CN101803572A - Method for cultivating jerusalem artichoke hybrid seeds and obtaining jerusalem artichoke planting hybrid strains and tuber - Google Patents
Method for cultivating jerusalem artichoke hybrid seeds and obtaining jerusalem artichoke planting hybrid strains and tuber Download PDFInfo
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Abstract
The invention relates to a method for cultivating jerusalem artichoke hybrid seeds and obtaining jerusalem artichoke planting hybrid strains and tuber. The method comprises the following steps that: the jerusalem artichoke hybrid seeds are organized to be cultivated and jerusalem artichoke seedlings are obtained. The method of the invention overcomes the defects that the jerusalem artichoke hybrid seeds have low seed setting rate, are not easy to sprout at conventional conditions, are difficult to obtain effective individual plants, the appraisal cycle of the method applied to the farmland from the seeds to obtaining the hybrid tubers which is affected by the growth period is long. The method of the invention can effectively keep the jerusalem artichoke hybrid mutation, greatly shortens the cycle from the mutation into the farmland appraisal, can provide quite many optional mutation sources for jerusalem artichoke hybridization, speed up the selection process of hybrid breeding, and have good application prospect in jerusalem artichoke hybridization.
Description
Technical field
The present invention relates to a kind of method that artichoke hybrid seeds obtains jerusalem artichoke crossbreed plant and stem tuber of cultivating.
Background technology
Jerusalem artichoke (Helianthus tuberosus L.) has another name called Jerusalem artichoke, and Jerusalem artichoke, ginger are not peppery, originates in the North America, in China various places cultivation is arranged all, is composite family Helianthus herbaceos perennial.Jerusalem artichoke happiness temperature, salt tolerant, anti-lean, resistance and high-yielding is high density energy-source plant and novel economizer crop, forage crop.
Jerusalem artichoke is a capitulum, and inflorescence is little, and flower quantity is many.(A represents capitate part rip cutting, B tubular flower, C ligulate flower to the structure of inflorescence as shown in Figure 1.1 receptacle of inflorescence, 2 involucres, 3 tubular flowers, 4 ligulate flowers, 5 bracts, 6 ovarys, 7 pappus, 8 corolla pipes expand part, 9 petals, 10 synantherous stamens, 11 column caps), flower cluster shape is arranged.Petal: 5 pieces, Colaesce becomes tubulose or ligule mutually, and it is asexual that ligulate flower is, faint yellow, and be born in the inflorescence edge; Tubular flower is a hermaphrodite flower, and is born in the middle of the inflorescence.Stamen: 5 pieces, filigree disconnected from each other and flower pesticide edge mutual Colaesce formation sky tubular, i.e. synantherous stamen.Stamen, is sprinkling upon pollen grain in " tube " of synantherous stamen when flower pesticide is ripe early than the gynoecium maturation on the same tubular flower, when treating the growth of gynoecium style, they " is pushed away " to go out outside the tube.Gynoecium: ovary is the next, and two carpels constitute, a Room, and one piece of anatropous ovule, substrate and is given birth to.One of style is stretched in the flower pesticide pipe, and top column cap 2 splits, but when gynoecium prematurity still, column cap does not open.At superior part of interlobule, Chang Sheng has a circle hair, cries " sweeping the powder hair ", and in the process of extending whenever the style growth, this pollen grain of " sweeping the powder hair " and stamen flower pesticide " can being spread " in the flower pesticide pipe " pushes away ", the insect pollination of being convenient to come to visit.It generally all is protandry that feverfew is spent, in the style elongation process pollen grain " pushed away " go out after, the column cap on top opens again accepts the pollen that other flower transmits, and accepts pollen and finishes fertilization.
The jerusalem artichoke florescence is the 8-9 month, and the tubiflorous open hour on its same capitulum are inconsistent, and along with the inflorescence growth, ligulate flower is open at first, and the tubular flower around is open subsequently, and is open by export-oriented central authorities successively, and follows the growth and the pollen loose powder of column cap.The immature tubular flower of stamen gynoecium as shown in Figure 2, the stamen top closure is not seen black flower pesticide silk.The gynoecium stamen begins ripe flower earlier as shown in Figure 3 all around, visible stamen black flower pesticide silk, and top visible yellow color pollen sheds.The gynoecium stamen begins ripe flower as shown in Figure 4 in the middle of the capitulum.
