CN114145197B - Method for improving resistance of wheat to scab - Google Patents
Method for improving resistance of wheat to scab Download PDFInfo
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- CN114145197B CN114145197B CN202111128163.4A CN202111128163A CN114145197B CN 114145197 B CN114145197 B CN 114145197B CN 202111128163 A CN202111128163 A CN 202111128163A CN 114145197 B CN114145197 B CN 114145197B
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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
- A01G22/20—Cereals
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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
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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
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
- A01N59/02—Sulfur; Selenium; Tellurium; Compounds thereof
Abstract
The invention discloses a method for improving gibberellic disease resistance of wheat, which relates to the technical field of plant growth regulation and control and comprises the following steps: (1) Preparing NaHS solution with the concentration of 0.1mM-1mM, and adding 0.1% -0.5% of attapulgite to prepare NaHS mixed solution; and (2) irrigating the prepared NaHS mixed solution to wheat. The invention has the beneficial effects that: (1) NaHS is used as an industrial byproduct, is cheap and easy to obtain, and is expected to be developed into a novel gibberellic disease prevention and control technical product. (2) The antibacterial agent has excellent antibacterial activity, and the coleoptile inoculation method result shows that: naHS treatment reduced the disease index by 97.60%.
Description
Technical Field
The invention relates to the technical field of plant growth regulation and control, in particular to a method for improving gibberellic disease resistance of wheat.
Background
Wheat is one of the main food crops, and the safe production of the wheat has important economic significance in the global range. However, many factors reduce the yield and quality of wheat, including abiotic and biotic stresses. Scab is evaluated as a head disease of wheat in China and even in the world, and is mainly expressed by early bleaching of wheat ears, so that the loss of yield and quality of wheat is caused, and huge economic loss is caused to agriculture all over the world every year. At the same time, gibberellic disease produces the mycotoxin Deoxynivalenol (DON) in cereals, and even asymptomatic wheat, which is infected to a low degree, can constitute a high level of emetic toxins, which are seriously detrimental to the health of humans and animals. Among them, fusarium graminearum is the most predominant pathogen causing head blight.
At present, the prevention and control of the gibberellic disease mainly depend on fungicides and variety breeding containing gibberellic disease resistance genes or loci. Although tens of thousands of wheat germplasm have been screened worldwide, fully resistant germplasm has not been discovered. The only genes that have been cloned at present are Fhb1 and Fhb7. In addition, commercial wheat breeding that combines desirable agronomic traits with a high level of gibberellic disease resistance remains a significant challenge. Resistance, agronomic and quality traits are controlled by multiple genes, and lines are obtained that are resistant to FHB, but either do not provide adequate protection for wheat in an environment particularly favorable to the disease or lack acceptability for single yield potential to meet the production needs of commercial wheat, thus requiring a comprehensive evaluation of the use of new resistant candidate varieties in production.
Application of a fungicide may provide some protection, for example, patent publication No. CN105248427A discloses a pesticide composition for controlling fusarium head blight through validamycin and triadimefon, but the cost of treatment and lack of a high-efficiency fungicide limit the use of chemical protection against fusarium head blight. Meanwhile, some fusarium graminearum strains show resistance to these chemicals in vitro tests, since triazole fungicides have long been used to control different pathogens on wheat. Therefore, the research and development of novel and efficient prevention and control technical products are urgently needed.
Hydrogen sulfide (H) 2 S) is a third gas signaling molecule discovered in recent years following NO and CO, and its function has been elucidated much in animals and plants. For example, patent publication No. CN107396935A discloses the use of sodium hydrosulfide as hydrogen sulfide donor for inhibiting the abscission of plant organs, but no research has shown that NaHS can improve the resistance of wheat to scab.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for improving the resistance of wheat to scab.
The invention solves the technical problems through the following technical means:
a method for improving resistance of wheat to scab comprises the following steps:
(1) Preparing NaHS solution with the concentration of 0.1mM-1mM, and adding 0.1% -0.5% of attapulgite to prepare NaHS mixed solution;
(2) And irrigating the prepared NaHS mixed solution to wheat.
