CN111280008A - Comprehensive resistance identification method for broad bean rust disease - Google Patents
Comprehensive resistance identification method for broad bean rust disease Download PDFInfo
- Publication number
- CN111280008A CN111280008A CN202010106381.7A CN202010106381A CN111280008A CN 111280008 A CN111280008 A CN 111280008A CN 202010106381 A CN202010106381 A CN 202010106381A CN 111280008 A CN111280008 A CN 111280008A
- Authority
- CN
- China
- Prior art keywords
- disease
- resistance
- broad bean
- rust
- inoculation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 201000010099 disease Diseases 0.000 title claims abstract description 201
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 201
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 title claims abstract description 84
- 240000006677 Vicia faba Species 0.000 title claims abstract description 67
- 235000010749 Vicia faba Nutrition 0.000 title claims abstract description 67
- 235000002098 Vicia faba var. major Nutrition 0.000 title claims abstract description 67
- 238000000034 method Methods 0.000 title claims abstract description 54
- 239000000463 material Substances 0.000 claims abstract description 122
- 241000196324 Embryophyta Species 0.000 claims abstract description 76
- 238000011081 inoculation Methods 0.000 claims abstract description 73
- 239000000725 suspension Substances 0.000 claims abstract description 28
- 238000009331 sowing Methods 0.000 claims abstract description 27
- 241000894006 Bacteria Species 0.000 claims abstract description 16
- 238000011835 investigation Methods 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 42
- 238000011156 evaluation Methods 0.000 claims description 30
- 238000005507 spraying Methods 0.000 claims description 25
- 208000015181 infectious disease Diseases 0.000 claims description 13
- 238000005406 washing Methods 0.000 claims description 12
- 238000004140 cleaning Methods 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 11
- 239000007921 spray Substances 0.000 claims description 10
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- 230000006378 damage Effects 0.000 claims description 9
- 206010015535 Euphoric mood Diseases 0.000 claims description 7
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 6
- 230000000172 allergic effect Effects 0.000 claims description 6
- 208000010668 atopic eczema Diseases 0.000 claims description 6
- 238000013461 design Methods 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 230000017074 necrotic cell death Effects 0.000 claims description 6
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 6
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 238000002791 soaking Methods 0.000 claims description 6
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 6
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 5
- 230000035945 sensitivity Effects 0.000 claims description 2
- 238000009395 breeding Methods 0.000 abstract description 5
- 230000001488 breeding effect Effects 0.000 abstract description 5
- 230000001580 bacterial effect Effects 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 238000012216 screening Methods 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 238000012214 genetic breeding Methods 0.000 abstract description 2
- 238000011160 research Methods 0.000 abstract description 2
- 208000035240 Disease Resistance Diseases 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 8
- 230000007613 environmental effect Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 206010022000 influenza Diseases 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 241000221198 Basidiomycota Species 0.000 description 1
- 241000544594 Uromyces viciae-fabae Species 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 239000002872 contrast media Substances 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
-
- 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
- A01G22/40—Fabaceae, e.g. beans or peas
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06Q—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES; SYSTEMS OR METHODS SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES, NOT OTHERWISE PROVIDED FOR
- G06Q10/00—Administration; Management
- G06Q10/06—Resources, workflows, human or project management; Enterprise or organisation planning; Enterprise or organisation modelling
- G06Q10/063—Operations research, analysis or management
- G06Q10/0639—Performance analysis of employees; Performance analysis of enterprise or organisation operations
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06Q—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES; SYSTEMS OR METHODS SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES, NOT OTHERWISE PROVIDED FOR
- G06Q50/00—Information and communication technology [ICT] specially adapted for implementation of business processes of specific business sectors, e.g. utilities or tourism
- G06Q50/02—Agriculture; Fishing; Forestry; Mining
Landscapes
- Business, Economics & Management (AREA)
- Human Resources & Organizations (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Strategic Management (AREA)
- Economics (AREA)
- Entrepreneurship & Innovation (AREA)
- General Physics & Mathematics (AREA)
- Educational Administration (AREA)
- Marketing (AREA)
- Development Economics (AREA)
- Theoretical Computer Science (AREA)
- Tourism & Hospitality (AREA)
- Physics & Mathematics (AREA)
- General Business, Economics & Management (AREA)
- Game Theory and Decision Science (AREA)
- Quality & Reliability (AREA)
- Operations Research (AREA)
- Agronomy & Crop Science (AREA)
- Animal Husbandry (AREA)
- Marine Sciences & Fisheries (AREA)
- Mining & Mineral Resources (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Primary Health Care (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Pretreatment Of Seeds And Plants (AREA)
Abstract
The invention relates to a method for identifying comprehensive resistance of broad bean rust disease, belonging to the technical field of agricultural breeding. The method comprises the four steps of collecting bacteria sources, preparing a mixed spore suspension, preparing and sowing broad bean materials, inoculating broad bean rust in a greenhouse at a seedling stage, setting temperature and humidity, and identifying the resistance of the broad bean rust in the greenhouse at an adult stage. The method is carried out in a controllable environment of a greenhouse, has good repeatability, and relatively more materials can be identified at a time; the adoption of multi-bacterial source inoculation is beneficial to screening materials with comprehensive resistance genes, and compared with the specific resistance of small seeds screened by using a single bacterial strain, the resistant materials obtained by the method have the advantages of better stability, adaptability and practicability; the method for inoculation in the seedling stage and investigation in the adult plant stage is similar to the field disease characteristics of broad bean rust, and well simulates the field disease process. The invention provides an efficient and stable method for rust resistance identification and genetic breeding research of broad bean materials.
Description
Technical Field
The invention belongs to the technical field of agricultural breeding, and particularly relates to a broad bean rust disease comprehensive resistance identification method.
