CN109526443A - A kind of rapid identification method of radish disease resistance - Google Patents
A kind of rapid identification method of radish disease resistance Download PDFInfo
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- CN109526443A CN109526443A CN201910031999.9A CN201910031999A CN109526443A CN 109526443 A CN109526443 A CN 109526443A CN 201910031999 A CN201910031999 A CN 201910031999A CN 109526443 A CN109526443 A CN 109526443A
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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
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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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Abstract
The invention belongs to plant resistance to environment stress identification technology fields, and in particular to a kind of rapid identification method of radish disease resistance.There is the radish of disease including collecting, obtain disease pollutant；Kind to be detected is planted in big Tanaka；When radish growth is to disease pollutant is inoculated between 3~8 leaf periods, 3~4 leaf phases, 7~8 leaf phases are each primary, and 7~8 leaf phases were inoculated with the 10th day investigation disease index after disease pollutant；Plant resistance to environment stress is determined according to disease index.Method of the invention increases the operation of artificial infection disease pollutant on the basis of natural Disease garden identification, has not only accelerated the probability of catching an illness of Infected Field, but also improves radish resistant variety screening accuracy rate.
Description
Technical field
The invention belongs to plant resistance to environment stress identification technology fields, and in particular to a kind of rapid identification method of radish disease resistance.
Background technique
The diseases such as black rot, black spot are radish planting Common Diseases, these diseases not only reduce radish yield and
Kind, and affect the income of grower.Disease is solved on the one hand to need to carry out disease-resistant disappear during radish planting
Work is killed, on the other hand then needs actively to cultivate radish disease-resistant varieties.
The prior art carries out new varieties anti-disease enzyme and is planted in sick nursery mostly, carries out resistance mirror using spontaneous induction
It is fixed, however at that time or local weather conditions and be centainly suitable for inducing disease, frequently resulting in identification work can not be successfully progress.For
Solve the problems, such as this, " research of radish black rot seedling restance identification method, Gan Caixia, hubei agricultural science, volume 49
12 phases " disclose a kind of radish black rot seedling stage indoors artificial inoculation identification method, by control inoculum density, temperature etc. because
Element can smoothly identify radish varieties disease resistance.But paper use is to sample separation identification from lesion to obtain for examination strain
, only tri- strains of HB1, WHHB1, JZHB1, then its disease-resistant variety identified also only resists the product of these three strains
Kind, resist the radish varieties of other microorganisms are different surely to be screened out.Furthermore the paper using artificial infection specified strain come
Induce disease, it is larger with the nursery of spontaneous induction identification mode difference, cause the strain of radish black rot not only these three,
It is also likely to be multiple strain comprehensive functions as a result, therefore, it is disease-resistant that the research method presence leakage of Gan Caixia et al. filters out radish
The case where property kind.
Summary of the invention
A kind of rapid identification method of radish disease resistance provided by the invention, increases on the basis of natural Disease garden identification
The operation of artificial infection disease pollutant, had not only accelerated the probability of catching an illness of Infected Field, but also improved the screening of radish resistant variety
Accuracy rate.
The rapid identification method for being to provide a kind of radish disease resistance of the invention, comprising the following steps:
S1 collects the radish with disease, is added in acetum, impregnates after radish is cut into radish fourth, filter, collects
Filtrate obtains disease pollutant；
S2 plants kind to be detected in big Tanaka, and planting patterns is as follows: being planted by the way of digging cave dibbling, every cave 1
~2 seeds, 10~20cm of spacing in the rows, 10~30cm of line-spacing；
S3, field management, when radish growth to inoculation disease pollutant, 3~4 leaf phases, 7~8 leaves between 3~8 leaf periods
Phase is each primary, and 7~8 leaf phases were inoculated with the 10th day investigation disease index after disease pollutant；
Disease investigation classification is as follows: 0 grade, disease-free；0.5 grade, there are several sick classes on blade face, and region of disease is less than 1/4 blade face
Product；Region of disease is more than or equal to 1/4 leaf area on 1 grade of leaf, and less than 3/8 leaf area；Region of disease is more than or equal to 3/8 leaf on 2 grades of leaves
Area, and less than 1/2 leaf area；Region of disease is more than or equal to 1/2 leaf area on 3 grades of leaves, and is less than or equal to 3/4 leaf area, 4 grades of leaves
Upper region of disease is greater than 3/4 leaf area；
Disease index=100 × ∑ (the diseased plant numbers at different levels × disease grade value)/(investigation total strain number × 4)
As disease index > 40, then kind to be detected is susceptible variety；It is as 40≤disease index < 20, then to be detected
Kind is disease tolerant variety；As 20≤disease index < 10, then kind to be detected is disease-resistant variety；When disease index≤10,
Then kind to be detected is high disease-resistant variety.