The jerusalem artichoke stem tuber is rich in synanthrin, accounts for the 70-80% of its stem tuber dry weight, is one of primary raw material of present China industrialization extraction synanthrin.Synanthrin is as a kind of functional ingredient of pure natural, double grading and function with soluble dietary fiber, oligosaccharide, it simultaneously also is the fat substitute of Bifidobacterium MF, excellence, be food to diabetes patient's safety, be nutritional supplement by more than 20 state approval in the world, be widely used in dairy products, beverage, low-fat low-calorie food, bake and bank up with earth in food, the health food, China Ministry of Public Health approved inulin, polyfructosan are as new resource food.Therefore the jerusalem artichoke industry has good development space in China; Simultaneously jerusalem artichoke still is good energy-source plant, also sees report with the jerusalem artichoke stem tuber as what raw material was produced bio-ethanol.The drought resisting of jerusalem artichoke is cold-resistant, anti-lean, the characteristic of salt tolerant alkali makes it have wide popularization space in northern China, can make full use of resource and abundant photo-thermal resource marginally such as northern large-area saline land, poor and barren land, side slope ground, wasteland, has good ecological benefits simultaneously.
At present China's jerusalem artichoke industry still is in the starting stage, in Qinghai, ground such as Ningxia, Gansu, Hubei form industry size.Aspects such as domestic research to jerusalem artichoke at present mainly concentrates on and introduces a fine variety, cultivation, physiology, ecology, downstream product processing and utilization.The development jerusalem artichoke hybrid variety breeding work of jerusalem artichoke industry seriously lags behind relatively, does not see the relevant report of jerusalem artichoke hybrid variety breeding as yet.For guaranteeing to answer the sustainable development of jerusalem artichoke industry, must carry out the seed selection of breed breeding work, the especially Hybrid of system, for the development of jerusalem artichoke industry provides good kind to guarantee.
In the production, jerusalem artichoke is based on vegetative propagation, and open naturally pollination ripening rate is extremely low, and the cross-fertile rate is lower; The jerusalem artichoke seed sowing or the sowing major part of germinateing can not normally germinate or germination rate extremely low.These characteristics of jerusalem artichoke make it utilize the possibility of natural variation seed selection new varieties extremely low, more can't satisfy the demand of breed breeding work, therefore must be by the more variation of the artificial creation of the means of artificial hybridization, and the acquisition that extremely low ripening rate of jerusalem artichoke and the lower germination rate of seed make hybrid seed, breeding are all extremely difficult, limited by the mode of artificial hybridization create, preserve, utilize variation may; A little less than the plant of the artichoke hybrid seeds breeding simultaneously growing way, be subject to influences such as disease, environment and dead, the variation that artificial hybridization is obtained is lost; And numerously become the land for growing field crops to identify time more than the minimum 2-3 of need of colony owing to expanded by the simple grain artichoke hybrid seeds by restriction that jerusalem artichoke breeding time and hybrid seed breeding plant stem tuber yield poorly, the cycle is long; Therefore must seek suitable method and as much as possible keep hybrid seed and make it in the shortest time, expand numerous bigger colony that obtains, be applied in the crossbreeding.
Summary of the invention
An object of the present invention is to provide a kind of method that artichoke hybrid seeds obtains the jerusalem artichoke seedling of cultivating.
Cultivation artichoke hybrid seeds provided by the present invention obtains the grow directly from seeds method of seedling of jerusalem artichoke, comprises the steps: that the tissue culture artichoke hybrid seeds obtains the jerusalem artichoke seedling.
In the said method, the method for described tissue culture comprises the steps:
1), obtains the artichoke hybrid seeds of sterilization with described artichoke hybrid seeds sterilization;
2) artichoke hybrid seeds with described sterilization is inoculated in the MS medium, handles 12-36h for 4 ℃, cultivates 10-30 days under temperature is the condition of 20 ℃-23 ℃ and lucifuge then, obtains jerusalem artichoke crossbreed seedling.
In the said method, in described step 2) after, comprise the step that following expansion is numerous: the described jerusalem artichoke seedling of growing directly from seeds is cut into the stem section of tool bud, the stem section of tool bud is transferred in the 1/2MS medium that adds α-Nai Yisuan, in temperature is that 20 ℃-25 ℃, light intensity 2500-3000lux and light application time are to cultivate under 14h/ days the condition, obtains the jerusalem artichoke seedling;
The described 1/2MS medium that adds methyl (NAA) of giving birth to is made up of 1/2MS medium and α-Nai Yisuan NAA, and the proportioning of 1/2MS medium and NAA is 1L: (0.05mg-0.15mg), be specially 1L: 0.1mg.
In the said method, described sterilization comprises the steps: described artichoke hybrid seeds peeling, place the mixed solution of washing agent and water to soak 1-2h, flowing water flushing 1-3h uses the ethanol water of 75% (volumn concentration) to soak 10-30s, aseptic water washing then, be 5% aqueous sodium hypochlorite solution sterilization 3-6min with available chlorine content again, aseptic water washing is used 0.1% (quality percentage composition) mercuric chloride solution sterilization 1-4min, aseptic water washing again; The volume ratio of washing agent and water is (0.5-4) in the mixed solution of described washing agent and water: 100.
Another object of the present invention provides a kind of method that artichoke hybrid seeds obtains the jerusalem artichoke plant of cultivating.
