CN110352852A - A kind of sweet potato propagation method - Google Patents
A kind of sweet potato propagation method Download PDFInfo
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- CN110352852A CN110352852A CN201910777512.1A CN201910777512A CN110352852A CN 110352852 A CN110352852 A CN 110352852A CN 201910777512 A CN201910777512 A CN 201910777512A CN 110352852 A CN110352852 A CN 110352852A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The present invention relates to plant breeding methods fields specifically, and the present invention provides a kind of sweet potato mating system, includes the following steps: the preparation of 1) detoxification test tube plantlet；2) the numerous culture of the expansion of detoxification test tube plantlet: detoxification test tube plantlet by multiple squamous subculture expand numerous；3) test tube seedling domestication hardening is obtained into sweet potato seedling；4) sweet potato seedling is subjected in protecting field field planting, obtains sweet potato strong sprout；5) by sweet potato strong sprout by 4 expansion numerous, obtained sweet potato sprout, the sweet potato sprout is planted in crop field, carries out Production of Large Fields, harvest sweet potato.Sweet potato mating system of the invention can be realized high production efficiency, save the purpose of production cost, shortens the seed production period 1 year, significantly reduces seed production cost.
Description
Technical field
The present invention relates to a kind of plant breeding methods, specifically, are related to a kind of sweet potato " 2 years two-stage methods " reproduction technique
Method.
Background technique
Sweet potato is important grain, feed, starch and raw materials of food processing, and the long-term cultivated area of Hubei Province sweet potato is 200
Ten thousand mu or so.As people's living standard steps up, the attention rate of health is increasingly increased, sweet potato has no longer been people in the past
Described " coarse food grain ", " disaster relief eke out a living grain ", but it is nutrition very abundant, complete, and there is important health care and prevent disease function
The food of energy.Except fresh food is outer, at present Hubei Province be engaged in sweet potato processing enterprise it is increasing, order purchase cultivated area is up to 100
Ten thousand mu or more, sweet potato converted products type is abundant, and the existing local characteristic to grow up on the basis of traditional " three powder " production produces
Product, and have emerging snack food, there are also the staple foods products such as sweet potato steamed bun, vermicelli.
However as the development of industrialization process, the mating problem of breeding becomes increasingly conspicuous in sweet potato production.Although excellent product
Kind is the material base of agricultural production, and promoting breeding area to expand rapidly popularization is cost-effective well stimulation, but sweet potato
There is its particularity with kind, such as: compared to other crops, sweet potato is with kind of demand in big, potato seed and seedling allocation and transportation difficulty, production
Breeding biology is generally and serious as frequently adjusting north and south disease caused by kind to mix generation phenomenon with mechanical admixture phenomenon
Deng these cause breeding and degenerate so that potentiality can not play, breeding mating the problems such as not catching up with industry demand.Therefore, have
Plan, system development kind prevent miscellaneous pure keeping, prevent degeneration work, and mating related agricultural technology, accelerate new breeding potato potato seed
Seedling breeding, is to give full play to breeding potentiality, realizes the requisite measure of industry upgrading synergy.
Traditional seed production technology uses " 3 years three-field systems ", and as Single-plant selection → plant garden → original seed farm original seed is raw
Production technology, the technology First Year are selected disease-free single plant and are marked, and range estimation primary election is carried out to it, and current year harvests potato kind, stores respectively
Hiding；Second year establishes garden of collecting seedling, and is selected in single plant potato wedge plant division nursery to last year, establishes plant garden, and identification overground part is special before sealing ridge
Sign, harvest time identify underground part feature, and all discoveries do not meet former breediness or seedling in spite of illness is rejected without exception；Third year will be upper
The plant garden potato kind that year is elected to mixes nursery, and sets Seedlings nursery, current year transplanting to appropriate area progress seed production.This method week
Phase is long, at high cost, does not meet the requirement of industry upgrading synergy.
Summary of the invention
The purpose of the present invention is realizing efficient abridged edition, shorten the seed production period 1 year, significantly reduce seed production at
This.
To achieve the goals above, the present invention provides a kind of sweet potato mating system, includes the following steps:
1) preparation of detoxification test tube plantlet；
2) the numerous culture of the expansion of detoxification test tube plantlet: detoxification test tube plantlet by multiple squamous subculture expand numerous；
3) test tube seedling domestication hardening is obtained into sweet potato seedling；
4) sweet potato seedling is subjected in protecting field field planting, obtains sweet potato strong sprout；
5) sweet potato strong sprout is numerous by 4 expansions, the sweet potato sprout is planted in crop field by obtained sweet potato sprout, is carried out
Production of Large Fields harvests sweet potato.
