CN106472182A - A kind of method that citrus yellow shoot disease is controlled on potted plant nursery stock - Google Patents
A kind of method that citrus yellow shoot disease is controlled on potted plant nursery stock Download PDFInfo
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- CN106472182A CN106472182A CN201610876911.XA CN201610876911A CN106472182A CN 106472182 A CN106472182 A CN 106472182A CN 201610876911 A CN201610876911 A CN 201610876911A CN 106472182 A CN106472182 A CN 106472182A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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Abstract
The invention discloses a kind of method that citrus yellow shoot disease is controlled on potted plant nursery stock, the antibacterial process of three phases is carried out respectively using T3 and T26 micromolecular compound, mode using pouring root and drip irrigation, coordinate normal rich water quality management, and combine real-time quantitative QPCR molecular detection technology, the former removal effect of monitoring HLB disease.The percentage rate of two groups of program removing cause of diseases respectively reaches 88.9% and 85.7%.This technology, for the domestic and international virus-free stock breeding introducing mandarin orange resource, for the popularization and application of important local characteristic citrus variety nursery stock, has important practical value, the sustainable development for Citrus Industry has important practical significance.
Description
Technical field
The invention belongs to the disease-resistant technical field of citrus yellow shoot disease, specifically, it is related to a kind of control Citrus chachiensis Hort. on potted plant nursery stock
The method of Fructus Citri tangerinae yellow twig.
Background technology
Citrus Huanglongbing pathogen (Citrus huanglongbing), is a kind of global destructiveness present in mandarin orange produces
Disease, at present, it has jeopardized China's vast mandarin orange main producing region, Fla., Jia Lifoniya state St. Paul,Brazil etc.
Important mandarin orange producing region.Plate umpire kind and the types such as main harm Citrus chachiensis Hort., Fructus Citri tangerinae, orange, Fructus Citri Limoniae, Fructus Citri grandiss class.Especially with any of several broadleaf plants Citrus chachiensis Hort., sugar orange,
The disease tolerance ability of the kinds such as Citrus chachiensis Hort., tangerine produced in fujian province, Citrus reticulata Blanco cv. Chachiensis is weaker, and this sick whole year can occur, susceptible kind typically occur uniform yellow,
Mottled yellow and nutritional deficiency shape yellow symptom, adult trees often the top yellow of part minority branch in full wafer orangery, the
Yellow branch expands Herb within 2 years, so that tree body is failed.The tied fruit of disease tree is little, and shape is strange, and areola is crooked, disease fruit epidermis light
Sliding, matt, sour in the mouth, it is yellowish green uneven that coat color has, and is changed into orange red, thus affecting Citrus near some kind fruit end
Yield, referred to as " red nose fruit ".This disease is very harmful to Orange Producing, and susceptible kind loses knot in the 3-5 obtaining after being ill
Fruit ability, in addition withered, and in recent years, onset area constantly expands, and is continuously increased to bacterium source, and epidemic situation diffusion velocity improves constantly,
Damage losses are extremely serious, become a War Torn sexually transmitted disease (STD) evil of Orange Producing.
Citrus huanglongbing pathogen (Candidatus Liberibacter asiaticus) is the Gram-negative of difficult culture
Antibacterial, is belonged to by Candidatus liberibacter in prokaryote Gracilicutes and causing, be a kind of bacterial disease.It is to be main by diaphorina citri
Communication media, does not also have effectively preventing measure currently for Citrus Huanglongbing pathogen, mainly excavates disease tree in field, in cultivation
Control wood louse, cultivate anosis nursery stock and strengthen quarantine to prevent it to infect and to spread.But still the generation of this disease can not be prevented.Citrus chachiensis Hort.
Fructus Citri tangerinae has once infected Candidatus liberibacter, the time of several years, and fruit is just basic to be destroyed.This phenomenon has seriously been dampened orchard worker and has been sent out
The enthusiasm that mandarin orange produces.In the last few years, the outbreak of communicable diseases that pathogenic microorganism led to is so that the research of novel antibacterial medicine becomes
Obtain extremely urgent.Since 2009, the examining order of citrus Huanglongbing pathogen microorganism full-length genome was close or has completed, this
Make us more deep for the understanding of disease, more efficiently measure can be taken for cause of disease, precisely be treated, this
Us are made more profoundly to recognize the importance of antibacterial drug target.
