CN114080962A - Bletilla striata seedling growth promoting method based on Piriformospora indica - Google Patents
Bletilla striata seedling growth promoting method based on Piriformospora indica 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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
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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
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
Abstract
The invention discloses a bletilla striata germchit growth promoting method based on Piriformospora indica, which comprises the following steps: sterilizing a substrate for transplanting and domesticating bletilla striata seedlings to obtain a sterilized seedling substrate; the preparation concentration is 25-50 g.L‑1The Indian pear-shaped spore bacterial liquid or the prepared concentration is 1-3 multiplied by 108cfu/mL of Piriformospora indica spore liquid; soaking or irrigating bletilla striata seedlings out of the bottle by using an Indian pyricularia indica bacterial liquid or an Indian pyricularia indica spore liquid; transplanting rhizoma Bletillae to the matrix, and planting until outplanting. The method of the invention can shorten the domestication period of the bletilla striata tissue culture seedlings and make the bletilla striata seedlings out of the nursery in advance.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a bletilla striata germchit growth promoting method based on Piriformospora indica.
Background
Bletilla striata (Thunb) Reichb. f.), the alias desmodium, rhizoma nardostachyos and radix cynanchi bungei, which are perennial herbaceous plants of Bletilla striata of the family orchidaceae, purple or pink flowers not only have high ornamental value, but also are one of the traditional Chinese medicines in China, and are recorded in pharmacopoeia of the people's republic of China, wherein dry tubers are mainly used as medicines, and the medicines can be harvested after being planted for 4-5 years.
The rhizoma bletillae dried tubers mainly contain bibenzyls, phenanthrenes and derivatives thereof, have the effects of astringing to stop bleeding, diminishing swelling and promoting granulation and the like, and are clinically used for externally treating traumatic hemorrhage, scalds, sore treatment and the like, and internally treating hematemesis, lung diseases, hemoptysis, chronic gastric ulcer, tumors and the like. Modern medicine proves that the bletilla has stronger antioxidation and anti-aging effects and can be used for health care, skin care and beauty maintenance. In addition, the bletilla striata can be used as a thickener to prepare cigarette butts of high-grade cigarettes, is industrially used for starching silk, slashing or coating raw materials, brewing and the like, and the bletilla striata polysaccharide gum can also be used as a natural coating preservative to be applied to fruit and vegetable preservation and used as a natural food thickener to be used for food processing.
Bletilla striata has wide application, but wild resources are endangered due to long-term mining, and are listed in secondary protective plants in Ming nations ' famous records of national intensive protective wild plants in 2021 in 9 th and 7 th days, and are also one of varieties cultivated intensively by ' pure six flavor ' and ' thoroughfare six flavor '. As the wild bletilla striata resources are listed in the 'national key protection wild plant record' secondary protection plants, the artificial cultivation of the bletilla striata seedlings by digging up the wild resources becomes impossible, the traditional division propagation coefficient is low, the variety degradation is easy to cause, the seedling production only depends on seed propagation or tissue culture, the stem tip tissue culture has long period and high cost, so that the bletilla striata seedlings are produced mostly in a seed sterile propagation mode, but the problems of long production period (generally more than 6 months), weak seedling growth, slow growth, environmental sensitivity, small flower, delayed flowering and the like of the bletilla striata tissue culture seedlings are found in the artificial cultivation, and the rapid development of industrialization and artificial cultivation of the bletilla striata seedlings is severely restricted. The invention provides a bletilla striata germchit growth promotion method based on Piriformospora indica to solve the problem of production of artificially cultivated bletilla striata germchits and promote healthy development of bletilla striata germchit industry.
