CN111345209A - Method for identifying field disease resistance of cassava to root rot - Google Patents
Method for identifying field disease resistance of cassava to root rot Download PDFInfo
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- CN111345209A CN111345209A CN202010229010.8A CN202010229010A CN111345209A CN 111345209 A CN111345209 A CN 111345209A CN 202010229010 A CN202010229010 A CN 202010229010A CN 111345209 A CN111345209 A CN 111345209A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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Abstract
The invention belongs to the technical field of agricultural plant protection, and particularly relates to a field disease resistance identification method of cassava to root rot; the identification method comprises the following steps: 1) inoculating root rot pathogenic bacteria to a PDA culture medium for culture for 7-10 days to obtain a culture medium with the root rot pathogenic bacteria; 2) preparing soil, planting cassava seed stems to be identified, and growing cassava root tubers; 3) punching the cassava root tuber growing on the cassava tree, and inoculating the culture medium with the root rot pathogenic bacteria in the step 1) to the punched position of the cassava root tuber; 4) taking out the inoculated potato blocks for anti-influenza identification. The method for identifying the disease resistance of cassava to the root rot can accurately and quickly screen out disease-resistant varieties, is simple to operate and easy to learn, has low requirements on equipment, does not influence the yield of the cassava, and does not pollute cassava field blocks.
Description
Technical Field
The invention belongs to the technical field of agricultural plant protection, and particularly relates to a field disease resistance identification method for root rot of cassava.
Background
Cassava (Manihot esculenta Crantz), a shrub-like perennial crop of the genus Manihot (Euphorbiaceae), has fleshy root and rich starch, and is one of the world's three tubers (cassava, potato, sweet potato).
The cassava root rot is one of important diseases which seriously affect the yield and the quality of the cassava, and the diseases are distributed in various planting areas in the world and can be caused by one or more pathogens. The main pathogens reported to cause this disease are botrydipnodia theobromae, Nattrassia magnifera and Fusarium spp, with the root rot pathogen being caused by Phytophthora palmita (Phytophthora palmivora) in the cassava producing areas of our country, causing a 30% yield loss and a 50% yield loss in Tamil Nadu areas of india. Guoguang et al reported that alum, flumorph-ethyl-aluminium, metalaxyl-propamocarb, etc. have better control effect on root rot of cassava. Because some cassava is planted in the barren hill, chemical prevention increases the labor intensity and prevention cost, increases pesticide residue and drug resistance of diseases, and therefore, the breeding and popularization of the cassava disease-resistant variety are the best method for solving the problem of the harm of the cassava to the root rot, and the economic benefit of the cassava industry development can be effectively improved. In the breeding process of the cassava disease-resistant variety, the method is particularly important for accurately and quickly screening the disease-resistant variety.
Disclosure of Invention
The invention aims to provide a field disease resistance identification method of cassava to root rot, and the identification method can accurately and quickly screen out disease-resistant cassava varieties.
In order to achieve the above object, the present invention provides the following technical solutions;
the invention provides a method for identifying field disease resistance of cassava to root rot, which comprises the following steps:
1) inoculating root rot pathogenic bacteria to a PDA culture medium for culture for 7-10 days to obtain a culture medium with the root rot pathogenic bacteria;
2) preparing soil, planting cassava seed stems to be identified, planting for 4-6 months, and growing cassava root tubers;
3) removing the surface covering soil of the cassava block roots in the step 2), selecting complete cassava block roots growing on cassava trees to be punched, inoculating the culture medium growing with the root rot pathogenic bacteria in the step 1) to the punched positions of the cassava block roots, sleeving a freshness protection bag on the inoculated cassava block roots, and covering the removed surface with the covering soil;
4) 7-10 days after inoculation, digging the surface covered soil, taking out the inoculated potato blocks, transversely cutting the roots of the cassava blocks from the inoculation position, and carrying out anti-infection identification on the grading standard and the resistance evaluation standard of the root rot by adopting cassava according to the disease incidence area;
there is no chronological restriction between the step 1) and the step 2).
Preferably, the root rot pathogenic bacteria in step 1) include pathogenic bacteria isolated from cassava root rot pathogenic tuberous roots.
Preferably, the root rot pathogen in step 1) comprises phytophthora palmae (phytophthora palmivora) of the genus phytophthora.
Preferably, the step 2) integrally comprises the following steps: sorting the moisture in the soil, and laying a black film; the width of the moisture seeds is 1m, and the height of the moisture seeds is 0.4-0.45 m.