Background
Broad bean rust disease is an important broad bean disease caused by basidiomycete broad bean monad rust (Uromyces viciae-fabae (pers.) Schroet), and mainly occurs in middle east and northern africa regions. The method is frequently generated in spring and autumn broad bean production areas and summer sowing anti-season broad bean production areas in China, and has serious harm to the production of autumn broad beans, particularly broad beans in southwest planting areas. Broad bean rust can cause 30 to 40 percent of yield reduction in general years, 70 to 80 percent of yield reduction in pandemic years, and even destroy the yield.
At present, the rust disease control method mainly comprises cultivation technology control, chemical agent control and biological control, and the broad bean infection caused by rust disease can be obviously reduced by controlling seeding density, reducing field humidity and mixed culture among different crops in cultivation. Chemical control and biological control with low cost and environmental protection have not achieved good progress so far.
In view of the harmfulness of the broad bean rust, the breeding of disease-resistant varieties and the screening of rust-resistant genes are used as effective ways for preventing and treating the broad bean rust in all major broad bean producing countries in the world. The establishment of a stable and efficient broad bean rust resistance identification system is an important prerequisite for rust resistance breeding and gene excavation. At present, reported broad bean rust identification methods mainly focus on a single strain seedling stage identification method in an incubator, a field adult stage natural disease garden identification method, an in vitro leaf identification method and the like. The identification method of single strain in the incubator has high repeatability and is slightly influenced by environmental factors, but the operation is complex and is not suitable for the resistance identification of a large amount of materials; meanwhile, the resistant materials screened by using a single strain are mostly small species specialized resistance, and the stability, the adaptability and the practicability are poor; the consistency of the seedling stage identification result and the adult stage identification result is different, and the comprehensive analysis needs to be carried out by combining the field adult stage identification result. The evaluation rule of the natural disease garden in the field plant growing period is greatly influenced by the field environment, and the repeatability and the accuracy of data are poor. The difference between the in vitro leaf identification method and the actual situation of the field environment is large, and the obtained material is difficult to put into the actual production. Therefore, establishing a set of high-efficiency, accurate and high-repeatability broad bean rust resistance identification system has important significance for improving the screening and application of broad bean rust resistance materials. Therefore, how to overcome the defects of the prior art is a problem to be solved urgently in the technical field of agricultural breeding at present.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides the method for identifying the comprehensive resistance of the broad bean rust, which is efficient and stable and is easy to popularize and apply.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a method for identifying the comprehensive resistance of broad bean rust comprises the following steps:
step (1), collecting bacteria sources and preparing a mixed spore suspension:
a) collecting a bacterium source: collecting rust spores on infected plants;
b) preparation of mixed spore suspension: collecting equal amount of rust spores collected from different places, mixing uniformly, pouring the obtained mixed spore powder into sterilized distilled water, adding 0.02-0.05% (v/v) Tween-20, and making into 4 × 105Spore. ml-1The mixed spore suspension of (a);
step (2), broad bean material preparation and sowing:
a) design of control material: setting disease-resistant and susceptible materials as a control group;
b) seed cleaning: before sowing, cleaning disease-resistant and disease-susceptible materials and seeds of materials to be identified, and removing the seeds which are shriveled, broken, germinated, susceptible, damaged by worms, mildewed and damaged by seed coats;
c) seed disinfection: washing seeds with clear water for many times, soaking the seeds in a sodium hypochlorite solution with the mass concentration of 0.5% for 10-20s, then immediately washing the seeds with the clear water for many times, and then immediately sowing the seeds;
and (3) inoculating broad bean rust in seedling stage in a greenhouse and setting temperature and humidity:
a) inoculation: inoculating disease-resistant and disease-susceptible materials and materials to be identified in 4-5 leaf stages of plants, humidifying greenhouse, disease-resistant and disease-susceptible materials and plants to be identified by spraying water before inoculation, ensuring that the surfaces of leaves are wet and the humidity of the greenhouse is kept between 80% and 100%, starting inoculation before 10 am or 2 pm, and performing spray inoculation on the disease-resistant and disease-susceptible materials and plants to be identified by adopting the mixed spore suspension obtained in the step (1);
b) temperature and humidity setting after inoculation:
1-21d after rust inoculation, controlling the temperature of the greenhouse at 20-25 ℃ in the daytime and 10-15 ℃ at night, and opening the greenhouse environment after 21d of inoculation so as to ensure the consistency of the internal environment and the external environment as much as possible;
spraying with water for 3-5 times a day from 1 st to 14 th days after inoculation to ensure plant surface moistening and greenhouse humidity stability at above 95%, spraying with water for 1 time a day from 15 th to 21 th days, and spraying without water after 22 th day;
step (4), identifying the resistance of the greenhouse broad bean rust in the adult plant stage:
a) and (4) disease investigation: after inoculation for 80-85d, grading the disease condition in the adult plant period;
b) grading the disease condition: adopting a six-level division method:
level 0: no visible infection;
level 1: the leaves only have allergic necrosis reaction and no sporophyte;
and 3, level: sporadic sporangium mass in the leaf; no damage and breakage, and the diameter of spore pile is less than 0.5 mm; occupies less than 5.0 percent of the leaf area;
and 5, stage: the spore pile on the leaf and the stem is common and large; the diameter of the spore pile is 0.51-1 mm; partial sporangial masses are broken; occupies 5.10 to 10.0 percent of the leaf area;
and 7, stage: the spore pile is common and large on the leaves and stems; the diameter is more than 1.1 mm; severe rupture of the sporangium; the small area of the leaf surface is faded and discolored; the area of the leaf is 10.1-25.0%;
and 9, stage: the spores are densely stacked and fully distributed, and account for more than 25.1 percent of the leaf area; large area of blade blackens or falls off, and plants die;
c) calculating the disease index: DI ∑ (number of diseased plants at each stage × the disease level value) × 100/(number of investigated total plants × highest level value);
d) evaluation criteria for resistance:
high Resistance (HR): disease index is 0-2.0;
anti (R): disease index is 2.1-15.0;
anti-Medium (MR): disease index is 15.1-40.0;
(iii) feeling (MS): disease index: 40.1-60.0;
infection (S): disease index is 60.1-80.0;
high feeling (HS): disease index is 80.1-100.0;
calculating disease indexes of the disease-resistant and disease-susceptible materials, determining the rust resistance of the disease-resistant and disease-susceptible materials according to resistance evaluation standards, and if the evaluation grades of the disease-resistant and disease-susceptible materials are the same as or different from the previous multi-year identification results by one grade but not reach significant differences, indicating that the identification is effective;
and then calculating the disease index of the material to be identified, and determining the rust resistance of the material according to the resistance evaluation standard.