Patent 201510921991.1 provides a method for rapidly producing bletilla striata germchit, liquid co-culturing bletilla striata seeds and bletilla striata mycorrhizal fungi until the bletilla striata seeds germinate, and culturing the germinated bletilla striata seeds with the germchit, wherein the bletilla striata mycorrhizal fungi is a mycorrhizal fungi strain YY-51 of ceriporida (Sebacinales sp.), the bletilla striata mycorrhizal fungi is preserved in the China general microbiological culture Collection center of China general microbiological culture Collection management Committee in 2015 at 7-31 days, the preservation address is No. 3 of the Ministry of China academy of sciences, the preservation number is GCMCC NO. 11104. The artificial bletilla striata seeds or seedlings prepared by the method are easy to preserve and transport, the complex operation process and high technical requirement in the traditional tissue culture process are avoided, a large amount of manpower and material resources are needed in the processes of multiple subculture, transplantation or field planting, and the production cost is low. Although only mycorrhizal fungi culture and with bletilla rhizome seed coculture and artifical seed embedding etc. need aseptic technique in this patent, but the preliminary treatment procedure of bletilla rhizome seed is complicated has improved the contaminated risk of seed, and bletilla rhizome seed and bletilla rhizome mycorrhizal fungi's liquid culture is changeed than solid culture and pollutes, this patent artifical seed is grown seedlings the matrix upper portion sprays the mixed solution that water one deck volume ratio is 1: 3: 5 coconut juice, banana juice and tomato juice formed, is the rich nutrient source of miscellaneous fungus in the matrix of growing seedlings, can lead to miscellaneous fungus to breed in the matrix in a large number, causes artifical seed rotten or influences seed germination and growth, has only described the survival rate of transplanting the field after 3 months in the artifical seed seeding nursery pot in addition, and has not explained the specification of the seedling of transplanting this moment, and the cycle of the size influence vigor and patent medicine of seedling.
Patent 201410265619.5 provides a culture method of oncidium: (1) culturing the tissue culture plantlets of the oncidium by adopting an MS culture medium; (2) culturing Piriformospora indica by using a solid PDA culture medium; (3) co-culturing oncidium and Piriformospora indica; (4) and (4) taking out the culture bottle of the tissue culture seedlings of oncidium. The culture method reduces the disease rate of the bottled seedlings of oncidium and improves the stress resistance.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a bletilla striata germchit growth promoting method based on Piriformospora indica, which can shorten the domestication period of bletilla striata tissue culture germchits and make the bletilla striata seedlings out of a nursery in advance.
In order to solve the technical problems, the invention provides a bletilla striata germchit growth promotion method based on Piriformospora indica, which comprises the following steps:
(1) sterilizing a substrate for transplanting and domesticating bletilla striata seedlings to obtain a sterilized seedling substrate;
the sterilized seedling culture substrate is respectively arranged in small plastic square pots with the caliber of 7cm for later use;
(2) the preparation concentration is 25-50 g.L-1The Indian pear-shaped spore bacterial liquid or the prepared concentration is 1-3 multiplied by 108cfu/mL of Piriformospora indica spore liquid for later use;
(3) any one of the following modes is selected:
the method A comprises the following steps: cleaning root agar of bletilla striata seedlings out of a bottle, sucking water on the surface of the roots, soaking the seedlings in a print pear spore liquid or an Indian pear spore liquid for 24 +/-2 hours (so that the roots of the seedlings are fully contacted with the Indian pear spores), taking out the seedlings, cleaning the Indian pear spore liquid or the Indian pear spore liquid adhered to the surfaces of the roots with clear water, and planting the seedlings in a sterilized seedling culture substrate;
the method B comprises the following steps: cleaning root agar of bletilla striata seedlings out of a bottle, sucking water on the surface of roots, directly planting the seedlings in a sterilized seedling substrate, irrigating with prepared piriformospora indica liquid or piriformospora indica spore liquid, and irrigating each plant with 10 +/-1 mL of piriformospora indica spore liquid or piriformospora indica spore liquid;
(4) transplanting rhizoma Bletillae to the matrix, and planting (watering and fertilizing according to conventional method) until outplanting.
