CN111187725A - Efficient separation method of sweet potato shell-rostratum - Google Patents
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Abstract
The invention relates to an efficient separation method for pathogenic bacteria of crop diseases and insect pests, in particular to an efficient separation method for sweet potato shell fungus longcoracoid. The method is scientific, simple in operation process and low in test condition requirement. Compared with the traditional separation method of the sweet potato shell fungus, the separation success rate is obviously improved. The method comprises the following steps: firstly, soaking filter paper in sterilized water to keep moisture of a disease sample to generate black pathogens; then, cutting up the black pathogens and the diseased tuber healthy junction, and inoculating the cut tuber into the crosscut sweet potato; and finally, after the burrs are formed on the potato blocks, picking up a single burr, putting the burr on a sweet potato agar medium (SPA-xu 32) added with lactic acid, and culturing for 3-5 days in an incubator at 25-28 ℃ to obtain the sweet potato shell-growth fungus.
Description
Technical Field
The invention relates to a method for separating pathogenic bacteria of plant diseases, in particular to a method for efficiently separating sweet potato shell-fusca longirostratum, which belongs to the field of agricultural plant protection.
Background
Sweet potato (sweet potato)Ipomoea batatas) The crop is widely planted in a plurality of countries and regions in the world, is one of important grain crops, and is a bottom line crop for ensuring grain safety. China is the biggest sweet potato producing country in the world, the planting area of the sweet potatoes accounts for 45% of the planting area of the sweet potatoes in the world, and the annual yield accounts for more than 75% of the total yield of the sweet potatoes in the world. However, the occurrence of sweet potato diseases, especially fungal diseases, is an important factor which hinders sweet potato production and affects sweet potato yield and quality. At present, the fungal diseases of the sweet potatoes in China are various, and mainly comprise sweet potato black spot, black nevus disease, scab disease, sweet potato dry rot, root rot, ulcer, vine cutting and other diseases caused by fusarium, wherein the black spot is one of three diseases in the potato area in the north of China, and is generally seriously damaged.
The black spot of sweet potato is caused by the fungus of the genus Dioscorea of the family Hymenomycetes, class Sclerotia, order Hymenochaetales, family Rhynchosia rostrata (Sterculiaceae, division of Hypocreales)Ceratocystis fimbriataEllis et Halsted), also called black scar disease, commonly called black furuncle, black sore and the like, was first discovered in the united states by Halsted in 1890, was introduced into japan in 1919, was introduced into gao county in liaining province in china from kanto island in japan in 1937, gradually spreading the damage from north to south, and the occurrence and damage of the disease were reported successively in the existing 26 provinces (city and autonomous region), and were more serious in north China, the yellow-Huai-river basin, the Yangtze river basin, the southern summer and the autumn potato region. The yield loss caused by the disease is about 5-10% every year, and the loss caused by serious harm is 20-50% or even higher. In addition, the diseased potato can generate furanterpene toxic substances such as Ipomoeaminodone (ipomoearone), and can cause poisoning and even death after being eaten by people and livestock; when the diseased potatoes are used as fermentation raw materials, yeasts and saccharifying enzyme bacteria can be poisoned, the fermentation process is delayed, and the yield and the quality of alcohol are reduced.
At present, the sweet potato black spot caused by the sweet potato caralluma longirostratum and the diseases (such as sweet potato canker and the like) caused by fusarium are frequently generated in the field and the storage period, and the two types of pathogenic bacteria are often mixed and infected to cause the symptoms similar to the disease of the sweet potato black spot. Therefore, the method can reduce the influence of fusarium, avoid using complicated disinfection-cleaning steps, remove the influence of the disinfection time on the separation frequency, and greatly improve the frequency of the successful separation of the sweet potato chayote shell fungus.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a high-efficiency separation method of sweet potato shell fungus longcoracoid. The method comprises the steps of cleaning and airing diseased sweet potatoes, disinfecting with 75% alcohol and then airing, cutting off small pieces at the boundary of diseased spots by a disinfected blade, putting the diseased sweet potatoes into a clean plastic lunch box with a cover, absorbing enough water by qualitative filter paper and applying the moisture on the surfaces of the diseased spots, and keeping the moisture for about 6 days until black pathogens are generated; cutting off black pathogens and diseased potato tuber at the diseased junction, and adding a proper amount of sterilized water to dilute to prepare spore mixed solution; cleaning potato blocks, sterilizing, and transversely cutting into potato wafers with the thickness of 0.5 cm-1 cm; clamping a potato wafer by using a pair of tweezers, putting the potato wafer into the spore mixed solution for more than 30 s, then putting the potato wafer into a sterilized culture dish, adding absorbent cotton which absorbs water to one side of the culture dish, covering the culture dish, and culturing for 6 d at the temperature of 25-28 ℃; the obvious thorn hair can be seen, the single thorn hair on the surface is picked by a sterilization forceps or a pick needle, and is placed on SPA-xu32 containing lactic acid, after the thorn hair is placed right for 10 min, the thorn hair is inverted and cultured for 3-5d, and the sweet potato shell fuscus alniformis is obtained.