Further, in the step (1), preferably, when the bacterial sources are collected, rust spores on infected plants in 3-4 diseased fields in the main producing area of the broad beans are collected respectively.
Further, it is preferable that in the step (2), the disease-resistant material is selected for a material whose resistance is evaluated as resistance (R) -High Resistance (HR); the disease-sensitive material was selected as a material whose resistance was evaluated as disease (S) -High Sensitivity (HS).
Further, it is preferable that in the step (2), the seeds are washed with clean water 3 times during the seed disinfection.
Further, in the step (2), it is preferable that the row spacing is 33cm, the plant spacing is 13cm, and the drill seeding is performed.
Further, it is preferable that 15 to 20 grains are sown per one part of the material.
Further, in the step (3), it is preferable that, in the inoculation, the mixed spore suspension obtained in the step (1) is applied to the plant to be inoculated for 2-4 times of spray inoculation.
Compared with the prior art, the invention has the beneficial effects that:
compared with the prior art, the method has the advantages that the environmental controllability in the greenhouse is high, the repeatability is good, the quantity of materials which can be identified at one time is relatively large, the necessary conditions required by the morbidity can be ensured, and the materials are fully attacked; the adoption of multi-bacterial source inoculation is beneficial to screening materials with comprehensive resistance genes, and compared with the small species specialized resistance screened by using a single bacterial strain, the resistant material obtained by the method has the advantages of better stability, adaptability and practicability; the method for inoculation in the seedling stage and investigation in the adult plant stage is similar to the field disease characteristics of broad bean rust, and well simulates the field disease process. Therefore, the method for the mixed strain greenhouse seedling stage inoculation and adult plant stage identification provides an efficient and stable method for rust resistance identification and genetic breeding research of broad bean materials.
Meanwhile, the method of the present invention is compared with a method for identifying a single strain in an incubator at a seedling stage, a method for identifying a natural disease nursery in a field planting stage and a method for identifying an isolated leaf, wherein the method is used for identifying the source of an inoculated strain, the inoculation time, the environmental treatment after inoculation, the investigation time, etc., and the results are shown in table 1.
TABLE 1
Detailed Description
The present invention will be described in further detail with reference to examples.
It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The materials or equipment used are not indicated by manufacturers, and all are conventional products available by purchase.
Example 1
The method for identifying the broad bean rust disease comprehensive resistance is characterized by comprising the following steps of:
step (1), collecting bacteria sources and preparing a mixed spore suspension:
a) collecting a bacterium source: collecting rust spores on infected plants;
b) preparation of mixed spore suspension: collecting equal amount of rust spores collected from different places, mixing, pouring the obtained mixed spore powder into sterilized distilled water, adding 0.02% (v/v) Tween-20, and making into 4 × 105Spore. ml-1The mixed spore suspension of (a);
step (2), broad bean material preparation and sowing:
a) design of control material: setting disease-resistant and susceptible materials as a control group;
b) seed cleaning: before sowing, cleaning disease-resistant and disease-susceptible materials and seeds of materials to be identified, and removing the seeds which are shriveled, broken, germinated, susceptible, damaged by worms, mildewed and damaged by seed coats;
c) seed disinfection: washing the seeds with clear water for multiple times, soaking the seeds in a sodium hypochlorite solution with the mass concentration of 0.5% for 10s, then immediately washing the seeds with the clear water for multiple times, and then immediately sowing the seeds;
and (3) inoculating broad bean rust in seedling stage in a greenhouse and setting temperature and humidity:
a) inoculation: inoculating disease-resistant and disease-susceptible materials and materials to be identified in 4-5 leaf stages of plants, humidifying greenhouse, disease-resistant and disease-susceptible materials and the plants to be identified by spraying water before inoculation, ensuring that the surfaces of leaves are wet and the humidity of the greenhouse is kept between 80% and 100%, starting inoculation before 10 am, and performing spray inoculation on the disease-resistant and disease-susceptible materials and the plants to be identified by adopting the mixed spore suspension obtained in the step (1);
b) temperature and humidity setting after inoculation:
1-21d after rust inoculation, controlling the temperature of the greenhouse at 20-25 ℃ in the daytime and 10-15 ℃ at night, and opening the greenhouse environment after 21d of inoculation so as to ensure the consistency of the internal environment and the external environment as much as possible;
spraying with water for 3-5 times a day from 1 st to 14 th days after inoculation to ensure plant surface moistening and greenhouse humidity stability at above 95%, spraying with water for 1 time a day from 15 th to 21 th days, and spraying without water after 22 th day;
step (4), identifying the resistance of the greenhouse broad bean rust in the adult plant stage:
a) and (4) disease investigation: after inoculation for 80 days, grading the disease condition in the adult plant period;
b) grading the disease condition: adopting a six-level division method:
level 0: no visible infection;
level 1: the leaves only have allergic necrosis reaction and no sporophyte;
and 3, level: sporadic sporangium mass in the leaf; no damage and breakage, and the diameter of spore pile is less than 0.5 mm; occupies less than 5.0 percent of the leaf area;
and 5, stage: the spore pile on the leaf and the stem is common and large; the diameter of the spore pile is 0.51-1 mm; partial sporangial masses are broken; occupies 5.10 to 10.0 percent of the leaf area;
and 7, stage: the spore pile is common and large on the leaves and stems; the diameter is more than 1.1 mm; severe rupture of the sporangium; the small area of the leaf surface is faded and discolored; the area of the leaf is 10.1-25.0%;
and 9, stage: the spores are densely stacked and fully distributed, and account for more than 25.1 percent of the leaf area; large area of blade blackens or falls off, and plants die;
c) calculating the disease index: DI ∑ (number of diseased plants at each stage × the disease level value) × 100/(number of investigated total plants × highest level value);
d) evaluation criteria for resistance:
high Resistance (HR): disease index is 0-2.0;
anti (R): disease index is 2.1-15.0;
anti-Medium (MR): disease index is 15.1-40.0;
(iii) feeling (MS): disease index: 40.1-60.0;
infection (S): disease index is 60.1-80.0;
high feeling (HS): disease index is 80.1-100.0;
calculating disease indexes of the disease-resistant and disease-susceptible materials, determining the rust resistance of the disease-resistant and disease-susceptible materials according to resistance evaluation standards, and if the evaluation grades of the disease-resistant and disease-susceptible materials are the same as or different from the previous multi-year identification results by one grade but not reach significant differences, indicating that the identification is effective; otherwise, re-identification is needed;
and then calculating the disease index of the material to be identified, and determining the rust resistance of the material according to the resistance evaluation standard.