Wherein, the step 1) includes taking sweet potato top stem with bud, carries out differentiation culture after cleaning, breaks up the item of culture
Part is the shoot apical meristem for taking 0.5-1mm of removing, is inoculated on differential medium, and at 28 DEG C, luminous intensity is (2000-3000)
Lux, the interior culture of daily illumination 10-18 hours environment.
The protecting field can be greenhouse or other can play the planting site with greenhouse identity function.
Further, the 0.5-1mm is 0.5mm, and described 10-18 hours is 16 hours, and the differential medium is MS+
1mg/L 6-BA and 0.5mg/L IAA, the MS+1mg/L 6-BA and 0.5mg/L IAA are to add 6-BA into MS culture medium
The culture medium obtained with IAA, in the MS+1mg/L 6-BA and 0.5mg/L IAA, the content of 6-BA is 1mg/L, and IAA's contains
Amount is 0.5mg/L.
In sweet potato mating system of the invention, in the step 2), squamous subculture is in MS culture medium, at 28 DEG C, light intensity
Degree is (2000-3000) lux, culture in daily illumination 10-18 hours environment.
In sweet potato mating system of the invention, in the step 2), the period of the squamous subculture is 7 days, the subculture
Incubation times are 6 times.
In sweet potato mating system of the invention, in the step 3), tamed in the bottle including test tube seedling, outside the bottle of test tube seedling
It cultivates in the nursery of domestication and test tube seedling.
In sweet potato mating system of the invention, it is to cut single plant that the numerous method of numerous each expansion is expanded in 4 times of the step 5)
Section obtains cuttings, and each cuttings contains a leaf segment or two leaf segments, and the cuttings is inserted into soil and is cultivated.
Further include the virus sampling observation of test tube seedling in sweet potato mating system of the invention, in the step 2): randomly selecting and
Viral diagnosis is carried out from more plants of test tube seedlings of same single plant.
Further, the method for the viral diagnosis is PCR detection method and serum detection method, and the virus of the detection is
SPCSV, Geminiviridae and SPVD.
It is long for the sweet potato propagation method period in the prior art, it is at high cost, do not meet asking for the requirement of industry upgrading synergy
Topic.To improve efficiency, the present invention improves the method, the method after improvement is known as sweet potato " 2 years two-stage methods ", i.e.,
Numerous for detoxic seedling+stem section expansion numerous → crop field autumn, the technology is numerous using the fast numerous expansion of indoor detoxic seedling pure keeping original silkworm egg combination stem section
Coefficient is grown, and the next year method of autumn Production of Large Fields potato wedge expanding propagation coefficient, can be realized high production efficiency, saves production
The purpose of cost shortens the seed production period 1 year, significantly reduces seed production cost.
Specific embodiment
The present invention will be described below by way of specific embodiments, but the present invention is not limited thereto；
Experimental method used in following embodiments is conventional method unless otherwise specified；
Reagent as used in the following examples, biomaterial etc., are commercially available unless otherwise specified.
Hubei Province potato 6 by the autonomous breeding of Grain Crop Institute of Hubei Academy of Agricultural Sciences.
Embodiment 1
In Grain Crop Institute of Hubei Academy of Agricultural Sciences, Wuhan City, Hubei China province tissue culture room and Ezhou summer
Big lake test site carries out following tests using Hubei Province potato 6:
1. the production of Sweetpotato Viruses Elimination seedling is numerous fastly with stem section
1) it sterilizes the preparation of explant stem section first arrival on March 15 April 15: taking No. 6 sweet potato tops of about 300 Hubei Province potatos successively
Hold stem with bud.Remove its blade and petiole, be trimmed to the stem with bud of about 3 ㎝ long, detergent clean the surface dust and soil,
Detergent is cleaned up with tap water.In superclean bench with 70% alcohol disinfecting 30s, then with containing 1-1.6% active oxygen
Liquor natrii hypochloritis rock impregnate 5-8min, finally use aseptic water washing, aseptic water washing number depend on processed stem
Sharp quantity, it is however generally that, flushing standard is while rinsing 30-50 stem apex, rinses 5-6 times, rinses and be lower than 30 stem apexs, rinses
3-4 times, obtain explant stem section.
2) first arrival on March 15 April 15, the differentiation of evoking adventive bud:, will with the scissors and tweezers of sterilizing in super-clean bench
Disinfection stem section tail portion in the second step that the explant stem section of surface sterilization refers to is cut, remaining stem with bud 2cm or so, stripping
Shoot apical meristem from 0.5mm or so is inoculated on (MS+1mg/L 6-BA+0.5mg/L IAA) culture medium, is placed on 28 DEG C,
Luminous intensity is daily illumination 16h in (2000-3000) lux culturing room, the differentiation of evoking adventive bud；Induction differentiation is opened after 15 days
Beginning forms adventitious bud.Induce breaking up the result shows that stem apex survival rate about 80% or so, it is indefinite to be each induced stem apex generation 1
Bud, it is about 240 that adventitious bud, which generates number,.
