Jute anthracnose resistance identification method

A technology for identification of resistance and anthracnose, applied in biochemical equipment and methods, measurement/inspection of microorganisms, etc., can solve problems such as identification methods of resistance to jute anthracnose, and achieve conditions that are conducive to rapid infection and onset Consistent, easy-to-use results

Inactive Publication Date: 2015-11-25
FUJIAN AGRI & FORESTRY UNIV
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Abstract

The invention discloses a jute anthracnose resistance identification method, and belongs to the technical field of identification of plant disease resistance. The identification method comprises steps of strain culture, wound formation, mycelium inoculation, bagging moisture preservation and disease spot measurement, and specifically the method comprises the following steps: acquiring a jute anthracnose strain by virtue of a tissue isolation method, culturing and purifying the strain, and removing upper leaves of a to-be-detected jute plant from petiole bases by virtue of a blade so as to form a wound; then inoculating the cultured jute anthracnose strain on the wound, spraying water to the inoculated part, covering the inoculated part with a preservative bag and moisturizing for 2-3 days, removing the bag and continuing to infect for 15 days; and finally, determining the resistance strength of the jute plant by measuring the size of diseases spots. The method has the advantages of simple operation process, short identification cycle, high inoculation efficiency and the like; and the method is quite high in application value on anthracnose resistance identification and screening of jute germplasm resources.

Application Domain

Microbiological testing/measurement

Technology Topic

Plant disease resistanceBiology +5

Examples

  • Experimental program(3)

Example Embodiment

[0015] Example 1
[0016] A method for identification of jute anthracnose resistance, its specific operation is as follows:
[0017] 1) Strain culture: The strain of Bacillus anthracis jute was obtained by tissue separation method, cultivated and purified on PSA (potato sucrose agar) medium;
[0018] 2) Wound formation: at 6:00 in the morning, cut off the upper 3 leaves at the base of the petiole with a blade to form a neat wound;
[0019] 3) Mycelia inoculation: use a hole puncher with a pore size of 5 mm to cut the bacterial block on the cultivated and purified jute anthracnose strain, pick the bacterial block with an inoculation needle, make the mycelium directly contact the wound on the jute plant, and Press the bacterial block on the wound for inoculation, and wrap it with tinfoil;
[0020] 4) Moisturizing in bags: Spray water on the inoculated part with a watering can, and cover it with a fresh-keeping bag. If there are no water droplets or moisture in the bag, continue to spray water in the bag with a watering can to moisturize;
[0021] 5) Lesion measurement: remove the fresh-keeping bag after bagging for 3 days, and measure the size of the lesion on the jute plant after 15 days of continuous infection. mm 2 ) are 0-8, 8.1-16.0, 16.1-24.0, 24.1-32.0, 32.1-40.0, which are divided into 0, 1, 2, 3 and 4 grades in turn;
[0022] 6) Resistance evaluation: Calculate the jute anthracnose disease index according to the disease classification, and determine the resistance strength of jute plants according to the obtained disease index: high resistance (0), resistance (0.1-10.0), moderate resistance (10.1-30.0 ), middle sense (30.1-50.0), sense (50.1-60.0) and high sense (above 60.0);
[0023] The calculation method of jute anthracnose disease index is as follows:
[0024] Disease index = 100 × ∑ (number of lesions at all levels × representative value at each level)/(total number of surveys × representative value at the highest level).

