CN108848975B - Method for improving cassava disease resistance through betaine treatment - Google Patents
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- CN108848975B CN108848975B CN201810602395.0A CN201810602395A CN108848975B CN 108848975 B CN108848975 B CN 108848975B CN 201810602395 A CN201810602395 A CN 201810602395A CN 108848975 B CN108848975 B CN 108848975B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a method for improving cassava disease resistance through betaine induction treatment, which comprises the following steps: continuously spraying exogenous substance betaine solution with the concentration of 25-400 mu M to the leaf surface of the cassava seedlings in any period of the growth stage for 5-14 days. The method is easy to implement, simple and convenient to operate, and quick in effect, can improve the disease resistance of the cassava to the bacterial wilt in a short time, and has great significance for maintaining the continuous and healthy development of the cassava industry.
Description
Technical Field
The invention belongs to the technical field of cassava disease resistance, and particularly relates to a method for improving the cassava disease resistance through short-time betaine treatment.
Background
Cassava (Manihot esculenta Crantz), belonging to the genus Manihot of the family Euphorbiaceae, is one of three potato crops (potato, sweet potato and cassava) in the world, has the reputation of underground granaries, starch king and special crops, is a tropical subtropical high-starch crop, is also a main food source of more than seven hundred million people in the world, and has wide application in the aspects of food, feed processing, industrial raw materials and the like. The cassava has the advantages of drought and barren resistance, easy growth, low requirement on land, relatively low production cost, high photosynthetic efficiency and the like, and is widely planted in the world. In recent years, the cassava planting area in China reaches more than 50 million hectares, and is second to rice, sweet potatoes, sugarcane and corn. The plants are planted in provinces, autonomous regions and direct prefectures such as Guangdong, Guangxi, Hainan, Jiangxi and Yunnan, Fujian, Sichuan, Chongqing, Guizhou, Hunan, Zhejiang and Taiwan, and are mainly concentrated in provinces such as Guangxi, Guangdong, Yunnan, Hainan and Jiangxi. According to statistics, the planting area and the yield of the Guangxi Zhuang autonomous region only account for more than 60 percent of the whole country.
Because the cassava is suitable for planting areas and has superior conditions such as light, heat, moisture and the like and rich biological diversity, the cassava is very beneficial to the generation and harm of various diseases, pests and weeds. Foreign cassava diseases are reported to include bacterial wilt, mosaic virus disease, brown spot, anthracnose, helminthosporium leaf spot, corynespora leaf spot, white spot, phytophthora root rot, powdery mildew, brown streak, frog skin disease and the like. The main diseases include mosaic virus disease, brown streak disease, powdery mildew and frog skin disease. At present, the diseases harmful to domestic cassava mainly comprise bacterial wilt, root rot and brown spot of cassava.
The cassava bacterial wilt disease is a bacterial disease caused by Xanthomonas axacuminata (cassava wilt pathogenic variety pv. manihotis) and is one of the internationally important quarantine diseases. With the continuous development of the cassava industry in China, germplasm and products are frequently exchanged with other countries, and worldwide diseases such as bacterial wilt and the like occur in main cassava planting areas in China and are very serious in the situation. In production, the disease is mainly prevented and controlled by methods such as seedling quarantine, biological prevention and control, medicament prevention and control and the like, and an effective prevention and control method is not available. Therefore, exogenous treatment is carried out on cassava to improve the disease resistance of the cassava in a short time so as to reduce the influence of diseases on the cassava industry and stably improve the cassava industry, and the method becomes a realistic problem to be solved for the continuous and healthy development of the cassava industry.
Betaine is a quaternary ammonium type water-soluble alkaloid, is very easy to dissolve in water, methanol, ethanol and ether, can generate trimethylamine through decomposition reaction of concentrated potassium hydroxide solution, has hygroscopicity and is very easy to deliquesce, and can release the trimethylamine. Betaine is used as an osmotic regulator and an enzyme protective agent, and accumulation of betaine enables a plurality of metabolic key enzymes to continuously keep activity under osmotic stress, and the integrity of cell membranes under stress is maintained to a certain extent. Researches on abiotic stress show that betaine has a certain effect on abiotic stress such as drought and salt, but the researches on the abiotic stress of plants are rarely reported.
How to further improve the disease resistance of cassava and enable the cassava to be well planted under the condition of relevant environmental stress is a hot spot of current research. At present, no method for rapidly improving the cassava disease resistance exists, so that the method has great significance for improving the cassava disease resistance by utilizing exogenous substances to treat the cassava so as to maintain the continuous and healthy development of the cassava industry.
Disclosure of Invention
In view of the defects of the prior art, the invention aims to provide a method for improving the disease resistance of plants, in particular to a method for improving the disease resistance of cassava through betaine induction treatment. The method is easy to implement, simple and convenient to operate, takes effect quickly, can improve the disease resistance of the cassava in a short time, and has great significance for maintaining the sustainable and healthy development of the cassava industry.