It is in cultivation link that mandarin orange infects the treatment of treelet of yellow twig and recovery technology, smooth to mandarin orange improved seeds
Put into production, produce the key link of high-quality fruits.As：In the LUGAN in China Fujian, the children on the ground such as the Citrus shatangju in Guangxi
Tree, is having just enter into the operation stage, is beginning to find to face the yellow of star or the tree of mottled symptom occurs.We adopt QPCR and PCR
Although Molecular Detection is carried out to such orchard it is found that some tree body symptoms can't observe with the naked eye, in fact, tree
In vivo with the presence of cause of disease ratio apparently higher than Symptomatic tree.
Introduce and increasing of exchanging, California, USA, Florida State, bar along with climate change and domestic and international citrus variety
The spreading rapidly of the main region yellow twig such as west, China's citrus yellow shoot disease infection is on the rise, and therefore, research is new, efficient, practical
Mandarin orange infect the treatment of treelet of yellow twig and the technology of getting well has important actual application value.
The existing preventing and treating with regard to citrus yellow shoot disease and detection are as follows：Inject agricultural tetracycline to be once used for preventing and treating HUANGLONG
Disease, in field relief of symptoms, recovers yield and has certain effect, but, high cost, in China Guangxi province, remove disease in time
Strain reduces cause of disease, and using the measure of joint defence, the control of citrus yellow shoot disease (HLB) creates good effect.In Brazil, by clear
Control wood louse except diseased plant with using insecticide, the preventing and treating of yellow twig has been achieved with certain achievement.In addition, it is strong from relevant antagonism
Kind, strengthen citrus psylla preventing and treating, promote nontoxic nursery stock, a kind Fructus psidii guajavae immaturus walks quickly and keeps away the gardening means such as wood louse, is also to subtract
Lack causing harm of yellow twig.In large-scale Newly built orchard, using virus-free nursery stock it is critical that method.At present, Citrus are yellow
Dragon disease PCR detection is mainly carried out according to 16S rDNA and β-operator design primer, and real-time quantitative PCR is also widely used for Citrus chachiensis Hort.
The detection of Fructus Citri tangerinae yellow twig, sensitivity is high, and accuracy is high, and for field, susceptible situation can rapidly monitor.
Mandarin orange infects the treatment of nursery stock of yellow twig and the technology of getting well has following importance：Governments at all levels are very heavy
Depending on introduction and the popularization of improved seeds, successively put into many funds and introduce all kinds of Citrus improved seeds from external.Time after plantation
Not long, tree body just will be carried out the operation stage, is just faced with the danger chopping at a tree.Except using virus-free nursery stock in cultivation
In, control wood louse to propagate outside these measures of cause of disease, mandarin orange infects the treatment of yellow twig treelet and gets well imperative, and it is right
Have great importance in the sustainable development protecting important mandarin orange resource and Citrus Industry.
Content of the invention
In view of this, the present invention is directed to above-mentioned problem, there is provided a kind of control citrus yellow shoot disease on potted plant nursery stock
Method, the present invention is the raw nursery stock of 2-4 to infection citrus yellow shoot disease or treelet carries out cause of disease removing and recovery process, can be to zero
The treelet that star infects citrus yellow shoot disease carries out cause of disease control.The antibacterial process of three phases is carried out using micromolecular compound, utilizes
Pouring root and the mode of drip irrigation, and coordinate normal rich water quality management, using real-time quantitative QPCR molecular detection technology, verify HLB disease
Former removal effect；Cultivation link in relatively low cost input, it is to avoid Citrus Orchard it may happen that heavy losses.
In order to solve above-mentioned technical problem, the invention discloses a kind of method that citrus yellow shoot disease is controlled on nursery stock, adopt
T3 or T26 micromolecular compound carries out the antibacterial process of three phases, using the mode of pouring root or drip irrigation, coordinates rich water quality management to strengthen
Tree vigo(u)r, and combine real-time quantitative QPCR molecular detection technology, the former removal effect of monitoring HLB disease.