Preferably, the rapid identification method of above-mentioned radish disease resistance, the mass fraction of the acetum are 1~3%, leaching
The bubble time is 5~20min.
Preferably, the mass ratio of the rapid identification method of above-mentioned radish disease resistance, radish fourth and acetum is 1:1~2.
Preferably, the rapid identification method of above-mentioned radish disease resistance, the method for being inoculated with disease pollutant are as follows: by disease dirt
Dilution after dye object dilutes 1000 times is sprayed on blade face, per acre 20~30L dilution.
Preferably, the rapid identification method of above-mentioned radish disease resistance, the radish fourth side length after radish is diced is 0.5~1cm.
Preferably, the rapid identification method of above-mentioned radish disease resistance, the tool that uses of step 1 filtering are mesh screen, and mesh screen
Bore dia be 0.5~2mm.
Preferably, the rapid identification method of above-mentioned radish disease resistance, S1 step is the following steps are included: by disease resistance radish product
Kind is used as type strain A；Using susceptible radish varieties as type strain B；
S2 step is further comprising the steps of: type strain A, type strain B and kind to be detected being planted in big Tanaka, and pressed
Arrangement plantation radish is repeated according to the sequence of a line type strain A, a line type strain B, a line kind to be detected；
S3 is further comprising the steps of: 7~8 leaf phases were inoculated with the 10th day Investigate incidence after disease pollutant；
Disease incidence=morbidity total strain number/investigation total strain number；
The disease incidence of type strain A is denoted as KA, and the disease incidence of type strain B is denoted as KB, and the disease incidence of kind to be detected is denoted as KD；
As KD≤KA, then kind to be detected is disease-resistant variety；As KD >=KB, then kind to be detected is susceptible variety.
Preferably, the rapid identification method of above-mentioned radish disease resistance, type strain A are identified according to claim 1 method
Disease index is 35~39, and type strain B is 40~45 according to the disease index that claim 1 method identifies；As KB < KD <
KA, then kind to be detected is susceptible variety.
Compared with prior art, the rapid identification method of radish disease resistance of the invention, has the advantages that
The present invention increases the operation of artificial infection disease pollutant on the basis of natural Disease garden identification, the bacterium being inoculated with
Strain has the radish of disease from itself, and when needing to identify which kind of is sick, then there is the radish of which kind of disease to prepare disease for selection
Pollutant, if the radish of selection has two or more diseases simultaneously, can firm kind simultaneously a variety of sick resistances, improve
Radish resistant variety screens accuracy rate.Method of the invention can also accelerate the probability of catching an illness of Infected Field.
It is the survival rate and proliferative speed for improving microorganism that the present invention, which uses the reason of acetum, studies have shown that using
When the acetum that mass fraction is 1~3% saves the rape Xanthomonas campestris for causing radish black rot, rape Huang unit cell after 3 days
The order of magnitude of bacillus is by 102Increase to 106；And when sterile water being used to save the rape Xanthomonas campestris for causing radish black rot, 3 days
The order of magnitude of rape Xanthomonas campestris is by 10 afterwards2Increase to 105。
Specific embodiment
The present invention is described in detail combined with specific embodiments below, but should not be construed as limitation of the invention.It is following
The test method of actual conditions is not specified in embodiment, operates usually according to normal condition, due to not being related to inventive point, thus it is not right
Its step is described in detail.
