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Inoculation method of Ustilago scitaminea

A technology for sugarcane smut and smut, applied in the agricultural field, can solve problems such as low accuracy, poor stability of identification results, and restrictions on the application of manual inoculation methods, and achieve the effects of convenient use and increased incidence

Inactive Publication Date: 2013-07-10
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing artificial inoculation method has a low incidence of inoculated varieties (including susceptible varieties), poor stability and low accuracy of identification results, which directly restrict the application of artificial inoculation methods

Method used

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  • Inoculation method of Ustilago scitaminea
  • Inoculation method of Ustilago scitaminea
  • Inoculation method of Ustilago scitaminea

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1. Collect the sugarcane smut whip in the sugarcane area of ​​Hainan, and collect the spores of the pathogen in a relatively cool, dry, and windless place, suspend the spores with sterile water, shake them with a vortex mixer, and adjust the spore suspension with a hemocytometer Concentration is 10 6 spores / ml, spare.

[0016] 2. Conventional artificial inoculation: soak the seedlings in a spore suspension (concentration of spore suspension 10 6 spores / ml), take it out after 10 minutes and put it in a cool place and cover it with plastic paper to moisturize overnight (16 hours).

[0017] Artificial inoculation method of the present invention: draw 20 microliters of spore suspensions with pipette gun and spot-connect on the surface of the seedling bud body, then gently wrap with moistened toilet paper and then put it in a cool place and cover it with plastic paper to keep it moist overnight ( 16 hours).

[0018] Wherein, the seedling adopts the conventional seedling R...

Embodiment 2

[0026] 1. Proliferate sugarcane smut half a year in advance to identify hosts NCO376 (highly resistant), NCO310 (resistant to race 2, susceptible to race 1), F134 (susceptible to race 2), F173 (highly susceptible) and main planters in Hainan sugarcane area Variety ROC22, spare.

[0027] 2. Collect sugarcane smut whip from different places and different cultivars in Hainan sugarcane area, collect smut spores, suspend the spores with sterile water, shake with a vortex mixer, and adjust the concentration of the spore suspension with a hemocytometer for 10 6 See Table 2 for spores / ml, number, sample species and number.

[0028] 3. Use a pipette gun to draw 20 microliters of spore suspension and apply it to the surface of the seedling bud, then gently wrap it with moistened toilet paper, put it in a cool place, cover it with plastic paper and keep it moist overnight (20 hours).

[0029] 4. Use the method in step 3 to inoculate the spore suspension on the identification host of su...

Embodiment 3

[0042] 1. Propagation of different strains SPAP1, SPAP2, ..., SPAP7 of sugarcane SPAP by transfecting ribosomal inactivation protein gene, spare.

[0043] 2. Collect sugarcane smut whips and collect smut spores in the sugarcane area of ​​Hainan. Suspend the spores with sterile water, oscillate with a vortex mixer, and adjust the concentration of the spore suspension to 10 with a hemocytometer 6 spores / ml, spare.

[0044] 3. Use a pipette gun to draw 20 microliters of spore suspension and place it on the surface of the seedling bud, then gently wrap it with moistened toilet paper, then put it in a cool place and cover it with plastic paper to moisturize overnight (24 hours), according to This method was used to inoculate different transgenic strains respectively; after that, they were planted in large plastic pots in greenhouses (mainly for isolation), and the non-transgenic strain variety ROC22 was used as a control.

[0045] 4. Evaluation. After 6 months of planting, the tot...

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Abstract

The invention discloses an inoculation method of Ustilago scitaminea. The method includes the steps of inoculating Ustilago scitaminea spore suspension to the surface of sugarcane buds by spot inoculation and keeping in the wet place for 16-24h. The pore suspension is 106 / ml in concentration. During moisture preservation, the inoculated sugarcane buds are wrapped with humid tissue and are then placed at a shade place after being covered with plastic paper. The inoculation method is convenient to use, incidence rate of susceptible varieties is increased effectively, up to more than 80%, sense resistance of the different sugarcane varieties to Ustilago scitaminea can be distinguished effectively, and accordingly good conditions are created for the intensive study on breeding for disease resistance to Ustilago scitaminea and the study on physiological race differentiation.

Description

technical field [0001] The invention belongs to the technical field of agriculture, and in particular relates to a method for inoculating sugarcane smut. Background technique [0002] Sugarcane smut is one of the important diseases in the production process of sugarcane, known as the "cancer" of sugarcane. The disease was first discovered in Natal, South Africa in 1877, and subsequently reported in most countries in Asia and Africa. After it was discovered in Argentina in America in 1940, the disease also occurred in the western hemisphere (Schenck S, Pear H M, Liu Z, et al. Genetic variation of Ustilago scitaminea pathotypes in Hawaii evaluated by host range and AFLP marker[J].Sugar Cane International, 2005, 23:15-19.). From the 1970s to the 1990s, Hawaii, Florida, Morocco, Australia and other countries and regions also reported the occurrence of the disease (Byther R S, Steiner G W, Wismer C A. New sugarcane diseases reported in Hawaii[J]. Sugarcane Pathologists'Newslet...

Claims

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Application Information

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IPC IPC(8): A01G7/06C12Q1/18
Inventor 熊国如伍苏然张雨良冯翠莲沈林波杨本鹏张树珍
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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