Preferably, the cassava is planted in the middle of the soil moisture in the step 2), and the planting distance is 0.8-1.2 m.
Preferably, after the inoculation in the step 3), moisturizing the inoculated part by using wet sterile absorbent cotton.
Preferably, in the step 3), the number of the holes punched is 2-3, and the distance between the two holes is 4-6 cm.
Preferably, the inoculation time in the step 3) is 8 days in the middle and last days of the month, the day of the inoculation is sunny, and the temperature in the daytime is above 28 ℃.
The invention has the beneficial effects that: the invention provides a method for identifying field disease resistance of cassava to root rot, which comprises the following steps: 1) inoculating root rot pathogenic bacteria to a PDA culture medium for culture for 7-10 days to obtain a culture medium with the root rot pathogenic bacteria; 2) preparing soil, planting cassava seed stems to be identified, and growing cassava root tubers; 3) punching the cassava roots, and inoculating the culture medium with the root rot pathogenic bacteria in the step 1) to the punched positions of the cassava roots; 4) taking out the inoculated potato blocks for anti-influenza identification. 1 cassava tree is inoculated with 5-20 bags of cassava root tubers with different numbers, the cassava root tuber 1 bag is selected for inoculation, and the inoculated cassava root tuber is wrapped by a fresh-keeping bag, so that the temperature and humidity required by pathogenic bacteria can be provided, and the soil can be prevented from being polluted by the pathogenic bacteria; during investigation, soil is only scraped to take out the inoculated cassava root blocks for anti-infection identification, so that the cassava tree and the yield are not affected. After investigation, the cassava root tubers inoculated with pathogenic bacteria are intensively destroyed, and the pathogenic bacteria cannot be transferred to soil to pollute the environment. The method for identifying the disease resistance of cassava to the root rot can accurately and quickly screen out disease-resistant varieties, is simple to operate and easy to learn, has low requirements on equipment, does not influence the yield of the cassava, and does not pollute cassava field blocks.
Detailed Description
The invention provides a method for identifying field disease resistance of cassava to root rot, which comprises the following steps:
1) inoculating root rot pathogenic bacteria to a PDA culture medium for culture for 7-10 days to obtain a culture medium with the root rot pathogenic bacteria;
2) preparing soil, planting cassava seed stems to be identified, planting for 4-6 months, and growing cassava root tubers;
3) removing the surface covering soil of the cassava block roots in the step 2), selecting complete cassava block roots growing on cassava trees to punch holes, inoculating the culture medium growing with the root rot pathogenic bacteria in the step 1) to the punched positions of the cassava block roots, covering a freshness protection bag on the cassava block roots, and covering the removed surface with the covering soil;
4) 7-10 days after inoculation, digging the surface covered soil, taking out the inoculated potato blocks, transversely cutting the roots of the cassava blocks from the inoculation position, and carrying out anti-infection identification on the grading standard and the resistance evaluation standard of the root rot by adopting cassava according to the disease incidence area;
there is no chronological restriction between the step 1) and the step 2).
Firstly, inoculating root rot pathogenic bacteria to a PDA culture medium for culture for 7-10 days to obtain a culture medium with the root rot pathogenic bacteria; the culture temperature is preferably 25 ℃ to 28 ℃. In the present invention, the root rot pathogen preferably includes Phytophthora palmae (Phytophthora palmivora) of the genus Phytophthora; the root rot pathogenic bacteria preferably comprise pathogenic bacteria separated from cassava root rot pathogenic tuberous roots, and the cassava root rot pathogenic tuberous roots come from the field; the method of isolation is preferably a fungal isolation method.
Soil preparation is carried out, cassava seed stems to be identified are planted, the cassava seed stems are planted for 4-6 months, and cassava root tubers grow; the soil preparation preferably comprises the following steps: sorting the moisture in the soil, and laying a black film; the width of the moisture seeds is preferably 1m, the height of the moisture seeds is preferably 0.4-0.45 m, and the moisture seeds with the width and the height are convenient to open the covering soil on the cassava block roots during inoculation; the cassava is preferably planted among the dead seedlings, the planting distance is preferably 0.8-1.2 m, and more preferably 1 m; the planting time is preferably 2 months; the cassava seed stems are preferably planted in an oblique inserting mode; in the specific implementation process of the method, cassava is normally managed in the late planting stage, and watering is further included if soil is dry; the normal management comprises topdressing, bud picking and pest control; the black film plays a role in preserving water in the early stage of planting and controlling grass in the late stage of planting in rainy season.
