CN107821159B - Method for simply identifying resistance of tobacco brown spot in artificial disease nursery - Google Patents
Method for simply identifying resistance of tobacco brown spot in artificial disease nursery Download PDFInfo
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Abstract
The invention discloses a method for simply identifying the resistance of tobacco brown spot in an artificial disease nursery, which comprises the steps of disease nursery selection, encrypted wire-type field layout, fertilizer control cultivation, low-concentration large-area spray inoculation to form the artificial disease nursery, moisture preservation induction and resistance evaluation. According to the invention, by utilizing the characteristics that the mature period of the leaves of the field tobacco plants is susceptible and the alternaria alternate is susceptible to attack at a proper temperature and high humidity, a proper ecological area with the mature period of the tobacco leaves, proper temperature and high humidity, convenient irrigation and drainage and good irrigation conditions is selected as a disease garden, and the encrypted strip type field layout and controlled fertilizer cultivation are adopted, so that the susceptibility of the leaves of the plants is increased, the diseases are effectively induced, and the accuracy and the reproducibility of the resistance identification of the alternaria alternate are improved; the method is characterized by mainly tracking and investigating the disease development process of the anti-infection (anti-infection and anti-infection) control variety, has the advantages of light and simple operation, low labor intensity, water and labor conservation, time conservation, cost conservation, high efficiency, accurate identification of the resistance phenotype of the alternaria alternate, and the like, and is a method worthy of popularization.
Description
Technical Field
The invention belongs to the technical field of tobacco disease-resistant breeding, and particularly relates to a method for simply identifying resistance of tobacco brown spot in an artificial disease garden.
Background
The recommended tobacco brown spot resistance identification method usually adopts a disease nursery identification method, is a disease resistance identification method formed in a tobacco field growth environment, and has two modes of artificial inoculation and natural induction of disease. The resource for resisting the alternaria alternate disease in the tobacco is obtained by the method through many years and many places of identification and screening, can objectively reflect the actual resistance level of the variety, but is easily influenced by weather conditions such as temperature and rainfall and the like and the field bacterial content, the annual fluctuation is large, and the reliable conclusion can be obtained only through repeated identification for many years. The accuracy and reproducibility of the identification result are seriously influenced by the change of the disease condition. In the national standard GB/T23224 tobacco variety disease resistance identification, the establishment of an artificial disease nursery is regulated from the aspects of culture and preparation of a brown spot inoculum, inoculation period, inoculation concentration and the like, but some technical operation details are fuzzy, human factors and environmental conditions have large influence in the implementation process, the identification result still has large fluctuation, and the working strength of disease investigation is large. Therefore, by utilizing the susceptibility of mature leaves of field tobacco plants and the disease characteristics of tobacco brown spot, the environment in which diseases are easy to occur is artificially created, the influence of environmental conditions on the identification result can be effectively avoided, and the labor intensity can be greatly reduced by improving the disease investigation mode.
Disclosure of Invention
The invention aims to provide a method for simply identifying the resistance of the tobacco brown spot in an artificial disease garden.
The invention aims to realize the method, which comprises the steps of disease nursery selection, encrypted wire type field layout, controlled fertilizer cultivation, low-concentration large-area spray inoculation to form an artificial disease nursery, moisture preservation induction and resistance evaluation, and specifically comprises the following steps:
A. selecting disease nursery: selecting a proper ecological area with proper temperature and humidity, convenient irrigation and drainage and good irrigation conditions in the tobacco leaf mature period as a disease nursery;
B. and (3) encryption of the line type field layout: b, reducing the plant spacing by 5-10 cm and the row spacing by 10-20 cm by using the disease nursery obtained in the step A and adopting a line-strip-type row-by-row layout, averagely increasing 100-500 tobacco plants per mu, arranging a protection row at the head of a field and not arranging a protection row at the edge of the field;
C. controlling fertilizer and cultivating: the total nitrogen and phosphorus of the tobacco fertilizer application is equivalent to that of the conventional cultivation, the potassium fertilizer application amount is reduced by 5-10% in the topdressing process, and the foliar potassium fertilizer is forbidden to be sprayed in the bud growth period, so that a fertilizer application mode with relative excess of nitrogen fertilizer and relative lack of potassium fertilizer is formed, the normal growth of tobacco plants can be ensured, and the susceptibility of plant leaves can be increased;
D. and (3) carrying out spray inoculation on a large surface with lower concentration to form an artificial disease nursery: after topping and removing the leaves, the lower leaves of the tobacco plant are transited from physiological maturity to process maturity, the leaves turn white and yellow, and the color of the leaves is 10% in the evening of sunny days or cloudy days4The middle and lower leaves of all materials are inoculated on the large surface by spraying of the conidium suspension of the alternaria alternata with lower concentration per milliliter until the leaf surface is fogged into beads and does not drip, and the living leaves drip to form an artificial disease garden with uniform concentration and uniform distribution of pathogenic bacteria;
E. moisture retention induction: the natural moisture retention can be induced in normal rainy season; when the inoculated soil does not rain, drip irrigation, shallow trench slow irrigation or micro-sprinkling irrigation is adopted for moisturizing, soil water saturation is preferably kept every day, and diseases are induced continuously for 5-7 days;
F. and (3) evaluating disease resistance: after the induction of moisture retention, the pathogenesis process of the anti-infection control varieties is intensively tracked and investigated, when the control varieties all reach the resistance evaluation index, the disease indexes of all the reference varieties are investigated, the resistance is evaluated according to the result, the resistance is divided by taking the red star disease index as the resistance identification index, and the disease-resistant and anti-resistant tobacco varieties are the resistant materials.
