants and can obviously improve the activity of total superoxide dismutase (T-SOD).
On the basis, the invention provides a method for promoting the growth of common andrographis herb and improving the content of active ingredients of the common andrographis herb by utilizing sesbania stem tumor nitrogen-fixing rhizobia, which is symbiotic cultivation of the sesbania stem tumor nitrogen-fixing rhizobia and the common andrographis herb.
Further, in the preferred embodiment of the present invention, the symbiotic cultivation is performed by using the sesbania stem cell azorhizobium liquid to perform seed soaking treatment on the andrographis paniculata seeds, or distributing the sesbania stem cell azorhizobium liquid around the root system of the andrographis paniculata plants (i.e. irrigation treatment). Researches show that the irrigation treatment of sesbania stem tumor nitrogen-fixing rhizobium can promote the physiological action of an antioxidant enzyme system of plant cells of andrographis paniculata plants and can obviously improve the activity of total superoxide dismutase (T-SOD).
Further, in a preferred embodiment of the present invention, the symbiotic cultivation is to distribute the liquid of sesbania stem-nodule azorhizobium azotobacter around the root system of the plant of Andrographis paniculata Nees.
Further, in a preferred embodiment of the present invention, the concentration of the liquid of the nodule bacteria azotobacter of sesbania is greater than or equal to 1.0 × 107one/mL.
Furthermore, in the preferred embodiment of the present invention, the concentration of the liquid of the nodule bacteria azotobacter of sesbania is 1.0 × 107～1.0×109one/mL.
Further, in a preferred embodiment of the present invention, the andrographis paniculata plant is young andrographis paniculata; the andrographis paniculata seedling is the time when the andrographis paniculata seeds grow to 2 cotyledons and are completely unfolded after germination is finished.
Further, in a preferred embodiment of the present invention, the method comprises the steps of: and (3) irrigating sesbania stem nodule nitrogen-fixing rhizobium liquid to the andrographis paniculata seedlings, distributing the sesbania stem nodule nitrogen-fixing rhizobium liquid around the root systems of the andrographis paniculata, and cultivating the andrographis paniculata in 4-7 months under natural conditions for 3-4 months.
Further, in a preferred embodiment of the present invention, the amount of the liquid of sesbania stem nodule azotobacter is 5-15 mL/time for one seedling of andrographis paniculata, and 2-3 times in total.
Further, in the preferred embodiment of the present invention, the planting density of Andrographis paniculata Nees is 667m211000 to 15000 strains.
Furthermore, in the preferred embodiment of the present invention, the planting density of Andrographis paniculata Nees is 667m215000 strains.
In addition, the invention also provides a fertilizer capable of promoting the growth of andrographis paniculata and improving the lactone content, which comprises sesbania stem tumor nitrogen-fixing rhizobium. The sesbania stem tumor nitrogen-fixing rhizobium can be the sesbania stem tumor nitrogen-fixing rhizobium bacterial liquid.
Preferably, the concentration of the sesbania stem tumor nitrogen-fixing rhizobium bacterial liquid in the fertilizer is 1.0 x 107～1.0×109The fertilizer is used per mL, and the using amount of the fertilizer is 195-200 liters per mu.
In addition, it is emphasized that the sesbania stem nodule azorhizobium bacteria solution can be added with auxiliary materials which do not have adverse effect on the growth of the andrographis paniculata, such as carriers, carbon sources, diluents, adsorbents, trace elements, nutrient elements and the like according to needs.
Compared with the prior art, the invention has the following beneficial effects:
(1) the invention adopts the technique of inoculating nitrogen-fixing rhizobia by the non-root nodule plant to induce sesbania stem nodule nitrogen-fixing rhizobia of leguminous plant to infect andrographis paniculata plant, thereby not only promoting the growth of andrographis paniculata, but also increasing the biomass, especially the biomass of overground part; more importantly, the content of the active ingredients of the common andrographis herb is greatly increased. The method has reasonable design and credible result, greatly improves the utilization rate of the land, and has stronger practicability and popularization value.