Has the advantages that: the invention has obvious improvement effect on the resistance of wheat scab by adjusting the concentration of NaHS solution, and can reduce the disease index by 97.60 percent. Has excellent antibacterial activity and can inhibit the germination of gibberellic disease spores in vitro.
The raw material of the hydrogen sulfide donor NaHS used in the invention is cheap and easy to obtain, and has the characteristics of high efficiency and easy use; the method is simple to operate, low in cost and very suitable for plant factories.
The attapulgite can increase the adhesion effect of the solution and play a role in properly slowly releasing H 2 And the influence of the attapulgite on the plant per se is reduced through the proportion.
Preferably, a NaHS mother liquor is prepared and then diluted to a NaHS solution having a concentration of 0.1mM to 1mM.
Preferably, the NaHS is taken as H 2 A donor of S.
Preferably, the NaHS begins to release H upon dissolution in deionized water 2 S。
Preferably, the preparation method of the NaHS mother liquor comprises the following steps: 3.0g NaHS was weighed and 500mL deionized water was added to make a 75mM NaHS stock solution.
Preferably, the NaHS mother liquor is stored protected from light and at 4 ℃.
Preferably, the NaHS mother liquor is used for a week and is reconstituted after a week.
Preferably, the formulated NaHS mixture is irrigated to the wheat roots.
Preferably, the NaHS mixture is irrigated in an amount just to flood the wheat seeds.
Preferably, the irrigation is continued for 4 days and then the water is regained.
Preferably, irrigation is performed 2 times a day, 12h apart.
Has the beneficial effects that: naHS is instantaneous release H 2 Donor of S to sustain H 2 S Environment, irrigating NaHS twice a day to supplement H 2 S。
The invention has the advantages that: the invention has obvious improvement effect on the resistance of wheat scab by adjusting the concentration of NaHS solution, and can reduce the disease index by 97.60 percent. Has excellent antibacterial activity and can inhibit the germination of gibberellic disease spores in vitro.
The raw material of the hydrogen sulfide donor NaHS used in the invention is cheap and easy to obtain, and has the characteristics of high efficiency and easy use; the method is simple to operate, low in cost and very suitable for plant factories.
NaHS is instantaneous release H 2 S donor, to maintain H 2 S Environment, irrigating NaHS twice a day to supplement H 2 S。
Drawings
FIG. 1 is a comparison of the occurrence of scab of coleoptile after treatment with NaHS mixed solution and a control solution in example 6 of the present invention;
FIG. 2 is a graph showing the results of measuring the disease index of wheat scab after treatment with NaHS mixed solution and control solution in example 6 of the present invention;
FIG. 3 is a graph showing the results of measuring the disease index of wheat scab after treatment with NaHS mixed solution, control solution and other mixed solutions in example 6 of the present invention;
FIG. 4 is a graph showing the comparison of the occurrence of head blight of wheat after treatment with NaHS mixed solution and control solution in example 6 of the present invention
FIG. 5 is a graph showing the results of the measurement of the biological amount of wheat in example 7 of the present invention;
FIG. 6 shows Fg spore germination status after treatment with NaHS mixed solution and control solution in example 13 of the present invention;
FIG. 7 is a graph showing Fg spore germination assay results after treatment with NaHS mixed solution and control solution in example 13 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Test materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Those skilled in the art who do not specify any particular technique or condition in the examples can follow the techniques or conditions described in the literature in this field or follow the product specification.
Example 1
(1) Wheat culture (coleoptile inoculation)
The wheat seeds (Bainong 207) used in this study were from the laboratory, surface sterilized with 1% sodium hypochlorite solution for 5min, and then repeatedly rinsed in deionized water. The seeds were allowed to germinate by standing at 23 ℃ for one day. The mixture was moved to a refrigerator at 4 ℃ and left for two days. Seeds of consistent size and germination were selected and randomly distributed on petri dishes (14.5 cm diameter. Times.2.7 cm depth, 50 seeds per dish) with hood (13.6 cm diameter. Times.11 cm height) to maintain high humidity. Then cultured in a tissue culture chamber at 25 ℃ under 16h light/8 h dark photoperiod. Planting in the field at the later stage, and planting in a greenhouse.
