CN103355109A - Method of detecting anti-aphid of soybean by utilizing soybean detached leaf - Google Patents
Method of detecting anti-aphid of soybean by utilizing soybean detached leaf Download PDFInfo
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- CN103355109A CN103355109A CN2013102940905A CN201310294090A CN103355109A CN 103355109 A CN103355109 A CN 103355109A CN 2013102940905 A CN2013102940905 A CN 2013102940905A CN 201310294090 A CN201310294090 A CN 201310294090A CN 103355109 A CN103355109 A CN 103355109A
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Abstract
The invention discloses a method of detecting anti-aphid of soybean by utilizing a soybean excised-leaf. According to the method, nutrition is supplied to the soybean excised-leaf through nutrient solution in a closed device, so as to breed soybean aphids for identifying the anti-aphid of the soybean. The method comprises the following main steps: the manufacturing technology of a soybean excised-leaf device; the preparation technology of soybean nutrient solution formula; the picking technology of soybean excised-leaf; the identification technology of the anti-aphid of the soybean; and the environment control technology of the soybean aphids. The method is simple and easy to operate, not limited by seasons, and able to identify the anti-aphid of the soybean largely and accurately.
Description
Technical field
The present invention relates to a kind of soybean varieties breeding method, be specifically related to a kind of method of utilizing excised leaf to detect aphis resistance.
Background technology
Soybean aphid is worldwide pest, and cultivating the resistant soybean kind is the effective way that solves this insect pest, and filial generation needs a large amount of resistances to detect and screening in the breeding.According to present Introduction of Literatures, the breeding of soybean aphid all is on whole plant, and whole plant breeding aphid has the shortcomings such as wayward.Yang Zhenyus etc. are commented in " Jilin agricultural science " the 29th volume in 2004 the 5th phase 3-6 page or leaf " research of Aphid-Resistant Wild Soybean in Northeast of China duplicate detection ": can carry out anti-aphid duplicate detection in climatic cabinate, but because soybean plant strain is larger, the method is not suitable for carrying out the aphid resistance detection of soybean varieties in enormous quantities.How to solve this difficulty significant to the breeding of Soybean Resistance aphid.
Summary of the invention
The present invention is directed to the deficiency that prior art exists, provide a kind of new soybean excised leaf device to detect the method for aphis resistance.The invention process is easy, can detect convenient, effectively, accurately aphid resistance of soybean.Mainly comprise: the manufacturing technology of soybean excised leaf device; The outfit technology of soybean nutritional formula of liquid; The stripped leaf of soybean is plucked technology; Soybean aphid Resistance Identification technology; The soybean aphid environment controlling technique.This invention has solved the Soybean Resistance aphid and has identified the technological difficulties that are subjected to environmental influence, reaches aphid resistance a large amount of under the laboratory condition that can control, the precise Identification soybean.The present invention has set up the method that a kind of soybean excised leaf device detects the soybean aphid resistance.
For achieving the above object, the present invention is by the following technical solutions:
A kind of method of utilizing the soybean excised leaf to detect aphid resistance of soybean may further comprise the steps:
1) plant to be planted grows to the 2-4 compound leaf during phase, gather the compound leaf that has just launched, be close to the cutting of plant stem with scalpel, the compound leaf sheet that will have whole petioles inserts rapidly and is equipped with in the excised leaf device of nutrient solution, described device is the container of end sealing, described interior of the container is divided into upper and lower two-layer by dividing plate, described nutrient solution is loaded into the lower floor of described interior of the container, and described nutrient solution is used for providing the growing nutrient of excised leaf;
2) on the positive blade of 2-4 aptery aphid of inoculation in described device;
3) described device is positioned over 23-25 ℃ culturing room, intensity of illumination 300 μ molm
-2s
-1, control daytime/night is 18/6h, the test for identification time is 13 days.
Preferably, described device is glass tube (diameter 10cm, high 15-17cm), described dividing plate is that diameter * thickness is the cystosepiment of 9cm * 2cm, be drilled with the aperture of 0.5cm * 0.5cm on the described cystosepiment, preferred described nutrient solution height is 3-5cm approximately, and a preferred described not end of sealing seals with sealed membrane.
