CN110073957A - A kind of system and method carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana - Google Patents
A kind of system and method carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana Download PDFInfo
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- CN110073957A CN110073957A CN201910314487.3A CN201910314487A CN110073957A CN 110073957 A CN110073957 A CN 110073957A CN 201910314487 A CN201910314487 A CN 201910314487A CN 110073957 A CN110073957 A CN 110073957A
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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
- A01G31/00—Soilless cultivation, e.g. hydroponics
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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
- A01G31/00—Soilless cultivation, e.g. hydroponics
- A01G31/02—Special apparatus therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- G01N2333/37—Assays involving biological materials from specific organisms or of a specific nature from fungi
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- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
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Abstract
The invention discloses a kind of system and method for carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana.The filling substrate in cultivation tray, banana seedlings to be identified are planted in matrix again, then several cultivation trays with banana seedlings are put into outer basin, solution is added in outer basin, it is added with soil fungi (Fusariun oxysporun f.sp.cubense) in the solution, it is cultivated, disease registration identification then is carried out to banana seedlings.Many dish for cultivating with banana seedlings can be disposably put into the disease resistance screening that large-scale research of fusarium wilt disesase of banana is carried out in the nutrient solution of outer basin by the present invention, without being individually inoculated with pathogen to banana seedlings one by one, enormously simplify process, improve efficiency, and more standardize, avoid the influence of artificial enchancement factor.
Description
Technical field:
The invention belongs to disease resistance of plant research fields, and in particular to a kind of to utilize cultivation technique without soil to banana sickle-like bacteria
The system and method that the disease resistance of wilt disease carries out Rapid identification.
Background technique:
Banana (Musa nana Lour.) belongs to Musaceae banana, and refers to its fruit, and torrid areas is planted extensively.
Banana flavor is fragrant, is rich in nutrition, and plant is large-scale draft, issues, is formed by leaf sheath lower part 3~6 meter (10~20 high from rhizomes
Ruler) false bar；For leaf oblong to ellipse, some is up to 3~3.5 meters (10~11.5 ruler), 65 centimeters wide (26 cun), and 10
~20 pieces of fasciation stem tops.Spike is sagging, is extracted out by false bar top, spends majority, faint yellow；Infructescence is curved to hang down, and as a result 10~20
String, about 50~150.It is withered after plant result, continue to breed by the sucker that rhizomes is grown, each stock can live for many years.It is former
Is produced from the Asia southeast, there is cultivation in Taiwan, Hainan, Guangdong, Guangxi etc..
Fusarium wilt is one of important disease of banana, is by soil fungi (Fusariun oxysporun
F.sp.cubense caused by).Plant is once susceptible, if control measure is not in time, it will disease is caused to spread rapidly.Sickle
Knife bacterium usually causes plant root-rot, stem rot, brown foot rot, the multiple diseases symptom such as spend corruption occur.Sickle-like bacteria often infects host's plant
The fibrovascular system of object destroys the conducting tissue of plant, and generates toxin in plant growth and development metabolic process and make to endanger
Object causes plant wilt dead, influences the yield and quality of banana.
Existing some control measures, such as chemical prevention, waterflooding are lain fallow, crop rotation and use organic matter improve soil etc.
Fusarium wilt cannot effectively be controlled.At present generally believe breeding resistant variety be control the disease most efficient method it
One.And establishing simple and effective Resistance Identification system is the key that carry out banana blight breeding for disease resistance.
Current identification system is to be inoculated with pathogen to culture medium in the culture of rootage stage under isolated culture condition mostly
In, then screen banana disease-resistant variety.But it has disadvantages that:
1, its needs is individually inoculated with pathogen into culture medium in each culture vessel, time-consuming and laborious, Large-scale Screening
It is relatively difficult.
2, it is affected by human factors bigger.
Summary of the invention:
The object of the present invention is to provide one kind can be simple and easy to do, is conducive to Large-scale Screening, and can reduce artificially with
The system and method for carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana of machine factor.
The system for carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana of the invention,
It is characterised in that it includes outer basin and cultivation tray, several cultivation trays are placed in the outer basin, cultivation basin bottom part is equipped with hole,
Matrix is housed, plantation there are banana seedlings to be identified in the matrix, and solution is housed in the outer basin in cultivation tray.
The method for carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana of the invention,
Characterized by comprising the following steps:
The filling substrate in cultivation tray, then banana seedlings to be identified are planted (as in culture of rootage basal growth in matrix
2-3 weeks test tube seedling), then several cultivation trays with banana seedlings are put into outer basin, solution are added in outer basin, molten
It is added with soil fungi (Fusariun oxysporun f.sp.cubense) in liquid, is cultivated, then banana seedlings are carried out
Disease registration identification.