It is divided into simple bud to carry out stem section Fiber differentiation by bud number adventitious bud obtained in step 2, and carries out multiple subculture
Culture obtains test tube seedling, and makes a mark to the stem section from different simple buds.Wherein, the culture medium that stem section Fiber differentiation uses for
MS culture medium, condition of culture are 28 DEG C, and luminous intensity is (2000-3000) lux.On April 15 or so worked as test tube seedling to May 15
It is grown successively to 7-8cm, starts squamous subculture, 1 time per week, amount to 4 times, breeding coefficient 34, at this point it is possible to obtain test tube seedling
19440 (y=240*3 of number4) strain.Continue squamous subculture twice, obtain 174960 (y=of test tube seedling 15-May 30 May
19440×32) strain.
1 stem apex of table strips length screening
| Stem apex length | Stem apex quantity | The malicious seedling quantity of band | Viral recall rate |
| 1mm | 20 | 1 | 5% |
| 0.5mm | 20 | 0 | 0% |
2 screening of medium formula of table
The processing screening of 3 light application time of table
| Lighting process | Growing way |
| 10h/d | Slow growth, blade is partially yellow, and stem is thin and delicate |
| 14h/d | Growth is normal, but slowly |
| 16h/d | Growth is most fast, and blade is emerald green, and stem thickness is strong |
| 18h/d | Growth is normal, fast speed, easy early ageing |
2. the virus sampling observation of test tube seedling: in April 15 to 240 plants during April 29, randomly selected from same single plant
Test tube seedling carries out viral diagnosis as follows.
Sweet potato test tube seedling viral diagnosis system
1) PCR detection method
PCR detection is the detection means mainly taken in current production, predominantly detects and seriously threatens sweet potato production
SPCSV, Geminiviridae, SPVD virus, should take different detection methods for above-mentioned three kinds of viral different characteristics.
SPCSV is long linear viral section (Closteroviridae), and Ampelo-virus (Crinivirus) is uniquely to invade
The long linear viral for contaminating sweet potato, is propagated in a manner of semi-durable by aleyrodid, and host range is mainly Convolvulaceae, Solanaceae and Amaranthaceae,
SPCSV can make sweet potato underproduction 15-88%.According to article, " serology and molecule of Viruses On Sweet Potato In China type are examined for the detection of SPCSV
Survey, Qiao Qi etc., plant physiology journal is published in for 2012 " in record, detected by the way of RT-PCR, it is specific to walk
It suddenly is first with total RNA extraction reagent box, to extract the total serum IgE in Sweet Potato Leaf, synthesized using specific reverse primer reverse transcription
First chain of cDNA extends target fragment using forward primer PCR
Geminivirdae is a kind of single stranded circle DNA virus, is had a very wide distribution, and is endangered in the world increasingly ferocious
Rampant, in Africa, Mediterranean, Southeast Asia, Central and South America and Caribbean prevalence at disaster, host includes that plurality of cereals is made
Object (wheat, potato etc.), industrial crops (cotton, tobacco, tomato and soybean etc.), rate of propagation is fast, in various crop and miscellaneous
There is geminivirus infection on grass, and spreads quickening from south to north in China.Geminivirdae can lead to sweet potato underproduction 11-
86%.Geminivirdae is a kind of DNA virus, extracts the genomic DNA in tissue first with genome extraction kit,
PCR extension is carried out by template of genomic DNA, primer and PCR reaction condition are shown in that " Chinese sweet potato geminivirus infection C4 gene molecule becomes
Record in different and analysis of genetic polymorphisms, Zhang Chengling, 2017, Jiangsu's agriculture journal ".
SPVD is sweet potato complex virus, is caused by raw infect altogether of SPFMV and SPCSV association, as long as therefore in the sample simultaneously
It both detects, then it is assumed that infected SPVD, the sweet potato variety for infecting SPVD shows as leaf curling, deformity, blade chlorisis, bright
The symptoms such as arteries and veins and plant dwarfing.SPVD starts to occur mainly in East Africa, West Africa and South America, influences greatly, sternly on yield of sweet potato
90% or more production loss, or even total crop failure can be caused when weight, are one of most serious diseases on sweet potato, are had become influence at present
The major obstacle of China epidemic-stricken area sweet potato production and the potential threat of Sweet Potato Industry development, ruin for mainly by the sweet potato that aleyrodid is propagated
Venereal disease of going out viral disease SPVD, replacing detoxification kind and the protection of sweet potato seedling stage is the important measures of the pre- disease prevention in epidemic-stricken area.SPFMV and
SPCSV is RNA virus, according to article " serology and Molecular Detection of Viruses On Sweet Potato In China type, Qiao Qi etc., plant physiology
Journal is published in for 2012 " in record, detected by the way of RT-PCR.
