CN103891618A - Method for breeding two-season seed potato - Google Patents
Method for breeding two-season seed potato Download PDFInfo
- Publication number
- CN103891618A CN103891618A CN201410146076.5A CN201410146076A CN103891618A CN 103891618 A CN103891618 A CN 103891618A CN 201410146076 A CN201410146076 A CN 201410146076A CN 103891618 A CN103891618 A CN 103891618A
- Authority
- CN
- China
- Prior art keywords
- potato
- seed
- test
- tube plantlet
- seasons
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Abstract
The invention discloses a method for breeding a two-season seed potato, relates to a plant regeneration method using a tissue culture technique, and is used for solving the technical problems that the potato in the Taihang Mountains area is too many, disordered and miscellaneous in plant variety, the seed potato in one-season planting areas is severe in disease and the quality of the seed potato is degraded, the cost in allocating and transporting the seed potato is high, the storage time is long and the great rot loss is caused. The method for breeding the two-season seed potato mainly comprises the following steps: selecting the seed potato, sterilizing the stem tip, performing scaled production of test-tube plantlets, performing soilless culture on breeder seeds, planting the breeder seeds in spring, and planting one-class seeds in autumn. By adopting the method, the terrain and climate advantages are sufficiently utilized, the diseases of the seed potato are alleviated, the utilization rate of the seed potato is improved, the on-site supply of the seed potato is achieved, the production cost is low, the yield of a product is high, and the quality is high. The method is applied to the field of seed potato breeding.
Description
Technical field
The present invention relates to a kind of plant regeneration method of tissue culture technique of passing through, particularly relate to a kind of method of breeding seed potato of cultivating of organizing.
Background technology
Potato, as important grain, vegetables dual-purpose crop, should make again the raw material of industry, can with various crop intercropping, meanwhile, the efficiency of water application of potato is 8 times of wheat and maize, is adapted at nonirrigated farmland, Wild jujube in Taihang Mountain Area and water resource and lacks area plantation.Therefore, potato is the staple crops of western Wild jujube in Taihang Mountain Area, Hebei province always, is the Main Economic source of local farmers.
Due to Wild jujube in Taihang Mountain Area immiseration, potato produces backward in technique, and potato planting is the situation of " many, assorted " for many years.Varieties of plant is diversification general layout, take " Guangdong white, little Bai bell " as main, with without flower, gram new, brave first-class kind, is wherein mainly " Guangdong is white ".Peasant is certainly to reserve seed for planting as main, remain post sales little potato and plant to plant then, also there are a small amount of peasant and local high height above sea level region peasant to change kind, the problems such as variety hybriding degradation, disease are serious, cause thus quality, the output of commodity potato to decline year by year, yampi ratio less than 40%, average annual output only maintains 1000～1500 kilograms every mu.The generation of calendar year 2001～2002 year late blight, makes the potato underproduction serious, and average yield only has 500 kilograms of left and right.
Nourishing and generating is the main modes of reproduction of potato.Potato nutritional when breeding susceptible viral is contaminated, and in plant internal breeding, transhipment with accumulate in lump stem, along with transmitting from generation to generation, virus endangers and increases the weight of year by year, generally can the underproduction more than 50%.Research is found, nearly more than the 30 kinds of virus infections potatos of vegetative kingdom, and is caused deterioration of variety.
Common Potyvirus has PVX, PVY, PLRV, PVS, PVA, PVM and potato spindle tuber viroid (PSTVd), generally there is PVY, PSTVd virus in potato planting district of Hebei province, becomes the restriction of Wild jujube in Taihang Mountain Area High Yield Potato high-quality because of the factor.Can the underproduction when PVY virus is serious more than 80%; PSTVd has the infectivity of height, very easily carries out juice propagation by contact, farm implements, clothing and cutting knife etc.Therefore, cultivating antiviral kind and virus-elimination seedlings is to alleviate virus disease, improves the important measures of potato yield and quality, but the application of virus-free seed potato is at that time blank substantially.
On the Northeast, Inner Mongolia Autonomous Region and our province Zhangjiakou, dam, Chengde, regional Yi Jizuo district is potato commodity potato main producing region, is also the main grown place of potato seed, the producing region that Cong Yijizuo district of Ye Zengshi Taihang mountainous areas of hebei province allocates and transports seed potato on a small quantity.Due to potato seed cannot with commodity potato isolation implant, and continuous cropping for many years, causes disease serious, deterioration of variety speed adds that allocation and transportation potato seed road is far away, cost is high, the storage time is long, rotten loss is large, effect is unsatisfactory.If can change " season is done " is " two seasons were done ", carrying out virus-free seed potato at local spring and autumn breeds, the breeding phase is avoided rainy season, the storage time of potato seed will be shortened greatly, reduce the disease of potato seed and rot to consume, reduce costs, make the lifting of potato seed quality, cost, promote Development of Potato Industry.
Summary of the invention
The present invention is in order to solve Wild jujube in Taihang Mountain Area potato planting kind " many, random, assorted "; Seed potato producing region, Yi Jizuo district continuous cropping for many years, causes disease serious, the potato seed quality deterioration causing; And allocation and transportation potato seed road is far away, cost is high, the storage time is long, the large technical problem of loss of rotting, and two seasons of a kind of potato that provide are made the mating system of potato seed.