After growing the cassava root tuber and obtaining the culture medium with root rot pathogenic bacteria, removing the surface covering soil of the cassava root tuber, selecting the complete cassava root tuber growing on a cassava tree to punch holes, inoculating the culture medium with the root rot pathogenic bacteria to the punched positions of the cassava root tuber, covering a freshness protection bag on the cassava root tuber, and covering the removed surface with the covering soil; after the inoculation, preferably moisturizing the inoculated part by using wet sterile absorbent cotton, in the specific implementation process of the invention, the sterile absorbent cotton is covered at the perforated hole which is inoculated with bacteria, and sterile water is sprayed on the sterile absorbent cotton until the sterile absorbent cotton is wet; the number of the punched holes is preferably 2-3, and the distance between the two holes is preferably 4-6 cm, and more preferably 5 cm; the diameter of the hole is preferably 6 mm; the holes are arranged in a line along the longitudinal direction of the cassava; the inoculation time is preferably in the middle and last ten days of 8 months, the inoculation weather is preferably sunny, and the temperature is preferably above 28 ℃ in the daytime, so that a suitable temperature condition can be provided for pathogenic bacteria; the inoculation number of each cassava variety to be identified is preferably 3-5, and 1 cassava root is selected from each cassava tree for punching inoculation.
According to the invention, the freshness protection package is sleeved on the cassava root tuber, so that the temperature and humidity required by pathogenic bacteria are provided, and the soil is prevented from being polluted by the pathogenic bacteria; the invention has no special requirements on the freshness protection package, and the freshness protection package can be sold on the market conventionally.
In the method, 7-10 days, preferably 8-9 days after inoculation, the inoculated potato blocks are taken out, the cassava block roots are transversely cut from the inoculation position, and resistance identification is carried out by adopting cassava to carry out grading standard and resistance evaluation standard on root rot according to the disease incidence area; in the specific implementation process of the invention, the grading standard and the resistance evaluation standard of the root rot are carried out by referring to the prior art (see [ Chunchangya, et al. 32 cassava germplasms for the resistance evaluation and the agronomic character analysis of the phytophthora root rot [ J ]. plant protection, 2017,43(01): 148-.
In the specific implementation process of the method, 5-20 bags of cassava root tubers with different numbers are grafted on 1 cassava tree, the cassava root tubers with 1 bag are selected for inoculation, and the inoculated cassava root tubers are wrapped by a fresh-keeping bag, so that the temperature and humidity required by pathogenic bacteria can be provided, and the soil can be prevented from being polluted by the pathogenic bacteria; during investigation, soil is only scraped to take out the inoculated cassava root blocks for anti-infection identification, so that the cassava tree and the yield are not affected. After investigation, the cassava root tubers inoculated with pathogenic bacteria are intensively destroyed, and the pathogenic bacteria cannot be transferred to soil to pollute the environment.
The method for identifying the field disease resistance of cassava to root rot provided by the present invention is described in detail below with reference to examples, but these should not be construed as limiting the scope of the present invention.
Example 1 identification method of field disease resistance of cassava to root rot
The phytophthora palmae root rot resistance identification test is carried out on 6 cassava varieties according to the following steps:
1) pathogen isolation and culture preparation: collecting diseased potato blocks from field, separating out root rot pathogenic bacteria by fungus separation method, and culturing on potato glucose agar culture medium (PDA) for 7 days;
2) land block arrangement: 2, at the beginning of the month, 1.0m wide furrow and 0.4-0.45 m high furrow are prepared in the field according to the terrain, and a black film is laid for later use;
3) planting the ginseng test cassava varieties: planting prepared seed stems of cassava varieties such as No. 5, No. 7, No. 8, No. 9, No. 11 and No. 124 in the middle of the dead furrow in an inclined inserting mode, watering, managing cassava normally in the later period, and inoculating the cassava after planting in 8 months;
4) and (3) inoculation identification: digging soil on the surface of cassava block roots to expose complete undamaged cassava block roots, putting a prepared small block of culture medium with pathogenic bacteria on the cassava with punched holes, longitudinally punching 2-3 holes on each 1 packet of cassava for inoculation, wherein the distance between one hole and one hole can be determined according to the length of the cassava and is generally about 5cm, inoculating a PDA culture medium without bacteria on each 1 packet of cassava as a contrast, covering the inoculation position with sterile absorbent cotton, spraying sterile water for moisture preservation, finally sleeving a freshness protection bag on the inoculated cassava block roots, covering the original soil with the cassava block roots sleeved with the freshness protection bag, and repeating 3 times for 3 varieties;
5) investigation and evaluation: during investigation, inoculated cassava roots are taken out independently, the cassava roots are cut transversely from an inoculation position, and resistance identification is carried out on the cassava variety to be tested by adopting a cassava root rot grading standard and a resistance evaluation standard according to the disease incidence area by referring to the prior art (the resistance evaluation and the agronomic character analysis of 32 cassava germplasms to phytophthora root rot [ J ] plant protection, 2017,43(01): 148-.