As an improvement of the bletilla striata germchit growth promotion method based on Pilus indica, the method comprises the following steps of (2):
the preparation method of the Indian pyriform spore liquid comprises the following steps:
adding water (sterilized water) into mycelia blocks obtained after the Piriformospora indica is cultured on a PDB culture medium, and scattering to prepare the mycelia blocks with the concentration of 25-50 g.L-1The bacterial liquid (broken up by a high-speed homogenizer);
the Piriformospora indica spore liquid is as follows:
adding physiological saline into Piriformospora indica cultured in PDA culture medium to prepare the Piriformospora indica with the concentration of 1-3 multiplied by 108cfu/mL spore fluid (spore concentration was calculated using a cytometer).
As a further improvement of the bletilla striata germchit growth promotion method based on piriformum indica of the invention: in the step (4), fertilizer can be irrigated after 30d of the bletilla striata transplanting matrix, and then the fertilizer is irrigated every 15d until the bletilla striata transplanting matrix is outplanted, wherein the fertilizer is N: p: k is 20: 10: 20, the irrigation concentration is 2 liters of water per 1 g, and the volume of the fertilizer for each irrigation is the same and is 40 mL.
As a further improvement of the bletilla striata germchit growth promotion method based on piriformum indica of the invention: in the step (1), the seedling substrate is sterilized at the high temperature of 121 ℃ for 60 min; or using the commercially accepted soil disinfectant, namely the dazomet, to operate according to a conventional disinfection method.
In step (4) of the present invention: after the rhizoma bletillae is transplanted with the matrix, spraying or watering the leaf surfaces within 15d according to the dryness and wetness of the leaves and the matrix; after new roots grow out, the fertilizer can be applied, and then the fertilizer is applied at certain intervals until the seedlings are outplanted, which is a conventional technology.
In general: the bottle-out and bletilla striata seedlings require that the pseudobulb reaches 5mm and the plant height reaches 8 cm.
The invention realizes the growth promotion of the white and the seedlings by adding Piriformospora indica, and the seedlings can be transplanted to a field after being transplanted and domesticated for 120 days.
The invention adopts Piriformospora indica with proper concentration to realize the growth promotion effect on white and seedlings.
Preferably, the method comprises the following steps: the technical scheme of the invention utilizes 25-50 g.L-1Soaking rhizoma bletilla seedling in Indian Pyricularia piriformis bacterial solution for 24h or 2 × 108L-110mL of Indian pyricularia spore liquid is poured into each strain, which is favorable for establishing the symbiotic relationship between Indian pyricularia and bletilla striata seedlings, the increase multiple of the fresh weight of 120d bletilla striata seedlings and the increase multiple of the diameters of pseudo bulbs respectively reach 3.777 +/-0.761-4.353 +/-0.112 and 1.522 +/-0.384-1.627 +/-0.208, the average height of the overground part of 120d reaches more than 21.10 +/-0.69-21.15 +/-1.66 cm, the average diameter of the pseudo bulbs reaches 15.582 +/-1.609-16.348 +/-0.790 mm, the average number of roots of each strain reaches 12.6 +/-0.7-12.8 +/-1.8, and the standard of first-class seedlings of bletilla striata seedlings nursery seedlings (the diameter of the pseudo bulb is more than or equal to 15mm, the height of the plant is more than or equal to 12cm, the robust plant, the root needs to be complete, and the root needs no mildew and bletilla striata normal acclimation period of at least 180 d) according to the mass classification standard of the bletilla striata 2019/DB 53/T2019. The method lays a good foundation for shortening the production period of the bletilla striata germchit and improving the quality of the bletilla striata germchit in future.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:
in the following examples:
the seedling culture substrate is as follows: lambert seedling substrate, model LM-2, manufactured by Lambert PEAT INC.
The PDB medium is conventional PDB medium (containing 200 g.L)-1Potato and cane sugar 20 g.L-1)，
The PDA culture medium is conventional PDA culture medium (containing 200 g.L)-1Potato and cane sugar 20 g.L-1Agar 10 g. L-1)，
The Piriformospora indica can be used as a Piriformospora indica strain in a paper of 'preliminary study on the disease resistance, growth promotion and stress resistance of Brassica oleracea and mechanism thereof by Piriformospora indica' (Master thesis of Zhejiang university, Sunji, 2010).
The culture dish is a conventional culture dish.