Wherein the growth of 1 grade-plantlet is normal, without any disease symptom；
The vanelets of 2 grades-vacation basal part of stem are withered, but the color of false stem itself does not change；
The small fine jade in the 3 grades-false dimmed region of stem color or equal to the 1/2 of entire false stem height；
4 grades-vacation dimmed the region of stem color is more than the 1/2 of entire false stem height；
5 grades-flavescence or withered upper leaf the piece number are less than or equal to the 1/2 of plantlet top total leaf number；
6 grades-flavescence or withered upper leaf the piece number are more than the 1/2 of plantlet top total leaf number；
7 grades-whole strain banana seedlings withered death.
The solution is can also to add soil fungi (Fusariun wherein for various nutrient solutions
oxysporun f.sp.cubense)。
It is preferred that the matrix is perlite.
Several cultivation trays with banana seedlings are put into outer basin by the present invention, are had in outer basin and are contained soil fungi
The nutrient solution of (Fusariun oxysporun f.sp.cubense), nutrient solution penetrate into cultivation by the hole below cultivation tray
In the matrix for training basin, and the root of banana seedlings is infiltrated, nutrient solution is absorbed by Banana Root, to meet the required liquid manure of banana seedlings growth
Supply, and the soil fungi (Fusariun oxysporun f.sp.cubense) in nutrient solution also infects banana seedlings, according to quilt
The growing state for infecting banana seedlings can identify banana seedlings to the disease resistance of research of fusarium wilt disesase of banana.
It can be seen that many dish for cultivating with banana seedlings can be disposably put into the nutrient solution of outer basin by the present invention
The disease resistance screening for carrying out large-scale research of fusarium wilt disesase of banana, without being individually inoculated with cause of disease to banana seedlings one by one
Bacterium enormously simplifies process, improves efficiency, and more standardize, avoids the influence of artificial enchancement factor.
Detailed description of the invention:
Fig. 1 is the disease resistance progress Rapid identification of the invention using cultivation technique without soil to research of fusarium wilt disesase of banana
The structural schematic diagram of system；
Wherein 1, outer basin；2, cultivation tray；3, nutrient solution；4, banana seedlings；5, perlite；6, root；7, hole.
Fig. 2 is perlite potting system, A: seed death of monks or nuns bacterium perlite of taking root；B and growth of seedling situation after C:15 days；
Fig. 3 is perlite potting system；A: rhizosphere perlite eluent microscopic findings；Bacterium colony on B:PCEA culture medium
Growing state；
Fig. 4 be Brazilian Banana and resist it is withered No. 5 inoculation after disease index；
Fig. 5 is Brazilian (A) and to resist the leaf growth situation of withered No. 5 (B) after inoculation；
Fig. 6 is to be inoculated with Brazilian (A) and resist withered No. 5 (B) change of height situations.
Specific embodiment:
The following examples are further illustrations of the invention, rather than limiting the invention.
Embodiment 1:
As shown in Figure 1, being carried out using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana for the present embodiment is fast
The system of speed identification, including outer basin 1 and cultivation tray 2 are placed with 9 cultivation trays 2 in the outer basin 1, and cultivation basin bottom part is equipped with hole
7, matrix (perlite 5) is housed in cultivation tray, plantation there are banana seedlings 4 to be identified in the matrix, in the outer basin
In be equipped with solution.The solution is can to add soil fungi (Fusariun wherein for nutrient solution 3
oxysporun f.sp.cubense)。
The side for carrying out Rapid identification to the disease resistance of research of fusarium wilt disesase of banana using cultivation technique without soil of the present embodiment
Method, comprising the following steps:
The filling substrate perlite 5 in cultivation tray 2, then banana seedlings 4 to be identified are planted (as in training of taking root in matrix
2-3 weeks test tube seedling of basal growth is supported, for by Brazilian any of several broadleaf plants and resisting withered No. 5), then the cultivation tray 2 with banana seedlings is put into outer
In basin 1, nutrient solution 3 is added in outer basin, the liquid level of nutrient solution borders on nethermost Banana Root, after preculture 2 weeks, 2 weeks
Soil fungi Fusarium oxysporum (Fusariun oxysporun f.sp.cubense) is added in nutrient solution, makes sickle-like bacteria spore
Concentration be 105A/ml is cultivated (regular water and fertilizer management) in the culturing room that temperature is 25 ± 2 DEG C, after inoculation 15 days,
Rhizosphere perlite is taken whether to carry out elution microscopically observation with the presence of Fusarium oxysporum.Root is cut into segment and is inoculated into PCEA training
It supports on base, 28 DEG C are cultivated 5-8 days, have seen whether bacterium colony growth.Periodic statistical blade disease grade, the number of blade and plant height.Often
A repetition nine times.Not to be inoculated with Fusarium oxysporum as blank control.