2) serological detection method
It is NCM-ELISA method using the optimum method of Serologic detection sweet potato viruses, this method is fine using nitric acid
That ties up plain membrane as carrier is immunized enzyme-linked reaction technology, has the characteristics that high specificity, method are easy, quick, is convenient for great amount of samples
Detection.The reagent used includes: Tris, sodium chloride (NaCl), sodium azide (NaN3), sodium sulfite (Na2SO3), defatted milk
Powder, Triton-100, magnesium chloride (MgCl2·6H2O), antiserum, goat-anti exempt from IgG alkaline phosphatase conjugate, nitrogen blue tetrazole
(NBT), 5 bromo- 4 chloro- 3 indoylphosphates (BCIP), N-N dimethylformamide, nitrocellulose filter etc..
Buffer is as follows: TBS buffer 0.02mol/L, Tris Base 4.84g, sodium chloride 0.5mol/L,
The sodium azide 0.4g that 58.44g mass fraction is 0.0001, is dissolved in 1995ml distilled water, and with HCl tune pH value to 7.5,
Distilled water is settled to 2000ml；Extraction buffer, sodium sulfite (Na2SO3) 1g adds TPS buffer 500ml；Block buffer takes off
Rouge milk powder 2.4g, Triton-1002.4ml add TPS buffer 120ml；T-TBS buffer (washing is used), 0.5ml Tween-
20, add TBS buffer 1000ml；Antibodies buffer, skimmed milk power 4.8g add TBS buffer 240ml；AP buffer (matrix),
0.1mol/L Tris Base6.05g, 0.1mol/L sodium chloride 2.92g, magnesium chloride 0.51g, mass fraction is 0.0001 nitrine
Change sodium 0.05g to be dissolved in 450ml distilled water, HCl tune pH to 9.5 is settled to 500ml；NBT and BCIP stoste preparation (NBT and
BCIP is extremely toxic substance, must be careful when taking, and wears gloves).NBT stoste, NBT 40mg add volume fraction to be 0.7 N-N bis-
Methyl nitrosourea 1.2ml is mixed, and is kept in dark place and (is wrapped up with dark-colored bottle or with aluminium foil) at 4 DEG C；BCIP stoste, BCIP 20mg
Add the N-N dimethylformamide 1.2ml that volume fraction is 0.7 to mix, is kept in dark place at 4 DEG C and (with dark-colored bottle or uses Aluminium Foil Package
It wraps up in).
Sample acquisition and extraction, the sample that need to be detected is entangled with polybag with the test tube or the metal pen cap of diameter 1cm every modeling
Material bag stays in bag from the roundleaf piece that diameter is 1cm is depressed on sample, takes out extra blade, and into sack plus 1ml extracting is slow
Fliud flushing is gently rolled with stick, and broken leaf texture and buffer are mixed, and is stood 30-40min at 4 DEG C, is taken clarified juice point sample.
When point sample, nitrocellulose filter is layered on filter paper, carefully draws supernatant in bag with clean sterilizing pipette
Liquid 20-30 μ l, drop is on film in grid, and often plus a sample changes a pipette water dropper, until going to film separately after being fully completed
On one clean filter paper, color development treatment is carried out after film is dry.
When color development treatment, point sample film is impregnated with Block buffer 30ml, vibration is incubated for 60min, discards confining liquid；Use 30ml
Antibodies buffer and appropriate sweet potato viruses antiserum mix (Specific amounts is according to antiserum operation instruction), and point sample film is immersed and is incubated for
Overnight.It discards primary antibody buffer within 2nd day, is washed 4 times with T-TBS buffer 30ml oscillation, each 3-5min.Point sample film is transferred to
In secondary antibody buffer added with enzyme label, oscillation incubation 60min discards secondary antibody buffer, is washed with T-TBS buffer 30ml oscillation
It washs 4 times, each 3-5min, is finally washed 1 time with AP liquid 30ml.Now match NBT/BCIP substrate solution, AP buffer 100ml is added
300 μ l of BCIP is added dropwise while stirring and mixes by NBT stoste 300ml.Point sample film is put into 25ml NBT/BCIP substrate buffer solution
Colour developing, vibration are incubated for 30-40min, terminate reaction.With distillation washing film 3 times, each 3min.Sampling point is reacted into blue purple
The positive shows the sample with virus, and color is deeper, and viral level is higher.