In order to achieve the above object, the present inventors have creatively induced cassava seedlings with a foreign substance, betaine, to obtain a method for improving the disease resistance of cassava in a short time. Specifically, the invention adopts the following technical scheme:
a method for improving the disease resistance of cassava through betaine induction treatment, wherein the method comprises the following steps: continuously spraying betaine solution with the concentration of 25-400 mu M to the leaf surface of the cassava seedlings in any period of the growth stage for 5-14 days.
Further preferably, the method for improving the cassava disease resistance by betaine induction treatment as described above, wherein the height of cassava seedlings when the betaine solution is sprayed is 15-30 cm.
Further preferably, the method for improving the disease resistance of cassava by betaine induction treatment as described above, wherein the concentration of the betaine solution is 50 μ M to 200 μ M.
Further preferably, the method for improving the cassava disease resistance through betaine induction treatment is as described above, wherein the time for continuously spraying the betaine solution is 7-9 days and 2 times/day.
Still further preferably, the method for improving the disease resistance of cassava through betaine induction treatment further comprises the steps of fertilizing, deep ploughing, leveling and planting the seedling growing land before seedling growing, and watering from 1 st to 40 th days after seedling growing; wherein:
the fertilizing step is that enough base fertilizers are applied at one time, including decomposed farmyard manure 2000-;
the planting step adopts a stem cuttage method, the cuttage density is 5 per square meter from early spring to late spring, 2/3 with the length of seed stems and the ground form an angle of 15-45 degrees and are buried in planting ditches, 1/3 of the seed stems expose surface soil, after cuttage, the seeds are pressed to enable the stems to contact with the soil, and the stems are watered thoroughly to keep the soil moist;
watering for 500-; 300-40 d watering, 600L/mu/day, and irrigating by rainwater under natural conditions after 40 days.
Further preferably, the method for improving the disease resistance of cassava through betaine induction treatment is as described above, wherein the cassava is deeply turned for 15-20 cm after fertilization, and then a seedling raising land is leveled.
Tests of the present invention show that by continuously spraying different low-concentration betaine solutions in the range of 50 μ M to 200 μ M (molar mass concentration, i.e. 5.857-23.43mg betaine per litre water) for a short period of time (7 days) the treated cassava material was found to be more resistant to diseases than the untreated cassava material. As shown in the attached drawing, when the cassava is not treated, the bacteria surviving on the cassava multiply faster, the bacteria number is more, when the cassava is infected for 6 days, the log value of the bacteria number reaches about 6.9, and the bacteria number of the cassava can reach about 5.3 after the cassava is treated by low-concentration betaine solution of 50-200 mu M (molar mass concentration, namely 5.857-23.43mg of betaine is contained in each liter of water); that is, the bacteria count of the cassava materials treated with the low-concentration betaine solution of 50 μ M to 200 μ M (molar mass concentration, i.e. containing 5.857 to 23.43mg betaine per liter of water) is about 3/4 of the bacteria count of the cassava materials not treated. Based on the research results, the invention also provides the application of betaine in the aspect of cassava resistance physiological induction technology, namely: the betaine is applied to improving the disease resistance of cassava. Further preferably, wherein said disease is bacterial wilt.
Compared with the prior art, the method for improving the cassava disease resistance through betaine induction treatment has the following advantages and progresses: the method is easy to implement, simple and convenient to operate, and quick in effect, can improve the disease resistance of the cassava to the bacterial wilt in a short time, and has great significance for maintaining the continuous and healthy development of the cassava industry.
Drawings
FIG. 1 is a schematic diagram showing the effect of continuous spraying of betaine solutions of different concentrations on the resistance of cassava to bacterial blight bacteria of cassava after 7 days of induction treatment.
The cassava material growing normally for 20 days is divided into four parts on average, the first part is sprayed with clear water for 7 days, the second part is sprayed with 50 mu M betaine (with molar mass concentration, namely, each liter of water contains 5.857mg of betaine) solution for 7 days, the third part is sprayed with 100 mu M betaine (with molar mass concentration, namely, each liter of water contains 11.715mg of betaine) solution for 7 days, and the fourth part is sprayed with 200 mu M betaine (with molar mass concentration, namely, each liter of water contains 23.43mg of betaine) solution for 7 days. Four material injections were inoculated Xam and samples were taken for bacterial counts.
Detailed Description
The following is a specific example related to the present invention, and further description is made on the technical scheme of the present invention, but the protection scope of the present invention is not limited to the example. All changes, modifications and equivalents that do not depart from the spirit of the invention are intended to be included within the scope thereof.
Example 1: test example for improving cassava disease resistance by betaine Induction treatment
A. Planting plants:
fertilizing the seedling raising field, deeply turning for 15-20 cm, and leveling. Applying enough base fertilizer once every 667m23000kg of decomposed farmyard manure (including pig manure, cow manure and sheep manure) is applied to 2000-one area, 40-60kg of compound fertilizer (containing high-concentration potassium sulfate and nitrate nitrogen, the total nutrient is not less than 45%) is added to each mu, and 50-60kg of ammonium bicarbonate is applied to each mu. Leveling the bed surface and preparing the seedbed.