Preferably, described compound leaf is the soybean compound leaf, described nutrient solution is soybean nutritional liquid, and the prescription of preferred described soybean nutritional liquid (grams per liter water) is: sucrose 10-12g, ammonium sulfate 0.237g, magnesium sulfate 0.537g, nitrate of lime 1.26g, potassium sulphate 0.25g, potassium phosphate 0.35g, b diammonium disodium edta 20mg, ferrous sulfate 15mg; Boric acid 6mg, manganese sulphate 4mg, zinc sulphate 1mg, copper sulphate 0.5mg, ammonium molybdate 0.5mg.
The invention has the advantages that: can allow aphid one controlled, the sealing environment in breed; The outfit technology of soybean nutritional formula of liquid can make soybean keep normal growth; Gather the stripped leaf of suitable soybean and can make test material be in unified growth time, and soybean aphid is quick in the tender Soybean Leaves growth of children; Soybean aphis resistance authenticate technology can the accurate statistics aphid breeding quantity, identify the aphid resistance of distinguishing soybean varieties; The control technology of aphid cultivating environment condition, the growth of regulation and control aphid.Identify the aphid resistance of soybean varieties with the inventive method, can in a large number, fast, accurately distinguish the difference between aphid resistance of soybean between kind.
Description of drawings
Fig. 1 is the structure chart of excised leaf device.
Used soybean sense aphid kind Williams82 is the soybean kind that openly checks order among the present invention, and anti-aphid kind Dowling can obtain by buying pattern in seeds company for openly having had the kind of Rag1 gene.
Embodiment
In the embodiment of the invention, utilize the device of soybean excised leaf detection aphid resistance of soybean as shown in the figure, the glass pipe shaft 1 that comprises end sealing, if foraminate foam dam 2, in the glass tube 1, foam dam 2 belows are nutrient solution 3, and the aperture of the petiole of soybean excised leaf 3 on foam dam 2 inserts and immerse in the nutrient solution 3, aphid 6 is seeded on the soybean leaves, with sealed membrane 4 sealed glass tubes.
To anti-aphid kind P203 and the sense aphid kind Williams82 hybridization that field resistance test screening obtains, F
2The method that generation utilizes soybean excised leaf device to detect aphid resistance is identified.Each F is adopted in test
2Individual plant repeats for 3 times, the random placement.
1) manufacturing technology of soybean excised leaf device.Device glass pipe shaft (diameter 10cm, high 15cm), in a diameter * thickness is arranged is the cystosepiment of 9cm * 2cm, be drilled with the aperture (such as Fig. 1) of 0.5cm * 0.5cm on the cystosepiment, the nutrient solution height is 3-5cm approximately, device seals with sealed membrane.
2) the outfit technology of soybean nutritional formula of liquid.Nutrient solution prescription (grams per liter water) is: sucrose 10g, ammonium sulfate 0.237g, magnesium sulfate 0.537g, nitrate of lime 1.26g, potassium sulphate 0.25g, potassium phosphate 0.35g, b diammonium disodium edta 20mg, ferrous sulfate 15mg, boric acid 6mg, manganese sulphate 4mg, zinc sulphate 1mg, copper sulphate 0.5mg, ammonium molybdate 0.5mg.
3) the stripped leaf of soybean is plucked technology.The sowing soybean varieties, plant to be planted grew to for 2 compound leaf phases, gathered the compound leaf that has just launched, and was close to the cutting of soybean stem with scalpel, had the compound leaf sheet of whole petioles, inserted rapidly in the excised leaf device.
4) soybean aphid Resistance Identification technology.Inoculate 2 aptery aphids on positive blade with writing brush.
5) soybean aphid environment controlling technique.The culturing room of 23 ℃ of soybean excised leaf device laying temperatures, intensity of illumination 300 μ molm
-2s
-1, control daytime/night is 18/6h.The test for identification time is 13 days.
Qualification result: the breeding quantity of adding up each individual plant soybean aphid, the Aphed population of the resistant strain that screening obtains is 3 times of initial Aphed population, sense aphid kind Aphed population is more than 100 times of initial Aphed population, and the soybean varieties aphid resistance shows significant difference.