Cultivation artichoke hybrid seeds provided by the present invention obtains the method for jerusalem artichoke plant, comprises the steps:
A) cultivate artichoke hybrid seeds according to the method described above, obtain jerusalem artichoke crossbreed seedling;
B) described jerusalem artichoke crossbreed seedling is carried out hardening and transplanting, obtain jerusalem artichoke crossbreed plant.
In the said method, after described step b), comprise the step of described jerusalem artichoke plant being carried out field cultivation.
Another object of the present invention provides a kind of method that artichoke hybrid seeds obtains jerusalem artichoke crossbreed stem tuber of cultivating.
Cultivation artichoke hybrid seeds provided by the present invention obtains the method for jerusalem artichoke crossbreed stem tuber, comprises the steps: to obtain according to above-mentioned arbitrary described method the jerusalem artichoke plant of field growing, continues field cultivation to gathering in the crops the jerusalem artichoke stem tuber.
Above-mentioned cultivation artichoke hybrid seeds obtains grow directly from seeds seedling, above-mentioned cultivation artichoke hybrid seeds of jerusalem artichoke and obtains the application of method in the species of jeruselem artichoke seed selection that the method for jerusalem artichoke plant, above-mentioned cultivation artichoke hybrid seeds obtain jerusalem artichoke crossbreed stem tuber and also belong to protection scope of the present invention.
The inventive method success artichoke hybrid seeds is cultivated into whole plant, transplant in field cultivation and gathered in the crops stem tuber.Experiment showed, that the inventive method carries out hybrid seed breeding on medium, the seed pollution rate is low, the percentage of seedgermination height, and seed is to land for growing field crops plant breeding coefficient height; Cycle is short, gathers in the crops more stem tuber from beginning to cultivate hybrid seed to field, only needs 8-12 month, has realized the efficient breeding of artichoke hybrid seeds.The inventive method has overcome that the artichoke hybrid seeds ripening rate is low, be difficult for germinateing, be difficult to obtain effective plant individuality and owing to be subjected to the influence of breeding time to be applied to land for growing field crops long defective qualification cycle from seed to obtaining the crossbreed stem tuber under the normal condition.The inventive method can effectively keep jerusalem artichoke hybridization variation, greatly shorten variation and enter the cycle that identify in the land for growing field crops, can be the jerusalem artichoke crossbreeding more alternative source of variation is provided, quicken the seed selection process of crossbreeding, in the jerusalem artichoke crossbreeding, have a good application prospect.
Description of drawings
Fig. 1 is the anatomical structure figure of jerusalem artichoke inflorescence.
Fig. 2 is a stamen gynoecium prematurity flower, and the stamen top closure is not seen black flower pesticide silk.
Fig. 3 is that the gynoecium stamen begins ripe flower all around, visible stamen black flower pesticide silk, and top visible yellow color pollen sheds.
Fig. 4 begins ripe flower for gynoecium stamen in the middle of the capitulum.
Fig. 5 cuts the inflorescence that applies glue after intermediate tubular is spent.
Fig. 6 is the suitableeest pollination pollen.
Fig. 7 is the suitableeest pollination column cap.
Fig. 8 is an artichoke hybrid seeds.
The jerusalem artichoke seed that Fig. 9 germinates down for aseptic condition.
Figure 10 is root and the seedling of fast numerous stage jerusalem artichoke seedling.
Jerusalem artichoke tissue cultivating seedling between Figure 11 cultivates.
Figure 12 is the jerusalem artichoke tissue cultivating seedling of transplanting in the nutritive cube kind.
Figure 13 is the jerusalem artichoke seedling of land for growing field crops kind.
Figure 14 is a jerusalem artichoke crossbreed stem tuber.
Embodiment
Employed experimental technique is conventional method if no special instructions among the following embodiment.
Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.
The acquisition of embodiment 1, jerusalem artichoke artificial hybridization seed
Select the jerusalem artichoke of purple stem tuber to carry out artificial hybridization in the present embodiment.
One, cross method
(1) castration of jerusalem artichoke female parent
1, the selection of inflorescence to be castrated
Tubular flower cluster shape in the jerusalem artichoke inflorescence is arranged, and its characteristics of blooming are by open successively toward the centre all around.Before morning, the temperature rises, gynoecium style in the jerusalem artichoke inflorescence on same tubular flower and stamen flower pesticide silk can ramp and both growth rate there are differences, the elongation of stamen flower pesticide silk is very fast and prior to the gynoecium maturation; Gynoecium style growth rate is slow, thereby forms certain distance between pistil stigma and the stamen top.Select the tubulous stamen of floral disc periphery edge to be about to ripe but not loose powder, gynoecium style not elongation as yet according to these characteristics, the still immature inflorescence of intermediate tubular flower is castrated as female parent.The inflorescence of the developmental condition of inflorescence after Fig. 2 and before Fig. 3 of selecting during castration.