Detoxic seedling is detected by both the above method, can accomplish that viral recall rate is 100% substantially.
3. the hardening of Sweetpotato Viruses Elimination test tube seedling:
Establish mature Sweetpotato Viruses Elimination test tube seedling hardening system
June 15 to July 15 hardening in batches is carried out to Sweetpotato Viruses Elimination seedling, this time is the best hardening time, specific to refine
Steps are as follows for seedling:
1) domestication in bottle
I.e. before test tube seedling bottle outlet, triangular flask is placed under stronger illumination, opens sealed membrane, increases ventilation, bottleneck adds
PE gloves are covered, or so 1 week of domestication in bottle.
2) it is tamed outside bottle
Test tube seedling is removed from culture bottle, container for plant growth is arrived in field planting, by one section of moisturizing growth phase, also known as " outside bottle
Domestication ".Test tube seedling has certain adaptability to natural environment after domestication in the bottle of certain phase, can be by it
It is transplanted to the space of the outer moisturizing of bottle.
Concrete operation method are as follows: test tube seedling is removed from triangular flask gently, cannot overexert, prevent from pulling apart seedling root.Such as
Fruit culture medium too dry (water content is lower than 20%), can first soak and impregnate 1 day seedling taking again.Attach to the culture of test tube seedling root
Base should clean up, in order to avoid bacterium of mildewing.One hand gently pinches the rhizome top of seedling when cleaning, and another hand gently rubs seedling root, will
Agar attached thereto and loose callus clean up；If root is too long to cut one section, auxin (50mg/L is dipped in
NAA seed plate) or after root-inducing powder is moved into.
Transplanting medium is advisable with loose, draining and good air permeability person, such as frog stone, perlite, turf, fertile soil, most
Well using the good composite interstitial substance of the physicochemical property of thorough disinfection.It, should be in cleaning, the condition of temperature control after test tube seedling is implanted into seedbed
Lower growth, air humidity are big, it is however generally that air humidity controls 28-30 DEG C of temperature in 70-80% or so.The humidity of matrix
Should not be too high, general moisture content is 30-40%, and water supply in media, which excessively understands poor aeration, influences long root, is easy to cause rotten seedling.For
The seedling just transplanted, it should be shaded in short term.After 1-3d is grown, gradually reinforce illumination, makes seedling slowly reform of nature
Environmental condition.After transplanting about 1-2 weeks, suitable foliage spray is carried out, it is preferred to use 1/4MS or 1/2MS culture medium a great number of elements
Mixed liquor, by one month domestication hardening.
3) land for growing field crops are transplanted
By in the nursery for taming 157464 plants of sweet potato transplantation of seedlings to 3 mu survived that hardening obtains, grows three months, obtain
157464 plants of sweet potato seedlings survived.
Select loose fertile, row irrigation convenience, the good plot of sunshine as nursery the sweet potato seedling after hardening.It transplants in seedling
The seedling on garden ground should ensure that 3 months growth periods, manage with delicacy, so that diameter increment and twig are mature.As long as test tube seedling
Healthy and strong, early period, hardening was appropriate, and balled transplanting, general survival rate is all 90% or so.
4, the plantation of seedling:
1) winter greenhouse is colonized: before the Frost's Descent, i.e., the climing point of the sweet potato seedling survived 157464 plants before lunar calendar September 26 carries
Be inserted in 3 mu of winter greenhouses to cultivate and took off canopy to 1 day April in the coming year and obtain 141718 plants of sweet potato strong sprouts, specific steps include: clip
The climing point of hardening 3 months detoxification strong sprouts (the sweet potato seedling that 157464 plants i.e. above survive), is planted, takes straight cutting
Or oblique cutting dense planting, covering film heat, and promote every plant to take root 10 or more, grow 2-3 piece young leaves, accomplish fluently overwintering basis.It takes outside
Big arch shed is built, middle space increases lid plastic film, increases day accumulated temperature.
2) Seedlings nursery expands numerous
3) Production of Large Fields
2nd year use single, double leaf segment breeding method: by collect seedling garden cultivation strong sprout, the seedling of one leaf segment of single plant clip or
The seedling of two leaf segments, dense planting are planted into seed-plot, are enhanced field management, and are amounted to and are expanded numerous 4 times April 1 coming year to July 1.