The invention selects a proper ecological area with proper temperature and humidity, convenient irrigation and drainage and good irrigation conditions in the tobacco leaf mature period as a disease nursery, adopts the encrypted wire-type field layout, controls fertilizer cultivation, sprays and inoculates all materials on a large surface with lower concentration to form an artificial disease nursery with uniform germ concentration and germ distribution, induces diseases by moisture preservation, has the advantages of light and simple operation, low labor intensity, water and labor conservation, time conservation, cost conservation, high efficiency, accurate identification of the resistance phenotype of the alternaria alternata, and the like, and is worthy of popularization.
Compared with the prior art, the invention has the following beneficial effects:
1. according to the invention, by utilizing the susceptibility of leaves of field tobacco plants in the mature period and the characteristic that the tobacco alternaria alternate is easy to attack at a proper temperature and high humidity, the rain season beneficial to the easy attack of the alternaria alternate is artificially simulated, and the rain season is naturally moisturized when the leaves are inoculated and do not rain, and when the leaves are inoculated and do not rain, the high-humidity environment meeting the disease occurrence requirement is moisturized by drip irrigation, shallow trench slow irrigation or micro-spray irrigation, so that the disease is effectively induced, the influence of environmental conditions on the identification result is avoided, and the accuracy and the reproducibility of the resistance identification of the tobacco alternaria alternate are improved;
2. compared with the traditional field identification method, the environmental conditions for controlling the onset of the disease, the base number of pathogenic bacteria and the field distribution are more strictly regulated, and the distance effect of the onset center is overcome, so that the results of different batches are more reliable, and the comparability and the repeatability are stronger;
3. the invention relates to inheritance and development of a traditional field identification method, which adopts an encrypted line type layout to replace an induction line surrounding type layout, replaces spray inoculation induced materials with all materials inoculated by spray inoculation (namely, surface induction is used for inducing the substitution line), intensively tracks and investigates the disease development process of anti-infection (anti-infection and anti-infection) control varieties (all materials of a standard method), investigates disease indexes of all reference varieties when the control varieties reach resistance evaluation indexes, and evaluates resistance according to the result. Compared with the traditional field identification method, the field utilization rate of the land is improved, the work load of disease investigation close to 2/3 is reduced, the method can be finished within 20 days from inoculation to identification completion, and the identification investigation time of 1/3-1/2 is shortened.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to be limiting in any way, and any modifications or alterations based on the teachings of the present invention are intended to fall within the scope of the present invention.