According to a conventional method, such as under isolated culture condition, in culture of rootage stage inoculation pathogen into culture medium, then
Screening banana disease-resistant variety is to above-mentioned Brazilian any of several broadleaf plants and resists withered No. 5 progress disease evaluation.
The nutrient solution is Hoagland ' s (Gram suddenly) nutrient solution.
The culture and preparation step of the Fusarium oxysporum are as follows: Fusarium oxysporum is seeded to nothing on superclean bench
On the CLA culture medium of bacterium, sealed membrane is sealed, and is cultivated 7-10 days at 27 DEG C.With sterile water wash media surface, and use liquid relief
Rifle gently scrapes surface so that more spores are eluted.Draw 1ml spore suspension dilute certain multiple, with specification be 25 lattice ×
The blood counting chamber of 16 lattice observes spore quantity under an optical microscope, determines spore mother liquid concentration, uses for subsequent inoculations.Meter
Calculate spore concentration formula are as follows: cell number/ml=80 small lattice inner cell number/80 × 400 × 10000 × extension rate (i.e. 1:
50000).The carnation blade agar medium (CLA) is to obtain blade from the carnation plant of healthy growth, is cleaned
Repeatedly guarantee that persticide residue is low on blade.Blade is cut into the segment of 5 × 5mm, 45-55 DEG C dries 2 hours or so, works as blade
Holding green should be appointed after dried.Blade is placed in aluminium pot (high 5cm, diameter 5cm), with the Co of 2.5 Megarads60Gamma is penetrated
Line sterilizes.On a sterile work bench, the carnation blade that 4-5 piece sterilizes is placed into and fills temperature and is cooled to 50 DEG C or so
Carnation blade agar medium is produced on the culture dish of 2% water agar of mass fraction.
The classification of blade disease grade is as follows:
Wherein the growth of 1 grade-plantlet is normal, without any disease symptom；
The vanelets of 2 grades-vacation basal part of stem are withered, but the color of false stem itself does not change；
The small fine jade in the 3 grades-false dimmed region of stem color or equal to the 1/2 of entire false stem height；
4 grades-vacation dimmed the region of stem color is more than the 1/2 of entire false stem height；
5 grades-flavescence or withered upper leaf the piece number are less than or equal to the 1/2 of plantlet top total leaf number；
6 grades-flavescence or withered upper leaf the piece number are more than the 1/2 of plantlet top total leaf number；
7 grades-whole strain banana seedlings withered death.
Disease index=∑ (state of an illness rank × state of an illness rank plant number)/(state of an illness superlative degree is worth × and it is total to handle plant
Number) × 100%
Disease incidence=disease plant number/processing plant sum × 100%
The death rate=death plant number/processing plant sum × 100%
As a result:
After seedling kind enters perlite, growing state is good (Fig. 2).Micronutrient element is contained only in perlite, by pouring
Mineral element needed for applying nutrient solution supplement plant strain growth, blade and root system can normal growths.Seedling is extracted, can be observed
It is vigorous to its root growth.Rhizosphere perlite is cleaned after inoculation 15 days and observes result such as Fig. 3 A under the microscope, is being eluted
It can be seen that sickle-shaped microorganism in liquid.Root system is inoculated into cultivation results such as Fig. 3 B on PCEA culture medium.PCEA culture medium
To separate sickle-like bacteria special culture media, being placed at root system in culture dish has bacterium colony growth, and not placing is to observe at root system
There is bacterium colony appearance.
Blade disease classification standard to the Brazilian any of several broadleaf plants of inoculation and resists withered No. 5 progress blade disease etc. in Pathogenic measurement
Grade statistics, calculates disease index result such as Fig. 4, and after data show Brazilian any of several broadleaf plants inoculation in figure, the 15th day to the 25th day is disease
The period that grade sharply increases, after inoculation the 30th day it is unobvious with blade disease index variation tendency at 35 days, so later
Disease rank can be counted within 30 days after being inoculated in qualification test.
Brazilian any of several broadleaf plants and after resisting withered No. 5 inoculations, carries out periodic logging to blade and plant height.As a result such as Fig. 5 and Fig. 6.Figure
5A is Brazilian any of several broadleaf plants leaf growth situation, when Brazilian any of several broadleaf plants is inoculated with 4.6 to inoculation the 35th day of the processing group number of blade since inoculation
9.5, and 9.0 when 4.5 that the blank group number of blade starts since inoculation were by inoculation 35 days, (inoculation is sharp for Foc processing group
Fusarium oxysporum group) and blank group the number of blade within the time period without significant difference, it can be observed how, resist withered No. 5 Foc in Fig. 5 B
Processing group and the blank group number of blade are interior without significant difference at inoculation 0-35 days, therefore can not be to leaf in later inoculation experiments
The piece number is counted.Fig. 6 A is Brazilian any of several broadleaf plants plant height growing state, and Brazilian any of several broadleaf plants inoculation processing group plant height is since inoculation
17.3cm to inoculation the 35th day when 22.4cm, and blank group plant height since inoculation start 17.5cm to be inoculated with 35 days when
The plant height of 25.2cm, Foc processing group and blank group resists withered No. 5 Foc to handle within the time period without significant difference in Fig. 6 B
Group and blank group plant height can not carry out plant height in inoculation 0-35 days without significant difference, therefore in later inoculation experiments
Statistics.
