CN108949899A - The identification method of Physiological Race of Phytophthora infestans and its application - Google Patents
The identification method of Physiological Race of Phytophthora infestans and its application Download PDFInfo
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- CN108949899A CN108949899A CN201810912673.2A CN201810912673A CN108949899A CN 108949899 A CN108949899 A CN 108949899A CN 201810912673 A CN201810912673 A CN 201810912673A CN 108949899 A CN108949899 A CN 108949899A
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- phytophthora infestans
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
Abstract
The invention belongs to Resistant breeding technical fields, identification method and its application more particularly, to a kind of Physiological Race of Phytophthora infestans.The identification method of Physiological Race of Phytophthora infestans is the following steps are included: sick nursery field is arranged in (a) in the present invention；(b) potato host material is cultivated on culture medium, then carries out water planting, form Rooted Cuttings；(c) the sick nursery field by water planting transplantation of seedlings into step (a)；(d) disease survey is carried out to the potato in step (c), records the type of late blight of potato biological strain.The present invention selects Infected Field to carry out experimental study, scientific and effective and environmental-friendly advantage is taken into account, whole experiment process is of less demanding to professional technique, operation is simple and feasible, experiment latter half is moved to crop field to identify, experiment effect is more significant, shortens experiment duration, while improving experiment accuracy.
Description
Technical field
The present invention relates to Resistant breeding technical fields, more particularly, to a kind of Physiological Race of Phytophthora infestans
Identification method and its application.
Background technique
The late blight of potato is a kind of typical aeroborne disease as caused by phytophthora infestans, encounters suitable temperature and humidity disease
Quickly spread.Pathogen is usually bred with vegetative manner；When two kinds of mating type bacterial strains (A1 and A2) exist simultaneously,
It is bred also by zoogamy organ egg spore.With the migration of A2 mating type, it is small to there is the stronger physiology of many virulence
Kind, the disease resistance of late blight of potato germ is enhanced, the generation and prevalence of disease have been aggravated.
According to incompletely statistics, the biological strain of phytophthora infestans is easy to happen variation and tends to be complicated all over the world
The probability and type difference changed, and occurred in different potato main producing regions.In order to identify the physiology of phytophthora infestans
Small seed type, the division of late disease bacteria biological strain at present, using it is a set of be respectively provided with R0, R1, R2, R3, R4, R5, R6, R7,
12 standard differential hosts of R8, R9, R10 and R11 carry out identification of living body, anti-on the above differential host according to late disease bacteria
It should determine biological strain.In the experimentation of identification Physiological Race of Phytophthora infestans, phytophthora infestans pass through
It is carried out obtained by isolation and purification culture after sick sample sampling, differential host is usually to cultivate ability after 1-3 months in tissue cultural seedlings of free room
Leaf-cutting is carried out, and 3-5 repetition need to be arranged in experiment every time, and guarantees to test pollution-free, existing phytophthora infestans physiology is small
It is high that the identification technology of kind has the disadvantages that identification technology requires, and cumbersome, larger workload takes a long time.
In consideration of it, the present invention is specifically proposed.
Summary of the invention
It is an object of the present invention to provide a kind of identification method of Physiological Race of Phytophthora infestans, this method is utilized
Infected Field carries out experiment, and operation is simple and feasible, shortens experiment duration, and experiment effect is more significant.
It is a further object to provide the identification methods of Physiological Race of Phytophthora infestans as described above to exist
Application in the identification of late disease bacteria biological strain.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
A kind of identification method of late blight of potato biological strain, includes the following steps:
(a) sick nursery field is set；
(b) potato host material is cultivated on culture medium, then carries out water planting, form Rooted Cuttings；
(c) the sick nursery field by water planting transplantation of seedlings into step (a)；
(d) disease survey is carried out to the potato in step (c), records the type of late blight of potato biological strain.
Preferably, the sick nursery field in the step (a) is that the late blight of potato is often sent out or high-incidence representative field.
Preferably, in the step (b), totally 12 kinds of potato host material, respectively R0, R1, R2, R3, R4, R5, R6,
R7, R8, R9, R10 and R11.
Preferably, the culture medium is MS culture medium.
Preferably, culture medium temperature is 22-28 DEG C.
Preferably, in the step (b), 3-5 then moves to water planting room after the leaf phase.
Preferably, in the step (b), the water planting time is 10-15 days；
Preferably, in the step (c), 7-10 plants of the water planting transplantation of seedlings of each differential host.