The invention discloses a method for simply identifying the resistance of tobacco brown spot in an artificial disease garden, which utilizes the susceptibility of leaves of a field tobacco plant in the mature period and the characteristic that the tobacco brown spot is easy to attack under the condition of suitable temperature and high humidity, selects a suitable ecological area with the mature period of tobacco leaves, suitable temperature and high humidity, convenient irrigation and good irrigation condition as the disease garden, adopts the layout of an encrypted strip type field, controls fertilizer and cultivates, particularly reduces the using amount of potassium fertilizer capable of enhancing the plant resistance, increases the susceptibility of the leaves of the plant, inoculates all materials by large-area spray with lower concentration to form the artificial disease garden with uniform germ concentration and uniform germ distribution, manually simulates moisture preservation to be beneficial to the rainy season in which the brown spot is easy to attack, meets the high-humidity environment with the disease occurrence requirement by spray irrigation, shallow trench slow irrigation or micro-moisture preservation when the rain is not inoculated, effectively induces the disease, and avoids the influence of the environmental condition on the identification result, the accuracy and the reproducibility of the resistance identification of the alternaria alternate are improved; the disease development process of the anti-infection (anti-infection and anti-infection) control varieties is mainly tracked and investigated, when the control varieties all reach the resistance evaluation index, the disease indexes of all the reference varieties are investigated, and the resistance is evaluated according to the result, so that the disease investigation workload of about 2/3 is reduced. The method has the advantages of light and simple operation, low labor intensity, water and time conservation, cost conservation, high efficiency, accurate identification of the resistance phenotype of the alternaria alternate, and the like, and is worthy of popularization.
The method for simply identifying the resistance of the tobacco brown spot in the artificial disease nursery comprises the steps of disease nursery selection, encrypted wire strip type field layout, fertilizer control cultivation, low-concentration large-area spray inoculation to form the artificial disease nursery, moisture preservation induction and resistance evaluation, and specifically comprises the following steps:
A. selecting disease nursery: selecting a proper ecological area with proper temperature and humidity, convenient irrigation and drainage and good irrigation conditions in the tobacco leaf mature period as a disease nursery;
B. and (3) encryption of the line type field layout: b, reducing the plant spacing by 5-10 cm and the row spacing by 10-20 cm by using the disease nursery obtained in the step A and adopting a line-strip-type row-by-row layout, averagely increasing 100-500 tobacco plants per mu, arranging a protection row at the head of a field and not arranging a protection row at the edge of the field;
C. controlling fertilizer and cultivating: the total nitrogen and phosphorus of the tobacco fertilizer application is equivalent to that of the conventional cultivation, the potassium fertilizer application amount is reduced by 5-10% in the topdressing process, and the foliar potassium fertilizer is forbidden to be sprayed in the bud growth period, so that a fertilizer application mode with relative excess of nitrogen fertilizer and relative lack of potassium fertilizer is formed, the normal growth of tobacco plants can be ensured, and the susceptibility of plant leaves can be increased;
D. and (3) carrying out spray inoculation on a large surface with lower concentration to form an artificial disease nursery: after topping and removing the leaves, the lower leaves of the tobacco plant are transited from physiological maturity to process maturity, the leaves turn white and yellow, and the color of the leaves is 10% in the evening of sunny days or cloudy days4The leaf blades of the middle and lower parts of all the materials are inoculated on the large surface by spraying the conidium suspension of the alternaria alternata with lower concentration per milliliter until the leaf surface is beaded and does not drip, and the living leaf blades drip, so that the uniform concentration and the uniform distribution of pathogenic bacteria are formedThe artificial disease nursery;
E. moisture retention induction: the natural moisture retention can be induced in normal rainy season; when the inoculated soil does not rain, drip irrigation, shallow trench slow irrigation or micro-sprinkling irrigation is adopted for moisturizing, soil water saturation is preferably kept every day, and diseases are induced continuously for 5-7 days;
F. and (3) evaluating disease resistance: after the induction of moisture retention, the pathogenesis process of the anti-infection control varieties is intensively tracked and investigated, when the control varieties all reach the resistance evaluation index, the disease indexes of all the reference varieties are investigated, the resistance is evaluated according to the result, the resistance is divided by taking the red star disease index as the resistance identification index, and the disease-resistant and anti-resistant tobacco varieties are the resistant materials.
The suitable temperature and humidity of the tobacco leaves in the maturation period in the step A are that the temperature in the maturation period of the tobacco leaves is 15-30 ℃, the temperature difference between day and night is more than 8 ℃, the daily average temperature is 20-25 ℃, the rainfall in the maturation period of the tobacco leaves is abundant, the rainfall amount is 300-500 mm, and the relative humidity of the field is more than 70%.
And D, preparing the alternaria alternate conidium suspension from an aqueous solution consisting of 1.00-2.00% of glucose, 1.00-2.00% of glycerol and 0.25-0.50% of Tween 80.
And E, performing drip irrigation, shallow trench slow irrigation or micro-sprinkling irrigation, wherein the shallow trench slow irrigation is semi-trench water infiltration wetting irrigation, and the drip irrigation and the micro-sprinkling irrigation are irrigation settings used in modern agricultural fields.