Embodiment 1
A kind of rapid identification method of radish disease resistance, comprising the following steps:
S1 collects the radish with black rot evil, and addition mass fraction is molten for 1% acetic acid after radish is cut into radish fourth
In liquid, the radish fourth side length (maximum width meter) after radish is diced is 0.5~0.7cm, the mass ratio of radish fourth and acetum
For 1:2,5min is impregnated, filtering, filtering the tool used is mesh screen, and the bore dia of mesh screen is 1mm, collects filtrate, obtains disease
Evil pollutant；
Kind to be detected is appeared green turnip (known black rot resistance, and the kind comprehensive disease resistance is strong) plantation in big by S2
Tanaka, planting patterns are as follows: being planted by the way of digging cave dibbling, 2, every cave seed, spacing in the rows 10cm, line-spacing 10cm；
S3, field management, when radish growth to inoculation disease pollutant, 3~4 leaf phases, 7~8 leaves between 3~8 leaf periods
Phase is each primary, and the method for being inoculated with disease pollutant is as follows: the dilution after 1000 times of disease pollutant dilution is sprayed on blade face,
20L dilution per acre；
7~8 leaf phases were inoculated with the 10th day investigation disease index after disease pollutant；
Disease investigation classification is as follows: 0 grade, disease-free；0.5 grade, there are several sick classes on blade face, and region of disease is less than 1/4 blade face
Product；Region of disease is more than or equal to 1/4 leaf area on 1 grade of leaf, and less than 3/8 leaf area；Region of disease is more than or equal to 3/8 leaf on 2 grades of leaves
Area, and less than 1/2 leaf area；Region of disease is more than or equal to 1/2 leaf area on 3 grades of leaves, and is less than or equal to 3/4 leaf area, 4 grades of leaves
Upper region of disease is greater than 3/4 leaf area；
Total 100 plants of the investigation of the present embodiment, wherein 0 grade 24 plants, 0.5 grade 40 plants, 1 grade 17 plants, 2 grades 13 plants, 3 grades 5 plants, 4
1 plant of grade.
Disease index=100 × ∑ (the diseased plant numbers at different levels × disease grade value)/(investigation total strain number × 4)=100 × (24 × 0+
40 × 0.5+17 × 1+13 × 2+5 × 3+1 × 4)/(100 × 4)=20.5
It is identified through this method, green turnip of appearing is disease-resistant variety, consistent with result generally acknowledged at present.
Embodiment 2
A kind of rapid identification method of radish disease resistance, comprising the following steps:
S1 collects the radish with black rot evil, and addition mass fraction is molten for 3% acetic acid after radish is cut into radish fourth
In liquid, the radish fourth side length (maximum width meter) after radish is diced is 0.8~1cm, and the mass ratio of radish fourth and acetum is
1:1 impregnates 20min, and filtering, filtering the tool used is mesh screen, and the bore dia of mesh screen is 2mm, collects filtrate, obtains disease
Pollutant；
S2, by kind all round victory to be detected, red flesh, 91-1 radish (known resisting turnip mosaic virus, downy mildew, but not
Black rot resistance) in big Tanaka, planting patterns is as follows: being planted by the way of digging cave dibbling, 1, every cave seed, spacing in the rows for plantation
15cm, line-spacing 30cm；
S3, field management, when radish growth to inoculation disease pollutant, 3~4 leaf phases, 7~8 leaves between 3~8 leaf periods
Phase is each primary, and the method for being inoculated with disease pollutant is as follows: the dilution after 1000 times of disease pollutant dilution is sprayed on blade face,
30L dilution per acre；
7~8 leaf phases were inoculated with the 10th day investigation disease index after disease pollutant；
Disease investigation classification is as follows: 0 grade, disease-free；0.5 grade, there are several sick classes on blade face, and region of disease is less than 1/4 blade face
Product；Region of disease is more than or equal to 1/4 leaf area on 1 grade of leaf, and less than 3/8 leaf area；Region of disease is more than or equal to 3/8 leaf on 2 grades of leaves
Area, and less than 1/2 leaf area；Region of disease is more than or equal to 1/2 leaf area on 3 grades of leaves, and is less than or equal to 3/4 leaf area, 4 grades of leaves
Upper region of disease is greater than 3/4 leaf area；
Total 100 plants of the investigation of the present embodiment, wherein 0 grade 15 plants, 0.5 grade 14 plants, 1 grade 16 plants, 2 grades 19 plants, 3 grades 21 plants, 4
15 plants of grade.
Disease index=100 × ∑ (the diseased plant numbers at different levels × disease grade value)/(investigation total strain number × 4)=100 × (15 × 0+
14 × 0.5+16 × 1+19 × 2+21 × 3+15 × 4)/(100 × 4)=46.0
It is identified through this method, all round victory red flesh 91-1 radish is susceptible variety, consistent with result generally acknowledged at present.