(2) Wheat plant selection (ear inoculation)
Selecting a wheat variety Bainong 207 plant with good growth condition in a wheat flowering stage, transferring the plant to a light culture box, and culturing at 25 ℃ in 16h light/8 h dark light period.
(3) Preparation of Fusarium graminearum spore suspension
Fusarium graminearum is derived from laboratory identified and preserved species, and is identical to (Tang et al, 2020) transformed organisms culture for a novel flow type, cleistogamous while the Crop Journal,8 (2), 313-326, is used. Spore suspensions were made with reference (Buhrow, cram et al 2016) to the Exogenous Abscisic Acid and Gibberellic Acid Electron manipulation on fungal grandium grandistudy introduction in wheat. Phytopathology (2016) 106 (9), 986-996 with minor modifications. The spores were filtered through four layers of gauze, and the spore concentration was adjusted to 3.2X 10 by hemocytometer 7 Conidia/ml (0.1% tween 80) for wheat inoculation as follows, noting that the suspension inoculation time should be within 2h.
(4) Coleoptile inoculation
After the wheat of step (1) had grown for 2 days, the wheat coleoptile was inoculated with head blight, with specific reference (Wu, li et al 2005) Comparative pathology of Fusarium graminearum isolates from China renewable by wheat straw science (2005) 160 (1), 75-83 and with minor modifications. The coleoptiles of wheat seedlings were cut 1-2mm away from the tops, and 2. Mu.l of F.graminearum spore suspension was added to the tops of the remaining seedlings. Note that coleoptiles requiring inoculation should not be disrupted by the leaf.
(5) Preparing NaHS mixed liquid
Weighing 3.0g NaHS (purchased from Wolk), adding 500mL deionized water to prepare 75mM mother liquor, further diluting with deionized water to obtain 0.5mM NaHS solution with working concentration, and adding 0.1-0.5% attapulgite to prepare NaHS mixed liquor.
(6) Irrigation method
Experimental groups: and (4) after the scab is inoculated on the coleoptile wheat in the step (4), irrigating the root of the wheat by the prepared NaHS mixed solution until the wheat seeds are just submerged. The irrigation was continued for 4 days, 2 times per day, 12h apart. Rehydration took place after 4 days.
Example 2
The present embodiment is different from embodiment 1 in that: the working concentration of NaHS solution was 0.1mM.
Example 3
This embodiment is different from embodiment 1 in that: the working concentration of NaHS solution was 0.35mM.
Example 4
This embodiment is different from embodiment 1 in that: the working concentration of NaHS solution was 1.0mM.
Example 5
The comparative example differs from example 1 in that: inoculating the F.graminearum spore suspension to the wheat head part in the step (2) by using a syringe, and inoculating 4 wheat heads to each wheat plant, wherein the inoculation volume is 10 mu l.
And (5) after the scab is inoculated to the coleoptile wheat in the step (4), irrigating the root of the wheat by using a prepared 0.5mM NaHS mixed solution.
Comparative example 1
The comparative example differs from example 1 in that: irrigation of 0.5mM Na 2 And (S) mixing the solution.
Comparative example 2
This comparative example differs from example 1 in that: irrigated with 0.5mMNaHSO 3 And (4) mixing the solution.
Comparative example 3
This comparative example differs from example 1 in that: irrigate 0.5mM Na 2 SO 3 And (3) mixing the solution.
Comparative example 4
This comparative example differs from example 1 in that: the 0.5mM NaAC mixture was irrigated.
Comparative example 5
This comparative example differs from example 1 in that: irrigation of 0.5mM NaHSO 4 And (3) mixing the solution.