Using water as medium in hydroponic culture systems, imposing nutrient solution enables banana seedlings uniformly to absorb various mineral elements.By
The microorganism rich in soil, rhizospheric environment is complicated when earth culture is inoculated with, and effect of inoculation can be made to be affected.With the treasure that sterilizes
Zhu Yan is matrix, keeps inoculation environment more consistent.It periodically pours and applies mineral element needed for nutrient solution provides normal plants.
In conclusion the present invention is to the disease resistance evaluation of research of fusarium wilt disesase of banana and the knot of conventional method disease resistance evaluation
Fruit is consistent (conventional method disease resistance evaluation as the result is shown Brazilian any of several broadleaf plants not anti-blight, No. 5 anti-blights of resistance), it was demonstrated that according to this
The method of invention is identified, quick, simplicity, the susceptible variety for accurately distinguishing research of fusarium wilt disesase of banana and disease-resistant product are capable of
Kind.
Claims (6)
1. a kind of system for carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana, feature
It is, including outer basin and cultivation tray, several cultivation trays is placed in the outer basin, cultivation basin bottom part is equipped with hole, in cultivation tray
In matrix is housed, plantation has banana seedlings to be identified in the matrix, and solution is housed in the outer basin.
2. system according to claim 1, which is characterized in that the solution is various nutrient solutions.
3. system according to claim 1, which is characterized in that the matrix is perlite.
4. a kind of method for carrying out Rapid identification using disease resistance of the cultivation technique without soil to research of fusarium wilt disesase of banana, feature
It is, comprising the following steps:
The filling substrate in cultivation tray, then banana seedlings to be identified are planted in matrix, several are then had into banana seedlings
Cultivation tray is put into outer basin, and solution is added in outer basin, is added with soil fungi (Fusariun oxysporun in the solution
F.sp.cubense), cultivated, disease registration identification then is carried out to banana seedlings.
5. according to the method described in claim 4, it is characterized in that, the solution is for various nutrient solutions.
6. according to the method described in claim 4, it is characterized in that, the matrix is perlite.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111642338A (en) * | 2020-05-26 | 2020-09-11 | 广东省农业科学院果树研究所 | Method for promoting growth of banana seedlings by inoculating arbuscular mycorrhizal fungi |
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| CN101643772A (en) * | 2009-09-09 | 2010-02-10 | 广东省农业科学院果树研究所 | Method for quickly evaluating disease resistance of banana fusarium wilt |
| CN202043495U (en) * | 2011-02-28 | 2011-11-23 | 中国热带农业科学院南亚热带作物研究所 | Banana seedling raising container for artificial inoculation of fusarium wilt pathogen |
| CN102771326A (en) * | 2012-08-08 | 2012-11-14 | 东莞市香蕉蔬菜研究所 | Method for rapidly identifying and screening anti-blight banana seedling |
| CN109006000A (en) * | 2018-08-07 | 2018-12-18 | 广西壮族自治区农业科学院 | Method for identifying and screening disease-resistant seedlings of banana wilt |
-
2019
- 2019-04-18 CN CN201910314487.3A patent/CN110073957A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101643772A (en) * | 2009-09-09 | 2010-02-10 | 广东省农业科学院果树研究所 | Method for quickly evaluating disease resistance of banana fusarium wilt |
| CN202043495U (en) * | 2011-02-28 | 2011-11-23 | 中国热带农业科学院南亚热带作物研究所 | Banana seedling raising container for artificial inoculation of fusarium wilt pathogen |
| CN102771326A (en) * | 2012-08-08 | 2012-11-14 | 东莞市香蕉蔬菜研究所 | Method for rapidly identifying and screening anti-blight banana seedling |
| CN109006000A (en) * | 2018-08-07 | 2018-12-18 | 广西壮族自治区农业科学院 | Method for identifying and screening disease-resistant seedlings of banana wilt |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111642338A (en) * | 2020-05-26 | 2020-09-11 | 广东省农业科学院果树研究所 | Method for promoting growth of banana seedlings by inoculating arbuscular mycorrhizal fungi |
| CN111642338B (en) * | 2020-05-26 | 2022-02-18 | 广东省农业科学院果树研究所 | Method for promoting growth of banana seedlings by inoculating arbuscular mycorrhizal fungi |
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