In the anti-infection control variety in the step F, the net leaf yellow is an anti-disease control, G28 is an anti-resistance control, and G140 is an infection control.
In the step F, the disease resistance identification index is disease resistance when the disease index is less than 25.0; the disease index is resistant when the disease index is 25.1-50.0, the disease index is sensitive when the disease index is 50.1-75.0, and the disease index is sensitive when the disease index is more than 75.0.
The invention is further illustrated by the following specific examples:
example 1
Identification of field brown spot resistance of area test variety
Separating alternaria alternata germs from typical diseased spots of field tobacco leaves, verifying the alternaria alternata germs as pathogenic bacteria by a Ke-style rule, investigating the diseases by taking the leaves as a unit, carrying out classification and investigation according to national standard GB/T23222 tobacco plant diseases and insect pests, arranging 2 rows of protection rows at the field, removing 2-3 tobacco plants at the field during investigation, selecting 5 inoculated tobacco plants, and investigating all the leaves. The control disease-resistant variety is net leaf yellow, the resistant variety is G28, the susceptible variety is G140, and the materials to be tested are 65 regional test varieties.
The method comprises the following specific steps:
A. selecting disease nursery: selecting Yuxi research in an ecological area suitable for being used as a disease garden, wherein the temperature of the tobacco leaves in the mature period of the whole year is 15-30 ℃, the temperature difference between day and night is more than 8 ℃, the average daily temperature is 20-25 ℃, the rainfall in the mature period of the tobacco leaves is abundant, the rainfall amount is 300-500 mm, and the relative humidity of the field is more than 70%;
B. and (3) encryption of the line type field layout: b, using the disease nursery obtained in the step A, arranging the strips in rows, reducing the plant spacing by 5 cm and the row spacing by 10 cm, wherein the plant spacing is 50 cm and the row spacing is 110 cm (the normal plant spacing is 55 cm and the row spacing is 120 cm), averagely increasing more than 200 tobacco plants per mu, arranging a protection row at the head of the field and not arranging a protection row at the edge of the field;
C. controlling fertilizer and cultivating: the total nitrogen and phosphorus of the tobacco fertilizer application is equivalent to that of the conventional cultivation, the potassium fertilizer application amount is reduced by 10% during top dressing, and the leaf surface potassium fertilizer is not sprayed when the bud growth period is started, so that a fertilizer application mode with relative excess of nitrogen fertilizer and relative lack of potassium fertilizer is formed, the normal growth of tobacco plants can be ensured, and the susceptibility of plant leaves can be increased;
D. and (3) carrying out spray inoculation on a large surface with lower concentration to form an artificial disease nursery: after topping and removing the leaves, the lower leaves of the tobacco plant are transited from physiological maturity to process maturity, the leaves turn white and yellow, and the color of the leaves is 10% in the evening of sunny days or cloudy days4The middle and lower leaves of all materials are inoculated on the large surface by spraying of the conidium suspension of the alternaria alternata with lower concentration per milliliter until the leaf surface is fogged into beads and does not drip, and the living leaves drip to form an artificial disease garden with uniform concentration and uniform distribution of pathogenic bacteria; the alternaria alternate conidium suspension is prepared from an aqueous solution consisting of 2.00% of glucose, 2.00% of glycerol and 0.50% of tween 80;
E. moisture retention induction: the natural moisture retention can be induced in normal rainy season; when the inoculated soil does not rain, drip irrigation is adopted to preserve moisture, soil water saturation is kept every day, and diseases are induced for 5-7 days continuously;
F. and (3) evaluating disease resistance: after the moisture preservation is induced, the pathogenesis process of anti (anti and anti) infection control varieties is intensively tracked and investigated, when the control varieties all reach the resistance evaluation index, the disease indexes of all the reference varieties are investigated, the resistance is evaluated according to the result, the resistance is divided by taking the red star disease index as the resistance identification index, wherein the anti-disease and anti-infection tobacco varieties are resistance materials; the resistance identification index of the red star disease is disease resistance when the disease index is less than 25.0; the disease index is resistant when the disease index is 25.1-50.0, the disease index is sensitive when the disease index is 50.1-75.0, and the disease index is sensitive when the disease index is more than 75.0.