Comparative example 6
This comparative example differs from example 1 in that: irrigation of 0.5mM Na 2 SO 4 And (4) mixing the solution.
Comparative example 7
This comparative example is control 1, which differs from example 1 in that: the irrigation solution is prepared by mixing 0.1-0.5% of attapulgite with deionized water.
Example 6
The onset, disease index measurement and results of wheat (which had been inoculated with head blight: coleoptile and ear (example 5)) treated in control 1, example 2, example 3, example 4, comparative example 1, comparative example 2, comparative example 3, comparative example 4, comparative example 5 and comparative example 6.
And (3) determining disease indexes: the disease symptoms seen in the stem base 7 days after inoculation with gibberellic disease in the different treatment groups were scored, the stem base disease score (DIS) being the product of lesion length (mm) and lesion color (lesion color scale: 0, no lesion; 1, very slight brown necrosis; 2, mild/moderate brown necrosis; 3, extensive brown necrosis; 4, extensive black necrosis). Assay reference (Nicholson, simpson et al 1998) Detection and quantification of fungal culmorum and fungal graminearin assays using PCR assays, physical and Molecular Plant Pathology (1998) 53 (1), 17-37. The relevant experiments were repeated 3 more times.
The experimental results are as follows: as can be seen from FIG. 1, the stem base and the leaf of the control group had severe scab, the leaf had yellow rot and the stem base had black necrosis, and after the treatment with NaHS, the development of scab was significantly inhibited even at a lower working concentration, and the degree of scab in wheat was lower as the working concentration increased. As can be seen in FIG. 2, the effect was best for treatment with 0.5mM and 1.0mM NaHS, which reduced the disease index by 97.60% and 98.13%, respectively.
FIG. 3 shows the result is H 2 S or HS - Other compounds than NaHS promote the resistance of wheat to head blight, but the effect is significantly lower than NaHS.
However, as can be seen from FIG. 1, at a concentration of 1.0mM, the wheat leaves showed fragment yellowing, indicating H 2 Too high S content is not beneficial to the growth of plants. Therefore, the concentration and the content of NaHS need to be controlled during treatment. The results in FIG. 4 show that treatment of wheat inoculated with head scab with NaHS also significantly inhibited the development of head scab.
Example 7
The effect of the treatment of control 1, example 2, example 3, example 4, comparative example 1, comparative example 2, comparative example 3, comparative example 4, comparative example 5 and comparative example 6 on the biomass of wheat (inoculated with head blight) was determined.
The measuring method comprises the following steps: after 1 week of NaHS treatment (first 4 consecutive days of treatment, last 3 days of rehydration), the whole wheat plants of the different treatment groups were individually dried to constant weight at 65 ℃ and then the dry weight of the wheat was determined, each treatment consisting of three biological replicates.
And (3) measuring results: as can be seen from FIG. 5, the bio-accumulation of wheat stressed by gibberellic disease was increased by treatment with low concentrations of NaHS. The low concentration of NaHS was shown to treat wheat without stress.
Example 8
Plate bacteriostasis experiment
PDA medium was purchased from Biotech and prepared according to the instructions. A fg. Spore suspension was prepared according to example 1, step (3), and four aliquots of spore suspension (4 μ L) were placed in petri dishes 9cm in diameter, which were stored in sealed 1L containers.
H 2 S, fumigating: 100mL of NaHS mixed solution with working concentration of 0.5mM was placed in the bottom of the sealed container at 28 ℃The fungus is fumigated at a relative humidity of 90-95%. After 24h, the microscope was observed and photographed.
Example 9
This embodiment is different from embodiment 8 in that: the working concentration of NaHS was 0.01mM.
Example 10
This embodiment is different from embodiment 8 in that: the working concentration of NaHS was 0.05mM.
Example 11
This embodiment is different from embodiment 8 in that: the working concentration of NaHS was 0.075mM.
Example 12
This embodiment is different from embodiment 8 in that: the working concentration of NaHS was 0.1mM.