Two seasons of potato of the present invention are made the mating system of potato seed, mainly comprise the following steps:
One, seed selection: choose output relatively the highest, increase yield significantly, suitable high-yield culturing, resting stage is shorter, the anti-Major Diseases of holding concurrently, the kind of good stress resistance is as culture materials;
Two, stem apex detoxify: get the whole potato of the health of selected kind in step 1 and carry out vernalization, bud is through 38 ℃ of thermal treatment 14d, carry out conventional surface sterilization, under aseptic condition, peel off the growing point of 1～2 leaf primordium, be seeded on sterile MS solid culture medium, in 21～25 ℃, illumination 3000～4000Lx cultivates 3 months, be that 3～5 or height are 5～10cm to test-tube plantlet blade number, obtain detoxification test tube plantlet; Adopting double antibody enzyme-linked immunosorbent assay and RT-PCR technology to carry out virus to gained test-tube plantlet identifies;
Three, the large-scale production of test-tube plantlet: after test-tube plantlet is virus-free after testing, cultivate 3～4 weeks, when test-tube plantlet grows to 6～7 joint, single-unit segment is expanded numerous; Detoxification test tube plantlet is cut into the stem section with an axillalry bud, be inoculated in and be equipped with in the triangular flask that expands breeding culture medium; In the triangular flask of each 100 milliliters, can inoculate 10～25 sections; After inoculation, 5d is taken root left and right, and young shoot can grow up to the seedling of 2-4 sheet leaf from axil sends about 10 days, and 10～15d can be transplanted to greenhouse; Within 3～4 weeks, form the plantlets of 6～7 joints, again can cutting section propagation;
Four, soilless culture breeder's stock (micro potato): the 1. domestication of test-tube plantlet: will expand numerous test-tube plantlet in step 3 in the situation that not abroaching, move to greenhouse from incubator and carry out hardening; The hardening of uncapping in the time that test-tube plantlet has the leaflet of 5～7 strong greens, can transplant after 3～4d; 2. the test-tube plantlet after hardening is moved in culture matrix, more than cultivating 60d, when breeder's stock quality is 2～5g, can gather in the crops for the first time; Then immediately seedling is heeled in, watering, fertilizing, the diseases prevention of spray medicine, carries out after-crop after 2 weeks;
Five, spring original seed production: the breeder's stock that adopts spring little shed coverage mode to gather in the crops in region of no relief sowing step 4, at the beginning of by the end of May or 6 months, results, obtain original seed;
Six, autumn one-level kind production: at the beginning of by the end of July to 8 month by after the original seed breaking dormancy of gathering in the crops in step 5, in Wild jujube in Taihang Mountain Area plantation, at the beginning of by the end of October to 11 month, results one-level kind.
Wherein, in step 2, adopt double antibody enzyme-linked immunosorbent assay can detect PVX, PVY, PLRV, PVS, PVA and PVM virus; Adopt RT-PCR technology can detect PSTVD and PVY virus.
Optionally, in described step 1, the kind of described culture materials is early No. 5, rheum officinale or Zheng's potato.
Optionally, in described step 3, when inoculation test-tube plantlet, can inoculate multiple single stem segments for every bottle.
Optionally, in described step 3, when described test-tube plantlet is cultivated, the light source in each incubator is two thermo-optical fluorescent tubes, and spectral composition is " blue light+ruddiness "; Expanding breeding culture medium is the improved culture medium that " agar and deionized water " in conventional MS solid culture medium is changed into " 3% common white sugar and running water ".
Optionally, in the expansion breeding culture medium described in described step 3, add IAA or IBA.
Optionally, in the expansion breeding culture medium described in described step 3, add IAA or the 0.5mg/L IBA of 0.5mg/L.
Optionally, the culture matrix described in described step 4 is pine needle humus, expanded chicken manure and vermiculite, and stromal thickness is 16～18cm, and the 5cm of matrix lower floor is pine needle humus, and every square meter adds expanded chicken manure 2.5kg.
Optionally, in described step 4, in the cultivating process of breeder's stock, fertilising can adopt organic and inorganic fertilizer to join and execute.
Optionally, in described step 4, in the cultivating process of breeder's stock, described organic and inorganic fertilizer is joined to execute as fertilizer or bio-bacterial manure coordinate compound potassium sulfate fertilizer and is used.
Optionally, in described step 4, fertilizer is chicken manure, and bio-bacterial manure is take chicken manure as carrier.
Optionally, in described step 4, after breeder's stock results, potato piece is deposited in to place nice and cool, that ventilate, have scattered light, ground is take cement surface and floor tile as best.
Optionally, in described step 5, the time of sowing breeder's stock is March 10.
Optionally, the thickness of sowing described in described step 5 is 4000～5000 plants/acre.
Optionally, in described step 5, before results, 5d stops watering, and should avoid soil overdrying or excess Temperature that original seed is burnt by the sun simultaneously;
Its object is promote potato skin aging, the damage to epidermis while reducing results.
Optionally, in described step 6, the method for breaking dormancy for 10～15 mgs/kg of gibberellin spray or 1% thiourea solution soak seed 1 hour.
The present invention has following beneficial effect:
1, the present invention selects Potato Cultivars early ripe phase of No. 5, rheum officinale and Zheng's potato, little Bai bell whiter than local main breed Guangdong shifts to an earlier date 10～15 days, and product quality is all improved.Early 2500 kgs/acre of the fresh potato average yields of rheum officinale, contrast the white average volume increase 25.0% in Guangdong, contrast little Bai bell volume increase 38.89%; 2700 kgs/acre of No. 5 fresh potato average yields of Zheng's potato, than the white volume increase 24.5% in contrast Guangdong, contrast little Bai bell volume increase 38.3%, all reach significance level.The early yellow meat of No. 5 Calusena lansium of rheum officinale and Zheng's potato, quality is better than contrast, the anti-Major Diseases of holding concurrently, good stress resistance.
2, the present invention adopts RT-PCR detection technique system to detect PVY and PSTVd virus, has realized quick, accurate, sensitive, the specific detection of seedling detoxification.
3, two seasons of potato of the present invention are made potato seed mating system, and in test-tube plantlet cultivation, in each incubator, light source is two thermo-optical fluorescent tubes, make illumination energy reduce by 20～30%, and the emergence rate of test-tube plantlet improves more than 10%, and improves growth conditions.
4, two seasons of potato of the present invention are made potato seed mating system, and in test-tube plantlet cultivation, the spectral composition of light source is " blue light+ruddiness ", cultivate the test-tube plantlet emergence rate obtaining and reach more than 90%, leaf look dark green, and plant height is little, and joint number is more, growing way stalwartness, overall merit is good.
5, two seasons of potato of the present invention are made potato seed mating system, test-tube plantlet adds IAA and IBA in cultivating in medium, test-tube plantlet seedling stem that cultivation obtains is thick, the number of blade, long, the maximum leaf of root are long is all greater than contrast, wherein stem difference thick, the number of blade reaches significance level, show as Subculture Time short, expand numerous coefficient high.