Further, the method comprises the following steps：
1) from January start to process, with 30 μM of triclosan reagent, the raw nursery stock of 2-4 or treelet are carried out at pouring root
Reason, consumption is 1.5L/ strain, fills once every 20 days, totally 3 times, crosses sampling in 20 days, makees QPCR analysis, determines HLB detoxification efficiency；?
During process, keep the normal rich water quality management of nursery stock；
2) start the triclosan reagent with 50 μM～75 μM from early March, step 1 be poured into the consumption of 1.5L/ strain) process after
The raw nursery stock of 2-4 or treelet root, nursery stock is big, and growing way strengthens person suitably increases TCL concentration, is up to 75 μM, fills one every 20 days
Secondary, totally 3 times, after third time is processed, cross 20d sampling and make QPCRR analysis, detect HLB virus elimination rate；
3) start the triclosan reagent with 50 μM from early May, 1.5L/ strain is poured into step 2) process after the raw nursery stock of 2-4
Or treelet root, filled once every 20 days, totally three times, after third time is processed, make QPCR analysis every sampling in 20 days, detect HLB detoxification
Rate.
Further, the method comprises the following steps：
1) from January start to process, add Mn with 30 μM of Sigma reagent 2699802SO4Reagent 15mM, gives birth to 2-4
Nursery stock or treelet carry out root irrigation, consumption be 1.5L/ strain, every 20 days fill once, totally 3 times；Enter as QPCR after 20d sampling
Row analysis, detects HLB detoxification glue effect；During processing, keep the normal rich water quality management of nursery stock；
2) start from early March to add Mn with 50 μM of Sigma reagent 2699802SO4Reagent 15mM, 1.5L/ strain transfusion bottle drips
Reagent is dripped to step 1 by the mode filling) root of the raw nursery stock of the 2-4 that processes or treelet, the speed of drip irrigation is adjusted to
1.04ml/min, 1.5L/ strain dripped off at 24 hours.Every one week, once, each 1.5L/ strain dripped off drip irrigation at 24 hours again；Altogether
Drip irrigation 3 times, crosses 10d sampling and is analyzed as QPCR, detect HLB detoxification glue effect；
3) start from early May to add Mn with 50 μM of Sigma reagent 2699802SO4Reagent 15mM, 1.5L/ strain is poured into step
2) reagent is dripped to the raw nursery stock of 2-4 or treelet with the mode of transfusion bottle drip irrigation by the raw nursery stock of 2-4 after processing or treelet root
Root, the speed of drip irrigation is adjusted to 1.04ml/min, 1.5L/ strain dripped off at 24 hours, every one week, drip irrigation again, drip altogether
Fill 3 times, after crossing 10d, through QPCR analysis, QPCR analysis, the detoxification efficiency of identification HLB are made in sampling.
Compared with prior art, the present invention can obtain including following technique effect：
The raw treelet of mandarin orange 2-4 of the technology of the present invention energy mandarin orange nursery stock and fragmentary infection yellow twig (HLB) carries out yellow twig
Cause of disease control.The antibacterial process of three phases is carried out using T3 and T26 micromolecular compound, using the mode of pouring root and drip irrigation,
Coordinate normal rich water quality management, and combine real-time quantitative QPCR molecular detection technology, the former removal effect of monitoring HLB disease.Two groups
The percentage rate that program removes cause of disease respectively reaches 88.9% and 85.7%.This technology is for important local characteristic citrus variety
Popularization and application, and introduce mandarin orange resource anosis poisoning have important practical value, for the sustainable development of Citrus Industry
Exhibition has important practical significance.
The present invention is the result of several reagent comprehensive effects, and the wherein single product implementing the present invention might not be same
When reach all the above technique effect.