Embodiment 3
A kind of rapid identification method of radish disease resistance, comprising the following steps:
S1 collects the radish with blackspot disease, and addition mass fraction is molten for 4% acetic acid after radish is cut into radish fourth
In liquid, the radish fourth side length (maximum width meter) after radish is diced is 1.1~1.3cm, the mass ratio of radish fourth and acetum
For 1:3,15min is impregnated, filtering, filtering the tool used is mesh screen, and the bore dia of mesh screen is 0.5mm, collects filtrate, is obtained
Disease pollutant；
S2, by kind all round victory to be detected, red flesh, 91-1 radish (known resisting turnip mosaic virus, downy mildew, but not
Anti- black spot) in big Tanaka, planting patterns is as follows: being planted by the way of digging cave dibbling, 2, every cave seed, spacing in the rows for plantation
20cm, line-spacing 20cm；
S3, field management, when radish growth to inoculation disease pollutant, 3~4 leaf phases, 7~8 leaves between 3~8 leaf periods
Phase is each primary, and the method for being inoculated with disease pollutant is as follows: the dilution after 1000 times of disease pollutant dilution is sprayed on blade face,
25L dilution per acre；
7~8 leaf phases were inoculated with the 10th day investigation disease index after disease pollutant；
Disease investigation classification is as follows: 0 grade, disease-free；0.5 grade, there are several sick classes on blade face, and region of disease is less than 1/4 blade face
Product；Region of disease is more than or equal to 1/4 leaf area on 1 grade of leaf, and less than 3/8 leaf area；Region of disease is more than or equal to 3/8 leaf on 2 grades of leaves
Area, and less than 1/2 leaf area；Region of disease is more than or equal to 1/2 leaf area on 3 grades of leaves, and is less than or equal to 3/4 leaf area, 4 grades of leaves
Upper region of disease is greater than 3/4 leaf area；
Total 100 plants of the investigation of the present embodiment, wherein 0 grade 10 plants, 0.5 grade 16 plants, 1 grade 21 plants, 2 grades 28 plants, 3 grades 16 plants, 6
9 plants of grade.
Disease index=100 × ∑ (the diseased plant numbers at different levels × disease grade value)/(investigation total strain number × 4)=100 × (10 × 0+
16 × 0.5+21 × 1+28 × 2+16 × 3+9 × 4)/(100 × 4)=42.2
It is identified through this method, all round victory red flesh 91-1 radish is susceptible variety, consistent with result generally acknowledged at present.
Embodiment 4
A kind of rapid identification method of radish disease resistance, comprising the following steps:
S1 collects the radish with black rot evil, and addition mass fraction is molten for 1% acetic acid after radish is cut into radish fourth
In liquid, the radish fourth side length (maximum width meter) after radish is diced is 0.5~0.7cm, the mass ratio of radish fourth and acetum
For 1:1,30min is impregnated, filtering, filtering the tool used is mesh screen, and the bore dia of mesh screen is 0.4mm, collects filtrate, is obtained
Disease pollutant；
Disease resistance radish varieties are appeared into green turnip as type strain A；By susceptible radish varieties all round victory, red flesh, 91-
1 radish is as type strain B；
S2, by type strain A, type strain B and kind to be detected like promise spring and summer sapphire fruit green turnip (known black rot resistance,
And the kind comprehensive disease resistance is strong) it plants in big Tanaka, and it is to be detected according to a line type strain A, a line type strain B, a line
The sequence of kind is repeated in arrangement plantation radish；Planting patterns is as follows: being planted by the way of digging cave dibbling, 2, every cave kind
Son, spacing in the rows 25cm, line-spacing 35cm；
S3, field management, when radish growth to inoculation disease pollutant, 3~4 leaf phases, 7~8 leaves between 3~8 leaf periods
Phase is each primary, and the method for being inoculated with disease pollutant is as follows: the dilution after 1000 times of disease pollutant dilution is sprayed on blade face,
20L dilution per acre；
7~8 leaf phases were inoculated with the 10th day investigation disease index after disease pollutant；
Disease investigation classification is as follows: 0 grade, disease-free；0.5 grade, there are several sick classes on blade face, and region of disease is less than 1/4 blade face
Product；Region of disease is more than or equal to 1/4 leaf area on 1 grade of leaf, and less than 3/8 leaf area；Region of disease is more than or equal to 3/8 leaf on 2 grades of leaves
Area, and less than 1/2 leaf area；Region of disease is more than or equal to 1/2 leaf area on 3 grades of leaves, and is less than or equal to 3/4 leaf area, 4 grades of leaves
Upper region of disease is greater than 3/4 leaf area；
Total 100 plants of the investigation of the present embodiment.
The disease incidence of type strain A=morbidity total strain number/investigation total strain number=75%
The disease incidence of type strain B=morbidity total strain number/investigation total strain number=85%
The disease incidence of kind to be measured=morbidity total strain number/investigation total strain number=71%
It is identified through this method, happiness promise spring and summer sapphire fruit green turnip is disease-resistant variety, consistent with result generally acknowledged at present.
It should be noted that described in the embodiment of the present invention 1~4 and " field management " mode it is as follows: take conventional
Fertilising, the operation such as weeding, plough, pour water.