The invention is realized by the following technical scheme: an efficient separation method of sweet potato shell fungus longrosthornia is carried out according to the following steps:
step (1), moisturizing a diseased sample: collecting a potato block sample with the disease of the black spot of the sweet potato from the field, cleaning, disinfecting the surface of 75% alcohol, cutting off a small block at the boundary of the disease and the health by adopting a sterilization blade, applying a filter paper sheet wetted by sterilization water on the surface of the disease tissue for moisturizing, and carrying out moisturizing culture for 4-7 days at the temperature of 25-28 ℃ to see that black pathogens are generated on the surface of the potato block for later use;
step (2), potato block expanding preparation: taking fresh sweet potato blocks of the same variety as a disease sample or 100-inch ropes, cleaning, airing, disinfecting the surface with 75% alcohol, and transversely cutting the sweet potato into sweet potato slices with the thickness of 0.5 cm-1 cm by using a sterilization knife, namely expanding and propagating the sweet potato blocks for later use;
step (3), spore mixed liquid preparation: cutting off the black pathogens in the step (1), chopping into a sterilized culture dish, adding 10 ml of sterilized water into each 1g of sample, mixing uniformly, and standing for 10 min to prepare a spore mixed solution;
step (4), potato block expanding and propagating inoculation: placing the potato pieces prepared in the step (2) into spore mixed liquor for at least 30 s by using sterilization forceps, taking out and placing into a sterilization culture dish, adding absorbent cotton with enough moisture on one side of the culture dish for moisturizing, and culturing for 6 d at 25-28 ℃ after the culture dish is covered;
step (5), preparation of culture medium: preparing a sweet potato agar culture medium, namely an SPA-xu32 culture medium, from Xushu 32 potato pieces, sterilizing at 121 ℃ for 20min, cooling to 50 ℃, adding 5ml of lactic acid with the volume fraction of 25% into each 1000 ml of the SPA-xu32 culture medium, uniformly mixing, and pouring into a flat plate to prepare the SPA culture medium containing the lactic acid for later use;
step (6), germ separation: and (4) expanding and propagating the potato blocks, inoculating, moisturizing and culturing for 6 d to see obvious burrs, picking a single burr on the surface by using a sterilization forceps or a pick needle, placing the single burr on the lactic acid-containing SPA culture medium prepared in the step (5), rightly placing for 10 min, and then inversely culturing for 3-5d to obtain the sweet potato shell-growth bacterium
The preferred scheme of the efficient separation method of the sweet potato Rhinoceros fuscus as the invention comprises the following steps: the sweet potato agar culture medium, namely SPA-xu32 culture medium, is prepared by cleaning and peeling Xushu 32 potato blocks, collecting 200 g, slicing, adding 1000 ml of water, boiling for 20min, filtering with gauze, slightly cooling, adding water to 1000 ml, collecting 150 ml, subpackaging in a triangular flask containing 1.5 g of agar powder, plugging, wrapping, sterilizing at 120 deg.C for 20min, taking out, cooling and storing for later use.
The preferred scheme of the efficient separation method of the sweet potato Rhinoceros fuscus as the invention comprises the following steps: uniformly mixing a lactic acid solution with a volume fraction of 25% with a sweet potato agar culture medium according to a volume ratio of 1:200 to obtain the sweet potato agar culture medium containing lactic acid.