Brief description
Accompanying drawing described herein is used for providing a further understanding of the present invention, constitutes the part of the present invention, this
Bright schematic description and description is used for explaining the present invention, does not constitute inappropriate limitation of the present invention.In the accompanying drawings：
Fig. 1 is the QPCR analysis of Stage1-1 stage yellow twig control effect of the present invention, and wherein, 1 is negative control, 2
For positive control, 3-21 is detected sample, 2-ΔΔtIt is negative sample that relative expression quantity is less than 1, as healthy sample；
Fig. 2 is the QPCR analysis of Stage1-2 stage yellow twig control effect of the present invention, and wherein, 1 is negative control, and 2 are
Positive control, 3-9 is detected sample, 2-ΔΔtIt is negative sample that relative expression quantity is less than 1, as healthy sample；
Fig. 3 is the QPCR analysis of Stage1-3 stage yellow twig control effect of the present invention, and wherein, 1 is negative control, and 2 are
Positive control, 3-9 is detected sample, 2-ΔΔtIt is negative sample that relative expression quantity is less than 1, as healthy sample；
Fig. 4 is the QPCR analysis of Stage2-1 stage yellow twig control effect of the present invention, and wherein, 1 is negative control, and 2 are
Positive control, 3-9 is detected sample, 2-ΔΔtIt is negative sample that relative expression quantity is less than 1, as healthy sample；
Fig. 5 is the QPCR analysis of Stage2-2 stage yellow twig control effect of the present invention, and wherein, 1 is negative control, and 2 are
Positive control, 3-9 is detected sample, 2-ΔΔtIt is negative sample that relative expression quantity is less than 1, as healthy sample；
Fig. 6 is the QPCR analysis of Stage3-3 stage yellow twig control effect of the present invention, and wherein, 1 is negative control, and 2 are
Positive control, 3-9 is detected sample, 2-ΔΔtIt is negative sample that relative expression quantity is less than 1, as healthy sample.
Specific embodiment
To describe embodiments of the present invention below in conjunction with embodiment in detail, thereby to the present invention how application technology handss
Section is solving technical problem and to reach realizing process and fully understanding and implement according to this of technology effect.
Embodiment 1
The present invention provides a kind of method controlling citrus yellow shoot disease on nursery stock, comprises the following steps：
1) proceed by process from first beginning of the month, with 30 μM of TCL (triclosan) the reagent nursery stock raw to 2-4 or treelet
Carry out root irrigation, consumption is 1.5L/ strain, fills once every 20 days, totally 3 times.During processing, keep nursery stock normal rich water pipe
Reason, after the 3rd pouring root, crosses sampling in 20 days, makees QPCR analysis, determine HLB detoxification efficiency.
2) start the TCL reagent with 50 μM from early March, 1.5L/ strain is poured into step 1) process after the raw nursery stock of 2-4 or children
Usage tree root, filled once every 20 days, 3 times totally, after third time is processed, crosses 20d sampling and makees QPCRR analysis, detect HLB virus elimination rate.
3) start the TCL reagent with 50 μM from early May, 1.5L/ strain is poured into step 2) process after the raw nursery stock of 2-4 or children
Usage tree root, filled once every 20 days, three times totally, after third time is processed, makees QPCR analysis every sampling in 20 days, detects HLB virus elimination rate.
Embodiment 2
The present invention also provides a kind of method controlling citrus yellow shoot disease on nursery stock, comprises the following steps：
1) proceed by process from first beginning of the month, add Mn with 30 μM of T26 (Sigma reagent 269980)2SO4Reagent 15mM,
The strain of pouring root 1.5L/, is poured into the raw nursery stock of 2-4 or treelet root, filled once every 20 days, totally 3 times.Make QPCR after 20d sampling
It is analyzed, detect HLB detoxification glue effect；During processing, keep the normal rich water quality management of nursery stock；
2) start from early March to add Mn with 50 μM of T262SO4The mode of reagent 15mM, 1.5L/ strain transfusion bottle drip irrigation will be tried
Agent drips to step 1) process after the raw nursery stock of 2-4 or treelet root, the speed of drip irrigation is adjusted to 1.04ml/min,
1.5L/ strain dripped off at 24 hours.Every one week, once, each 1.5L/ strain dripped off drip irrigation at 24 hours again；Drip irrigation 3 times altogether, mistake
10d sampling is analyzed as QPCR, detects HLB detoxification glue effect；
3) start from early May to add Mn with 50 μM of T262SO4Reagent 15mM, 1.5L/ strain drip irrigation, to root, is dripped with transfusion bottle
Reagent is dripped to step 2 by the mode filling) process after the raw nursery stock of 2-4 or treelet root, the speed of drip irrigation is adjusted to
1.04ml/min, 1.5L/ strain dripped off at 24 hours, every one week, drip irrigation again, and drip irrigation 3 times altogether.After crossing 10d, through QPCR analysis,
QPCR analysis, the detoxification efficiency of identification HLB are made in sampling.