Preferably, in the step (d), the type of Physiological Race of Phytophthora infestans is with R0 morbidity for control, certain
Kind R differential host, which falls ill, corresponds to certain small seed type of r germ；
The potato leaf state of an illness is divided into 2 grades: 1 grade: susceptible, >=20% leaf area is fallen ill；0 grade: disease-resistant, < 20%
Leaf area morbidity.
The identification method of Physiological Race of Phytophthora infestans as described above is in late disease bacteria Race Identification
Using.
Compared with the prior art, the invention has the following beneficial effects:
(1) present invention in Physiological Race of Phytophthora infestans identification method, select Infected Field carry out experimental study,
Scientific and effective and environmental-friendly advantage is taken into account, whole experiment process is of less demanding to professional technique, and operation is simple and feasible, will
Experiment latter half moves to crop field and is identified, experiment effect is more significant, shortens experiment duration, accelerates experiment progress,
Experiment accuracy is improved simultaneously.
(2) identification method of Physiological Race of Phytophthora infestans has in late blight Race Identification in the present invention
Great meaning can more accurately and efficiently identify the type of different regions late disease bacteria biological strain.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person according to normal conditions or manufactures.
A kind of identification method of late blight of potato biological strain, includes the following steps:
(a) sick nursery field is set；
(b) potato host material is cultivated on culture medium, then carries out water planting, form Rooted Cuttings；
(c) the sick nursery field by water planting transplantation of seedlings into step (a)；
(d) disease survey is carried out to the potato in step (c), records the type of late blight of potato biological strain.
Infected Field is selected to carry out experimental study in the present invention, whole experiment process is of less demanding to professional technique, operation
Experiment latter half is moved to crop field and identified by simple possible, and experiment effect is more significant, shortens experiment duration, is accelerated
Experiment progress, while improving experiment accuracy.
Preferably, the sick nursery field in the step (a) is that the late blight of potato is often sent out or high-incidence representative field.
Sick nursery field select Liaoning, Jilin, Heilungkiang, Shandong, Henan, Hebei, Shaanxi, Jiangxi, the Inner Mongol, Sichuan, Yunnan,
The high-incidence representative field of the late blight of potato in this 14 provinces of Guizhou, Hubei and Chongqing；
The sick nursery field selection late blight of potato is often sent out or high-incidence representative field, experiment effect are more significant.
Preferably, in the step (b), totally 12 kinds of potato host material, respectively R0, R1, R2, R3, R4, R5, R6,
R7, R8, R9, R10 and R11.
Preferably, the culture medium is MS culture medium.
Preferably, culture medium temperature is 22-28 DEG C.
Preferably, it is, for example, 22 DEG C, 23 DEG C, 24 DEG C, 25 DEG C, 26 DEG C, 27 DEG C that culture medium temperature is typical but non-limiting
Or 28 DEG C.
Under the conditions of suitable temperature, the healthier growth of potato set ability.
Preferably, in the step (b), 3-5 then moves to water planting room after the leaf phase.
It is typical but non-limiting, water planting room is then moved to after 3 leaf phases, 4 leaf phases or 5 leaf phases.
Preferably, in the step (b), the water planting time is 10-15 days；
Preferably, it is, for example, 10 days, 11 days, 13 days, 14 days or 15 days 12 days that the water planting time is typical but non-limiting.
Preferably, in the step (c), 7-10 plants of the water planting transplantation of seedlings of each differential host.
The typical but non-limiting water planting transplantation of seedlings strain number of each differential host is, for example, 7 plants, 8 plants, 9 plants or 10 plants.
Preferably, in the step (d), the type of Physiological Race of Phytophthora infestans is with R0 morbidity for control, certain
Kind R differential host, which falls ill, corresponds to certain small seed type of r germ, according to the biological strain of phytophthora infestans unified in the world
Naming method is named (Black et al.1953)；
The potato leaf state of an illness is divided into 2 grades: 1 grade: susceptible, >=20% leaf area is fallen ill；0 grade: disease-resistant, < 20%
Leaf area morbidity.
Statistical result: 0 indicates 0 grade, and 1 indicates 1 grade
One Physiological Race of Phytophthora infestans type Control table of table
When R0 shows as susceptible, if R1 shows as susceptible, R2-R11 shows as disease-resistant, the then potato to be identified
Late disease bacteria is r1 type Physiological Race of Phytophthora infestans.
When R0 shows as susceptible, if R1 and R8 show as it is susceptible, other differential hosts show as it is disease-resistant, then should wait reflect
Other phytophthora infestans are r1.8 type Physiological Race of Phytophthora infestans.
When R0 shows as susceptible, if R1, R3 and R11 show as it is susceptible, other differential hosts show as it is disease-resistant, then should
Phytophthora infestans to be identified are r1.3.11 type Physiological Race of Phytophthora infestans.