Embodiment 2
The condition for changing Sweetpotato Viruses Elimination seedling production and the stem section adventitious buds differentiation in numerous stage fastly in embodiment 1 compares its disease
Malicious recall rate, planting percent and growing state are as shown in table 1- table 3:
The different stem apexs of table 1 strip length and its viral recall rate
| Stem apex length | Stem apex quantity | The malicious seedling quantity of band | Viral recall rate |
| 1mm | 20 | 1 | 5% |
| 0.5mm | 20 | 0 | 0% |
The different culture medium prescription of table 2 and its corresponding planting percent
| Medium component | Stem apex number | Seedling quantity | Planting percent |
| MS+0.5mg/LIBA+1.0mg/L6-BA | 20 | 10 | 50% |
| MS+1.0mg/LIAA+0.5mg/L6-BA | 20 | 16 | 80% |
The different light application times of table 3 handle corresponding growing state
| Lighting process | Growing way |
| 10h/d | Slow growth, blade is partially yellow, and stem is thin and delicate |
| 14h/d | Growth is normal, but slowly |
| 16h/d | Growth is most fast, and blade is emerald green, and stem thickness is strong |
| 18h/d | Growth is normal, fast speed, easy early ageing |
In the differentiation cultivation stage of adventitious bud it can be seen from table 1-3, using " removing 0.5mm or so in embodiment 1
Shoot apical meristem, be inoculated on (MS+1mg/L 6-BA+0.5mg/L IAA) culture medium, be placed on 28 DEG C, luminous intensity is
In (2000-3000) lux culturing room, daily illumination 16h, the mode of the differentiation of evoking adventive bud has best cultivate effect,
Viral recall rate 0%, planting percent is up to 80%, and the speed of growth is most fast, and blade is emerald green, and stem thickness is strong.
By in embodiment 12 years two-part sweet potato mating systems and 3 years three-field system sweet potatoes in the prior art breeding side
The data comparison of method is as shown in table 4.
The data comparison of table 4 two years two-part sweet potato mating systems and 3 years three-field system sweet potato mating systems
In conjunction with table 1-4, it can be seen that due to 2 years two-part sweet potato mating systems of the invention, using stem apex as explant
Body carries out cultivation detoxic seedling, and in test tube seedling reproductive process by the way of multiple squamous subculture, test tube seedling expand it is numerous,
Expanding numerous coefficient is 3n, wherein n is the number of squamous subculture, so that the seedling amount greatly increased, shortens to obtain identical quantity
The growing-seedling period of seedling amount.Compared with prior art, the potato seed cost of unit kilogram reduces by 26.51%, and breeding coefficient expands 278
Again, cultivation age limit has and becomes within 3 years 2 years, reduces the time of one third, while can guarantee purity and disease-free insect infection.Its
In, the calculation that cost compares is as follows:
A: " 2 years two-stage methods " seed production technical units potato seed cost calculation:
A1: " 2 years two-stage methods " seed production technology totle drilling cost calculates
A11: test tube seedling production cost (mid-March-is by the end of May)
(1) test tube seedling throughput rate in unit period
Test tube seedling and propagating coefficient 3n, generation seedling amount is y=a × 3n, wherein n indicates that the number of squamous subculture, a indicate initial
Seedling number, the seedling number after y expression expansion is numerous.On 15-April 15 March, 5kg potato wedge vernalization is taken, obtains 240 stem apexs；On March 15-4
The moon 15,240 stem apexs are renewable at 240 plants of test tube seedlings；15-April 29 April (1 week subculture 1 time, y=240 × 32),
Obtain 2160 plants of test tube seedling；29-May 15 April (2 week subculture 2 times, y=2160 × 32), obtain test tube seedling 19440
Strain；15-May 27 May (2 week subculture 2 times, y=19440 × 32), obtain 174960 plants of test tube seedling.
Through 3 half a month stem apex detoxifications and fast numerous, 729 times of breeding amount expansion.
(2) cost calculation
Detoxification test tube plantlet production and fast numerous cost: every bottle 1.1 yuan of culture medium cost, single plant subculture 6 times in unit period are single
Strain culture medium cost 1.1 × 6=6.6 member obtains 174960 plants of test tube seedling altogether, therefore culture medium cost is 6.6 yuan × 174960
Strain=1154736 yuan；
Wage for workmen's expenditure: everyone monthly 3000 yuan, 2 people work 4 months, and wage expenditure is 3000 yuan × 2 people × 4
Month=24000 yuan；
Water power expenditure: laboratory water power monthly pays 1500 yuan, and 4 months, water power expenditure was 1500 yuan × 4 months=6000
Member；
Fast numerous period internal loss consumables cost: triangular flask 174960 are needed in the detoxifying fast breeding period, each triangular flask 4
Member, the triangular flask proportion of goods damageds are 30%, and the loss of triangular flask consumptive material is 209952 yuan total；
Detoxifying fast breeding period domestic demand aluminium-foil paper volume 5,300 yuan of every volume, the loss of aluminium-foil paper consumptive material are 1500 yuan total；
Consumptive material expenditure amounts to 209952+1500=211452 member.