Illustrate the technique effect of the present invention with reference to specific experimental data：
Citrus Huanglongbing pathogen Real-time PCR detects foundation：
1st, DNA extraction and QPCR detection
CTAB method extracts the genomic DNA of Citrus material to be measured, and extracting method is with reference to document (Huang et such as Huang
al.,2000).Detection DNA concentration, and within being diluted to 100ng/ μ L, as the template of QPCR detection.Special base used
Because Plant cytochrome lysyloxidase gene COX, primer is COX+/COX-, the document such as sequence information reference Li (Li et al.,
2006), fragment length 68bp；Genes of interest amplimer is A04+/A04-, the document such as sequence information reference Hu Hao (Hu Hao etc.,
2006), amplified production is the distinguished sequence of the former ribosomal protein gene rplJ/rplL of Asian type Citrus HLB disease, fragment length
87bp, primer sequence is shown in Table 1.QPCR reaction system and program are shown in Table 2 and table 3 respectively.
Specific target sequence design primer (table 1) using Asia bast bacillus ribosomal protein gene.Light of dashing forward in real time is fixed
Amount PCR kit:2XSYBRGreen qPCR Mix, purchased from border biological gene Science and Technology Ltd. of village ally.PCR instrument (the U.S.
Bio-Rad company), DYY-5 type voltage stabilization and current stabilization electrophresis apparatuses (Liuyi Instruments Plant, Beijing), gel imaging system (Segrate company),
Nucleic acid-protein detector (Amersham company), prominent Fluorescent Quantitative PCR instrument quantitative real time PCR Instrument Light Cycler 480.Every kind of
The detection processing is repeated 3 times.
The primer being related in table 1 experiment and its sequence
Table 2 fluorescent quantitation reaction system
Table 3 fluorescent quantitation reaction system program
2nd, test method
Method one：Because the remolding sensitivity PCR sensitivity of QPCR analysis result is higher, therefore, only list QPCR's here
Testing result, the results are shown in Table 5 to table 7.
First stage processes (stage1-1)：Proceed by process from first beginning of the month, with 30 μM of TCL (triclosan) reagent,
1.5L/ strain pouring root, the raw Seedling of 2-4 or treelet, filled once every 20 days, 3 times, after 20d totally.In each stage, keep nursery stock normal
Rich water quality management.
As shown in table 5 and Fig. 1, cross 20d sampling make QPCR analysis, as long as QPCR test positive, result be considered sun
Property.This result of study shows：19 plants of positive Seedlings, 12 plants become negative after treatment, and the raw treelet virus elimination rate of 3-4 is
63.61%.
Second stage processes (stage1-2)：Proceed by from early March.With 50 μM of TCL reagent, 1.5L/ strain drip irrigation arrives
Root, the raw treelet of 3-4, filled once every 20 days, totally 3 times, after third time is processed, cross 20d sampling and make QPCRR analysis, such as table 6
With shown in Fig. 2, result shows：14 plants of positive Seedlings, 10 plants switch to feminine gender after treatment.The raw treelet feminine gender Seedling ratio of 3-4 is equal
For 71.4%.
Phase III processes (stage1-3)：Proceed by from early June, with 50 μM～75 μM of TCL reagent, nursery stock is big,
Growing way strengthens person suitably increases TCL concentration, is up to 75 μM, 1.5L/ strain drip irrigation to root, fills once every 20 days, totally three times, the 3rd
After secondary process, cross 20d sampling and make QPCRR analysis, as shown in table 7 and Fig. 3, result shows：11 plants of raw positive treelets of 3-4
In, 10 plants switch to feminine gender, and virus elimination rate reaches 88.9%.2 years raw Seedlings, 4 plants of positives switch to feminine gender, and virus elimination rate is 100%.