The invention has the advantages that: the method cuts off a small border of pathogenic bacteria before moistening, and is beneficial to the growth of pathogenic bacteria; the qualitative filter paper sheet is used for preserving moisture, so that the moisture can be ensured, and the foreign bacteria can be generated due to too large moisture; the interface between the black pathogen and the diseased potato tuber after moisturizing is made into spore mixed solution, which can ensure the quantity of the pathogenic bacteria; after the black pathogens generated by moisture preservation comprise the alternaria alternata and part of mixed bacteria, the sweet potato black spot bacteria amount is large, the growth speed is higher than that of the mixed bacteria after the secondary inoculation, and the sweet potato black spot bacteria are easy to separate; the sweet potato is inoculated and cut into pieces, so that the sweet potato pieces are easy to catch diseases, and the amount of formed spores is large; the formed spinuloid substance is a typical ascocarp of sweet potato black spot germ, and is easy to observe and pick; the culture medium is prepared by using the potato blocks of the variety Xushu 32, sugar does not need to be added, and the culture medium is more beneficial to the growth of the sweet potato caragana fusca compared with the culture medium prepared by other potato blocks, and belongs to a specific culture medium for separating the sweet potato caragana fusca. The method is simple to operate and easy to learn, has low requirements on experimental equipment, and can be used for identifying each plant protection unit.
Drawings
FIG. 1 is a schematic view of steps 1-6 of a high-efficiency separation method of sweet potato shell fungus longrosthornia. Wherein a is sample moisture preservation, b is a black pathogen obtained after moisture preservation, c is spore mixed liquid preparation, d is inoculated on the potato wafer, e is culture after inoculation, and f is the obtained sweet potato carapace fungi.
Example 1
A method for efficiently separating sweet potato shell fungus longrostone comprises the following steps: in the research institute, 15 suspected sweet potato black spot disease samples were collected from 5 prefectural regions such as Shandong Weifang, Jining, Anhui Sixian, Jiangsu Xuzhou and brush cloud, and the method was carried out according to the following steps (fig. 1):
(1) and moisturizing the diseased sample: collecting three suspected sweet potato black spot samples in each area, cleaning the samples by 15 parts, disinfecting the surfaces by 75% alcohol, cutting off a small block at the junction of disease and health by adopting a sterilization blade, wherein the size of the small block is about 0.5 cm by 0.3 cm, applying a filter paper sheet wetted by sterilization water on the surface of the diseased tissue of each sweet potato block for moisturizing, and culturing the samples for 6 days at the temperature of 25-28 ℃ to see that black pathogens are generated on the surfaces of the sweet potato blocks for later use;
(2) preparing potato blocks for propagation: taking sweet potato of the same variety as the disease sample, namely, Xushu 32 fresh sweet potato, cleaning and airing potato blocks, sterilizing the surface of the sweet potato and a kitchen knife by using 75% alcohol, and transversely cutting the sweet potato into sweet potato slices (approximate to a circle) with the thickness of about 0.8 cm by using the kitchen knife for later use;
(3) and preparing a pathogenic spore mixed solution: cutting off black pathogenic substance and pathogenic junction, mincing, placing into a sterilized culture dish, adding 10 ml of sterilized water into 1g of tissue, mixing, standing for 10 min, and making into spore mixture;
(4) and expanding and propagating potato block inoculation: placing the cut Xushu 32 potato round blocks into pathogenic spore liquid for 30 s by using a sterilization forceps, taking out, placing into a sterilization culture dish, adding absorbent cotton which absorbs water to one side of the culture dish, covering the culture dish, and culturing for 7 d at the temperature of 25-28 ℃;
(5) and preparing a culture medium: preparing sweet potato Agar (SPA-xu 32) culture medium from Xuchi potato 32 potato pieces, sterilizing at 121 deg.C for 20min, cooling to 50 deg.C, adding 5ml of lactic acid with volume fraction of 25% into each 1000 ml of SPA-xu32 culture medium, mixing, and pouring into flat plate.
(6) And pathogen separation: and (3) inoculating the expanded potato blocks, performing moisture-preserving culture for about 5 days to see obvious burrs, picking a single burr on the surface by using a sterilization forceps or a pick needle, placing the burr on the SPA-xu32 containing lactic acid, rightly placing for 10 min, and performing inverted culture for 3-5 days to obtain the sweet potato shell fusca.
The sweet potato agar culture medium is prepared by cleaning and peeling 32 potato blocks of slow potato, collecting 200 g of the sweet potato blocks, slicing, adding 1000 ml of water, boiling for 20min, filtering with gauze, slightly cooling, supplementing water to 1000 ml, collecting 150 ml, subpackaging in a triangular flask containing 1.5 g of agar powder, plugging, wrapping, sterilizing at 120 ℃ for 20min, taking out, cooling and storing for later use; uniformly mixing a lactic acid solution with a volume fraction of 25% with a sweet potato agar culture medium according to a volume ratio of 1:200 to obtain the sweet potato agar culture medium containing lactic acid.