Example 2
The method for identifying the broad bean rust disease comprehensive resistance is characterized by comprising the following steps of:
step (1), collecting bacteria sources and preparing a mixed spore suspension:
a) collecting a bacterium source: collecting rust spores on infected plants;
b) preparation of mixed spore suspension: collecting equal amount of rust spores collected from different places, mixing, pouring the obtained mixed spore powder into sterilized distilled water, adding 0.05% (v/v) Tween-20, and making into 4 × 105Spore. ml-1The mixed spore suspension of (a);
step (2), broad bean material preparation and sowing:
a) design of control material: setting disease-resistant and susceptible materials as a control group;
b) seed cleaning: before sowing, cleaning disease-resistant and disease-susceptible materials and seeds of materials to be identified, and removing the seeds which are shriveled, broken, germinated, susceptible, damaged by worms, mildewed and damaged by seed coats;
c) seed disinfection: washing seeds with clear water for multiple times, soaking the seeds in a sodium hypochlorite solution with the mass concentration of 0.5% for 20s, then immediately washing the seeds with the clear water for multiple times, and then immediately sowing the seeds;
and (3) inoculating broad bean rust in seedling stage in a greenhouse and setting temperature and humidity:
a) inoculation: inoculating disease-resistant and disease-susceptible materials and materials to be identified in 4-5 leaf stages of plants, humidifying greenhouse, disease-resistant and disease-susceptible materials and the plants to be identified by spraying water before inoculation, ensuring that the surfaces of leaves are wet and the humidity of the greenhouse is kept between 80% and 100%, starting inoculation after 2 pm, and performing spray inoculation on the disease-resistant and disease-susceptible materials and the plants to be identified by adopting the mixed spore suspension obtained in the step (1);
b) temperature and humidity setting after inoculation:
1-21d after rust inoculation, controlling the temperature of the greenhouse at 20-25 ℃ in the daytime and 10-15 ℃ at night, and opening the greenhouse environment after 21d of inoculation so as to ensure the consistency of the internal environment and the external environment as much as possible;
spraying with water for 3-5 times a day from 1 st to 14 th days after inoculation to ensure plant surface moistening and greenhouse humidity stability at above 95%, spraying with water for 1 time a day from 15 th to 21 th days, and spraying without water after 22 th day;
step (4), identifying the resistance of the greenhouse broad bean rust in the adult plant stage:
a) and (4) disease investigation: after inoculation for 85d, grading the disease condition in the adult plant period;
b) grading the disease condition: adopting a six-level division method:
level 0: no visible infection;
level 1: the leaves only have allergic necrosis reaction and no sporophyte;
and 3, level: sporadic sporangium mass in the leaf; no damage and breakage, and the diameter of spore pile is less than 0.5 mm; occupies less than 5.0 percent of the leaf area;
and 5, stage: the spore pile on the leaf and the stem is common and large; the diameter of the spore pile is 0.51-1 mm; partial sporangial masses are broken; occupies 5.10 to 10.0 percent of the leaf area;
and 7, stage: the spore pile is common and large on the leaves and stems; the diameter is more than 1.1 mm; severe rupture of the sporangium; the small area of the leaf surface is faded and discolored; the area of the leaf is 10.1-25.0%;
and 9, stage: the spores are densely stacked and fully distributed, and account for more than 25.1 percent of the leaf area; large area of blade blackens or falls off, and plants die;
c) calculating the disease index: DI ∑ (number of diseased plants at each stage × the disease level value) × 100/(number of investigated total plants × highest level value);
d) evaluation criteria for resistance:
high Resistance (HR): disease index is 0-2.0;
anti (R): disease index is 2.1-15.0;
anti-Medium (MR): disease index is 15.1-40.0;
(iii) feeling (MS): disease index: 40.1-60.0;
infection (S): disease index is 60.1-80.0;
high feeling (HS): disease index is 80.1-100.0;
calculating disease indexes of the disease-resistant and disease-susceptible materials, determining the rust resistance of the disease-resistant and disease-susceptible materials according to resistance evaluation standards, and if the evaluation grades of the disease-resistant and disease-susceptible materials are the same as or different from the previous multi-year identification results by one grade but not reach significant differences, indicating that the identification is effective; otherwise, re-identification is needed;
and then calculating the disease index of the material to be identified, and determining the rust resistance of the material according to the resistance evaluation standard.
In the step (1), when the bacteria source is collected, rust spores on infected plants in 3 disease fields in the main production area of the broad beans are respectively collected.
In the step (2), the disease-resistant material is selected as a material with resistance evaluation as resistance (R); the susceptible material was selected for materials evaluated for susceptibility (S) for resistance.
In the step (2), the seeds are washed with clear water for 3 times during seed disinfection.
In the step (2), when sowing, sowing is carried out according to the row spacing of 33cm and the plant spacing of 13cm in a drill mode.