When R0 shows as susceptible, if R2, R4, R6 and R10 show as it is susceptible, other differential hosts show as it is disease-resistant,
Then the phytophthora infestans to be identified are r2.4.6.10 type Physiological Race of Phytophthora infestans.
When R0 shows as susceptible, if R1, R2, R3, R5, R7 and R9 show as susceptible, other differential hosts are shown as
Disease-resistant, then the phytophthora infestans to be identified are r1.2.3.5.7.9 type Physiological Race of Phytophthora infestans.
By parity of reasoning, can identify more than 1,000 kinds of Physiological Race of Phytophthora infestans using identification method of the invention.
It is preferably carried out in scheme in one kind, the identification method of late blight of potato biological strain specifically:
(a) sick nursery field is arranged: in potato main producing region, the selection late blight of potato is often sent out or high-incidence representative field；
(b) water planting is numerous fastly: training after the detoxification test tube plantlet of R0-R11 totally 12 kinds of standard differential hosts is sheared indoors in MS
It supports and is cultivated on base, cultivation temperature is 22-28 DEG C, and the 3-5 leaf phase moves back to water planting room and carries out cuttage and quick-propagation, is grown 10-15 days
Afterwards, it takes root for forming water planting strong sprout；
(c) Rooted Cuttings are into sick nursery: the Rooted Cuttings of differential host being transplanted to the sick nursery field in step (a), each identification is posted
7-10 plants of main transplanting；
(d) disease survey: it is control with R0 morbidity in late blight of potato epizootic modeling, investigates all of different main producing regions
The incidence of differential host R1, R2, R3, R4, R5, R6, R7, R8, R9, R10 and R11, with certain R differential host morbidity pair
Certain small seed type of r germ is answered, is named according to the biological strain naming method of phytophthora infestans unified in the world
(Black et al.1953), while recording the biological strain type of phytophthora infestans；
The potato leaf state of an illness is divided into 2 grades: 1 grade: susceptible, >=20% leaf area is fallen ill；0 grade: disease-resistant, < 20%
Leaf area morbidity.
Statistical result: 0 indicates 0 grade, and 1 indicates 1 grade.
Physiological Race of Phytophthora infestans type Control table is as shown in Table 1.
The identification method of Physiological Race of Phytophthora infestans as described above is in late disease bacteria Race Identification
Using.Infected Field is selected to carry out experiment, experiment effect is more significant, shortens experiment duration, accelerates experiment progress, simultaneously
Improve experiment accuracy.
Embodiment 1
A kind of identification method of Physiological Race of Phytophthora infestans, comprising the following steps:
(a) sick nursery field is set: selecting the horse in Liaoning, Jilin, Heilungkiang, Shandong, Henan, this 7 provinces of Hebei and Shaanxi
The high-incidence representative field of bell potato late blight；
(b) water planting is numerous fastly: training after the detoxification test tube plantlet of R0-R11 totally 12 kinds of standard differential hosts is sheared indoors in MS
It supports and is cultivated on base, culture medium temperature is 22 DEG C, and 3 leaf phases moved back to water planting room and carry out cuttage and quick-propagation, raw after growth 10 days
Root forms water planting strong sprout；
(c) Rooted Cuttings are into sick nursery: the Rooted Cuttings of differential host being transplanted to the sick nursery field in step (a), each area is every
A differential host transplants 10 plants；
(d) disease survey: in late blight of potato epizootic modeling, it is control with R0 morbidity, investigates all mirror of different regions
The incidence of other host R1, R2, R3, R4, R5, R6, R7, R8, R9, R10 and R11 are fallen ill with certain R differential host and are corresponded to
Certain small seed type of r germ, is named according to the biological strain naming method of phytophthora infestans unified in the world
(Black et al.1953), while recording the biological strain type of phytophthora infestans；
The potato leaf state of an illness is divided into 2 grades: 1 grade: susceptible, >=20% leaf area is fallen ill；0 grade: disease-resistant, < 20%
Leaf area morbidity.
Statistical result: 0 indicates 0 grade, and 1 indicates 1 grade.