Control separation method: 1. the method adopts a tissue isolation method to separate the sweet potato caralluma longirostratum and comprises the following steps: cutting the diseased tissue at the juncture of the diseased part into pieces with size of 5 mm x 3 mm, and sterilizing with 75% alcohol for 5 s; sterilizing with 5% sodium hypochlorite for 8 min, washing with sterilized water for 3-4 times, inoculating to sweet potato agar culture medium containing lactic acid under aseptic condition, and culturing at 25-28 deg.C. 2. Adopting a conventional spore dilution method, and comprising the following steps: moisturizing according to the step (1) to generate black pathogens, picking the pathogens into a 1.5 ml centrifuge tube with a proper amount of sterilized water, sucking 20 ul of the pathogens, coating the pathogens on a lactic acid-containing sweet potato agar culture medium, culturing at 25-28 ℃ for 3-5 days, observing the formation of the pathogens, transferring the suspected sweet potato longrostral shell fungi into a culture dish of the lactic acid-containing sweet potato agar culture medium again, culturing for 7 days, observing the spore morphology through a microscope, and identifying the pathogens.
Table 1 comparison of three separation methods
| Sample (I) | Variety of sweet potato | Method for producing a composite material | Tissue isolation method | Spore dilution method |
| Weifang 1 | Impact rope 100 | + | + | + |
| Weifang 2 | Impact rope 100 | + | - | - |
| Weifang 3 | Jishu 25 | + | - | + |
| Jining 1 | Longshu No. 9 | + | - | - |
| Jining 2 | Mercury potato 19 | + | - | - |
| Jining 3 | Tobacco potato 25 | + | + | + |
| Xuzhou 1 | Tobacco potato 25 | + | - | + |
| Xuzhou 2 | Longshu No. 9 | + | - | - |
| Xuzhou 3 | Mercury potato 19 | + | - | - |
| Pouring cloud 1 | Xushu 32 | + | - | - |
| Irrigation cloud 2 | Xushu 32 | + | - | |
| Irrigation cloud 3 | Mercury potato 19 | + | - | - |
| Sixian 1 | Mercury potato 19 | + | + | + |
| Sixian 2 | Xushu 32 | + | - | - |
| Sixian 3 | Xushu 32 | + | - | - |
Note: + represents the acquisition of sweetpotato black spot germ, and-represents the non-acquisition of sweet potato black spot germ
And (4) conclusion: as can be seen from Table 1, 15 samples of the suspected sweet potato black spot disease collected this time are obtained, and 15 sweet potato longrostral shell strains are separated by the method, with a success rate of 100%; the efficiency of successfully separating out pathogenic bacteria by a conventional tissue separation method is low, 3 parts of pathogenic bacteria are separated out totally, the success rate is only 20%, and 100% of samples are separated to obtain fusarium; after the samples were moisturized, spores were picked and 5 were successfully isolated with a success rate of 45.6%.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described with reference to the following examples.