It should be noted that when the present invention provides numberical range, it should be appreciated that except non-present invention is otherwise noted, every number
Being worth any one numerical value between two endpoints and two endpoints of range can be selected.Unless otherwise defined, make in the present invention
All technical and scientific terms are identical as the normally understood meaning of those skilled in the art of the present technique.Although this hair has been described
Bright preferred embodiment, once a person skilled in the art knows basic creative concepts, then can be to these embodiments
Make other change and modification.So the following claims are intended to be interpreted as including preferred embodiment and falls into the present invention
All change and modification of range.
Obviously, various changes and modifications can be made to the invention without departing from essence of the invention by those skilled in the art
Mind and range.In this way, if these modifications and changes of the present invention belongs to the range of the claims in the present invention and its equivalent technologies
Within, then the present invention is also intended to include these modifications and variations.
Claims (8)
1. a kind of rapid identification method of radish disease resistance, which comprises the following steps:
S1 collects the radish with disease, is added in acetum, impregnates after radish is cut into radish fourth, and filter is collected in filtering
Liquid obtains disease pollutant；
S2 plants kind to be detected in big Tanaka, and planting patterns is as follows: being planted by the way of digging cave dibbling, every cave 1~2
Grain seed, 10~20cm of spacing in the rows, 10~30cm of line-spacing；
S3, field management, when radish growth is to disease pollutant is inoculated between 3~8 leaf periods, 3~4 leaf phases, 7~8 leaf phases are each
Once, the 10th day investigation disease index after 7~8 leaf phases inoculation disease pollutant；
Disease investigation classification is as follows: 0 grade, disease-free；0.5 grade, there are several sick classes on blade face, and region of disease is less than 1/4 leaf area；1
Region of disease is more than or equal to 1/4 leaf area on grade leaf, and less than 3/8 leaf area；Region of disease is more than or equal to 3/8 leaf area on 2 grades of leaves,
And less than 1/2 leaf area；Region of disease is more than or equal to 1/2 leaf area on 3 grades of leaves, and is less than or equal to 3/4 leaf area, 4 grades of Ye Shangfa
Lesion is greater than 3/4 leaf area；
Disease index=100 × ∑ (the diseased plant numbers at different levels × disease grade value)/(investigation total strain number × 4)
As disease index > 40, then kind to be detected is susceptible variety；As 40≤disease index < 20, then kind to be detected
For disease tolerant variety；As 20≤disease index < 10, then kind to be detected is disease-resistant variety；When disease index≤10, then to
Detection kind is high disease-resistant variety.
2. the rapid identification method of radish disease resistance according to claim 1, which is characterized in that the matter of the acetum
Measuring score is 1~3%, and soaking time is 5~20min.
3. the rapid identification method of radish disease resistance according to claim 2, which is characterized in that radish fourth and acetum
Mass ratio be 1:1~2.
4. the rapid identification method of radish disease resistance according to claim 3, which is characterized in that inoculation disease pollutant
Method is as follows: the dilution after 1000 times of disease pollutant dilution being sprayed on blade face, per acre 20~30L dilution.
5. the rapid identification method of radish disease resistance according to claim 1, which is characterized in that radish dice after radish
Fourth side length is 0.5~1cm.
6. the rapid identification method of radish disease resistance according to claim 5, which is characterized in that step 1 filtering uses
Tool is mesh screen, and the bore dia of mesh screen is 0.5~2mm.
7. the rapid identification method of radish disease resistance according to claim 1, which is characterized in that S1 step includes following step
It is rapid: using disease resistance radish varieties as type strain A；Using susceptible radish varieties as type strain B；
S2 step is further comprising the steps of: type strain A, type strain B and kind to be detected being planted in big Tanaka, and according to one
Row type strain A, a line type strain B, a line kind to be detected sequence be repeated in arrangement plantation radish；
S3 is further comprising the steps of: 7~8 leaf phases were inoculated with the 10th day Investigate incidence after disease pollutant；
Disease incidence=morbidity total strain number/investigation total strain number；
The disease incidence of type strain A is denoted as KA, and the disease incidence of type strain B is denoted as KB, and the disease incidence of kind to be detected is denoted as KD；Work as KD
≤ KA, then kind to be detected is disease-resistant variety；As KD >=KB, then kind to be detected is susceptible variety.
8. the rapid identification method of radish disease resistance according to claim 7, which is characterized in that type strain A is according to right
It is required that the disease index that 1 method identifies is 35~39, type strain B is according to the disease index that claim 1 method identifies
40~45；As KB < KD < KA, then kind to be detected is susceptible variety.
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