(2) The invention relates to a method for treating common andrographis herb by sesbania stem nodule nitrogen-fixing rhizobia, which is one of effective ways for solving the problems of yield and quality reduction of common andrographis herb and the like, provides technical support for deeply researching the growth promoting mechanism of the nitrogen-fixing rhizobia on common andrographis herb, cultivating high-yield and high-quality organic green medicinal materials of common andrographis herb, provides reference for the sustainable development of standardized planting of the medicinal materials of common andrographis herb, and provides a new idea and a new way for the research and development of special bacterial fertilizers for common andrographis herb.
(3) The invention also realizes a new application of the sesbania stem tumor nitrogen-fixing rhizobium, and provides a new thought and direction for the development and application of the sesbania stem tumor nitrogen-fixing rhizobium.
(4) Compared with the traditional chemical and agricultural measures, the method has the characteristics of good effect, no secondary pollution, low operation cost and the like.
Detailed Description
The present invention is further illustrated by the following specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
Chinese rhizobium freudenreichizus, Rhizobium radiobacter (purchased from Guangdong province microorganism collection center), and sesbania stem nodule Azorhizobium ORS 571 were provided by Liuhua Wei teacher of northwest agriculture and forestry science and technology university, and Andrographis paniculata seeds were produced in Xingheng county of Yulin city of Guangxi province and identified as mature seeds of Andrographis paniculata (Burm.f.) Nees by professor Dujiu in the institute of medicinal and botanical science and research of Guangzhou Chinese medicinal university.
Example 1 Effect of sesbania Stem tumor azorhizobium on growth of Andrographis paniculata plants
1. Method of producing a composite material
(1) The andrographis paniculata test seeds germinate in a culture dish, and after germination is finished (2 leaves are fully unfolded), andrographis paniculata seedlings are obtained; transplanting the andrographis paniculata seedlings to flowerpots, wherein each pot contains 1 plant and 75 pots, and the seedlings are divided into 5 groups, and each group contains 15 pots;
(2) after transplanting, the concentration of the seed was 1.0X 10 at 1, 3 and 5 days61.0X 10 units/mL71.0X 10 units/mL81.0X 10 units/mL9Irrigating 5mL of sesbania stem tumor nitrogen-fixing rhizobium liquid for each plant, setting blank groups, and replacing the blank groups with sterile water;
(3) after the bacterial liquid treatment for 3d, watering (tap water) every afternoon, based on the fact that the soil is completely watered;
(4) after sesbania stem tumor nitrogen-fixing rhizobia is treated for 100 days, randomly selecting 9 andrographis paniculata plants in each concentration group, and measuring plant height, leaf number (true leaves) and branch number;
(5) collecting herba Andrographitis plants, washing with flowing water to remove silt from root, air drying, counting lateral roots, and measuring fresh weight and dry weight of root length, aerial parts and underground parts.
2. Results
The results are shown in tables 1 and 2.
(1) As shown in Table 1, the concentration of 1.0X 10 of the root nodule of Sesbania sessilifolia after treating Andrographis paniculata seedlings for 100 days7The plant height, leaf number and branch number of the concentration treatment group per mL are respectively and remarkably higher than those of the blank group in the level (P < 0.05), and the experimental result shows that the plant height, the leaf number and the branch number of the concentration treatment group per mL are respectively and remarkably higher than those of the blank group7The concentration treatment group per mL has the most obvious growth promoting effect on the overground part of the common andrographis herb.
(2) As shown in Table 2, after sesbania stem tumor nitrogen-fixing rhizobia of different concentration groups is infected for 100 days, the root length, the number of lateral roots and the dry weight of underground parts of each concentration group have no obvious difference, which indicates that the infection of the bacterial liquid has no obvious influence on the underground parts of the andrographis paniculata plants. And 1.0X 107Concentration treatment group of one/mL and 1.0X 109The fresh and dry aerial weight of the aerial parts of the individual/mL concentration treatment group were both significantly higher at the level (P < 0.05) than the blank group; indicating 1.0X 107Concentration treatment group of one/mL and 1.0X 109The concentration treatment group per mL has obvious growth promoting effect on the overground part of the common andrographis herb.