Example 1 growth promoting Effect of addition of Piriformospora indica on white and tissue culture seedlings
(1) And (3) disinfection of the transplanting matrix: sterilizing the seedling substrate at high temperature, cooling to room temperature (sterilizing at 121 deg.C for 60min, and cooling to room temperature), and packaging in small square pots with diameter of 7 cm; as a sterilized seedling raising substrate for later use;
(2) preparation of Piriformospora indica/spore liquid
Preparing an Indian pyriform spore liquid:
culturing Piriformospora indica in culture dish containing PDA solid culture medium at 25 + -2 deg.C with photoperiod of 12 h.d-110-15 days; the edge of the colony of the Piriformospora indica cultured on the PDA solid culture medium is punched by a stainless steel puncher with the diameter of 6mm to obtain Piriformospora indica blocks with the diameter of 6mm and containing vigorous hyphae.
300mL of PDB culture medium is filled in a culture container, 3 Piriformospora indica blocks with the diameter of 6mm are added, the mixture is placed in a shaking table for shaking culture, the culture temperature is 27 +/-1 ℃, the culture speed is 180rpm, the mixture is cultured for 15-20 days, and the dark culture is carried out; centrifuging the bacterial liquid at 6000rpm for 5min, taking a hypha block positioned at the lower layer obtained by centrifugation, adding sterilized water according to a material-liquid ratio of 25-50 g/L, and scattering by using a high-speed homogenizer to prepare the bacterial liquid (namely, the concentration of the bacterial liquid is 25-50 g/L);
② preparation of Indian pyriform spore liquid
Culturing Piriformospora indica in culture dish containing PDA solid culture medium at 25 + -2 deg.C with photoperiod of 12 h.d-1And 10-15 days.
Then adding 10-15 mL of physiological saline containing 0.5% (volume%) Tween into the Piriformospora indica cultured in the PDA solid culture dish, and scraping the surface of a bacterial colony by using a medicine spoon with a thinner edge to enable spores to be separated from hyphae; filtering with gauze 20min after ultrasonic separation to remove impurities, calculating spore concentration with cell counting plate, and diluting to required concentration as required.
(3) Preparing bletilla striata seedlings: cleaning root agar of bletilla striata tissue culture seedlings with pseudobulb being more than or equal to 5mm and plant height being more than or equal to 8cm, wherein the pseudobulb and the plant height meet the bottle-out standard, and sucking water on the surface of roots for later use;
description of the drawings: the culture method of rhizoma Bletillae tissue culture seedling refers to published pseudo bulb induction test in rhizoma Bletillae bottle, and the formula of the culture medium is MS +30 g.L-1Sucrose +6-BA 1.0 mg.L-1+KT 0.5mg·L-1+NAA0.2rng·L-1And agar until bletilla striata tissue culture seedling (pseudobulb is more than or equal to 5mm, plant height is more than or equal to 8cm) which can be taken out of the bottle is obtained, and the number of roots is generally 3-4.
(4) Addition of Piriformospora indica:
mode a (addition mode is soaking):
soaking the roots of the bletilla striata seedlings in the step (3) in the Piriformospora indica bacterial liquid or the spore liquid prepared in the step (2) for a period of time (about 24 hours), taking out the bletilla striata seedlings, washing the Piriformospora indica bacterial liquid or the spore liquid adhered to the surfaces of the roots with clear water, and planting the Piriformospora indica bacterial liquid or the spore liquid in a sterilized seedling culture substrate;
the concentration of the Piriformospora indica liquid is respectively 25, 50 and 100 g.L-1(ii) a The spore liquid concentration is 2 × 107、2×108、2×109L-1；
Mode B (addition mode is irrigation):
directly planting the roots of the bletilla striata seedlings in the step (3) in a sterilized seedling raising substrate, irrigating with the Indian pyricularia liquid or spore liquid prepared in the step (2), and irrigating 10mL of the Indian pyricularia liquid or spore liquid for each plant;
the concentration of the Piriformospora indica liquid is 50 g.L-1The concentration of spore liquid is 2 × 108L-1。
(5) Transplanting, domesticating and managing bletilla striata seedlings:
and (4) carrying out the following conventional treatment on the bletilla striata seedlings transplanted into the seedling culture substrate obtained in the step (4):
spraying or watering the leaf surfaces within 15 days according to the dryness and wetness of the leaves and the matrix; spraying or watering can be carried out according to published technical research on germination characteristics of rhizoma bletillae radix rehmanniae and hardening of tissue culture seedlings;
after new roots grow out, the fertilizer can be applied, and then the fertilizer is applied at certain intervals until the seedlings are out of nursery. In the step, fertilizer can be irrigated after 30d of rhizoma bletillae transplanting matrix, and then the fertilizer is irrigated once every 15d until the rhizoma bletillae is outplanted, wherein the fertilizer is N: p: the mass ratio of K is 20: 10: 20 (mass ratio), the irrigation concentration is 2 liters of water per 1 g, and the volume of each irrigation fertilizer is the same and is 40 mL/time.