6, two seasons of potato of the present invention are made potato seed mating system, the culture matrix of cultivation breeder's stock selects the abundant pine needle humus in Wild jujube in Taihang Mountain Area, expanded chicken manure to mix with vermiculite, compared with the cultivation matrix vermiculite generally adopting in producing, perlite, the peat composed of rotten mosses etc., 5.1 of micro potato individual plant knot potato numbers, average single seed reaches 8.02g, cultivate and increase by 1.9 than traditional cultivation matrix, average simple grain is brought up again high 4.61g; And make full use of local resources, substitute rare peat composed of rotten mosses resource, reduced substrate culture cost, increase the benefit.
7, two seasons of potato of the present invention are made potato seed mating system, adopt organic and inorganic fertilizer to join and execute to improve the quick-acting and slow supply of matrix nutrient, matrix micro-ecological environment be better than single to fertilize, improve the resistance of micro potato.Compared with the control, micro potato single-strain grain number and the simple grain of executing compound potassium sulfate fertilizer are joined in bio-bacterial manure increases heavily respectively 24.9% and 15.6%; Chicken dung and compound potassium sulfate fertilizer join to execute increases respectively 13.8% and 31.1%.
8, two seasons of potato of the present invention are made potato seed mating system, and changing " once results " is " twice results ", has increased the harvesting frequency of breeder's stock, and potato seed yield potentiality is fully excavated, and has improved matrix plant recovery of nutrient, reduces soilless culture cost simultaneously.Once results change after twice results, and potato seed survival rate does not change, but breeder's stock individual plant piece number increases by 64%, and monolithic is brought up again high by 54.7%, and total block data increases by 62.7%, and average simple grain does not heavily have difference, and the breeder's stock size of results adds evenly.
9, two seasons of potato of the present invention are made potato seed mating system, autumn, one-level production of seed stock carried out in Wild jujube in Taihang Mountain Area, make full use of the advantage that high altitude mountainous area summer climate is nice and cool, the temperature difference is large, rainfall is many, illumination is sufficient, assurance potato seed is emerged smoothly, is produced, and kind property is not degenerated.
10, two seasons of potato of the present invention are made potato seed mating system, changing " 1 year generation " is " 1 year two generation ", mountain area and Plain combine and have improved potato seed quality, shorten breeding cycle, potato seed has been avoided rainy season during breeding, alleviate potato seed disease, improved the availability of potato seed, realized potato seed and supplied on the spot.Adopt the potato seed that this technology is produced to increase production 500-520 kg/acre than native breed.
11, two seasons of potato of the present invention are made potato seed mating system, one-level production of seed stock in autumn in good time early sowing in mountain area (July 25) than Plain Er Jizuo district autumn sowing (August 5) emerge soon, seedling is neat, seedling is strong, individual plant knot potato increases by 40.0%, 50g ties potato number above increases by 80.0%, monolithic potato heavily increases by 47.4%, potato seed output reaches 1500 kgs/acre, volume increase 47.3%.If mountain area and region of no relief are sowing on August 5, mountain area potato seed is emerged early 8 days, and early 12 days harvest time, individual plant knot potato number increase by 20%, 50g ties potato number above increases by 30.0%, and monolithic potato heavily increases by 20.0%, potato seed volume increase 19.6%
Accompanying drawing explanation
Fig. 1 is 5% polyacrylamide gel electrophoresis RT-PCR testing result; In figure, 1,2,4 is sample, 3 negative contrasts;
Fig. 2 is 5% polyacrylamide gel electrophoresis RT-PCR testing result; 1,2,4 positive samples in figure, 3 negative contrasts;
Fig. 3 is 5% polyacrylamide gel electrophoresis RT-PCR testing result; In figure, 1 is blank, 2 positive contrasts, and 3～11 for detecting sample.
Embodiment
Adopt following examples and contrast verification test to be further detailed the present invention.
Two seasons of the present embodiment potato are made the mating system of potato seed, mainly comprise the following steps:
One, seed selection: with white, the little Bai Lingwei check variety in local main breed Guangdong, introduce early rheum officinale, No. 5, Zheng's potato, the Atlantic Ocean, early great Bai, No. 25 Potato Cultivars of middle potato, breeding time, productivity, quality and commodity, the resting stage index of investigation kind, select output relatively the highest, increase yield significantly, suitable high-yield culturing, resting stage is shorter, double anti-Major Diseases, the kind of good stress resistance is as culture materials.
Two, stem apex detoxify: get selected kind in step 1 early the whole potato of health of No. 5, rheum officinale and Zheng's potato carry out vernalization, bud is through 38 ℃ of thermal treatment 14d, carry out conventional surface sterilization, under aseptic condition, peel off the growing point of 1～2 leaf primordium, be seeded on sterile MS solid culture medium, in 23 ℃, illumination 3500Lx cultivates 3 months, be that 3～5 or height are 5～10cm to test-tube plantlet blade number, obtain detoxification test tube plantlet; Adopting double antibody enzyme-linked immunosorbent assay and RT-PCR technology to carry out virus to gained test-tube plantlet identifies.
Three, the large-scale production of test-tube plantlet: after test-tube plantlet is virus-free after testing, cultivate about 3～4 weeks, in the time that test-tube plantlet grows to 6 joint, single-unit segment is expanded numerous; Detoxification test tube plantlet is cut into the stem section with an axillalry bud, be inoculated in and be equipped with in the triangular flask that expands breeding culture medium; In the triangular flask of each 100 milliliters, inoculate 20 sections; After inoculation, 5d is taken root left and right, and young shoot grows up to the seedling of 2-4 sheet leaf from axil sends about 10 days, and 12d is transplanted to greenhouse; Within 3～4 weeks, form the plantlet of 6～7 joints, continue cutting section propagation.
Receive with Shang Haijia a group training special light source, the two Philip thermo-optical fluorescent tubes that company produces respectively, carry out comparative trial with three common fluorescent tubes as light source, the relatively impact of Different Light on test-tube plantlet.