TABLE 1 sesbania Stem tumor Nitrogen-fixing Rhizobium influence on overground part growth of Andrographis paniculata (n ═ 3)
Note: the data in the table are mean ± sd, with different lower case letters on the same column indicating significant differences (P < 0.05).
TABLE 2 sesbania Stem tumor Nitrogen-fixing Rhizobium influence on aerial and underground parts of Andrographis paniculata (n ═ 3)
Note: the data in the table are mean ± sd, with different lower case letters on the same column indicating significant differences (P < 0.05).
Example 2 Effect of sesbania Stem tumor Nitrogen fixing Rhizobium on the accumulation of effective ingredients of Andrographis paniculata
1. Method of producing a composite material
Referring to the pharmacopoeia of 2015 edition, the content of andrographolide is determined by high performance liquid chromatography.
(1) Control solution: accurately weighing appropriate amount of andrographolide control substance and dehydroandrographolide control substance, adding methanol to obtain mixed solution containing andrographolide control substance 0.1228mg/mL and dehydroandrographolide 0.1482mg/mL per l mL, and filtering with 0.45 μm microporous membrane.
Chromatographic conditions are as follows: Hydrosphere-C18 column (250 mm. times.4.6 mm, 5 μm), mobile phase: (methanol: water 52: 48), column temperature: 25 ℃, detection wavelength: the detection wavelength of andrographolide is 225nm, and the detection wavelength of dehydroandrographolide is 254 nm;
(2) preparation of a test solution: the whole plant of the andrographis paniculata sample plant of example 1 above was collected, and each treatment component was divided into root, stem, leaf and aerial part groups. Drying in the shade for 48h, baking in an oven at 60 ℃ for 10h, pulverizing with a pulverizer, sieving with a sieve of four sizes, sieving with about 0.5g, precisely weighing, placing in a conical flask with a plug, precisely adding 25mL of 40% methanol, weighing, soaking for 1h, ultrasonically treating (power 250W, frequency 33kHz) for 30min, cooling, weighing again, supplementing the lost weight with 40% methanol, shaking up, and filtering. Precisely measuring 10mL of the subsequent filtrate, eluting on a neutral alumina column (200-300 meshes, 5g, the inner diameter of 1.5cm) by using 15mL of methanol, collecting eluent, putting the eluent into a 50mL measuring flask, adding the methanol to the scale, and shaking uniformly.
(3) The determination method comprises the following steps: flow rate: 0.8 mL/min; sample introduction amount: 5 mu L, the flow rate is 0.8 mL/min; detection time: 60 min; the number of theoretical plates: calculated according to andrographolide and dehydroandrographolide, the content of andrographolide is not lower than 2000.
2. Results
As can be seen from Table 3, the yield of Andrographis paniculata seedlings treated with the sesbania stem nodosa azorhizobium inoculant solution can be significantly improved without applying fertilizers. Wherein, 1.0 is multiplied by 109The yield increase effect of each concentration treatment group is the best, and each 667m2The yield of the andrographolide can be improved by 112.32% to the maximum.
TABLE 3 influence of sesbania stem tumor nitrogen-fixing rhizobium treatment on the yield per mu of andrographis paniculata (n is 3)
The method of the invention can not only improve the growth of the overground part of the andrographis paniculata plant, but also improve the contents of andrographolide and dehydroandrographolide.
Example 3 Effect of different Rhizobium soaking and watering treatments on seed Germination and plant growth of Andrographis paniculata
1. Materials and apparatus
(1) Experimental Material
Chinese rhizobium freudenreichizus, Rhizobium radiobacter (purchased from Guangdong province microorganism collection center), and sesbania stem nodule Azorhizobium ORS 571 were provided by Liuhua Wei teacher of northwest agriculture and forestry science and technology university, and Andrographis paniculata seeds were produced in Xingheng county of Yulin city of Guangxi province and identified as mature seeds of Andrographis paniculata (Burm.f.) Nees by professor Dujiu in the institute of medicinal and botanical science and research of Guangzhou Chinese medicinal university.