The concrete selection florescence is in as the jerusalem artichoke inflorescence of next stage castrates, this in period inflorescence structure such as following (1) and (2): tubular flower around the tubular flower note that floral disc periphery edge in the jerusalem artichoke inflorescence is open is earlier done, all tubular flowers notes around will removing the tubular flower are made intermediate tubular and are spent; Around tubular flower when having structure described in following (1), the intermediate tubular in the inflorescence spends nature that structure described in following (2) is just arranged.
(1) tubiflorous all around structure: stamen is a synantherous stamen, is made of filigree, flower pesticide, and filigree separates, and the mutual Colaesce of flower pesticide becomes tubulose, and flower pesticide is loose powder not; Gynoecium is made of column cap, style and ovary, and style and column cap are positioned at flower pesticide inner bottom part centre and style does not extend, and the top of column cap and described flower pesticide is at a distance of 2mm-3mm; The top of described flower pesticide is away from an end of floral disc in the flower pesticide.
At this moment, tubiflorous all around stamen extends, and the filigree that can see black grows and middle rudimentary Xiao Hua is about 2-3mm, but its top still is a closed state, does not see that yellow pollen sheds, just flower pesticide full maturity not as yet; The also not elongation of same with it pistil stigma of taking for a short time this moment, the pistil stigma top has (2-3mm) with a certain distance from pistillate flower pencil top.
Flower pesticide is exactly the pollen load of tunicle bag, pollen prematurity (flower pesticide is loose powder not) during castration, the pollen that sheds so flower pesticide can't break.And pollen is attached to the flower pesticide inside pipe wall, directly removes the flower pesticide pipe and can remove all pollen;
Have only this period, the top of column cap and flower pesticide is just at a distance of 2mm-3mm, and along with the growth column cap of gynoecium style can be more and more near the stamen top, last bursting stamen flower pesticide top is stretched out column cap and released pollen simultaneously.
(2) intermediate tubular flower: stamen is a synantherous stamen, and stamen is made of filigree and flower pesticide, and filigree separates, and tubulose flower pesticide is to become tubulose by the mutual Colaesce of independent flower pesticide, but tubular flower flower pesticide, filigree still in growth, are not seen elongation; Ovary in the gynoecium, style and column cap are in the growth course, and also differentiation is incomplete.
2, castrate
(1) tubiflorous all around castration:
With 1/3 place that tweezers are clamped flower pesticide, perpendicular to the direction of described tubiflorous floral disc described stamen is dragged, leave gynoecium and expose column cap; 1/3 place of described flower pesticide from flower pesticide away from an end of floral disc to the floral disc telegoniometer.
The also elongation of gynoecium style in this in period, the just stamen filigree of elongation has certain distance between the two, therefore can not injure gynoecium, and removing what expose behind the stamen is the column cap of gynoecium.
(2) castration of intermediate tubular flower:
Cut the intermediate tubular flower with scissors at 1/2 to 1/4 place of intermediate tubular flower; 1/2 to 1/4 place of described intermediate tubular flower is from intermediate tubular flower and floral disc link meter.This step will be noted grasping and wipes out the position, wipes out and too much easily causes inflorescence death, the very few pollen contamination that easily causes.In the practical operation, have from 1/2 place, have from 1/4 place, falling between of having, statistics shows does not all have significant difference in 1/2 to 1/4 scope.
Dip in little writing brush and to get glue and spread upon edge of a knife position, to suppress its flower pesticide regrowth and to suppress loose powder (Fig. 5); In operating process, avoid glue to touch column cap to be pollinated.The method can effectively prevent slow growing tubular flower loose powder, reduces and castrates number of times, and the column cap that has exposed after having avoided castrating is subjected to the pollution of the pollen of same inflorescence.
Thoroughly remove stamen above-mentioned steps (1) and (2), prevents selfing, produces selfed seed, prevents that just the pollen of oneself from giving oneself pollination.
So far, the inflorescence of castrating, with it as female parent.
3, bagging
For inflorescence and other pollen hybridization after preventing to castrate, the inflorescence after will castrating subsequently puts the sulfuric acid paper bag, and allows gynoecium continue to grow.
(2) selection of male parent
Choose do not bloom and with the inflorescence of maternal flower synchronization, put the template hybridization bag, as the male parent inflorescence.The tubular flower loose powder time that flower synchronization specifically is meant male parent meets with the maternal back column cap duration of run of castrating.So long as consistent the getting final product of flowering time of the flower after male parent flower flowering time and maternal the castration, the inflorescence or the middle inflorescence all around of male parent that it doesn't matter are because male parent only provides pollen.
(3) pollination
Select the female parent and the male parent of following period and state to carry out artificial pollination:
Male parent: column cap has just exposed pistillate flower pencil top and pollen just has been ejected flower pesticide (Fig. 6) in the tubular flower of male parent.
Female parent: after maternal the castration, the style elongation is that the best is accepted the pollen time (Fig. 7) in the time of division of column cap front end and sliver flattening; This moment, pollination face was exposed, the abundant paulin exhibition of top mastoid process, and content is full transparent, and awarding property of column cap is best, and is easier to be solid.