(6) After the bletilla striata seedlings are transplanted for 120 days, 15 plants are randomly selected for each group of treatment, the indexes such as fresh weight, pseudobulb diameter, overground part height, root number and the like are measured, and the increase multiple of each index is calculated respectively. The results obtained are shown in Table 1 below.
TABLE 1
The method takes cultured bletilla striata tissue culture seedlings as materials, and promotes the growth of the bletilla striata seedlings by screening proper addition concentration and mode of the piriformospora indica. Addition mode of Piriformospora indicaThe bacterial liquid addition concentration in A is 25-50 g.L-1Most suitably, through the technical method, the fresh weight growth multiple and the pseudobulb diameter growth multiple of the bletilla striata tissue culture seedlings which are transplanted and domesticated for 120d reach 3.777 +/-0.761-4.353 +/-0.112 and 1.522 +/-0.384-1.627 +/-0.208 respectively; spore liquid concentration 2X 10 in Piriformospora indica addition mode B8L-1Watering the common bletilla pseudobulb seedlings, wherein the fresh weight growth multiple and the pseudobulb diameter growth multiple of the common bletilla pseudobulb seedlings respectively reach 4.299 +/-0.263 and 1.714 +/-0.089, and the addition concentration of the common bletilla pseudobulb seedlings and bacterial liquid is 25-50 g.L-1The growth promoting effect is equivalent; the diameters of pseudo bulbs of the bletilla striata seedlings cultured for 120 days by the method can reach 15mm, the heights of the plants can reach more than 20cm, and the standards of first-class seedlings of the bletilla striata seedlings out of the nursery are met by referring to the quality grading standards of DB 53/T901-containing 2019 bletilla striata seedlings (the diameters of the pseudo bulbs are more than or equal to 15mm, the plant heights are more than or equal to 12cm, and the pseudo bulbs are robust, complete in root hairs and free of mildew and rot).
Blank comparative example, growth of bletilla striata tissue culture seedling without addition of Piriformospora indica
Bletilla striata seedlings were immersed in sterile water for 24 hours as in example 1, transplanted into a sterilized seedling substrate, acclimatized, managed and index-measured as in example 1, and the results are shown in table 1 above.
The fresh weight growth multiple and the pseudobulb diameter growth multiple of the transplanted bletilla striata tissue culture seedling of 120d are 2.351 +/-0.157 and 1.150 +/-0.257 respectively, the average height of the overground part is 15.88 +/-0.53 cm, the average diameter of the pseudobulb reaches 11.800 +/-1.022 mm, the average root number of a single plant reaches 8.6 +/-0.7, and the number is obviously lower than that of a treatment group added with Piriformospora indica.
Comparative example 1, addition of Pyricularia indica was carried out by changing the step (4) of example 1 as follows:
directly planting the roots of the bletilla striata seedlings in the step (3) in a sterilized seedling culture substrate, and embedding a 6mm Piriformospora indica block in the substrate at the position 0.5-1 cm away from the roots of each bletilla striata seedling; the rest is equivalent to embodiment 1.