To anti-aphid kind P203 and the sense aphid kind Williams82 hybridization that field resistance test screening obtains, F
2The method that generation utilizes soybean excised leaf device to detect aphid resistance is identified.Each F is adopted in test
2Individual plant repeats for 3 times, the random placement.
1) manufacturing technology of soybean excised leaf device.Device glass pipe shaft (diameter 10cm, high 16cm), in a diameter * thickness is arranged is the cystosepiment of 9cm * 2cm, be drilled with the aperture (such as Fig. 1) of 0.5cm * 0.5cm on the cystosepiment, the nutrient solution height is 3-5cm approximately, device seals with sealed membrane.
2) the outfit technology of soybean nutritional formula of liquid.Nutrient solution prescription (grams per liter water) is: sucrose 10g, ammonium sulfate 0.237g, magnesium sulfate 0.537g, nitrate of lime 1.26g, potassium sulphate 0.25g, potassium phosphate 0.35g, b diammonium disodium edta 20mg, ferrous sulfate 15mg, boric acid 6mg, manganese sulphate 4mg, zinc sulphate 1mg, copper sulphate 0.5mg, ammonium molybdate 0.5mg.
3) the stripped leaf of soybean is plucked technology.The sowing soybean varieties, plant to be planted grew to for 3 compound leaf phases, gathered the compound leaf that has just launched, and was close to the cutting of soybean stem with scalpel, had the compound leaf sheet of whole petioles, inserted rapidly in the excised leaf device.
4) soybean aphid Resistance Identification technology.Inoculate 3 aptery aphids on positive blade with writing brush.
5) soybean aphid environment controlling technique.The culturing room of 24 ℃ of soybean excised leaf device laying temperatures, intensity of illumination 300 μ molm
-2s
-1, control daytime/night is 18/6h.The test for identification time is 13 days.
Qualification result: the breeding quantity of adding up each individual plant soybean aphid, the Aphed population of the resistant strain that screening obtains is 3 times of initial Aphed population, sense aphid kind Aphed population is more than 100 times of initial Aphed population, and the soybean varieties aphid resistance shows significant difference.
To anti-aphid kind P203 and the sense aphid kind Williams82 hybridization that field resistance test screening obtains, F
2The method that generation utilizes soybean excised leaf device to detect aphid resistance is identified.Each F is adopted in test
2Individual plant repeats for 3 times, the random placement.
1) manufacturing technology of soybean excised leaf device.Device glass pipe shaft (diameter 10cm, high 17cm), in a diameter * thickness is arranged is the cystosepiment of 9cm * 2cm, be drilled with the aperture (such as Fig. 1) of 0.5cm * 0.5cm on the cystosepiment, the nutrient solution height is 3-5cm approximately, device seals with sealed membrane.
2) the outfit technology of soybean nutritional formula of liquid.Nutrient solution prescription (grams per liter water) is: sucrose 12g, ammonium sulfate 0.237g, magnesium sulfate 0.537g, nitrate of lime 1.26g, potassium sulphate 0.25g, potassium phosphate 0.35g, b diammonium disodium edta 20mg, ferrous sulfate 15mg, boric acid 6mg, manganese sulphate 4mg, zinc sulphate 1mg, copper sulphate 0.5mg, ammonium molybdate 0.5mg.
3) the stripped leaf of soybean is plucked technology.The sowing soybean varieties, plant to be planted grew to for 4 compound leaf phases, gathered the compound leaf that has just launched, and was close to the cutting of soybean stem with scalpel, had the compound leaf sheet of whole petioles, inserted rapidly in the excised leaf device;
4) soybean aphid Resistance Identification technology.Inoculate 4 aptery aphids on positive blade with writing brush.
5) soybean aphid environment controlling technique.The culturing room of 25 ℃ of soybean excised leaf device laying temperatures, intensity of illumination 300 μ molm
-2s
-1, control daytime/night is 18/6h.The test for identification time is 13 days.
Qualification result: the breeding quantity of adding up each individual plant soybean aphid, the Aphed population of the resistant strain that screening obtains is 3 times of initial Aphed population, sense aphid kind Aphed population is more than 100 times of initial Aphed population, and the soybean varieties aphid resistance shows significant difference.