Take conventional MS+7% agarose medium as contrast, respectively process medium and be the common white sugar of MS(3% and running water) medium, be provided with 8 hormon processing, repeat for three times.Process 1:MS+0.1mg/LNAA, process 2:MS+0.3mg/LNAA, process 3:MS+0.2mg/LIAA, process 4:MS+0.5mg/LIAA, process 5:MS+0.7mg/LIAA, process 6:MS+0.2mg/LIBA, process 7:MS+0.5mg/LIBA, process 8:MS+0.7mg/LIBA); Investigation plant height, stem are thick, the number of blade, radical, long, the maximum leaf of root are long, Ye Se and transplanting survival rate, the relatively impact of different culture media on test-tube plantlet.
Four, soilless culture breeder's stock (micro potato): the 1. domestication of test-tube plantlet: will expand numerous test-tube plantlet in step 3 in the situation that not abroaching, move to greenhouse from incubator and carry out hardening; The hardening of uncapping in the time that test-tube plantlet has the leaflet of 6 strong greens, transplants after 4d; 2. the test-tube plantlet after hardening is moved in culture matrix, more than cultivating 60d, when breeder's stock quality is 4g, gather in the crops for the first time; Then immediately seedling is heeled in, watering, fertilizing, the diseases prevention of spray medicine, carries out after-crop after 2 weeks.
Be mixed into culture matrix with pine needle humus, expanded chicken manure and vermiculite, add vermiculite matrix as contrast take the peat composed of rotten mosses.If two processing: process 1,5 centimetres of matrix lower floors are the peat composed of rotten mosses, and every square meter adds 2.5 kilograms of expanded chicken manures; Process 2,5 centimetres of matrix lower floors are pine needle humus, and every square meter adds 2.5 kilograms of expanded chicken manures.Survival rate, plant height, individual plant piece number, individual plant piece weight, total block data, output, culture matrix more of the present invention and the impact of traditional culture matrix on breeder's stock of test-tube plantlet in investigation different substrates.
If 4 processing are respectively and process A: bio-bacterial manure (take chicken manure as carrier), treatments B: common Chicken dung, processing C: bio-bacterial manure+compound potassium sulfate fertilizer, processing D: common chicken manure+compound potassium sulfate fertilizer; Contrast is compound potassium sulfate fertilizer, observes the proterties such as process of growth middle period look, growing way, resistance (to the adaptive capacity of high temperature, lack of water), fertilizer efficiency stability, investigation individual plant knot potato grain number, individual plant knot potato grams.Relatively the present invention and the impact of common fertilizer on breeder's stock.
If contrast experiment, A: adopt method of the present invention to carry out results twice to breeder's stock; B: adopt conventional method only once to gather in the crops breeder's stock; The impact of more different harvesting approaches on breeder's stock output and quality.
If two processing, repeat for three times; Process 1: little shed plantation, process 2: mulching planting.Each processing two row (20 meters are long, 24 square metres), density is 8000 plants/acre, and other cultivation management is consistent with potato land for growing field crops.The relatively little shed plantation and the impact of mulching planting on breeder's stock seeding stage, output and harvest time in the present invention.
Five, spring original seed production: the breeder's stock that adopts spring little shed coverage mode to gather in the crops in region of no relief sowing step 4, at the beginning of by the end of May or 6 months, results, obtain original seed.
Stripping and slicing time test: establish 2 processing: process 1: the rear stripping and slicing of results in spring; Process 2: stripping and slicing before planted in fall; Preservative treatment test: establish 4 processing: process 1: talcum powder+bactericide (agricultural streptomycin and thiophanate methyl; Process 2: talcum powder; Process 3: clear water; Process 4: contrast, do not do preservative treatment.
100 potato seeds of each processing in above two groups of tests, establish three repetitions.The potato piece recording after stripping and slicing or the preservative treatment 15 days ratio of rotting.Stripping and slicing time in contrast the present invention and method for anticorrosion treatment and conventional method affect potato piece corrosion-resistant.
Six, autumn one-level kind production: at the beginning of by the end of July to 8 month by after the original seed breaking dormancy of gathering in the crops in step 5, in Wild jujube in Taihang Mountain Area plantation, at the beginning of by the end of October to 11 month, results one-level kind.
If three processing: process the 1:7 month 25 sowing; Process the 2:7 month 30 sowing; Process the 3:8 month 5 sowing, repeat for three times, each two row (20m is long, 24m2) that are treated to, mainly investigate seeding stage, output and harvest time, and other cultivation management is consistent with potato land for growing field crops.The impact that in contrast the present invention, date of seeding and normal sowing dates are bred one-level kind.
Comparative test result:
1, Potato Cultivars
Screening varieties result of the test shows (in table 1): from output, quality and resistance index analysis, 5 Potato Cultivars of introduction are all better, wherein relatively the highest with the average yield of No. 5, Zheng's potato, two kinds of rheum officinale morning, increase yield significantly, suitable high-yield culturing.Early 2500 kgs/acre of the fresh potato average yields of rheum officinale, contrast the white average volume increase 25.0% in Guangdong, contrast little Bai bell volume increase 38.89%, 2700 kgs/acre of No. 5 fresh potato average yields of Zheng's potato, than the white volume increase 24.5% in contrast Guangdong, contrast little Bai bell volume increase 38.3%, all reach significance level.The early yellow meat of No. 5 Calusena lansium of rheum officinale and Zheng's potato, quality is better than contrast, the anti-Major Diseases of holding concurrently,
Good stress resistance.
Be applicable to vegetative period that Er Jizuo district's potato seed breeds, resting stage take whether and as evaluation index, compared with check variety, early 10～15 days in advance ripe phase of No. 5, rheum officinale and Zheng's potato, resting stage is shorter, and marketing quality improves.