Claims (3)
1. A high-efficiency separation method of sweet potato shell fungus longrosthornia is characterized in that: the method comprises the following steps:
step (1), moisturizing a diseased sample: collecting a potato block sample with the disease of the black spot of the sweet potato from the field, cleaning, disinfecting the surface of 75% alcohol, cutting off a small block at the boundary of disease and health by adopting a sterilization blade, applying a filter paper sheet wetted by sterilization water on the surface of disease tissues for moisturizing, and carrying out moisturizing culture for 4-7 days at the temperature of 25-28 ℃ to see that black pathogens are generated on the surface of the potato block for later use;
step (2), potato block expanding preparation: taking fresh sweet potato blocks of the same variety as a disease sample or 100-inch ropes, cleaning, airing, disinfecting the surface with 75% alcohol, and transversely cutting the sweet potato into sweet potato slices with the thickness of 0.5 cm-1 cm by using a sterilization knife, namely expanding and propagating the sweet potato blocks for later use;
step (3), spore mixed liquid preparation: cutting off the black pathogens in the step (1), chopping into a sterilized culture dish, adding 10 ml of sterilized water into each 1g of sample, mixing uniformly, and standing for 10 min to prepare a spore mixed solution;
step (4), potato block expanding and propagating inoculation: placing the potato pieces prepared in the step (2) into spore mixed liquor for at least 30 s by using sterilization forceps, taking out and placing into a sterilization culture dish, adding absorbent cotton with enough moisture on one side of the culture dish for moisturizing, and culturing for 6 d at 25-28 ℃ after the culture dish is covered;
step (5), preparation of culture medium: preparing a sweet potato agar culture medium, namely an SPA-xu32 culture medium, from Xushu 32 potato pieces, sterilizing at 121 ℃ for 20min, cooling to 50 ℃, adding 5ml of lactic acid with the volume fraction of 25% into each 1000 ml of the SPA-xu32 culture medium, uniformly mixing, and pouring into a flat plate to prepare the SPA culture medium containing the lactic acid for later use;
step (6), germ separation: and (4) inoculating and moisturizing the expanded potato blocks for 6 days to see obvious burrs, picking a single burr on the surface by using a sterilization forceps or a pick needle, placing the single burr on the lactic acid-containing SPA culture medium prepared in the step (5), rightly placing for 10 min, and then inversely culturing for 3-5 days to obtain the sweet potato shell fusca.
2. The efficient separation method of sweet potato hull-longum as claimed in claim 1, wherein: the sweet potato agar culture medium, namely SPA-xu32 culture medium, is prepared by cleaning and peeling Xushu 32 potato blocks, collecting 200 g, slicing, adding 1000 ml of water, boiling for 20min, filtering with gauze, slightly cooling, adding water to 1000 ml, collecting 150 ml, subpackaging in a triangular flask containing 1.5 g of agar powder, plugging, wrapping, sterilizing at 120 deg.C for 20min, taking out, cooling and storing for later use.
3. The efficient separation method of sweet potato hull-longum as claimed in claim 1 or 2, characterized in that: uniformly mixing a lactic acid solution with a volume fraction of 25% with a sweet potato agar culture medium according to a volume ratio of 1:200 to obtain the sweet potato agar culture medium containing lactic acid.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824791A (en) * | 2005-12-27 | 2006-08-30 | 云南农业大学 | Fast identification process of plant pathogenic bacterium pathogenicity |
| CN103333944A (en) * | 2013-07-08 | 2013-10-02 | 江苏徐州甘薯研究中心 | Identification method of sweet potato soft rot resistance |
| CN106635839A (en) * | 2016-12-26 | 2017-05-10 | 中国农业大学 | Method for separating ceratocystis spp from soil |
| CN106635830A (en) * | 2016-11-22 | 2017-05-10 | 江苏省农业科学院 | Culture method of alternaria brassicicola used for identifying resistance to black spot of sweet potatoes |
| CN107815437A (en) * | 2017-09-14 | 2018-03-20 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of method of sweet potato black rot pathogen rapid, high volume production spore |
| CN109507368A (en) * | 2018-11-16 | 2019-03-22 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of accurate Resistance Identification method of seedling stage sweet potato black rot |
-
2020
- 2020-03-17 CN CN202010184746.8A patent/CN111187725A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1824791A (en) * | 2005-12-27 | 2006-08-30 | 云南农业大学 | Fast identification process of plant pathogenic bacterium pathogenicity |
| CN103333944A (en) * | 2013-07-08 | 2013-10-02 | 江苏徐州甘薯研究中心 | Identification method of sweet potato soft rot resistance |
| CN106635830A (en) * | 2016-11-22 | 2017-05-10 | 江苏省农业科学院 | Culture method of alternaria brassicicola used for identifying resistance to black spot of sweet potatoes |
| CN106635839A (en) * | 2016-12-26 | 2017-05-10 | 中国农业大学 | Method for separating ceratocystis spp from soil |
| CN107815437A (en) * | 2017-09-14 | 2018-03-20 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of method of sweet potato black rot pathogen rapid, high volume production spore |
| CN109507368A (en) * | 2018-11-16 | 2019-03-22 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | A kind of accurate Resistance Identification method of seedling stage sweet potato black rot |
Non-Patent Citations (2)
| Title |
|---|
| 甘薯黑斑病菌快速大量产孢方法研究: "王波，等", 《安徽农业科学》 * |
| 禹阳等: "甘薯黑斑病抗性鉴定中黑斑病菌培养方法研究", 《江苏农业科学》 * |
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Application publication date: 20200522 |