Example Embodiment

[0025] Example 2
[0026] A method for identification of jute anthracnose resistance, its specific operation is as follows:
[0027] 1) Strain culture: The strain of Bacillus anthracis jute was obtained by tissue separation method, cultivated and purified on PSA (potato sucrose agar) medium;
[0028] 2) Wound formation: at 7:00 in the morning, cut off the upper 3 leaves at the base of the petiole with a blade to form a neat wound;
[0029] 3) Mycelia inoculation: use a hole puncher with a pore size of 5 mm to cut the bacterial block on the cultivated and purified jute anthracnose strain, pick the bacterial block with an inoculation needle, make the mycelium directly contact the wound on the jute plant, and Press the bacterial block on the wound for inoculation, and wrap it with tinfoil;
[0030] 4) Moisturizing in bags: Spray water on the inoculated part with a watering can, and cover it with a fresh-keeping bag. If there are no water droplets or moisture in the bag, continue to spray water in the bag with a watering can to moisturize;
[0031] 5) Lesion measurement: Remove the fresh-keeping bag after bagging for 2 days, and measure the size of the lesion on the jute plant after 15 days of continuous infection. mm 2 ) are 0-8, 8.1-16.0, 16.1-24.0, 24.1-32.0, 32.1-40.0, which are divided into 0, 1, 2, 3 and 4 grades in turn;
[0032] 6) Resistance evaluation: Calculate the jute anthracnose disease index according to the disease classification, and determine the resistance strength of jute plants according to the obtained disease index: high resistance (0), resistance (0.1-10.0), moderate resistance (10.1-30.0 ), middle sense (30.1-50.0), sense (50.1-60.0) and high sense (above 60.0);
[0033] The calculation method of jute anthracnose disease index is as follows:
[0034] Disease index = 100 × ∑ (number of lesions at all levels × representative value at each level)/(total number of surveys × representative value at the highest level).

Example Embodiment

[0035] Example 3
[0036] A method for identification of jute anthracnose resistance, its specific operation is as follows:
[0037] 1) Strain culture: The strain of Bacillus anthracis jute was obtained by tissue separation method, cultivated and purified on PSA (potato sucrose agar) medium;
[0038] 2) Wound formation: At 8:00 in the morning, cut off the upper 4 leaves at the base of the petiole with a blade to form a neat wound;
[0039] 3) Mycelia inoculation: use a hole puncher with a pore size of 5 mm to cut the bacterial block on the cultivated and purified jute anthracnose strain, pick the bacterial block with an inoculation needle, make the mycelium directly contact the wound on the jute plant, and Press the bacterial block on the wound for inoculation, and wrap it with tinfoil;
[0040]4) Moisturizing in bags: Spray water on the inoculated part with a watering can, and cover it with a fresh-keeping bag. If there are no water droplets or moisture in the bag, continue to spray water in the bag with a watering can to moisturize;
[0041] 5) Lesion measurement: remove the fresh-keeping bag after bagging for 3 days, and measure the size of the lesion on the jute plant after 15 days of continuous infection. mm 2 ) are 0-8, 8.1-16.0, 16.1-24.0, 24.1-32.0, 32.1-40.0, which are divided into 0, 1, 2, 3 and 4 grades in turn;
[0042] 6) Resistance evaluation: Calculate the jute anthracnose disease index according to the disease classification, and determine the resistance strength of jute plants according to the obtained disease index: high resistance (0), resistance (0.1-10.0), moderate resistance (10.1-30.0 ), middle sense (30.1-50.0), sense (50.1-60.0) and high sense (above 60.0);
[0043] The calculation method of jute anthracnose disease index is as follows:
[0044] Disease index = 100 × ∑ (number of lesions at all levels × representative value at each level)/(total number of surveys × representative value at the highest level).
[0045] 144 jute germplasms were identified for anthracnose resistance by the method described in Example 1, wherein 32 parts of disease-resistant germplasms were identified, 16 parts of susceptible germplasms, 59 parts of moderately resistant germplasms, and 37 parts of moderately sensitive germplasms , compared with the results of natural disease in the field, the coincidence rate was over 98%.
[0046] Compared with the existing acupuncture inoculation method, the method provided by the invention has the advantages of susceptibility, rapid onset, and easy identification; compared with the spore spray method, the method provided by the invention has the advantages of being basically not affected by climatic conditions and having high reliability. It has great application value in the identification and screening of anthracnose resistance in jute germplasm resources, and can provide a theoretical basis for the selection of new jute varieties resistant to anthracnose and QTL positioning.

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