(2) Laboratory apparatus and reagent
Plant SOD reagent box (Nanjing institute of bioengineering, lot number: 20181030); tryptone (national group chemical Co., Ltd., batch No. 20151202); yeast extract powder (Guangdong Huanji Biotech Co., Ltd., lot: 3205048), other reagents were all analytical grade.
JJ300 type electronic balance (guangzhou xiang instrument electromechanical devices limited); constant temperature cultivation shaker (Shanghai-Hengchun scientific instruments Co., Ltd.); QF-PRX-160 Intelligent phytotron (Shanghai georgen industries, Inc.); GXX-9023MBE electric heating air blast drying oven (Shanghai Boxun industries, Ltd.) medical equipment factory; TG6-W minim high-speed centrifuge (Changshan appearance centrifuge, Inc.); model 755B ultraviolet spectrophotometer (shanghai precision scientific instruments ltd).
2. Experimental methods
(1) Rhizobium seed soaking treatment
The andrographis paniculata test seeds are placed in culture bottles, 250 granules are placed in each bottle, and the total number of the bottles is 4. Taking rhizobia bacteria liquid in logarithmic phase, counting the number of bacteria (counting by swimming bacteria) by using a blood cell counting method, and diluting to 1.0 multiplied by 107Adding each/mL into the above 4 culture flasks, and adding distilled waterTreatment as a control, soaking for 24 h. Placing 2 layers of gauze and 1 layer of filter paper at the bottom of each culture dish, grouping according to different treatments, and adding 6mL of distilled water into each culture dish to saturate the gauze and the filter paper in a liquid absorption manner. And transferring the soaked common andrographis herb seeds into culture dishes for germination experiments, uniformly placing 50 seeds in each culture dish for 5 times, weighing the culture dishes and the seeds together, recording the weight A, and culturing in a constant-temperature culture box with the temperature of 28 ℃, the humidity of 60 percent and the light of 12 h/dark of 12 h. Adding water to the original weight every 12h, and counting the germinated seeds every 24h until no germinated seeds are germinated after 2 d. After the germination experiment is completed, the radicle length of the seeds is measured, and the vitality index is calculated. And randomly selecting 20 seedlings from each concentration group, and planting the seedlings into a seedling tray.
Germination percentage (%) (number of seeds normally germinated at the end of germination/number of seeds for feeding) × 100
Germination potential (%) - (number of seeds normally germinated after d/number of seeds for feeding) × 100
Germination index (germination number after seed soaking d/seed supply number)
Vigor index ═ germination index × germinating seed length (radicle length + hypocotyl length)
(2) Rhizobium irrigation treatment
Soaking andrographis paniculata test seeds in sterile water for 24h, transferring the seeds to culture dishes with corresponding labels and 2 layers of wet filter paper laid on the corresponding labels, placing the culture dishes in a plant incubator for culturing, adding a proper amount of distilled water every day to keep the filter paper wet, recording the germination number of the seeds until no seeds germinate for 2 days continuously, transplanting the seeds to a seedling tray for culturing, and dividing the seeds into 4 groups of 20 seeds.
Taking rhizobium bacterial liquid in logarithmic phase, diluting to 1.0 multiplied by 107each/mL, the roots of the andrographis paniculata seedlings on the seedling tray which is watered with the corresponding label at 1, 3 and 5 days respectively, each seedling is 5mL, and the blank group is replaced by sterile water.
(3) Detection of plant copper-zinc superoxide dismutase (Cu-Zn superoxide dismutase) as physiologically active substance of andrographis paniculata seedlings
After the rhizobia is irrigated, the seedlings are cultured for 40d in a seedling tray, the whole plant of the andrographis paniculata seedlings of different treatment groups is taken, 3 parts of each 0.2g is accurately weighed, and the content of total superoxide dismutase and copper-zinc superoxide dismutase (CuZn-SOD) of the plants is determined by adopting a Nanjing kit building method.