Generally be to castrate about 4 of that afternoons to the next morning just can pollinate, awarding property of column cap is best at this moment.Use little writing brush to cling pollen, on the column cap of choosing, pollinate, put the sulfuric acid paper bag after pollination finishes, put on label.
(4) cultivation after the pollination
After pollination finished 4-7 days, remove the sulfuric acid paper bag, normal field management obtains hybrid seed.
The form of artichoke hybrid seeds as shown in Figure 8.
MS medium: production code member HB8469 manufacturer: Qingdao GaoKeYuan Hai Bo Bioisystech Co., Ltd.Mercury chloride: analyze pure (AR), mercury chloride content is no less than 99.5%, the manufacturer: reagent Manufacturing Co., Ltd is learned in permanent Xinghua, Tianjin.
Hypochlorous acid is received solution (available chlorine content is 10.0%), manufacturer: Tianjin BASF chemical industry Co., Ltd, credit number: Tianjin Q/AG3-333-2001.With the available chlorine content of buying is that 10% liquor natrii hypochloritis uses distilled water diluting, obtains available chlorine content and be 5% liquor natrii hypochloritis.With available chlorine content is that 5% liquor natrii hypochloritis experimentizes.
Washing agent is commercially available common carving board liquid detergent.
All experimentize among following each embodiment with 200 hybrid seeds.
The cultivation of embodiment 2, artichoke hybrid seeds
One, tissue culture
1, seed disinfection
Artichoke hybrid seeds is removed the peel, place the mixed solution (volume ratio of washing agent and water is 2: 100) of washing agent and water to soak 1.5h, flowing water flushing 3h, use the ethanol water of 75% (volumn concentration) to soak 20s then, aseptic water washing 3 times is 5% the liquor natrii hypochloritis 4min that sterilizes with available chlorine content again, aseptic water washing 3 times, use 0.1% (quality percentage composition) mercuric chloride solution sterilization 2min again, aseptic water washing 3 times.
Seed aseptic filter paper suck dry moisture with after the sterilization is inoculated in the MS medium, after 3 days, and the pollution rate of statistics seed.The seed that grows bacterial plaque is thought to pollute.
Pollution rate (%)=(seed number of seed number/inoculation occurring polluting) * 100%
The result who repeats for 3 times is: average pollution rate is 7.5%.
2, obtain complete seedling
Seed aseptic filter paper suck dry moisture with after step 1 sterilization is inoculated in the MS medium, handles 24h for 4 ℃, under being the condition of 22 ℃ and lucifuge, temperature cultivated 20 days then, and the artichoke hybrid seeds that the obtains seedling of growing directly from seeds, and add up percentage of seedgermination (Fig. 9); The MS medium is formed as shown in table 1.Under seed directly germinates and grows is exactly the seedling of growing directly from seeds.
Germination rate (%)=(chitting piece number/inoculation seed number) * 100%
The result who repeats for 3 times is: average percentage of seedgermination is 75%.
3, young seedling and propagating
The artichoke hybrid seeds that step 2 the is obtained seedling of growing directly from seeds is cut into the stem section of tool bud, be inoculated in the 1/2MS medium that adds α-Nai Yisuan (NAA), in temperature is that 23 ℃, light intensity 3000lux and light application time are to cultivate 20 days under 14h/ days the condition, obtain group training seedling (Figure 10 and Figure 11, Figure 10 A is a root, Figure 10 B is group training seedling), and statistics tissue cultivating seedling rooting rate.Expand the seedling major part that obtains in numerous step and all have root, but be not with root on a small quantity, the quantity of the seedling of this experiment statistics band root is calculated the tissue cultivating seedling rooting rate.
Add the 1/2MS medium of α-Nai Yisuan (NAA) and be made up of α-Nai Yisuan and 1/2MS medium, the proportioning of 1/2MS medium and NAA is 1L: 0.1mg.
Tissue cultivating seedling rooting rate (%)=(the stem hop count of taking root/inoculation stem hop count) * 100%
The result who repeats for 3 times is: average group training seedling rooting rate is 95%.
In each above-mentioned stage, what the stage 2 obtained is seedling by the direct seedling that obtains that germinates of seed.By seedling segment that the stage 2 is grown directly from seeds, the plant that cuttage grows up to the medium is the clone plant in stage 3; By the stage 2, the tissue culture in 3 two stages of stage can grow up to a strain seedling with a seed sprouting, and transfer on medium by cut-out, make the stem section of an one band bud just can develop into a complete plant, thereby make a strain plant breed into tens strains even tens strain plant in the short period of time.Because this tens strain or tens strain plant come from same seed in other words from same seedling, therefore be the clone body that comes from the grain seed.And can only obtain a plant for seed of traditional seminal propagation.