20 seeds were sown per portion of material.
In the step (3), during inoculation, the mixed spore suspension obtained in the step (1) is adopted for carrying out spray inoculation treatment for 2 times on a plant to be inoculated.
Example 3
The method for identifying the broad bean rust disease comprehensive resistance is characterized by comprising the following steps of:
step (1), collecting bacteria sources and preparing a mixed spore suspension:
a) collecting a bacterium source: collecting rust spores on infected plants;
b) preparation of mixed spore suspension: collecting equal amount of rust spores collected from different places, mixing, pouring the obtained mixed spore powder into sterilized distilled water, adding 0.03% (v/v) Tween-20, and making into 4 × 105Spore. ml-1The mixed spore suspension of (a);
step (2), broad bean material preparation and sowing:
a) design of control material: setting disease-resistant and susceptible materials as a control group;
b) seed cleaning: before sowing, cleaning disease-resistant and disease-susceptible materials and seeds of materials to be identified, and removing the seeds which are shriveled, broken, germinated, susceptible, damaged by worms, mildewed and damaged by seed coats;
c) seed disinfection: washing the seeds with clear water for multiple times, soaking the seeds in a sodium hypochlorite solution with the mass concentration of 0.5% for 15s, then immediately washing the seeds with the clear water for multiple times, and then immediately sowing the seeds;
and (3) inoculating broad bean rust in seedling stage in a greenhouse and setting temperature and humidity:
a) inoculation: inoculating disease-resistant and disease-susceptible materials and materials to be identified in 4-5 leaf stages of plants, humidifying greenhouse, disease-resistant and disease-susceptible materials and the plants to be identified by spraying water before inoculation, ensuring that the surfaces of leaves are wet and the humidity of the greenhouse is kept between 80% and 100%, starting inoculation before 10 am, and performing spray inoculation on the disease-resistant and disease-susceptible materials and the plants to be identified by adopting the mixed spore suspension obtained in the step (1);
b) temperature and humidity setting after inoculation:
1-21d after rust inoculation, controlling the temperature of the greenhouse at 20-25 ℃ in the daytime and 10-15 ℃ at night, and opening the greenhouse environment after 21d of inoculation so as to ensure the consistency of the internal environment and the external environment as much as possible;
spraying with water for 3-5 times a day from 1 st to 14 th days after inoculation to ensure plant surface moistening and greenhouse humidity stability at above 95%, spraying with water for 1 time a day from 15 th to 21 th days, and spraying without water after 22 th day;
step (4), identifying the resistance of the greenhouse broad bean rust in the adult plant stage:
a) and (4) disease investigation: after inoculation for 80-85d, grading the disease condition in the adult plant period;
b) grading the disease condition: adopting a six-level division method:
level 0: no visible infection;
level 1: the leaves only have allergic necrosis reaction and no sporophyte;
and 3, level: sporadic sporangium mass in the leaf; no damage and breakage, and the diameter of spore pile is less than 0.5 mm; occupies less than 5.0 percent of the leaf area;
and 5, stage: the spore pile on the leaf and the stem is common and large; the diameter of the spore pile is 0.51-1 mm; partial sporangial masses are broken; occupies 5.10 to 10.0 percent of the leaf area;
and 7, stage: the spore pile is common and large on the leaves and stems; the diameter is more than 1.1 mm; severe rupture of the sporangium; the small area of the leaf surface is faded and discolored; the area of the leaf is 10.1-25.0%;
and 9, stage: the spores are densely stacked and fully distributed, and account for more than 25.1 percent of the leaf area; large area of blade blackens or falls off, and plants die;
c) calculating the disease index: DI ∑ (number of diseased plants at each stage × the disease level value) × 100/(number of investigated total plants × highest level value);
d) evaluation criteria for resistance:
high Resistance (HR): disease index is 0-2.0;
anti (R): disease index is 2.1-15.0;
anti-Medium (MR): disease index is 15.1-40.0;
(iii) feeling (MS): disease index: 40.1-60.0;
infection (S): disease index is 60.1-80.0;
high feeling (HS): disease index is 80.1-100.0;
calculating disease indexes of the disease-resistant and disease-susceptible materials, determining the rust resistance of the disease-resistant and disease-susceptible materials according to resistance evaluation standards, and if the evaluation grades of the disease-resistant and disease-susceptible materials are the same as or different from the previous multi-year identification results by one grade but not reach significant differences, indicating that the identification is effective; otherwise, re-identification is needed;
and then calculating the disease index of the material to be identified, and determining the rust resistance of the material according to the resistance evaluation standard.
In the step (1), when the bacteria source is collected, rust spores on infected plants in 4 disease fields in the main production area of the broad beans are respectively collected.
In the step (2), the disease-resistant material is selected to be a material with High Resistance (HR) evaluation; the susceptible material was selected for materials whose resistance was evaluated as high-sensitivity (HS).
In the step (2), the seeds are washed with clear water for 3 times during seed disinfection.
In the step (2), when sowing, sowing is carried out according to the row spacing of 33cm and the plant spacing of 13cm in a drill mode.
15 seeds were sown per portion of material.
In the step (3), during inoculation, the mixed spore suspension obtained in the step (1) is adopted for carrying out spray inoculation treatment for 4 times on a plant to be inoculated.
Examples of the applications
A method for identifying the comprehensive resistance of broad bean rust comprises the following steps:
step (1), collecting bacteria sources and preparing a mixed spore suspension:
collecting 3-4 pieces of serious disease fields in the main production area of broad beans in the current year of the test, and mixing;
a) collecting a bacterium source: collecting susceptible plants in areas with serious diseases in the main production area of the broad beans every year, and collecting rust spores on the susceptible plants by using a brush;
b) preparation of mixed spore suspension: collecting equal amount of rust spores from different places, mixing to obtain mixed spore powder, pouring into sterilized distilled water, adding 0.02% Tween-20, and making into 4 × 105Spore. ml-1The mixed spore suspension of (a), for spray inoculation;
(2) preparing and sowing broad bean materials:
a) design of control material: setting disease-resistant and susceptible materials as a control group; b) seed cleaning: before sowing, removing seeds with small size, breakage, germination, disease, worm damage, mildew and seed coat damage; c) seed disinfection: washing the seeds with clear water for 3 times, soaking the seeds with 0.5% NaClO for 10-20s, and immediately washing with clear water for 3 times; d) sowing: and (3) immediately sowing after the seeds are disinfected, wherein the seeds are sowed in a drill mode according to the row spacing of 33cm and the plant spacing of 13cm, and 15-20 seeds are sowed in each material.