The results obtained show that: the Indian pyricularia piriformis clods are embedded in the roots of the bletilla striata seedlings, the plant growth is equivalent to that of a blank control, and no obvious growth promoting effect exists.
From comparison with comparative example 1, it can be seen that: embedding the Piriformospora indica blocks at the root of the bletilla striata seedling, which is time-consuming and labor-consuming, and the bacterial liquid soaking or spore liquid irrigating is convenient and quick, and is convenient to operate above production; secondly, the components and the nutritional components of the Lanbo seedling substrate are not suitable for the growth of Piriformospora indica, and hypha is difficult to contact the root of rhizoma bletillae, so the growth promoting effect is not obvious.
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (5)
1. A bletilla striata germchit growth promotion method based on Pilus indica is characterized by comprising the following steps:
(1) sterilizing a substrate for transplanting and domesticating bletilla striata seedlings to obtain a sterilized seedling substrate;
(2) the preparation concentration is 25-50 g.L-1The Indian pear-shaped spore bacterial liquid or the prepared concentration is 1-3 multiplied by 108cfu/mL of Piriformospora indica spore liquid;
(3) any one of the following modes is selected:
the method A comprises the following steps: cleaning root agar of bletilla striata seedlings out of a bottle, sucking water on the surface of the root, soaking the bletilla striata seedlings in a print degree piriformis liquid or a piriformis indicum spore liquid for 24 +/-2 hours, taking out the bletilla striata seedlings, cleaning the piriformis indicum liquid or the piriformis indicum spore liquid adhered to the surface of the root with clear water, and planting the bletilla striata seedlings in a sterilized seedling substrate;
the method B comprises the following steps: cleaning root agar of bletilla striata seedlings out of a bottle, sucking water on the surface of roots, directly planting the seedlings in a sterilized seedling substrate, irrigating with prepared piriformospora indica liquid or piriformospora indica spore liquid, and irrigating each plant with 10 +/-1 mL of piriformospora indica spore liquid or piriformospora indica spore liquid;
(4) transplanting rhizoma Bletillae to the matrix, and planting until outplanting.
2. The method of growth promotion of bletilla striata (P.indica) -based seedlings according to claim 1, wherein in step (2):
the preparation method of the Indian pyriform spore liquid comprises the following steps:
adding water into hypha blocks obtained after the Piriformospora indica is cultured on a PDB culture medium and scattering the hypha blocks to prepare the hypha blocks with the concentration of 25-50 g.L-1The bacterial liquid of (a);
the Piriformospora indica spore liquid is as follows:
adding physiological saline into Piriformospora indica cultured in PDA culture medium to prepare the Piriformospora indica with the concentration of 1-3 multiplied by 108cfu/mL spore liquid.
3. The method of growth promotion of bletilla striata based seedlings according to claim 2, wherein: in the step (4), fertilizer is poured after 30d of bletilla striata transplanting matrix, and then fertilizer is poured once every 15d until the bletilla striata is outplanted, wherein the fertilizer is N: p: k is 20: 10: 20, 2 liters of water is added to each 1 g of irrigation concentration, and 40mL of fertilizer is irrigated to each plant each time.
4. The method of growth promotion of bletilla striata based seedlings according to claim 3, wherein: in the step (1), the seedling substrate is sterilized at the high temperature of 121 ℃ for 60 min.
5. The method for growth promotion of bletilla striata seedlings based on Piriformospora indica according to any one of claims 1 to 4, wherein:
the method A comprises the following steps: using 25-50 g.L-1Soaking the Piriformospora indica liquid for 24 hours;
the method B comprises the following steps: 10mL of water is poured into each plant, and the concentration is 2 multiplied by 108L-1The Piriformospora indica spore liquid.
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|---|---|---|---|---|
| CN115226580A (en) * | 2022-06-16 | 2022-10-25 | 河北工业大学 | Method for preventing and treating wheat sheath blight biogenic sources and application |
| CN115226580B (en) * | 2022-06-16 | 2023-08-15 | 河北工业大学 | Biological source method for preventing and treating banded sclerotial blight of wheat and application thereof |
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