3 years screening varieties of table 1 are tested the general performance of each kind
2, the evaluation of RT-PCR detection technique system to PVY, PSTVd virus-virus
The detection and identification method of potato PVY, PSTVd virus is mainly enzyme-linked immunosorbent assay (ELISA), but is subject to the restriction of plant virus antiserum condition, and can not detect accurately and reliably PVY virus and PSTVd virus in potato tubers; Because viroids does not have protein coat, do not there is antigenicity in addition, can not detect with serological method.This research has been synthesized efficient special primer according to the nucleotide sequence of the main virus of potato, utilize synthetic primer, adopt the pcr amplification technology successfully PSTVd and PVY virus to be detected, set up the quick RT-PCR detection technique system of PVY, PSTVd virus.
20 batches of potato virus-free plantlet samples are carried out to PVY detection, feminine gender and positive control are set simultaneously, partial results shows detected detoxic seedling positive rate approximately 7% left and right (seeing Fig. 1～3), during the PVY RT-PCR technical system of setting up can be applicable to produce.High and the high specificity of this sensitivity, its selectivity and sensitivity are higher more than 10000 times than ELISA detection method, low to viral level in detoxic seedling especially, when serological method is difficult to detect, better effects if.Apply cost and detection time that this detection technique greatly reduces detection, improved the speed and the sensitivity that detect, lay a good foundation for further carrying out viral composite PCR detection research.
3, the light source that detoxification test tube plantlet is bred
The light source of traditional group training is generally fluorescent lamp, and infrared and far ratio is large, and energy is easily lost in environment, and plant luminous energy availability is low, and power consumption cost is high.By contrasting a group training special light source (Shang Haijia receive company produce), two thermo-optical fluorescent tubes (PHILIPS Co.), with the impact of three common fluorescent tubes on test-tube plantlet growth, result shows: at the test initial stage, three processing have no obvious difference, test-tube plantlet growth stalwartness, is showed no abnormal.But later stage, finds to adopt the test-tube plantlet growing way of special light source to die down gradually, shows as blade and diminishes, and stem slightly attenuates, and excessive growth etc. are the performance of illumination deficiency, continues to cultivate symptom and does not improve.The test-tube plantlet growing way of two Philip thermo-optical fluorescent tubes dies down slightly, but recovers by add appropriate growth regulator test-tube plantlet growth in medium.Result shows, Shang Haijia receives group training special light source that company produces and cannot meet the illumination needs of potato tissue culture seedling, two Philip thermo-optical fluorescent tubes coordinate growth regulator can guarantee the normal growth of test-tube plantlet, make illumination energy reduce by 20～30%, the emergence rate of test-tube plantlet improves more than 10%, and improves growth conditions
4, the spectral composition that detoxification test tube plantlet is bred
The impact of Different Light structure on test-tube plantlet growth, result shows (in table 2): the test-tube plantlet emergence rate of blue light+ruddiness processing reaches more than 90%, and leaf look dark green, and plant height is little, and joint number is more, growing way stalwartness, overall merit is good, as preferred spectral composition.
The impact of the different spectral compositions of table 2 on Potato Plantlets in vitro
5, the medium of detoxification test tube plantlet
Result of study shows (in table 3), and medium after improvement adds hormon to be processed test-tube plantlet growth is all had to obvious impact, and the each index of test-tube plantlet is thick all higher than contrast except stem, and wherein plant height, radical, root are long all contrasts that there were significant differences; From test-tube plantlet growing way, the test-tube plantlet that adds NAA is taken root early early stage, and root system development is fast, height of seedling, but seedling is slightly very thin, and successive propagation performance is general.Add the higher test-tube plantlet stem end callus that causes of NAA concentration occur morning and more.The test-tube plantlet stem that adds IAA and IBA is thick, the number of blade, long, the maximum leaf of root are long is all greater than contrast, and wherein stem difference thick, the number of blade reaches significance level, shows as Subculture Time short, expands numerous coefficient high.The test-tube plantlet Comprehensive Traits that MS+0.5mg/L IAA and MS+0.5mg/L IBA process is effective, and a generation expands that numerous coefficient is the highest, and transplanting survival rate is high, between the two without significant difference.Wherein, MS+0.5mg/L IAA increases by 22.8% than the test-tube plantlet plant height of contrast, and stem slightly increases by 21.8%, and the number of blade increases by 37.9%, radical increase by 16.6%, and root is long increases by 13.8%, and maximum leaf is long increases by 26.7%, and transplanting survival rate raising reaches 92%; MS+0.5mg/LIBA increases by 21.6% than the test-tube plantlet plant height of contrast, and stem slightly increases by 16.8%, and the number of blade increases by 29.3%, radical increase by 5.4%, and root is long increases by 14.1%, and maximum leaf is long increases by 26.3%, and transplanting survival rate raising reaches 91%; Comprehensive analysis, two formulas of MS+0.5mg/L IAA or MS+0.5mg/LIAA are best as the culture medium prescription effect of potato detoxicating cuvette seedling Fast-propagation.
The impact of the medium of table 3 hormon processing on test-tube plantlet growth
6, the culture matrix of breeder's stock soilless culture
The micro potato major traits of improvement matrix all contrasts effective, adds pine needle and produce micro potato in matrix, individual plant stem tuber number, individual plant stem tuber are heavy, stem tuber is total and output all higher than contrast, volume variance reaches significance level (in table 4).Compare with contrasting (conventional matrix, the peat composed of rotten mosses+vermiculite), 5.1 of the micro potato individual plant knot potato numbers of improvement matrix, average single seed reaches 8.02g, increases by 1.9 than contrast, and average simple grain is brought up again high 4.61g.Improvement matrix does not have notable difference to detoxic seedling survival rate, but improvement matrix makes full use of local resources, substitutes rare peat composed of rotten mosses resource, has reduced substrate culture cost, increases the benefit.