(3) Inoculation and temperature and humidity setting of broad bean rust in seedling stage in a greenhouse:
a) inoculation: inoculating in the leaf stage of 4-5 plants, humidifying the greenhouse and the materials by spraying before inoculation to ensure that the leaf surface is wet and the humidity of the greenhouse is kept between 80% and 100%, starting inoculation at 10 am or 2 pm, and performing spraying inoculation for 3 times on all the materials; b) temperature/humidity treatment after inoculation: 1-21 days after rust inoculation, controlling the temperature at 20-25 ℃ in the daytime and 10-15 ℃ at night in the greenhouse; spraying for 3-5 times every day from 1 st to 14 th days after inoculation to ensure that the surface of the plant is wet and the humidity of the greenhouse is stable at more than 95%, spraying for 1 time every day from 15 th to 21 th days, and spraying for no more than 22 th days; c) after inoculation for 21d, the greenhouse environment was left open to ensure as much consistency as possible between the internal environment and the external environment.
(4) Method for identifying resistance of greenhouse broad bean rust in adult plant stage
a) And (4) disease investigation: after inoculation for 80-85d, grading the disease condition in the adult plant period; b) the disease condition is classified by adopting a six-grade division method: level 0: no visible infection; level 1: the leaves only have allergic necrosis reaction and no sporophyte; and 3, level: sporadic sporangium mass in the leaf; no damage and breakage, and the diameter of spore pile is less than 0.5 mm; occupies less than 5.0 percent of the leaf area; and 5, stage: the spore pile on the leaf and the stem is common and large; the diameter of the spore pile is 0.51-1 mm; partial sporangial masses are broken; occupies 5.10 to 10.0 percent of the leaf area; and 7, stage: the spore pile is common and large on the leaves and stems; the diameter is more than 1.1 mm; severe rupture of the sporangium; the small area of the leaf surface is faded and discolored; the area of the leaf is 10.1-25.0%; and 9, stage: the spores are densely stacked and fully distributed, and account for more than 25.1 percent of the leaf area; the leaves turn black or fall off in a large area, and the plants die. c) Disease Index (DI) ═ Σ (number of diseased plants at each stage × Disease level value) × 100/(number of total investigated plants × highest level value); d) evaluation criteria for resistance: high Resistance (HR): disease index is 0-2.0; anti (R): disease index is 2.1-15.0; anti-Medium (MR): disease index is 15.1-40.0; (iii) feeling (MS): disease index: 40.1-60.0; the disease index of susceptible (S) is 60.1-80.0; high feeling (HS): the disease index is 80.1-100.0.
(5) Repeatability/feasibility analysis of broad bean rust comprehensive resistance identification method
The test is provided with double controls, and anti-infection materials (respectively named as K0772 and K90315) are used as CK1、C K2In the year 2014-2015, a control group 12 is set, and in the years 2015-2016 and 2016-2017, a control group 21 is set, so that 3 groups of data are provided.
a) Identification result of anti-influenza control Material
The disease index and disease resistance evaluation are shown in table 2: the disease resistance indexes of the disease resistance control K0772 in all years are all between 2.1 and 15.0, and the disease resistance evaluation is resistance (R); the disease index of the disease control K90315 over the years is 74.7, 80.0 and 81.0, and the disease resistance evaluation is respectively feeling (S), feeling (S) and high feeling (HS). The annual identification results of the contrast materials are basically the same and are consistent with the identification results of the predecessors, which shows that the established rust resistance identification method for seedling stage inoculation and adult stage identification can stably distinguish the rust resistance difference of the broad beans.
TABLE 2 index of disease over years for anti-influenza control material
Note: 2014-2015 year setting a control group 12; 21 groups are set in 2015-2016 and 2016-2017 years; "-" indicates nothing.
CK was performed using SPSS12.0 software1、CK2The results of the T-test for disease indices between different years are shown in table 3: according to P>0.05, then the P (2-t aied) value of the row of Equal varians assigned is selected, P<0.05, the requirement for the P (2-tall) value of the Equalariaces not assembled row is selected. K0772 (CK)1) The P (2-tailed) values of the three comparison groups were 0.584. 0.423 and 0.350, both greater than 0.05; k90315 (CK)2) The P (2-tailed) values for the three comparison groups were 0.473, 0.528 and 0.389, respectively, all greater than 0.05. There was no significant difference in disease index between the different years for the 2 control materials.
TABLE 3 disease index T test of different years of infection and disease resistance
Note: "-" indicates nothing.
b) Resistance identification result of material repeatedly identified for many years
Of the 3 year old authentication material, 77 duplicate authentication materials (as in table 4) were provided and analyzed by Microsoft Excel and SPSS12.0 software. The annual disease index and disease resistance evaluation of the materials are shown in table 3, the results of the same materials in different years have certain difference, and the difference of the disease resistance grades is within one grade.
TABLE 4 evaluation of disease index and resistance of repeatedly identified materials
The analysis of variance and correlation between groups for 3-year-repeated data is shown in Table 5. The P (2-tailed) values for the 3 comparison groups were all greater than 0.05, 0.231, 0.895, and 0.339, respectively, indicating that the differences between the results of the 3 year repeat identification were not significant; meanwhile, the correlation coefficients among groups are respectively 0.385(P <0.01), 0.276(P <0.05) and 0.313(P <0.01), and the groups respectively show extremely remarkable, remarkable and extremely remarkable positive correlation. The repeated identification results are highly consistent between years, and the method has better stability and repeatability, and the mixed bacterial source is used for seedling inoculation, so that the evaluation of the rust resistance of the broad bean variety in the adult stage is feasible.