(4) Data analysis method
Statistical analysis is performed by using SPSS 18.0 software, and the metering data conforms to the use of normal distribution
Performing statistical description, and describing by using M (P25, P75) if the distribution does not conform to the normal distribution; two groups of measurement data conforming to normal distribution and homogeneity of variance are subjected to two independent sample t tests, and a plurality of groups of data are subjected to single-factor variance analysis tests; the two groups of measurement data which do not conform to normal distribution and variance homogeneity are compared by adopting non-parameter Mann-Whitney U test, and the multiple groups of measurement data are compared by adopting non-parameter Kruskal-Wallis H test; p values less than 0.05 are considered statistically different.
3. Results
(1) Effect of Rhizobium soaking on seed germination of Andrographis paniculata
By 1.0X 107Soaking seeds of the Chinese rhizobium freudenreichii, the sesbania stem nodule azotobacter and the Rhizobium radiobacter for 24 hours, transferring the seeds into a culture dish for germination, recording the number of the germinated seeds every day till no germinated seeds are continuously generated for 2 days, measuring the length of the radicle, and calculating the activity index of the radicle, wherein the experimental result is shown in table 4.
TABLE 4 Effect of different Rhizobium soaking treatments on cardiotropha seeds (n. 5)
Note: the data in the table are mean ± sd, with different lower case letters on the same column indicating significant differences (P < 0.05).
As shown in Table 4, 3 kinds of Rhizobium freudenreichii, sesbania stem nodule azotobacter and Rhizobium radiobacter, which have significant promoting effects on elongation of radicle of Andrographis paniculata and activity index of seed, are significantly higher than those of blank group at P < 0.05 level, wherein the stimulating effect of sesbania stem nodule bacterial seed soaking is the best.
(2) Influence of different rhizobia seed soaking treatment on growth of pistacia stratiotes seedlings
And (3) soaking different rhizobia, transplanting for 40d, randomly selecting 9 seedlings in each concentration group, measuring plant height, leaf number, leaf area, root length and fresh weight, and obtaining experimental results shown in tables 5 and 6.
TABLE 5 Effect of Rhizobium after soaking and transplanting 40d on the growth of seedlings of Andrographis paniculata (n-3)
Note: the data in the table are mean ± sd, with different lower case letters on the same column indicating significant differences (P < 0.05).
As can be seen from Table 5, after the seeds of Andrographis paniculata were treated by soaking seeds of Sinorhizobium freudenrz, Sinorhizobium sessiliflorum and Rhizobium radiobacter, and the leaves of the treated group of Sinorhizobium sessiliflorum and Rhizobium radiobacter were transplanted for 40 days, the leaf area of the treated group was significantly larger than that of the blank group, and the leaf area of the other treated groups was not significantly different from that. The results show that: after 3 kinds of rhizobia are soaked in the seed and transplanted for 40 days, the sesbania stem nodule nitrogen-fixing rhizobia treatment group has a good effect of promoting the growth of the andrographis paniculata plants.
(3) Influence of different rhizobia irrigating on superoxide dismutase of cardiotropha plantlets
After different rhizobia are irrigated and transplanted for 40 days, the activity of total superoxide dismutase (T-SOD) and copper-zinc superoxide dismutase (CuZn-SOD) of the plants is measured, and the experimental results are shown in table 6.
TABLE 6 Effect of different Rhizobium on SOD of seedlings of Andrographis paniculata (n. 3)
As can be seen from Table 6, the T-SOD and CuZn-SOD enzyme activities of the azotobacter nodule treatment group were the highest after the Andrographis paniculata seedlings were irrigated with 3 kinds of rhizobia, and after 40 days of transplantation, the T-SOD and CuZn-SOD enzyme activities of the azorhizobium nodule treatment group were significantly lower than those of the blank group. The result shows that the irrigation treatment of sesbania stem nodule nitrogen-fixing rhizobia can promote the physiological action of an antioxidant enzyme system of plant cells of andrographis paniculata plants, which indicates that the irrigation treatment of sesbania stem nodule nitrogen-fixing rhizobia can enhance the antioxidant capacity in the organism of andrographis paniculata seedlings, thereby enhancing the stress resistance and disease resistance of the organism.