The composition of table 1, MS medium
The 1/2MS medium is formed: the constituent in the 1/2MS medium is identical with the MS medium, and wherein, macroelement and microelement concentration are 1/2 of MS medium, and the concentration of sucrose, agar, molysite and organic principle is identical with the MS medium, and pH is 5.8.
MS, 1/2MS boil dissolving back branch to be filled in the 150ml triangular flask, and 115 ℃ then, high pressure steam sterilization 20min.
Two, hardening and transplanting
(1) hardening
Step 1 acquisition group training seedling is carried out hardening, and concrete hardening step is as follows: blake bottle (triangular flask) is sealed film open 1/3, took exercise 3 days, then triangular flask is sealed film and all open, took exercise 5 days.This hardening process is to make tissue cultivating seedling progressively adapt to external environment, improves the transplanting survival rate of tissue cultivating seedling.
(2) transplant
1, the tissue cultivating seedling after the hardening is taken out, clean the medium of root remnants, transplant to the nutritive cube that matrix is housed, be in temperature that 23 ℃, ambient humidity are 75%, light intensity 3000lux and light application time be to cultivate 7 days under 14h/ days the condition; Guaranteeing to water less under the moistening prerequisite of matrix but definitely avoiding arid in this process, and forbidding ponding and strong illumination in the nutritive cube.
2, again temperature be 23 ℃, ambient humidity be 70% and the natural lighting condition under cultivated 20 days, statistics is transplanted survival rate of plant; The plant that obtains as shown in figure 12.
Matrix used in the said process is made of turfy soil turfy soil and sandy soil: sandy soil=1: 1.5 (volume ratio).
In the said process, hardening is to make plant adapt to external environment gradually, and transplanting in nutritive cube is to make plant obtain adaptive training in culturing room.This hardening and transplanting can improve the transplanting survival rate of tissue cultivating seedling.
Transplant survival rate of plant (%)=(transplant survival plant number/nutritive cube is transplanted the plant number) * 100%
The result who repeats for 3 times is: average nutritive cube transplanting survival rate is 87%.
Three, field cultivation
The jerusalem artichoke plant band compost that step 2 obtains is transplanted in big Tanaka, water, shading (taking the sunshade net to cover) 5 days.After the field planting, early stage the duty maintenance ground moistening that waters; Plant survives suitable minimizing the in the back of growing up fully and waters.Statistics is transplanted survival rate of plant (%); Transplant that the October can be gathered in the crops a large amount of hybrid seed plant stem tubers then.The plant of field planting as shown in figure 13.The stem tuber of results as shown in figure 14.
Wherein, water and suitably 5 days processing of shading can promote surviving of seedling.
Transplant survival rate of plant (%)=(land for growing field crops survives plant number/field-transplanting plant number) * 100%
The result who repeats for 3 times is: average field-transplanting survival rate is 85%.
After experiment finishes, statistics reproduction coefficient and breeding cycle.
Reproduction coefficient: be defined as every hybrid seed by experiment after one, two and three the step, can the surviving and can produce the plant number of stem tuber of acquisition.
Breeding cycle: from seed disinfection to the time of gathering in the crops stem tuber.
The result who repeats for 3 times is: reproduction coefficient is 7, and be 10 months breeding cycle.
The cultivation of embodiment 3, artichoke hybrid seeds
One, tissue culture
1, seed disinfection
Artichoke hybrid seeds is removed the peel, place the mixed solution (volume ratio of washing agent and water is 0.5: 100) of washing agent and water to soak 1h, flowing water flushing 1h, use the ethanol water of 75% (volumn concentration) to soak 10s then, aseptic water washing 3 times is again with the aqueous sodium hypochlorite solution of available chlorine content 5% sterilization 3min, aseptic water washing 3 times, use 0.1% (quality percentage composition) mercuric chloride solution sterilization 1min again, aseptic water washing 3 times.
Seed aseptic filter paper suck dry moisture with after the sterilization is inoculated in the MS medium, after 3 days, and the pollution rate of statistics seed.
Pollution rate (%)=(seed number of seed number/inoculation occurring polluting) * 100%
The result who repeats for 3 times is: average pollution rate is 12%.
2, obtain complete seedling
With the seed aseptic filter paper suck dry moisture after step 1 sterilization, be inoculated in the MS medium, handle 12h for 4 ℃, under temperature is the condition of 20 ℃ and lucifuge, cultivated 10 days then, the artichoke hybrid seeds that the obtains seedling (having root and cotyledon) of growing directly from seeds, and statistics percentage of seedgermination; The MS medium is formed as shown in table 1.
Germination rate (%)=(chitting piece number/inoculation seed number) * 100%
The result who repeats for 3 times is: average percentage of seedgermination is 71%.