The Physiological Race of Phytophthora infestans type of 7, two Liaoning of table etc. different regions
Embodiment 2
A kind of identification method of Physiological Race of Phytophthora infestans, comprising the following steps:
(a) sick nursery field is set: selecting the horse in Jiangxi, the Inner Mongol, Sichuan, Yunnan, Guizhou, this 7 provinces of Hubei and Chongqing
The high-incidence representative field of bell potato late blight；
(b) water planting is numerous fastly: training after the detoxification test tube plantlet of R0-R11 totally 12 kinds of standard differential hosts is sheared indoors in MS
It supports and is cultivated on base, culture medium temperature is 28 DEG C, and 5 leaf phases moved back to water planting room and carry out cuttage and quick-propagation, raw after growth 15 days
Root forms water planting strong sprout；
(c) Rooted Cuttings are into sick nursery: the Rooted Cuttings of differential host being transplanted to the sick nursery field in step (a), each area is every
A differential host transplants 7 plants；
(d) disease survey: in late blight of potato epizootic modeling, it is control with R0 morbidity, investigates all mirror of different regions
The incidence of other host R1, R2, R3, R4, R5, R6, R7, R8, R9, R10 and R11 are fallen ill with certain R differential host and correspond to certain
The kind small seed type of r germ, is named according to the biological strain naming method of phytophthora infestans unified in the world
(Black et al.1953), while recording the biological strain type of phytophthora infestans；
The potato leaf state of an illness is divided into 2 grades: 1 grade: susceptible, >=20% leaf area is fallen ill；0 grade: disease-resistant, < 20%
Leaf area morbidity.
Statistical result 0 indicates 0 grade, and 1 indicates 1 grade.
The Physiological Race of Phytophthora infestans type of 7, three Jiangxi of table etc. different regions
Using the identification method in the present invention to the Physiological Race of Phytophthora infestans of 14 different regions such as Liaoning
Type identified, table two and table three it can be seen that the Physiological Race of Phytophthora infestans of different regions type.The present invention
The discrimination method of middle Physiological Race of Phytophthora infestans selects Infected Field to carry out experimental study, has taken into account scientific and effective and ring
The advantage of border close friend, it is simple and easy, shorten experimental period, accuracy with higher.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: its according to
So be possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features into
Row equivalent replacement；And these are modified or replaceed, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of identification method of Physiological Race of Phytophthora infestans, which comprises the steps of:
(a) sick nursery field is set；
(b) potato host material is cultivated on culture medium, then carries out water planting, form Rooted Cuttings；
(c) the sick nursery field by water planting transplantation of seedlings into step (a)；
(d) disease survey is carried out to the potato in step (c), records the type of late blight of potato biological strain.
2. the identification method of late blight of potato biological strain according to claim 1, which is characterized in that the step
(a) the sick nursery field in often sends out for the late blight of potato or high-incidence representative field.
3. the identification method of Physiological Race of Phytophthora infestans according to claim 1, which is characterized in that the step
(b) in, potato host material totally 12 kinds, respectively R0, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10 and R11.
4. the identification method of Physiological Race of Phytophthora infestans according to claim 1, which is characterized in that the culture
Base is MS culture medium.
5. the identification method of Physiological Race of Phytophthora infestans according to claim 4, which is characterized in that culture medium temperature
Degree is 22-28 DEG C.
6. the identification method of Physiological Race of Phytophthora infestans according to claim 1, which is characterized in that the step
(b) in, 3-5 then moves to water planting room after the leaf phase.
7. the identification method of Physiological Race of Phytophthora infestans according to claim 1, which is characterized in that the step
(b) in, the water planting time is 10-15 days.
8. the identification method of Physiological Race of Phytophthora infestans according to claim 1, which is characterized in that the step
(c) in, 7-10 plants of the water planting transplantation of seedlings of each differential host.
9. the identification method of Physiological Race of Phytophthora infestans according to claim 1-8, which is characterized in that
In the step (d), the type of Physiological Race of Phytophthora infestans is with R0 morbidity for control, certain R differential host morbidity
Certain corresponding small seed type of r germ；
The potato leaf state of an illness is divided into 2 grades: 1 grade: susceptible, >=20% leaf area is fallen ill；0 grade: disease-resistant, 20% blade face <
Product morbidity.
10. as the identification method of the described in any item Physiological Race of Phytophthora infestans of claim 1-9 is raw in late disease bacteria
Manage the application in Race identification.
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| CN111316918A (en) * | 2020-04-20 | 2020-06-23 | 福建农林大学 | Method for quickly water-culturing phytophthora solani physiological race resistance tissue culture seedling |
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| CN110628650A (en) * | 2019-10-17 | 2019-12-31 | 河南科技学院 | Phytophthora infestans culture medium RIS and method for detecting phytophthora infestans pathogenicity to plants |
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| CN111316918A (en) * | 2020-04-20 | 2020-06-23 | 福建农林大学 | Method for quickly water-culturing phytophthora solani physiological race resistance tissue culture seedling |
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| CN108949899B (en) | 2022-04-29 |
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