Table 4 is improved the impact of matrix formulations on micro potato major traits
7, the fertilizer practice of breeder's stock soilless culture
Chicken manure organic fertilizer or chicken manure are that the bio-bacterial manure of carrier coordinates micro potato growing way and the resistance of compound potassium sulfate fertilizer cultivation better, and single-strain grain number and simple grain are heavily significantly higher than contrast (in table 5), have set up soilless culture organic and inorganic fertilizer and have joined the technology of executing.Organic and inorganic fertilizer is joined to execute and can be improved the quick-acting and slow supply of matrix nutrient, matrix micro-ecological environment be better than single to fertilize, improve the resistance of micro potato.Compared with the control, micro potato single-strain grain number and the simple grain of executing compound potassium sulfate fertilizer are joined in bio-bacterial manure increases heavily respectively 24.9% and 15.6%; Chicken dung and compound potassium sulfate fertilizer join to execute increases respectively 13.8% and 31.1%.
The impact of table 5 different fertilizer kind on micro potato growth and potato kernel Characters
8, twice results of breeder's stock
By increasing the harvesting frequency of breeder's stock, potato seed yield potentiality is fully excavated, and has improved matrix plant recovery of nutrient, reduces soilless culture cost simultaneously.Once results change (in table 6) after twice results, and potato seed survival rate does not change, but breeder's stock individual plant piece number increases by 64%, and monolithic is brought up again high by 54.7%, and total block data increases by 62.7%, and average simple grain does not heavily have difference, and the breeder's stock size of results adds evenly.
The impact of the different harvesting approaches of table 6 on potato seed proterties
9, the little shed plantation of seed production in spring
Seed production was generally arranged in region of no relief and carried out spring, at the beginning of by the end of May or 6 months harvest times, breed to one-level potato seed in autumn and reserve enough time, broke potato seed resting stage.For guaranteeing original seed quality, realize early field planting of breeder's stock, original seed early harvest, avoid the high-incidence season of rainy season in later stage, high temperature and biography poison medium aphid, adopt little shed coverage mode to increase ground temperature, field planting potato primary stock ahead of time.Result of study showed (table 7), in 10 days March sowing date of breeder's stock spring, adopts little shed plantation, than conventional plastic mulching seedling-growing time 5 days in advance.End June 1 harvest date, little shed increases by 20%, 50g than the individual plant knot potato number of plastic mulching and ties above potato number increase by 30%, and individual plant piece is brought up again high by 32.6%, and per mu yield improves 32.7%.If produce 100 grams of potatos as standard take average individual plant, little shed plantation can shift to an earlier date 8～10 days than plastic mulching, for autumn sowing breaking dormancy is laid a good foundation.Meanwhile, little shed cropping pattern is simple, invests littlely, easily carries out industrialization development.
The impact of table 7 Different Ways of Planting on potato seed production in spring
10, potato breaking dormancy
After spring original seed results, spray or 1% thiourea solution, the 1 hour breaking dormancy of soaking seed with 10～15 mgs/kg of gibberellin.Whole potato vernalization is processed sowing and is germinateed soon, emerges fast, has rotten potato phenomenon after broadcasting.And vernalization processing germination is the not slowest, emerge late, broadcast rear rotten potato rate high.
11, potato cutting is anticorrosion
Gather in the crops spring after original seed breaking dormancy, carry out stripping and slicing preservative treatment.By large potato piece (single potato weighs more than 50 grams potato seed) stripping and slicing, each stripping and slicing has 1～2 eye, and weight is not less than 20～30 grams.After stripping and slicing due to summer and autumn temperature higher, potato seed easily rot.For reducing rotten potato rate, stripping and slicing time and preservative treatment technology are studied.Result shows (in table 8), and no matter from rotting rate and emergence rate, the rear stripping and slicing in time of results in spring is processed and significantly reduced original seed rotting rate, is controlled at 3.1%, and field emergence rate reaches 96.9%; Before planted in fall, the potato piece rotting rate of stripping and slicing controls to 6.1%, and field emergence rate reaches 93.9%.Talcum powder+bactericide, talcum powder, clear water after spring results and do not process four kinds of modes and reduce by 3.0%, 5.2%, 9.8% and 15% than the potato seed rotting rate of stripping and slicing in autumn respectively, also corresponding raising of emergence rate after potato planted in fall.
Different stripping and slicing time of table 8 potato original seed and preservative treatment are on potato piece rotting rate and the after planting impact of emergence rate
Note: carry out one-way analysis of variance between the different preservative treatments after gathering in the crops spring and before planted in fall.
Adopt talcum powder processing can absorb in time the moisture of the stripping and slicing surface of a wound, suppress miscellaneous bacteria existence, increase bactericide and can reduce rotten potato rate, realize the neat seedling of seedling strong.Under the rear stripping and slicing condition in time of results in spring, the treatment effect of talcum powder+bactericide is best, reduces by 6.9% than the potato seed rotting rate of not processing, and processes and reduces by 5.1% than clear water, and emergence rate is up to 96.9%; Before planted in fall, under stripping and slicing condition, the treatment effect of talcum powder+bactericide is best, reduces by 18.9% than the potato seed rotting rate of not processing, and processes and reduces by 12.9% than clear water.
12, Wild jujube in Taihang Mountain Area Autumn sowing period
After spring potato original seed results, stripping and slicing in time is also carried out preservative treatment, then utilizes that mountain area temperature cools, the weather conditions of suitable production of seed stock, carries out planted in fall production potato seed in Wild jujube in Taihang Mountain Area, has avoided the rainy unfavourable weather condition of region of no relief summer high temperature.Different sowing dates result of the test shows (in table 9), the in good time early sowing in mountain area (July 25) than Plain Er Jizuo district autumn sowing (August 5) emerge soon, seedling is neat, seedling is strong, individual plant knot potato increases by 40.0%, 50g ties potato number above increases by 80.0%, monolithic potato heavily increases by 47.4%, potato seed output reaches 1500 kgs/acre, volume increase 47.3%.Mountain area and region of no relief are sowing on August 5, and mountain area potato seed is emerged early 8 days, and early 12 days harvest time, individual plant knot potato number increase by 20%, 50g ties potato number above increases by 30.0%, and monolithic potato heavily increases by 20.0%, potato seed volume increase 19.6%.