TABLE 5 repeated identification of material disease index inter-annual variance and correlation analysis
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (7)
1. The method for identifying the broad bean rust disease comprehensive resistance is characterized by comprising the following steps of:
step (1), collecting bacteria sources and preparing a mixed spore suspension:
a) collecting a bacterium source: collecting rust spores on infected plants;
b) preparation of mixed spore suspension: collecting equal amount of rust spores collected from different places, mixing uniformly, pouring the obtained mixed spore powder into sterilized distilled water, adding 0.02-0.05% (v/v) Tween-20, and making into 4 × 105Spore. ml-1The mixed spore suspension of (a);
step (2), broad bean material preparation and sowing:
a) design of control material: setting disease-resistant and susceptible materials as a control group;
b) seed cleaning: before sowing, cleaning disease-resistant and disease-susceptible materials and seeds of materials to be identified, and removing the seeds which are shriveled, broken, germinated, susceptible, damaged by worms, mildewed and damaged by seed coats;
c) seed disinfection: washing seeds with clear water for many times, soaking the seeds in a sodium hypochlorite solution with the mass concentration of 0.5% for 10-20s, then immediately washing the seeds with the clear water for many times, and then immediately sowing the seeds;
and (3) inoculating broad bean rust in seedling stage in a greenhouse and setting temperature and humidity:
a) inoculation: inoculating disease-resistant and disease-susceptible materials and materials to be identified in 4-5 leaf stages of plants, humidifying greenhouse, disease-resistant and disease-susceptible materials and plants to be identified by spraying water before inoculation, ensuring that the surfaces of leaves are wet and the humidity of the greenhouse is kept between 80% and 100%, starting inoculation before 10 am or 2 pm, and performing spray inoculation on the disease-resistant and disease-susceptible materials and plants to be identified by adopting the mixed spore suspension obtained in the step (1);
b) temperature and humidity setting after inoculation:
1-21d after rust inoculation, controlling the temperature of the greenhouse at 20-25 ℃ in the daytime and 10-15 ℃ at night, and opening the greenhouse environment after 21d of inoculation so as to ensure the consistency of the internal environment and the external environment as much as possible;
spraying with water for 3-5 times a day from 1 st to 14 th days after inoculation to ensure plant surface moistening and greenhouse humidity stability at above 95%, spraying with water for 1 time a day from 15 th to 21 th days, and spraying without water after 22 th day;
step (4), identifying the resistance of the greenhouse broad bean rust in the adult plant stage:
a) and (4) disease investigation: after inoculation for 80-85d, grading the disease condition in the adult plant period;
b) grading the disease condition: adopting a six-level division method:
level 0: no visible infection;
level 1: the leaves only have allergic necrosis reaction and no sporophyte;
and 3, level: sporadic sporangium mass in the leaf; no damage and breakage, and the diameter of spore pile is less than 0.5 mm; occupies less than 5.0 percent of the leaf area;
and 5, stage: the spore pile on the leaf and the stem is common and large; the diameter of the spore pile is 0.51-1 mm; partial sporangial masses are broken; occupies 5.10 to 10.0 percent of the leaf area;
and 7, stage: the spore pile is common and large on the leaves and stems; the diameter is more than 1.1 mm; severe rupture of the sporangium; the small area of the leaf surface is faded and discolored; the area of the leaf is 10.1-25.0%;
and 9, stage: the spores are densely stacked and fully distributed, and account for more than 25.1 percent of the leaf area; large area of blade blackens or falls off, and plants die;
c) calculating the disease index: DI = (number of diseased plants at each stage × disease level value) × 100/(number of investigated total plants × highest level value);
d) evaluation criteria for resistance:
high Resistance (HR): disease index is 0-2.0;
anti (R): disease index is 2.1-15.0;
anti-Medium (MR): disease index is 15.1-40.0;
(iii) feeling (MS): disease index: 40.1-60.0;
infection (S): disease index is 60.1-80.0;
high feeling (HS): disease index is 80.1-100.0;
calculating disease indexes of the disease-resistant and disease-susceptible materials, determining the rust resistance of the disease-resistant and disease-susceptible materials according to resistance evaluation standards, and if the evaluation grades of the disease-resistant and disease-susceptible materials are the same as or different from the previous multi-year identification results by one grade but not reach significant differences, indicating that the identification is effective;
and then calculating the disease index of the material to be identified, and determining the rust resistance of the material according to the resistance evaluation standard.
2. The method for identifying the broad bean rust disease comprehensive resistance according to claim 1, wherein in the step (1), when the bacteria source is collected, rust spores on 3-4 infected plants in the diseased field in the main producing area of the broad bean are respectively collected.
3. The method for identifying the broad bean rust disease comprehensive resistance according to claim 1, wherein in the step (2), the resistance of the disease-resistant material is selected and evaluated as a material resistant to (R) -High Resistance (HR); the disease-sensitive material was selected as a material whose resistance was evaluated as disease (S) -High Sensitivity (HS).
4. The method for identifying the broad bean rust disease comprehensive resistance according to claim 1, wherein in the step (2), the seeds are washed with clear water for 3 times during seed disinfection.
5. The method for identifying the broad bean rust disease comprehensive resistance according to claim 1, wherein in the step (2), sowing is carried out according to the row spacing of 33cm, the plant spacing of 13cm and drill sowing.
6. The method for identifying the broad bean rust disease comprehensive resistance, according to claim 4, is characterized in that 15-20 seeds are sown in each material.