Method two：
First stage processes (stage2-1)：Proceed by process from first beginning of the month, with 30 μM of T26 (Sigma reagent
269980) add Mn2SO4Reagent 15mM, 1L/ strain pouring root, filled once every 20 days, 3 times totally.Carry out point as QPCR after 20d sampling
Analysis.As shown in table 8 and Fig. 4, result shows：30 plants of positive Seedlings, 15 plants become negative after treatment.In each stage, keep nursery stock
Normal rich water quality management.
Second stage processes (stage2-2)：Proceed by from early March.Add Mn with 50 μM of T262SO4Reagent 15mM,
To root, by the speed drip irrigation of 1.04ml/min, 1.5L/ involves a 2d in a criminal case to be completed 1.5L/ strain drip irrigation, every one week, drips 1.5L/ again
Strain, drip irrigation 3 times altogether.As shown in table 9 and Fig. 5, after 10d, through QPCRR analysis, 3-4 raw treelet virus elimination rate reaches 75%.
Phase III processes (stage2-3)：Proceed by from early May, add Mn with 50 μM of T262SO4Reagent 15mM,
1.5L/ strain drip irrigation, to root, is even dripped 2d and is completed, every one week, again with the speed drip irrigation of 1.04ml/min, 1.5L/ strain drip irrigation arrived
Root needs 2 day time, altogether drip irrigation 3 times.As shown in table 10 and Fig. 6, after 10d, through QPCR analysis, 7 plants of the raw treelet of 3-4, place
After reason, virus elimination rate reaches 85.7%.
3rd, experimental result：
Table 4 mandarin orange treelet yellow twig controls practical operation program and effect analyses
Note：Upper reagent mother solution all promotes dissolving with 100%DMSO, then is diluted to concentration with clear water.
Table 5 TCL adopts QPCR to detect the result of HLB after processing the treelet infecting citrus yellow shoot disease
Table 6 TCL adopts QPCR to detect the result of HLB after processing the treelet infecting yellow twig
Table 7 TCL adopts QPCR to detect the result of HLB after processing the treelet infecting yellow twig
Table 8 TCL adopts QPCR to detect the result of HLB after processing the treelet infecting yellow twig
Table 9 TCL adopts QPCR to detect the result of HLB after processing the treelet infecting yellow twig
Table 10 TCL adopts QPCR to detect the result of HLB after processing the treelet infecting yellow twig
Described above illustrate and describes some preferred embodiments of invention, but as previously mentioned it should be understood that inventing not
It is confined to form disclosed herein, be not to be taken as the exclusion to other embodiment, and can be used for various other combinations, modification
And environment, and can be carried out by the technology or knowledge of above-mentioned teaching or association area in invention contemplated scope described herein
Change.And the change that those skilled in the art are carried out and change without departing from the spirit and scope of invention, then all should weighed appended by invention
In the protection domain that profit requires.
Claims (3)
1. a kind of method of control citrus yellow shoot disease on potted plant nursery stock is it is characterised in that comprise the following steps：Using T3 or
T26 micromolecular compound carries out the antibacterial process of three phases, using the mode of pouring root or drip irrigation, coordinates normal rich water quality management,
And combine real-time quantitative QPCR molecular detection technology, the former removal effect of monitoring HLB disease.
2. according to claim 1 on potted plant nursery stock control citrus yellow shoot disease method it is characterised in that include below
Step：
1) from January start to process, with 30 μM of triclosan reagent, root irrigation is carried out to the raw nursery stock of 2-4 or treelet, use
Measure as 1.5L/ strain, filled once every 20 days, totally 3 times, cross sampling in 20 days, make QPCR analysis, determine HLB detoxification efficiency；In process phase
Between, keep the normal rich water quality management of nursery stock；
2) start the triclosan reagent with 50 μM～75 μM from early March, step 1 be poured into the consumption of 1.5L/ strain) process after 2-
4 years raw nursery stocks or treelet root, nursery stock is big, and growing way strengthens person suitably increases TCL concentration, is up to 75 μM, fills once every 20 days, altogether
3 times, after third time is processed, cross 20d sampling and make QPCRR analysis, detect HLB virus elimination rate；
3) start the triclosan reagent with 50 μM from early May, 1.5L/ strain is poured into step 2) process after the raw nursery stock of 2-4 or children
Usage tree root, filled once every 20 days, three times totally, after third time is processed, makees QPCR analysis every sampling in 20 days, detects HLB virus elimination rate.