3, young seedling and propagating
The artichoke hybrid seeds that step 2 the is obtained seedling of growing directly from seeds is cut into the stem section of tool bud, be inoculated in the 1/2MS medium that adds α-Nai Yisuan (NAA), in temperature is that 20 ℃, light intensity 2500lux and light application time are to cultivate 15 days under 14h/ days the condition, obtain group training seedling, and statistics tissue cultivating seedling rooting rate.
Add the 1/2MS medium of α-Nai Yisuan (NAA) and be made up of α-Nai Yisuan and 1/2MS medium, the proportioning of 1/2MS medium and NAA is 1L: 0.05mg.
Tissue cultivating seedling rooting rate (%)=(the stem hop count of taking root/inoculation stem hop count) * 100%
The result who repeats for 3 times is: average group training seedling rooting rate is 89%.
The MS medium is formed, the 1/2MS medium is formed all with identical described in the embodiment 1.
Two, hardening and transplanting
(1) hardening
Step 1 acquisition group training seedling is carried out hardening, and concrete hardening step is as follows: blake bottle (triangular flask) is sealed film open 1/3, took exercise 1 day, then triangular flask is sealed film and all open, took exercise 3 days.This hardening process is to make tissue cultivating seedling progressively adapt to external environment, improves the transplanting survival rate of tissue cultivating seedling.
(2) transplant
1, the tissue cultivating seedling after the hardening is taken out, clean the medium of root remnants, transplant to the nutritive cube that matrix is housed, be in temperature that 20 ℃, ambient humidity are 70%, light intensity 2000lux and light application time be to cultivate 5 days under 14h/ days the condition; Guaranteeing to water less under the moistening prerequisite of matrix but definitely avoiding arid in this process, and forbidding ponding and strong illumination in the nutritive cube.。
2, again temperature be 20 ℃, ambient humidity be 60% and the natural lighting condition under cultivated 15 days, statistics is transplanted survival rate of plant.
Matrix used in the said process is made of turfy soil turfy soil and sandy soil: sandy soil=1: 1.5.
In the said process, hardening is to make plant adapt to external environment gradually, and transplanting in nutritive cube is to make plant obtain adaptive training in culturing room.This hardening and transplanting can improve the transplanting survival rate of tissue cultivating seedling.
Transplant survival rate of plant (%)=(survive plant number/nutritive cube and transplant the plant number) * 100%
The result who repeats for 3 times is: average nutritive cube transplanting survival rate is 76%.
Three, field cultivation
The jerusalem artichoke plant band compost that step 2 obtains is transplanted in big Tanaka, water, shading (taking the sunshade net to cover) 3 days.After the field planting, early stage the duty maintenance ground moistening that waters; Plant survives suitable minimizing the in the back of growing up fully and waters.Statistics is transplanted survival rate of plant (%); Transplant that the October can be gathered in the crops hybrid seed plant stem tuber then.
Wherein, water and suitably 3 days processing of shading can promote surviving of seedling.
Transplant survival rate of plant (%)=(land for growing field crops survives plant number/field-transplanting plant number) * 100%
The result who repeats for 3 times is: average field-transplanting survival rate is 79%.
After experiment finishes, statistics reproduction coefficient and breeding cycle.
Reproduction coefficient: define every hybrid seed by experiment after one, two and three the step, can the surviving and can produce the plant number of stem tuber of acquisition.
Breeding cycle: from seed disinfection to the time of gathering in the crops stem tuber.
The result who repeats for 3 times is: reproduction coefficient is 5, and be 10 months breeding cycle.
The cultivation of embodiment 4, artichoke hybrid seeds
One, tissue culture
1, seed disinfection
Artichoke hybrid seeds is removed the peel, place the mixed solution (volume ratio of washing agent and water is 4: 100) of washing agent and water to soak 2h, flowing water flushing 3h, use the ethanol water of 75% (volumn concentration) to soak 30s then, aseptic water washing 3 times is 5% aqueous sodium hypochlorite solution sterilization 6min with available chlorine content again, aseptic water washing 3 times, use 0.1% (quality percentage composition) mercuric chloride solution sterilization 4min again, aseptic water washing 3 times.
Seed aseptic filter paper suck dry moisture with after the sterilization is inoculated in the MS medium, after 3 days, and the pollution rate of statistics seed.
Pollution rate (%)=(seed number of seed number/sterilization and inoculation occurring polluting) * 100%
The result who repeats for 3 times is: average pollution rate is 3%.
2, obtain complete seedling
Seed aseptic filter paper suck dry moisture with after step 1 sterilization is inoculated in the MS medium, handles 36h for 4 ℃, is cultivation 30 days under the condition of 23 ℃ and lucifuge in temperature then, grow directly from seeds seedling and add up percentage of seedgermination of the artichoke hybrid seeds that obtains; The MS medium is formed as shown in table 1.
Germination rate (%)=(chitting piece number/inoculation seed number) * 100%
The result who repeats for 3 times is: average percentage of seedgermination is 68%.