Viral diagnosis cost: on 15-April 29 April, randomly selecting the test tube seedling from same single plant and detected, inspection
Surveying strain number is 240 plants, and every plant of testing cost is 4 yuan, and virus detection reagent consumptive material pays 960 yuan.
Test tube seedling production cost is accumulative: 1154736+24000+6000+211452+960=1397148 member.
A2: hardening cost (the 15-7 month in June 15)
Matrix (Nutrition Soil and vermiculite): test tube seedling amounts to 174960 plants, single plant average growth area 0.01m2, test tube seedling refining
Seedling adds up cultivated area 1749.6m2, required matrix total amount 1749.6m2×2kg/m2, add up to 3499.2kg.Nutrition Soil cost is 2
Member/6998 yuan of kg × 3499.2kg=6998.4 member ≈.
Wage for workmen's expenditure: hardening is total to ask 15 person-times of work, everyone 100 yuan of each wage, wage expenditure be 100 yuan/people/
It is secondary × 15 person-times=1500 yuan；
Hardening cost is accumulative: 6998+1500=8498 member
Hardening survival rate 90%, survival seedling number is 174960 × 0.9=157464 plants after hardening
A3: the field planting of winter greenhouse (before the lunar calendar September 26 Frost's Descent-April 1 coming year)
Greenhouse depreciation cost: annual 1800 yuan per acre, totally 3 mu (total seedling number/60000 plant/acre=157464/60000 ≈ 3
Mu), greenhouse depreciation cost is 1800 yuan × 3 mu=5400 yuan；
Land rent: annual 600 yuan per acre, totally 3 mu, land rent is 600 yuan × 3 mu=1800 yuan；
Wage for workmen expenditure: contract entered by an individual take 1200 yuan/mu, amount to 3 mu, wage expenditure for 1200 yuan/mu × 3 mu=
3600 yuan；
Rich water takes: annual 400 yuan per acre, totally 3 mu, land rent is 400 yuan × 3 mu=1200 yuan；
It is accumulative that winter greenhouse is colonized cost: 5400+1800+3600+1200=12000 member
Survival rate 90% after passing the winter, survival seedling number is 157464 × 0.9=141718 plants after passing the winter
A4: Seedlings nursery expands numerous cost Production of Large Fields cost (1 day-July 1 April in the coming year)
Greenhouse depreciation cost: annual 1800 yuan per acre, the 4-6 month expand within every 15 days numerous 1 time, expand numerous coefficient 2m, generation seedling amount is y
=a × 2m, wherein m indicates to expand the number of numerous culture, and seedling amount at the beginning of 6 months is 141718 × 24=2267488 plants, 38 mu of Seedlings nursery are needed,
3 mu of nursery of passing the winter is subtracted, increases 35 mu of nursery newly.Greenhouse depreciation cost is 1800 yuan × 35 mu=63000 yuan；
Land rent: the 6-7 month, then expand numerous 1 time, total seedling amount is 2267488 × 2=4534976 plants；(compare 240 plants of original lists
Strain, breeding coefficient expand 18896 times；) nursery ground area becomes 4534976 plants/60000 plants/acre 76 mu of ≈；Per acre annual 600
Member, land rent are 600 yuan × 76 mu=45600 yuan；
Wage for workmen expenditure: contract entered by an individual take 1200 yuan/mu, amount to 76 mu, wage expenditure for 1200 yuan/mu × 76 mu=
91200 yuan.
Rich water takes: annual 400 yuan per acre, totally 76 mu, land rent is 400 yuan × 76 mu=30400 yuan.
Seedlings nursery expense is accumulative: 63000+45600+91200+30400=230200 member.
A5: Production of Large Fields
Mu number calculates: total 1134 mu of seedling number/4000 plant/acre=4534976 plant/4000 plant/acre ≈；Production of Large Fields includes ground
It grants contract fee 2 expenditures, annual 600 yuan per acre of land rent, 1600 yuan/mu of contract fee, 2200 yuan of average outgo per acre；
Production of Large Fields takes accumulative: 2200 yuan/mu × 1134 mu=2494800 yuan.
To sum up, " 2 years two-stage methods " totle drilling cost adds up to 1397148+8498+12000+230200+2494800=4142646
Member.
A6: " 2 years two-stage methods " seed production technology Gross Output calculates
Sweet potato total output: 1500kg/ mus × 1134 mu=1701000kg；
Compared to the first year, breeding amount expands 1701000kg/5kg=340200 times.
A7: " 2 years two-stage methods " unit production of seed stock cost calculation
Unit production of seed stock cost: totle drilling cost/total output=414,264,6/1,701,000 2.44 yuan/kg of ≈.