The experimental results showed that 4 resistant varieties accounted for 6.15%, 10 resistant varieties accounted for 15.39%, 37 resistant varieties accounted for 56.92%, and 14 susceptible varieties accounted for 21.54% of the 65 test materials (Table 1). The results show that most varieties tested have poor red star disease resistance, and the disease-resistant variety accounts for about 1/5.
TABLE 1 tobacco variety artificial disease nursery simplified identification of tobacco brown spot resistance results
Example 2
BC4F1Identification of resistance to field brown spot of strain
Separating alternaria alternata germs from typical diseased spots of field tobacco leaves, verifying the alternaria alternata germs as pathogenic bacteria by a Ke-style rule, investigating the diseases by taking the leaves as a unit, carrying out classification and investigation according to national standard GB/T23222 tobacco plant diseases and insect pests, arranging 2 rows of protection rows at the field, removing 2-3 tobacco plants at the field during investigation, selecting 5 inoculated tobacco plants, and investigating all the leaves. The contrast disease-resistant variety is leaf yellow, the resistant variety is G28, the susceptible variety is G140, and the to-be-detected material is 120 BC formed by Y3 and Beinhart1000-14F1And (5) strain.
The method comprises the following specific steps:
A. selecting disease nursery: selecting Yuxi research in an ecological area suitable for being used as a disease garden, wherein the temperature of the tobacco leaves in the mature period of the whole year is 15-30 ℃, the temperature difference between day and night is more than 8 ℃, the average daily temperature is 20-25 ℃, the rainfall in the mature period of the tobacco leaves is abundant, the rainfall amount is 300-500 mm, and the relative humidity of the field is more than 70%;
B. and (3) encryption of the line type field layout: b, using the disease nursery obtained in the step A, arranging the strips in rows, reducing the plant spacing by 10 cm and the row spacing by 20 cm, wherein the plant spacing is 45 cm and the row spacing is 100 cm (the normal plant spacing is 55 cm and the row spacing is 120 cm), increasing more than 480 tobacco plants per mu on average, arranging a protection row at the head of the field and not arranging a protection row at the edge of the field;
C. controlling fertilizer and cultivating: the total nitrogen and phosphorus of the tobacco fertilizer application is equivalent to that of the conventional cultivation, the potassium fertilizer application amount is reduced by 10% during top dressing, and the leaf surface potassium fertilizer is not sprayed when the bud growth period is started, so that a fertilizer application mode with relative excess of nitrogen fertilizer and relative lack of potassium fertilizer is formed, the normal growth of tobacco plants can be ensured, and the susceptibility of plant leaves can be increased;
D. and (3) carrying out spray inoculation on a large surface with lower concentration to form an artificial disease nursery: after topping and removing the leaves, the lower leaves of the tobacco plant are transited from physiological maturity to process maturity, the leaves turn white and yellow, and the color of the leaves is 10% in the evening of sunny days or cloudy days4The middle and lower leaves of all materials are inoculated on the large surface by spraying of the conidium suspension of the alternaria alternata with lower concentration per milliliter until the leaf surface is fogged into beads and does not drip, and the living leaves drip to form an artificial disease garden with uniform concentration and uniform distribution of pathogenic bacteria; the alternaria alternate conidium suspension is prepared from an aqueous solution consisting of 1.00% of glucose, 1.00% of glycerol and 0.25% of tween 80;
E. moisture retention induction: the natural moisture retention can be induced in normal rainy season; when the inoculated soil does not rain, the semi-ditch is used for slow irrigation and moisture preservation, soil water saturation is preferably kept every day, and diseases are induced continuously for 5-7 days;
F. and (3) evaluating disease resistance: after the moisture preservation is induced, the pathogenesis process of anti (anti and anti) infection control varieties is intensively tracked and investigated, when the control varieties all reach the resistance evaluation index, the disease indexes of all the reference varieties are investigated, the resistance is evaluated according to the result, the resistance is divided by taking the red star disease index as the resistance identification index, wherein the anti-disease and anti-infection tobacco varieties are resistance materials; the resistance identification index of the red star disease is disease resistance when the disease index is less than 25.0; the disease index is resistant when the disease index is 25.1-50.0, the disease index is sensitive when the disease index is 50.1-75.0, and the disease index is sensitive when the disease index is more than 75.0.
The experimental result shows that the 120 anti-brown spot strains in the test material have 4 strains accounting for 3.33 percent, the medium strains have 20 strains accounting for 16.67 percent, the medium strains have 73 strains accounting for 60.83 percent, and the medium strains have 23 strains accounting for 19.17 percent, which indicates that the number of the disease-resistant strains in the population is far lower than that of the disease-resistant strains, and the disease-resistant strains account for only 20 percent.