More than describe preferred embodiment of the present invention in detail.Should be appreciated that those of ordinary skill in the art need not creative work and just can design according to the present invention make many modifications and variations.Therefore, all those skilled in the art all should be in the determined protection domain by claims under this invention's idea on the basis of existing technology by the available technical scheme of logical analysis, reasoning, or a limited experiment.
Claims (8)
1. method of utilizing the soybean excised leaf to detect aphid resistance of soybean may further comprise the steps:
(1) plant to be planted grows to the 2-4 compound leaf during phase, gather the compound leaf that has just launched, be close to the cutting of plant stem with scalpel, the compound leaf sheet that will have whole petioles inserts rapidly and is equipped with in the excised leaf device of nutrient solution, described device is the container of end sealing, described interior of the container is divided into upper and lower two-layer by the foraminate dividing plate of brill, described nutrient solution is loaded into the lower floor of described interior of the container, and described nutrient solution is used for providing the growing nutrient of excised leaf;
(2) on the positive blade of 2-4 aptery aphid of inoculation in described device;
(3) be not positioned over 23-25 ℃ culturing room, intensity of illumination 300 μ molm after an end of sealing seals with sealed membrane described device
-2s
-1, the time at control daytime and night is respectively 18 and 6h, and the time is 13 days.
2. the method for claim 1, wherein described device is canals of stilling.
3. the method for claim 1, wherein described dividing plate is cystosepiment.
4. the method for claim 1, wherein, described nutrient solution is soybean nutritional liquid, and the constituent content of one liter of described soybean nutritional liquid is: sucrose 10-12g, ammonium sulfate 0.237g, magnesium sulfate 0.537g, nitrate of lime 1.26g, potassium sulphate 0.25g, potassium phosphate 0.35g, b diammonium disodium edta 20mg, ferrous sulfate 15mg, boric acid 6mg, manganese sulphate 4mg, zinc sulphate 1mg, copper sulphate 0.5mg and ammonium molybdate 0.5mg.
5. the method for claim 1, wherein the diameter of described aperture is 0.5cm * 0.5cm.
6. method as claimed in claim 2, wherein, described assembly dia is 10cm, height is 15-17cm.
7. method as claimed in claim 3, wherein, described cystosepiment diameter * thickness is 9cm * 2cm.
8. such as claim 6 or 7 described methods, wherein, described nutrient solution height is 3-5cm.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105165604A (en) * | 2015-10-15 | 2015-12-23 | 中国农业科学院作物科学研究所 | Method for identifying resistance of soybeans to soybean phytophthora blight and application of method in breeding |
| CN105993282A (en) * | 2016-06-03 | 2016-10-12 | 柳州市能兴科技有限公司 | Method for improving germination rate of soybean seeds |
| CN106613287A (en) * | 2017-01-23 | 2017-05-10 | 湖北省农业科学院粮食作物研究所 | Method for rapidly identifying aphid resistance of mung beans |
| CN116593656A (en) * | 2023-07-05 | 2023-08-15 | 扬州大学 | Device and method for evaluating and identifying aphid resistance of cucumber |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105165604A (en) * | 2015-10-15 | 2015-12-23 | 中国农业科学院作物科学研究所 | Method for identifying resistance of soybeans to soybean phytophthora blight and application of method in breeding |
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| CN105993282A (en) * | 2016-06-03 | 2016-10-12 | 柳州市能兴科技有限公司 | Method for improving germination rate of soybean seeds |
| CN106613287A (en) * | 2017-01-23 | 2017-05-10 | 湖北省农业科学院粮食作物研究所 | Method for rapidly identifying aphid resistance of mung beans |
| CN106613287B (en) * | 2017-01-23 | 2020-09-04 | 湖北省农业科学院粮食作物研究所 | Method for rapidly identifying aphid resistance of mung beans |
| CN116593656A (en) * | 2023-07-05 | 2023-08-15 | 扬州大学 | Device and method for evaluating and identifying aphid resistance of cucumber |
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| CN103355109B (en) | 2015-03-25 |
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