B " 3 years three-field systems " seed production technical units potato seed cost calculation:
B1 breeding coefficient
First Year: 1 mu, middle seed selection potato 750kg, winter storage loss 30%, remaining 525kg after winter is stored；Second year:
525kg potato seed can plant 17.5 mu of ground (30kg/ mus), 17.5 mu × 1500kg=26250kg of second year total output, winter storage loss
30%, remaining 18375kg after winter is stored.Third year: 18375kg potato wedge can plant 612.5 mu, third gross annual output amount 612.5
Mu × 1500kg=918750kg.
Expanded within 3 years numerous, 1225 times of breeding amount expansion.
B2 reproductive-cost
Cost of labor: worker 2, for each person every year 120000 yuan；Cost of labor is accumulative: 2 people × 120,000 yuan × 3 year=720000
Member.
Production of Large Fields expense: Production of Large Fields include contract fee 2 expenditures of granting, annual 600 yuan per acre of land rent, contract fee
1600 yuan/mu, per acre 2200 yuan of average outgo；Production of Large Fields takes accumulative: (1+17.5+612.5) × 2200=1388200
Member.
Winter storage cost: winter storage cost is calculated according to 0.25 yuan/kg, containing packaging, water power, potato cellar depreciation etc..Winter
Season storage cost is accumulative: (1500+52500+1837500) × 0.5=945750 member.
Totle drilling cost is total: 720000+1388200+945750=3053950 member.
B3: unit potato seed cost price
Unit production of seed stock cost: totle drilling cost/total output=3053950/918750=3.32 member/kg.
Claims (9)
1. a kind of sweet potato mating system, which comprises the steps of:
1) preparation of detoxification test tube plantlet；
2) the numerous culture of the expansion of detoxification test tube plantlet: detoxification test tube plantlet by multiple squamous subculture expand numerous；
3) test tube seedling domestication hardening is obtained into sweet potato seedling；
4) sweet potato seedling is subjected in protecting field field planting, obtains sweet potato strong sprout；
5) by sweet potato strong sprout by 4 expansion numerous, obtained sweet potato sprout, the sweet potato sprout is planted in crop field, progress crop field
Production harvests sweet potato.
2. being washed the method according to claim 1, wherein the step 1) includes taking sweet potato top stem with bud
Differentiation culture is carried out after net, the condition for breaking up culture is the shoot apical meristem for taking 0.5-1mm of removing, is inoculated into differential medium
On, at 28 DEG C, luminous intensity is (2000-3000) lux, culture in daily illumination 10-18 hours environment.
3. according to the method described in claim 2, described 10-18 hours is 16 it is characterized in that, 0.5-the 1mm is 0.5mm
The hour differential medium is MS+1mg/L 6-BA and 0.5mg/L IAA, the MS+1mg/L 6-BA and 0.5mg/L IAA
It is that the obtained culture medium of 6-BA and IAA is added into MS culture medium, in the MS+1mg/L 6-BA and 0.5mg/L IAA, 6-BA
Content be 1mg/L, the content of IAA is 0.5mg/L.
4. method according to claim 1 to 3, which is characterized in that in the step 2), squamous subculture is trained in MS
It supports in base, at 28 DEG C, luminous intensity is (2000-3000) lux, culture in daily illumination 10-18 hours environment.
5. method according to any one of claims 1-4, which is characterized in that in the step 2), the squamous subculture
Period is 7 days, and the Subculture times are 6 times.
6. any method in -5 according to claim 1, which is characterized in that in the step 3), the bottle including test tube seedling
Interior domestication, the bottle nursery of the domestication and test tube seedling culture outside of test tube seedling.
7. the method according to claim 1, wherein expanding the numerous method of numerous each expansion for 4 times of the step 5)
Obtain cuttings for single plant is cut section, each cuttings contains a leaf segment or two leaf segments, by the cuttings be inserted into soil in into
Row culture.
8. -7 any method according to claim 1, which is characterized in that further include the virus of test tube seedling in the step 2)
Sampling observation: the more plants of test tube seedlings from same single plant are randomly selected and carry out viral diagnosis.
9. according to the method described in claim 8, it is characterized in that, the method for the viral diagnosis is PCR detection method and serum
Detection method, the virus of the detection are SPCSV, Geminiviridae and SPVD.
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| CN114051927A (en) * | 2021-11-09 | 2022-02-18 | 延安市农业科学研究所 | Simplified breeding method for virus-free sweet potato seedlings |
| CN115644063A (en) * | 2022-11-11 | 2023-01-31 | 青岛农业大学 | Rapid propagation method of virus-free raw seedling of sweet potato |
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| CN115644063A (en) * | 2022-11-11 | 2023-01-31 | 青岛农业大学 | Rapid propagation method of virus-free raw seedling of sweet potato |
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