Claims (3)
1. A method for simply identifying the resistance of the alternaria alternate in an artificial disease nursery is characterized by comprising the steps of disease nursery selection, encrypted wire strip type field layout, fertilizer control cultivation, low-concentration large-area spray inoculation to form the artificial disease nursery, moisture preservation induction and resistance evaluation, and specifically comprises the following steps:
A. selecting disease nursery: selecting a proper ecological area with proper temperature and humidity, convenient irrigation and drainage and good irrigation conditions in the tobacco leaf mature period as a disease nursery; the suitable temperature and humidity of the tobacco leaves in the mature period are that the temperature of the tobacco leaves in the mature period is 15-30 ℃, the temperature difference between day and night is more than 8 ℃, the daily average temperature is 20-25 ℃, the rainfall is abundant in the mature period of the tobacco leaves, the rainfall is 300-500 mm, and the relative humidity of the field is more than 70%;
B. and (3) encryption of the line type field layout: b, reducing the plant spacing by 5-10 cm and the row spacing by 10-20 cm by using the disease nursery obtained in the step A and adopting a line-strip-type row-by-row layout, averagely increasing 100-500 tobacco plants per mu, arranging a protection row at the head of a field and not arranging a protection row at the edge of the field;
C. controlling fertilizer and cultivating: the total nitrogen and phosphorus of the tobacco fertilizer application is equivalent to that of the conventional cultivation, the potassium fertilizer application amount is reduced by 5-10% in the topdressing process, and the foliar potassium fertilizer is forbidden to be sprayed in the bud growth period, so that a fertilizer application mode with relative excess of nitrogen fertilizer and relative lack of potassium fertilizer is formed, the normal growth of tobacco plants can be ensured, and the susceptibility of plant leaves can be increased;
D. and (3) carrying out spray inoculation on a large surface with lower concentration to form an artificial disease nursery: after topping and removing the leaves, the lower leaves of the tobacco plant are transited from physiological maturity to process maturity, the leaves turn white and yellow, and the leaves are sunny or cloudyIn the evening of the day, the dosage is 104The middle and lower leaves of all materials are inoculated on the large surface by spraying of the conidium suspension of the alternaria alternata with lower concentration per milliliter until the leaf surface is fogged into beads and does not drip, and the living leaves drip to form an artificial disease garden with uniform concentration and uniform distribution of pathogenic bacteria; the alternaria alternate conidium suspension is prepared from an aqueous solution consisting of 1.00-2.00% of glucose, 1.00-2.00% of glycerol and 0.25-0.50% of tween 80;
E. moisture retention induction: the natural moisture retention can be induced in normal rainy season; when the inoculated soil does not rain, drip irrigation, shallow trench slow irrigation or micro-sprinkling irrigation is adopted for moisturizing, soil water saturation is preferably kept every day, and diseases are induced continuously for 5-7 days;
F. and (3) evaluating disease resistance: after the induction of moisture retention, the disease process of the anti-infection control varieties is intensively tracked and investigated, the net leaf yellow in the anti-infection control varieties is used as a disease-resistant control, G28 is used as an anti-infection control, G140 is used as a disease-susceptible control, when the control varieties all reach a resistance evaluation index, the disease indexes of all the reference varieties are investigated, the resistance is evaluated according to the result, the resistance is divided by using the red star disease index as a resistance identification index, wherein the disease-resistant and anti-resistant tobacco varieties are resistant materials.
2. The method for identifying alternaria alternate resistance of tobacco in an artificial disease nursery as claimed in claim 1, wherein the step E comprises drip irrigation, shallow trench slow irrigation or micro-irrigation, wherein the shallow trench slow irrigation is semi-trench water infiltration wetting irrigation, and the drip irrigation and the micro-irrigation are irrigation settings used in modern agricultural fields.
3. The method for identifying the resistance to alternaria alternate in a simplified manner in an artificial disease nursery as claimed in claim 1, wherein the identification of resistance to alternaria alternate in step F is characterized in that the disease resistance is a disease resistance when the disease index is less than 25.0; the disease index is resistant when the disease index is 25.1-50.0, the disease index is sensitive when the disease index is 50.1-75.0, and the disease index is sensitive when the disease index is more than 75.0.
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