3. according to claim 1 on potted plant nursery stock control citrus yellow shoot disease method it is characterised in that include below
Step：
1) from January start to process, add Mn with 30 μM of Sigma reagent 2699802SO4Reagent 15mM, the Seedling to 2-4 life
Wood or treelet carry out root irrigation, and consumption is 1.5L/ strain, fills once every 20 days, totally 3 times；Carry out point as QPCR after 20d sampling
Analysis, detects HLB detoxification glue effect；During processing, keep the normal rich water quality management of nursery stock；
2) start from early March to add Mn with 50 μM of Sigma reagent 2699802SO4Reagent 15mM, 1.5L/ strain transfusion bottle drip irrigation
Reagent is dripped to step 1 by mode) root of the raw nursery stock of the 2-4 that processes or treelet, the speed of drip irrigation is adjusted to 1.04ml/
Min, 1.5L/ strain dripped off at 24 hours.Every one week, once, each 1.5L/ strain dripped off drip irrigation at 24 hours again；Drip irrigation 3 altogether
Secondary, cross 10d sampling and be analyzed as QPCR, detect HLB detoxification glue effect；
3) start from early May to add Mn with 50 μM of Sigma reagent 2699802SO4Reagent 15mM, 1.5L/ strain is poured into step 2) place
Reagent is dripped to the root of the raw nursery stock of 2-4 or treelet by the raw nursery stock of the 2-4 after reason or treelet root with the mode of transfusion bottle drip irrigation
Portion, the speed of drip irrigation is adjusted to 1.04ml/min, and 1.5L/ strain dripped off at 24 hours, every one week, drip irrigation again, and drip irrigation 3 altogether
Secondary, after crossing 10d, through QPCR analysis, QPCR analysis, the detoxification efficiency of identification HLB are made in sampling.
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| CN103039309A (en) * | 2011-10-14 | 2013-04-17 | 刘素珍 | Method for preventing and treating vegetable morbidity in adult-plant stage |
| CN103460872A (en) * | 2013-07-30 | 2013-12-25 | 中国科学院新疆生态与地理研究所 | Method for regulating and controlling root system architecture of under-mulch-drip irrigation cotton in sand through irrigation and nitrogen application |
| CN103891558A (en) * | 2014-04-02 | 2014-07-02 | 华中农业大学 | Citrus scion pretreatment technology for promoting removal of composite infection pathogens |
| CN104823770A (en) * | 2015-05-14 | 2015-08-12 | 张家界市玉凰绿色农业有限责任公司 | Prevention and treatment method of tomato bacterial wilt |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102405722A (en) * | 2011-10-10 | 2012-04-11 | 广东省农业科学院土壤肥料研究所 | Method for applying fertilizer for banana through drop irrigation |
| CN103039309A (en) * | 2011-10-14 | 2013-04-17 | 刘素珍 | Method for preventing and treating vegetable morbidity in adult-plant stage |
| CN102577728A (en) * | 2012-03-27 | 2012-07-18 | 河南农业大学 | Cultivation waste water trickle irrigation method for wheat-corn crop rotation system in arid-semiarid regions |
| CN103460872A (en) * | 2013-07-30 | 2013-12-25 | 中国科学院新疆生态与地理研究所 | Method for regulating and controlling root system architecture of under-mulch-drip irrigation cotton in sand through irrigation and nitrogen application |
| CN103891558A (en) * | 2014-04-02 | 2014-07-02 | 华中农业大学 | Citrus scion pretreatment technology for promoting removal of composite infection pathogens |
| CN104823770A (en) * | 2015-05-14 | 2015-08-12 | 张家界市玉凰绿色农业有限责任公司 | Prevention and treatment method of tomato bacterial wilt |
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