The above detailed description is of the preferred embodiment for the convenience of understanding the present invention, but the present invention is not limited to the above embodiment, that is, it is not intended that the present invention necessarily depends on the above embodiment for implementation. It will be apparent to those skilled in the art that any modification of the present invention, equivalent substitutions of selected materials and additions of auxiliary components, selection of specific modes and the like, which are within the scope and disclosure of the present invention, are contemplated by the present invention.
Claims (8)
1. The application of sesbania stem tumor nitrogen-fixing rhizobia in promoting the growth of andrographis paniculata plants and/or improving the content of active ingredients of andrographis paniculata;
the promotion of the growth of the andrographis paniculata plants refers to the promotion of the growth of overground parts of the andrographis paniculata plants;
the effective component content of herba Andrographitis refers to total lactone content of herba Andrographitis.
2. The use of claim 1, wherein the andrographolide comprises andrographolide and dehydroandrographolide.
3. A method for promoting the overground part growth of andrographis paniculata plants and improving the total lactone content of andrographis paniculata by sesbania stem tumor nitrogen-fixing rhizobia is characterized in that sesbania stem tumor nitrogen-fixing rhizobia and andrographis paniculata are subjected to symbiotic cultivation; soaking the andrographis paniculata seeds by utilizing sesbania stem nodule azorhizobium bacteria liquid, or distributing the sesbania stem nodule azorhizobium bacteria liquid around the root system of the andrographis paniculata plants.
4. The method of claim 3, wherein the concentration of the liquid of the nodule bacteria azotobacter of sesbania is greater than or equal to 1.0 x 107one/mL.
5. The method of claim 4, wherein the concentration of the liquid of the nodule bacteria azotobacter of sesbania is 1.0 x 107～1.0×108one/mL.
6. The method of claim 3, wherein the Andrographis paniculata plants are young Andrographis paniculata Nees; the andrographis paniculata seedling is the time when the andrographis paniculata seeds grow to 2 cotyledons and are completely unfolded after germination is finished.
7. A method according to any one of claims 3 to 6, comprising the steps of: irrigating the andrographis paniculata seedlings with sesbania stem nodule nitrogen-fixing rhizobium liquid, distributing the sesbania stem nodule nitrogen-fixing rhizobium liquid around the root systems of the andrographis paniculata, and cultivating the andrographis paniculata in 4-7 months under natural conditions for 3-4 months; the dosage of the sesbania stem nodule azotobacter rhizobium bacterial liquid is 5-15 mL/time of one andrographis paniculata seedling, and the total dosage is 2-3 times.
8. A fertilizer capable of promoting the growth of overground parts of andrographis paniculata plants and increasing the content of total lactones of andrographis paniculata, which is characterized by comprising sesbania stem nodule nitrogen-fixing rhizobia.
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| EP3193615A1 (en) * | 2014-09-19 | 2017-07-26 | Taxon Biosciences, Inc. | Plant growth-promoting microbes, compositions, and uses |
| CN106106142A (en) * | 2016-06-27 | 2016-11-16 | 广州中医药大学 | A kind of method inducing Herba Andrographis polyploid |
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| CN107949280A (en) * | 2015-08-20 | 2018-04-20 | 孟山都技术公司 | For obtaining the high throughput method of seed treatment resistant one |
| CN107318433A (en) * | 2017-07-28 | 2017-11-07 | 周运德 | The method for improving ginsenoside Re's content in cultivation field seven |
| CN109122151A (en) * | 2018-08-10 | 2019-01-04 | 李妍 | It is a kind of to improve the cultural method for guaranteeing Radix Paeoniae Alba drug effect |
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