Cassava is planted by adopting a stem cuttage method. The planting is carried out in late spring (4 months bottom), and the planting density is 5 per square meter. Burying 2/3 with seed stem length in planting ditch at 15/20/25/30/35/40/45 degree with ground, exposing 1/3 of seed stem to surface soil, cutting, and pressing to make stem contact with soil. Watering thoroughly to keep soil moist. After the seed stem is cut at 15/20/25/30/35/40/45 degrees with the ground, the seed stem can germinate and sprout about a week.
B. Seedling stage management:
watering every 667m at the early stage of seedling culture2Watering for 500-1000L, and keeping the ground dry; 20 days before seedling, each 667m2Watering for 500-1000L; 20-40 days, each 667m2Watering for 300-600L.
C. And (3) betaine induction treatment:
selecting cassava seedlings (10-15 cm higher) with the size of 20d, continuously spraying different low-concentration betaine solutions (the molar mass concentration is that each liter of water contains 5.857-23.43mg of betaine) to the leaf surfaces for 50-200 mu M (twice a day) in 7 days; watering according to normal conditions or irrigating by rainwater according to natural conditions in other times; 20-40 days, each 667m2Watering for 300-; after 40 days, irrigating with rainwater according to natural conditions.
D. Inoculation Xam and statistics of bacterial count:
activating cassava bacterial wilt pathogen (xanthomonas carpi) stored in the laboratory, namely a wilting pathogenic variety of the xanthomonas carpi, which is separated from cassava leaves in teaching bases of the institute of agriculture and forestry, tropics, university of Hainan, on an LB (10 g of NaCl, 10g of tryptone and 5g of yeast extract per liter) plate, picking the single bacterium, dropping the single bacterium into 10mL of LB culture solution for culturing (28 ℃, 180rpm) for 12 hours, transferring the bacterium solution to a new LB culture solution for continuous culture until the concentration of the bacterium solution reaches OD (OD)6000.6. After centrifugation at 4000rpm for 10min at room temperature, the supernatant was discarded. With 10mM MgCl2Solution (molar mass concentration, i.e. containing 95.21mg MgCl per litre water)2) Suspend and dilute the bacterial solution to OD600About 0.2, injecting the bacterial liquid to mesophyll on both sides of the main vein by using an injector. At different time points, taking the injection point as the center of a circle, taking leaf discs with consistent sizes by using a puncher (the diameter is 1cm), sterilizing for 3min by using 75% alcohol, washing for 4 times by using sterile water, grinding into homogenate, diluting with sterile water in a gradient manner, dripping the homogenate on an LB (LB) flat plate, culturing overnight at 28 ℃, and counting the number of bacteria.
By continuously spraying the low-concentration betaine solution at 50 μ M to 200 μ M (molar mass concentration, i.e., 5.857 to 23.43mg betaine per liter of water) for the above short time (7 days), it was found that the treated cassava material was more resistant to diseases than the untreated cassava material. As shown in figure 1, when the bacteria survived on the cassava multiply faster and the number of bacteria is more, when the bacteria are infected for 6 days, the log value of the number of the bacteria reaches about 6.9, and the number of the bacteria after being treated by the low-concentration betaine solution of 50-200 mu M (the molar mass concentration is 5.857-23.43mg of betaine contained in each liter of water) reaches about 5.3; that is to say that the bacteria count of the cassava material treated with the low-concentration betaine solution of 50 μ M to 200 μ M (molar mass concentration, i.e. containing 5.857 to 23.43mg betaine per litre of water) is about 3/4 of the bacteria count of the cassava material not treated. Test results show that the betaine spraying treatment can improve the resistance of cassava to bacterial wilt.
Claims (3)
1. A method for improving the bacterial wilt resistance of cassava through betaine induction treatment, wherein the bacteria are xanthomonas carpi cassava wilting pathovar, is characterized by comprising the following steps: continuously spraying betaine solution with the concentration of 50-100 mu M to the leaf surfaces of the cassava seedlings for 7-9 days and 2 times/day, wherein the height of the cassava seedlings when the betaine solution is sprayed is 15-30 cm.
2. The method for improving the bacterial wilt resistance of cassava through betaine induction treatment according to claim 1, which further comprises the steps of fertilizing, deep ploughing, leveling and planting the seedling growing land before seedling growing, and watering at 1-40d after seedling growing; wherein:
the fertilizing step is that enough base fertilizers are applied at one time, including decomposed farmyard manure 2000-;
the planting step adopts a stem cuttage method, the cuttage density is 5 per square meter from early spring to late spring, 2/3 with the length of seed stems and the ground form an angle of 15-45 degrees and are buried in planting ditches, 1/3 of the seed stems expose surface soil, after cuttage, the seeds are pressed to enable the stems to contact with the soil, and the stems are watered thoroughly to keep the soil moist;
watering for 500-; 300-40 d watering, 600L/mu/day, and irrigating by rainwater under natural conditions after 40 days.
3. The method for improving the bacterial wilt resistance of cassava through betaine induction treatment according to claim 2, wherein the cassava is deeply turned over for 15-20 cm after fertilization, and then the seedling land is leveled.
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