Comparative example 8
This comparative example is control 2, which differs from example 14 in that: the fumigated solution is prepared by mixing 0.1% -0.5% of attapulgite with deionized water.
Example 13
And F g, spore germination condition and germination rate determination after fumigation of control group 2, example 8, example 9, example 10, example 11 and example 12.
Spore germination rate was determined according to (Fu, hu et al 2014) An anti-uniform roll of hydrogen sulfite on the posthardest pathogens Aspergillus niger and Penicillium italicum. PLoS One (2014) 9 (8), e104206. Related experiments were repeated 3 more times.
The experimental results are as follows: fig. 6 and 7 show that the effect of low concentration of NaHS on fg. Spore germination was small at 24h, 0.075mM NaHS half inhibited spore germination, and 0.1mM, 0.5mM NaHS completely inhibited spore germination. Indicating a high concentration of excess H 2 S has obvious inhibition effect on germination of fusarium graminearum spores. H 2 S has excellent bacteriostatic activity.
The above examples are only intended to illustrate the technical solution of the present invention, and not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (5)
1. A method for improving the resistance of wheat to scab is characterized in that: the method comprises the following steps:
(1) Preparing NaHS solution with the concentration of 0.1mM-1mM, and adding 0.1% -0.5% of attapulgite to prepare NaHS mixed solution;
(2) Irrigating the prepared NaHS mixed solution on the root of the wheat; naHS as H 2 A donor of S; the irrigation quantity of the NaHS mixed liquor is to submerge the wheat seeds; continuously irrigating for 4 days, 2 times per day, and 12h every time; rehydration took place after 4 days.
2. The method of increasing resistance of wheat to head blight as claimed in claim 1, wherein: preparing NaHS mother liquor, and then diluting the mother liquor into NaHS solution with the concentration of 0.1mM-1 mM.
3. The method of increasing resistance of wheat to head blight as claimed in claim 2, wherein: the preparation method of the NaHS mother liquor comprises the following steps: 3.0g NaHS was weighed and prepared into a 75mM NaHS stock solution by adding 500mL deionized water.
4. The method of increasing resistance of wheat to head blight as claimed in claim 2, wherein: and storing the NaHS mother liquor at 4 ℃ in a dark place.
5. The method of increasing resistance of wheat to head blight as claimed in claim 2, wherein: the NaHS mother liquor is used for a week and needs to be reconstituted after a week.
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| BRPI0705038B1 (en) * | 2007-12-28 | 2020-10-06 | Petróleo Brasileiro S/A - Petrobras | SEED TREATMENT PROCESS WITH SHALE WATER FOR PATHOGEN CONTROL |
| CN101385466B (en) * | 2008-10-15 | 2011-06-08 | 合肥工业大学 | New use of sodium hydrosulfide for promoting wheat seeds sprouting under heavy metal stress |
| US20140310832A1 (en) * | 2011-03-28 | 2014-10-16 | Commonwealth Scientific And Industrial Research Organisation | Stress tolerant wheat plants |
| JPWO2012161133A1 (en) * | 2011-05-20 | 2014-07-31 | 日産化学工業株式会社 | Substituted pyridazine compounds and agricultural and horticultural fungicides |
| EP2773183A4 (en) * | 2011-11-04 | 2015-04-15 | Arista Cereal Technologies Pty Ltd | High amylose wheat - ii |
| EP3013803B1 (en) * | 2013-06-26 | 2018-05-16 | Bayer Cropscience AG | N-cycloalkyl-n-[(fusedphenyl)methylene]-(thio)carboxamide derivatives |
| CN104584727B (en) * | 2015-01-22 | 2017-10-10 | 河南农业大学 | It is a kind of that Drought-resistance in Wheat, the method for salt stress-resistant are improved using NaHS as hydrogen sulfide donor |
| CN107372483A (en) * | 2017-07-28 | 2017-11-24 | 中国科学院合肥物质科学研究院 | A kind of wheat scab inhibitor and preparation method thereof |
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