The impact of table 9 different sowing dates in autumn on potato seed Growth and yield
Comprehensive analysis, region of no relief produces toxicity-removing white potato original seed can carry out one-level production of seed stock in Wild jujube in Taihang Mountain Area, not only make full use of mountain resource, has avoided rainfall and high temperature potato to be infected to the impact of virus and transmitted virus simultaneously.The potato seed disease that mountain area produces gently, kind property is good, potato piece is little, with the obvious advantage as original seed, can produce original seed in potato seed base, Wei Yijizuo district, and development Potato Industry, increases local farmers and take in.Therefore, region of no relief plantation in spring potato original seed, autumn is in the potato seed of Wild jujube in Taihang Mountain Area plantation one-level, built that Plain combines with Wild jujube in Taihang Mountain Area two seasons of toxicity-removing white potato to make potato seed breeding mode feasible, and the key issue of the each link of virus-free seed potato breeding of method solves, realize the batch production of toxicity-removing white potato potato seed.
Above-described embodiment is described the preferred embodiment of the present invention; not scope of the present invention is limited; design under the prerequisite of spirit not departing from the present invention; various distortion and improvement that those of ordinary skills make technical scheme of the present invention, all should fall in the definite protection domain of the claims in the present invention book.
Claims (10)
1. two seasons of potato are made a mating system for potato seed, it is characterized in that: mainly comprise the following steps:
One, seed selection: choose output relatively the highest, increase yield significantly, suitable high-yield culturing, resting stage is shorter, the anti-Major Diseases of holding concurrently, the kind of good stress resistance is as culture materials;
Two, stem apex detoxify: get the whole potato of the health of selected kind in step 1 and carry out vernalization, bud is through 38 ℃ of thermal treatment 14d, carry out conventional surface sterilization, under aseptic condition, peel off the growing point of 1～2 leaf primordium, be seeded on sterile MS solid culture medium, in 21～25 ℃, illumination 3000～4000Lx cultivates 3 months, be that 3～5 or height are 5～10cm to test-tube plantlet blade number, obtain detoxification test tube plantlet; Adopting double antibody enzyme-linked immunosorbent assay and RT-PCR technology to carry out virus to gained test-tube plantlet identifies;
Three, the large-scale production of test-tube plantlet: after test-tube plantlet is virus-free after testing, cultivate 3～4 weeks, when test-tube plantlet grows to 6～7 joint, single-unit segment is expanded numerous; Detoxification test tube plantlet is cut into the stem section with an axillalry bud, be inoculated in and be equipped with in the triangular flask that expands breeding culture medium; In the triangular flask of each 100 milliliters, inoculate 10～25 sections, cultivate 10～15d;
Four, soilless culture breeder's stock: the 1. domestication of test-tube plantlet: will expand numerous test-tube plantlet in step 3 in the situation that not abroaching, move to greenhouse from incubator and carry out hardening; The hardening of uncapping in the time that test-tube plantlet has the leaflet of 5～7 strong greens, cultivates 3～4d; 2. the test-tube plantlet after hardening is moved in culture matrix, more than cultivating 60d, when breeder's stock quality is 2～5g, gather in the crops for the first time; Then immediately seedling is heeled in, watering, fertilizing, the diseases prevention of spray medicine, carries out after-crop after 2 weeks;
Five, spring original seed production: the breeder's stock that adopts spring little shed coverage mode to gather in the crops in region of no relief sowing step 4, at the beginning of by the end of May or 6 months, results, obtain original seed;
Six, autumn one-level kind production: at the beginning of by the end of July to 8 month by after the original seed breaking dormancy of gathering in the crops in step 5, in Wild jujube in Taihang Mountain Area plantation, at the beginning of by the end of October to 11 month, results one-level kind.
2. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in described step 1, the kind of described culture materials is early No. 5, rheum officinale or Zheng's potato.
3. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in described step 3, when inoculation test-tube plantlet, can inoculate multiple single stem segments for every bottle.
4. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in described step 3, when described test-tube plantlet is cultivated, the light source in each incubator is two thermo-optical fluorescent tubes, and spectral composition is " blue light+ruddiness "; Expanding breeding culture medium is the improved culture medium that " agar and deionized water " in conventional MS solid culture medium is changed into " 3% common white sugar and running water ".
5. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in the expansion breeding culture medium described in described step 3, add IAA or IBA.
6. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: the culture matrix described in described step 4 is pine needle humus, expanded chicken manure and vermiculite, stromal thickness is 16～18cm, the 5cm of matrix lower floor is pine needle humus, and every square meter adds expanded chicken manure 2.5kg.
7. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in described step 4, in the cultivating process of breeder's stock, described fertilising is that fertilizer inorganic fertilizer is joined and executed.
8. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in described step 4, after breeder's stock results, potato piece is deposited in to place nice and cool, that ventilate, have scattered light, ground is take cement surface and floor tile as best.
9. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in described step 5, the time of sowing breeder's stock is March 10.