6) And after evaluation, the cassava root tuber is destroyed to prevent pathogenic bacteria from spreading.
Grading standard of cassava root rot:
0 grade, the ratio of the disease incidence area to the transverse cutting area of the potato blocks is less than 1/16;
grade 1, the ratio of the disease incidence area to the transverse cutting area of the potato blocks is greater than or equal to 1/16 and less than 1/8;
2, the ratio of the disease incidence area to the transverse cutting area of the potato blocks is greater than or equal to 1/8 and less than 1/4;
grade 3, the ratio of the disease incidence area to the transverse cutting area of the potato blocks is greater than or equal to 1/4 and less than 1/2;
4, the ratio of the disease incidence area to the transverse cutting area of the potato blocks is greater than or equal to 1/2 and less than 3/4;
and 5, stage: the ratio of the disease incidence area to the transverse area of the potato blocks is greater than or equal to 3/4.
Calculating disease index, dividing the disease resistance of cassava germplasm into 5 grades according to the disease index, wherein DI is less than or equal to 20: high Resistance (HR); DI is more than or equal to 20.1 and less than or equal to 40: disease resistance (R); DI is more than or equal to 40.1 and less than or equal to 60: (ii) a neutral feeling (MS); DI is more than or equal to 60.1 and less than or equal to 80: (ii) susceptibility to disease (S); 80.1 DI or less: high feeling (HS).
The test identification results are shown in table 1:
TABLE 1 cassava field root rot resistance identification results
And (4) conclusion: as seen from Table 1, among the 6 cassava varieties identified in the table, the level of resistance of No. 11 cassava in south China to phytophthora palmae root rot is "high resistance"; the resistance level of No. 8 cassava in south China to phytophthora palmae root rot is disease resistance; the resistance level of cassava of south China 124, south China No. 5 and south China No. 7 to phytophthora palmae root rot is 'susceptible'; the level of resistance of No. 9 cassava in south China to phytophthora palmae root rot is "high-grade".
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (8)
1. A method for identifying the field disease resistance of cassava to root rot comprises the following steps:
1) inoculating root rot pathogenic bacteria to a PDA culture medium for culture for 7-10 days to obtain a culture medium with the root rot pathogenic bacteria;
2) preparing soil, planting cassava seed stems to be identified, planting for 4-6 months, and growing cassava root tubers;
3) removing the surface covering soil of the cassava block roots in the step 2), selecting complete cassava block roots growing on cassava trees to be punched, inoculating the culture medium growing with the root rot pathogenic bacteria in the step 1) to the punched positions of the cassava block roots, sleeving a freshness protection bag on the inoculated cassava block roots, and covering the removed surface with the covering soil;
4) 7-10 days after inoculation, digging the surface covered soil, taking out the inoculated potato blocks, transversely cutting the roots of the cassava blocks from the inoculation position, and carrying out anti-infection identification on the grading standard and the resistance evaluation standard of the root rot by adopting cassava according to the disease incidence area;
there is no chronological restriction between the step 1) and the step 2).
2. The method of claim 1, wherein the root rot pathogens in step 1) include pathogens isolated from cassava root rot pathogenic tuberous roots.
3. The method of claim 1, wherein the root rot pathogen in step 1) comprises phytophthora palmae (phytophthora palmivora) of the genus phytophthora.
4. The method of claim 1, wherein the step 2) of integrating comprises the steps of: sorting the moisture in the soil, and laying a black film; the width of the moisture seeds is 1m, and the height of the moisture seeds is 0.4-0.45 m.
5. The identification method according to claim 4, wherein the cassava in the step 2) is planted in the middle of the soil moisture, and the planting distance is 0.8-1.2 m.
6. The method of claim 1, further comprising moisturizing the inoculated portion with moist sterile cotton wool after the inoculating in step 3).
7. The method according to claim 1, wherein the number of the holes punched in step 3) is 2 to 3 and the distance between the two holes is 4 to 6 cm.
8. The method according to claim 1, wherein the inoculation in step 3) is performed in the middle and last 8 months in sunny days at a temperature of 28 ℃ or higher in the daytime.
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Application publication date: 20200630 |