7. The method for identifying the broad bean rust disease comprehensive resistance, according to claim 1, is characterized in that in the step (3), during inoculation, the mixed spore suspension obtained in the step (1) is adopted for carrying out spray inoculation treatment for 2-4 times on a plant to be inoculated.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010106381.7A CN111280008A (en) | 2020-02-21 | 2020-02-21 | Comprehensive resistance identification method for broad bean rust disease |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010106381.7A CN111280008A (en) | 2020-02-21 | 2020-02-21 | Comprehensive resistance identification method for broad bean rust disease |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111280008A true CN111280008A (en) | 2020-06-16 |
Family
ID=71024764
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010106381.7A Pending CN111280008A (en) | 2020-02-21 | 2020-02-21 | Comprehensive resistance identification method for broad bean rust disease |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111280008A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114651689A (en) * | 2022-03-17 | 2022-06-24 | 福建农林大学 | Peanut bacterial wilt field initial flowering stage inoculation identification method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105409613A (en) * | 2015-12-05 | 2016-03-23 | 武汉市蔬菜科学研究所 | Seedling-stage root rot resistance identification method for vigna unguiculata |
| CN105779565A (en) * | 2016-04-14 | 2016-07-20 | 浙江省农业科学院 | Sesquipedalis germplasm rust disease resistance in-vitro identification method and rust fungus liquid used in method |
| CN109022538A (en) * | 2018-08-22 | 2018-12-18 | 湖南省蔬菜研究所 | A kind of method identified in watermelon anthrax resistance room |
| CN110100685A (en) * | 2019-05-24 | 2019-08-09 | 河南省农业科学院 | A method of using conidium inoculated identification Peanut Web Blotch Disease disease resistance |
| CN110199711A (en) * | 2019-06-20 | 2019-09-06 | 黑龙江省农业科学院耕作栽培研究所 | A kind of mould accurate identification method of Resistance To Root Rot Disease resource of big bean curd |
-
2020
- 2020-02-21 CN CN202010106381.7A patent/CN111280008A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105409613A (en) * | 2015-12-05 | 2016-03-23 | 武汉市蔬菜科学研究所 | Seedling-stage root rot resistance identification method for vigna unguiculata |
| CN105779565A (en) * | 2016-04-14 | 2016-07-20 | 浙江省农业科学院 | Sesquipedalis germplasm rust disease resistance in-vitro identification method and rust fungus liquid used in method |
| CN109022538A (en) * | 2018-08-22 | 2018-12-18 | 湖南省蔬菜研究所 | A kind of method identified in watermelon anthrax resistance room |
| CN110100685A (en) * | 2019-05-24 | 2019-08-09 | 河南省农业科学院 | A method of using conidium inoculated identification Peanut Web Blotch Disease disease resistance |
| CN110199711A (en) * | 2019-06-20 | 2019-09-06 | 黑龙江省农业科学院耕作栽培研究所 | A kind of mould accurate identification method of Resistance To Root Rot Disease resource of big bean curd |
Non-Patent Citations (3)
| Title |
|---|
| 宋治世 等: "《胡豆种植与加工利用》", 30 November 1986, 四川科学技术出版社 * |
| 赵振玲 等: "蚕豆叶片气孔及其抗锈病性研究", 《云南农业大学学报》 * |
| 骆平西 等: "蚕豆种质资源抗锈病鉴定研究", 《作物品种资源》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114651689A (en) * | 2022-03-17 | 2022-06-24 | 福建农林大学 | Peanut bacterial wilt field initial flowering stage inoculation identification method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Pornpienpakdee et al. | Improving the micropropagation efficiency of hybrid Dendrobium orchids with chitosan | |
| Roychowdhury et al. | Chemical mutagenic action on seed germination and related agro-metrical traits in M1 Dianthus generation | |
| EP2854513B1 (en) | Method for large scale generation of artificial seeds comprising symbiota | |
| WO2009143856A1 (en) | The method for potato virus eradication and creation of meristem clones with improved traits, virus-free potato meristem plant and virus-free potato | |
| CN104082055A (en) | Method for high-flux identification of ralstonia solanacearum resistance | |
| CN101317548A (en) | Cultivation method for Isolated microspore of cucumber | |
| Tuwo et al. | Improvement of orchid Vanda hybrid (Vanda limbata Blume X Vanda tricolor Lindl. var. suavis) by colchicines treatment in vitro | |
| CN113388526A (en) | Endophytic fungus FO-R20 and application thereof | |
| Percy et al. | Qualitative genetics and utilization of mutants | |
| CN107190011A (en) | The gene of the coding myosin of one cotton quality trait associations | |
| CN111280008A (en) | Comprehensive resistance identification method for broad bean rust disease | |
| Bouharmont | Application of somaclonal variation and in vitro selection to plant improvement | |
| CN112715355A (en) | Safe and efficient mutation method for bluegrass | |
| CN115287194B (en) | Medicinal wild rice endophytic fungi YYA21 and application thereof | |
| WU et al. | Identification of soybean resources of resistance to aphids | |
| JPH07274723A (en) | Bent grass and introduction of endophyte to bent grass | |
| KR102663790B1 (en) | Pseudoxanthomonas sp. JBCE485 and Variovorax paradoxus JBCE486 strins in promoting plant growth tolerance and resistance to salt stress and composition comprising the same | |
| CN109628322B (en) | Bletilla striata endophytic fungus 3-G2 and application thereof | |
| AU2021103557A4 (en) | Method for Quickly Identifying and Screening Plants Resistant to Fusarium Wilt and Application Thereof | |
| Avci et al. | Autotetraploid plant production in endemic Onobrychis elata with colchicine treatments | |
| Mullaney et al. | Resistance to Septoria nodorum in a durum wheat cultivar as determined by stage of host development | |
| Ashraf et al. | Conventional breeding of cotton | |
| Zouari et al. | Effect of liquid seaweed (Ulva rigida) extract on the growth and rooting of carob (Ceratonia siliqua L.) | |
| CN107365829B (en) | Method for screening black shank resistant germplasm of tobacco in laboratory | |
| Vishnupandi et al. | Effect of varying concentrations of colchicine on polyploid induction in Jasminum sambac (L.) Aiton |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200616 |
|
| RJ01 | Rejection of invention patent application after publication |