10. two seasons of a kind of potato according to claim 1 are made the mating system of potato seed, it is characterized in that: in described step 6, the method for breaking dormancy for 10～15 mgs/kg of gibberellin spray or 1% thiourea solution soak seed 1 hour.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410146076.5A CN103891618A (en) | 2014-04-11 | 2014-04-11 | Method for breeding two-season seed potato |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410146076.5A CN103891618A (en) | 2014-04-11 | 2014-04-11 | Method for breeding two-season seed potato |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103891618A true CN103891618A (en) | 2014-07-02 |
Family
ID=50983457
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410146076.5A Pending CN103891618A (en) | 2014-04-11 | 2014-04-11 | Method for breeding two-season seed potato |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103891618A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106034609A (en) * | 2016-05-30 | 2016-10-26 | 云南农业大学 | High-efficiency high-quality breeding method for planting potato |
| CN106134687A (en) * | 2015-04-22 | 2016-11-23 | 蒲建刚 | Two seasons breeding Early cropping potato potato seed new technique |
| CN106258975A (en) * | 2016-08-24 | 2017-01-04 | 遵义华富生物科技有限公司 | A kind of potato primary stock production method |
| CN111480580A (en) * | 2020-05-28 | 2020-08-04 | 严柯雯 | Method for cultivating virus-free potatoes in test tubes |
| CN111713368A (en) * | 2020-07-27 | 2020-09-29 | 胡智远 | Planting method for royaljelly |
| CN111713367A (en) * | 2020-07-27 | 2020-09-29 | 胡智远 | Method for planting Huangshan tribute chrysanthemum |
| CN111820130A (en) * | 2020-07-28 | 2020-10-27 | 南京农业大学 | Potato microtuber induction method based on LED energy-saving light source |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102804950A (en) * | 2012-08-21 | 2012-12-05 | 刘勇 | Method for breaking dormancy of potato seed tubers |
| CN103053418A (en) * | 2012-12-31 | 2013-04-24 | 湖北凯瑞百谷农业科技股份有限公司 | Potato microtuber manufacturing technology |
| CN103190264A (en) * | 2013-03-27 | 2013-07-10 | 四川省农业科学院作物研究所 | Potato stem tip detoxification method |
-
2014
- 2014-04-11 CN CN201410146076.5A patent/CN103891618A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102804950A (en) * | 2012-08-21 | 2012-12-05 | 刘勇 | Method for breaking dormancy of potato seed tubers |
| CN103053418A (en) * | 2012-12-31 | 2013-04-24 | 湖北凯瑞百谷农业科技股份有限公司 | Potato microtuber manufacturing technology |
| CN103190264A (en) * | 2013-03-27 | 2013-07-10 | 四川省农业科学院作物研究所 | Potato stem tip detoxification method |
Non-Patent Citations (6)
| Title |
|---|
| 屈冬玉等: "松针土作基质生产脱毒微型薯试验研究", 《马铃薯杂志》, vol. 13, no. 1, 31 December 1999 (1999-12-31), pages 17 * |
| 庞淑敏等: "马铃薯脱毒苗快繁低成本培养基改良试验", 《河南农业科学》, no. 12, 31 December 2004 (2004-12-31) * |
| 张海霞等: "马铃薯茎尖脱毒培养技术研究", 《吉林农业》, no. 10, 31 October 2013 (2013-10-31), pages 23 * |
| 沈清景等: "马铃薯脱毒原原种高产快繁技术研究", 《福建农业学报》, vol. 15, no. 4, 31 December 2000 (2000-12-31) * |
| 王培伦等: "二季作区脱毒马铃薯种薯繁殖技术规程", 《中国马铃薯》, vol. 15, no. 2, 31 December 2001 (2001-12-31) * |
| 王芳: "无土基质栽培生产脱毒马铃薯微型薯的关键技术", 《作物杂志》, no. 5, 31 December 2008 (2008-12-31) * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106134687A (en) * | 2015-04-22 | 2016-11-23 | 蒲建刚 | Two seasons breeding Early cropping potato potato seed new technique |
| CN106034609A (en) * | 2016-05-30 | 2016-10-26 | 云南农业大学 | High-efficiency high-quality breeding method for planting potato |
| CN106034609B (en) * | 2016-05-30 | 2019-03-29 | 云南农业大学 | A kind of high-efficiency high-quality mating system of seed potato |
| CN106258975A (en) * | 2016-08-24 | 2017-01-04 | 遵义华富生物科技有限公司 | A kind of potato primary stock production method |
| CN111480580A (en) * | 2020-05-28 | 2020-08-04 | 严柯雯 | Method for cultivating virus-free potatoes in test tubes |
| CN111713368A (en) * | 2020-07-27 | 2020-09-29 | 胡智远 | Planting method for royaljelly |
| CN111713367A (en) * | 2020-07-27 | 2020-09-29 | 胡智远 | Method for planting Huangshan tribute chrysanthemum |
| CN111820130A (en) * | 2020-07-28 | 2020-10-27 | 南京农业大学 | Potato microtuber induction method based on LED energy-saving light source |
| CN111820130B (en) * | 2020-07-28 | 2021-11-02 | 南京农业大学 | Potato microtuber induction method based on LED energy-saving light source |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103891618A (en) | Method for breeding two-season seed potato | |
| CN104838995B (en) | A kind of method for culturing seedlings of tomato water planting cuttage root-taking | |
| CN107155434A (en) | A kind of breeding method of multifunction manual woods | |
| CN106613823A (en) | Soilless culture substrate for tomato and preparation method thereof | |
| CN104472366A (en) | Tissue culture rapid-propagation method for improving salt tolerance of seedlings of southern ecotype jujubes | |
| CN107027579A (en) | A kind of method of peach nursery cutting propagation | |
| CN103918531A (en) | Solanaceous vegetable culture medium and preparation method thereof | |
| CN1324947C (en) | Potato seed breeding method | |
| CN107079697A (en) | The Yellow River and Huai He River area sunlight-heat reaources rationally utilize the double early double evening sequential cropping cultivation methods of wheat and corn | |
| CN107211891B (en) | Konjak callus direct seeding and rapid propagation technology | |
| CN101897284A (en) | High-yield cultivation method of hybrid rice | |
| CN104956889A (en) | Muskmelon plantation method | |
| CN105940901A (en) | Method for postponing bolting of Wuta-tsai | |
| CN105248229A (en) | Planting method for increasing Vc content of kiwi fruits | |
| CN102783410A (en) | Breeding method of high-quality and double-low rapes suitable for mechanization work and with high yield, and lodging resistance | |
| CN105145042B (en) | Virus-free basic potato seed soil incubation method | |
| CN103444520A (en) | Breeding technology of novel strawberry virus-free seedling | |
| AU2020101771A4 (en) | Method for in vitro cyclic preservation of Potato Germplasm | |
| CN104782371B (en) | A kind of ratooning rice for being suitable for southern ricefish system, which stores, educates cultural method | |
| CN104322369B (en) | A kind of method of lombardy poplar stem segment tissue culture fast breeding | |
| CN108029471A (en) | A kind of spinach growing nursery and culture method | |
| CN106386469A (en) | Resistance breeding method of zea mays | |
| CN106665366A (en) | Tissue culture method of dendrobium unicum | |
| CN105766288A (en) | Method for staged harvest and whole-year production of early-spring sweet potatoes | |
| CN112243856A (en) | Early-maturing late-japonica rice hybrid breeding method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140702 |