3, young seedling and propagating
The artichoke hybrid seeds that step 2 the is obtained seedling of growing directly from seeds is cut into the stem section of tool bud, be inoculated in the 1/2MS medium that adds α-Nai Yisuan (NAA), in temperature is that 25 ℃, light intensity 3000lux and light application time are to cultivate 25 days under 14h/ days the condition, obtain group training seedling, and statistics tissue cultivating seedling rooting rate.
Add the 1/2MS medium of α-Nai Yisuan (NAA) and be made up of α-Nai Yisuan and 1/2MS medium, the proportioning of 1/2MS medium and NAA is 1L: 0.15mg.
Tissue cultivating seedling rooting rate (%)=(the stem hop count of taking root/inoculation stem hop count) * 100%
The result who repeats for 3 times is: average group training seedling rooting rate is 80%.
The MS medium is formed, the 1/2MS medium is formed all with identical described in the embodiment 1.
Two, hardening and transplanting
(1) hardening
Step 1 acquisition group training seedling is carried out hardening, and concrete hardening step is as follows: blake bottle (triangular flask) is sealed film open 1/3, took exercise 5 days, then triangular flask is sealed film and all open, took exercise 5 days.This hardening process is to make tissue cultivating seedling progressively adapt to external environment, improves the transplanting survival rate of tissue cultivating seedling.
(2) transplant
1, the tissue cultivating seedling after the hardening is taken out, clean the medium of root remnants, transplant to the nutritive cube that matrix is housed, be in temperature that 25 ℃, ambient humidity are 80%, light intensity 3000lux and light application time be to cultivate 10 days under 14h/ days the condition; Guaranteeing to water less under the moistening prerequisite of matrix but definitely avoiding arid in this process, and forbidding ponding and strong illumination in the nutritive cube.
2, again temperature be 25 ℃, ambient humidity be 75% and the natural lighting condition under cultivated 25 days, statistics is transplanted survival rate of plant.
Matrix used in the said process is made of turfy soil turfy soil and sandy soil: sandy soil=1: 1.5.
In the said process, hardening is to make plant adapt to external environment gradually, and transplanting in nutritive cube is to make plant obtain adaptive training in culturing room.This hardening and transplanting can improve the transplanting survival rate of tissue cultivating seedling.
Transplant survival rate of plant (%)=(transplant survival plant number/nutritive cube is transplanted the plant number) * 100%
The nutritive cube transplanting survival rate is more than 85%;
The result who repeats for 3 times is: average nutritive cube transplanting survival rate is 89%.
Three, field cultivation
The jerusalem artichoke plant band compost that step 2 obtains is transplanted in big Tanaka, waters shading (taking the sunshade net to cover) 5 days.After the field planting, early stage the duty maintenance ground moistening that waters; Plant survives suitable minimizing the in the back of growing up fully and waters.Statistics is transplanted survival rate of plant (%); Transplant that the October can be gathered in the crops a large amount of hybrid seed plant stem tubers then.
Wherein, water and suitably 7 days processing of shading can promote surviving of seedling.
Transplant survival rate of plant (%)=(land for growing field crops survives plant number/field-transplanting plant number) * 100%
The result who repeats for 3 times is: average field-transplanting survival rate is 87%.
After experiment finishes, statistics reproduction coefficient and breeding cycle.
Reproduction coefficient: be defined as every hybrid seed by experiment after one, two and three the step, can the surviving and can produce the plant number of stem tuber of acquisition.
Breeding cycle: from seed disinfection to the time of gathering in the crops stem tuber.
The result who repeats for 3 times is: reproduction coefficient is 5, and be 11 months breeding cycle.
Claims (9)
1. cultivate the method that artichoke hybrid seeds obtains the jerusalem artichoke seedling for one kind, comprise the steps: that the tissue culture artichoke hybrid seeds obtains the jerusalem artichoke seedling.
2. method according to claim 1 is characterized in that: the method for described tissue culture comprises the steps:
1), obtains the artichoke hybrid seeds of sterilization with described artichoke hybrid seeds sterilization;
2) artichoke hybrid seeds with described sterilization is inoculated in the MS medium, handles 12-36h for 4 ℃, cultivates 10-30 days under temperature is the condition of 20 ℃-23 ℃ and lucifuge then, obtains the jerusalem artichoke seedling;
The time of described 4 ℃ of processing is specially 24h, 12h or 36h, and described temperature is specially 22 ℃, 20 ℃ or 23 ℃, and the time of described cultivation is 20 days, 10 days or 30 days.
3. method according to claim 1 and 2, it is characterized in that: in the described method, in described step 2) after, comprise the step that following expansion is numerous: the stem section that described jerusalem artichoke seedling is cut into the tool bud, the stem section of tool bud is transferred in the 1/2MS medium that adds α-Nai Yisuan, in temperature is that 20 ℃-25 ℃, light intensity 2500-3000lux and light application time are to cultivate under 14h/ days the condition, obtains the jerusalem artichoke seedling; Described temperature is specially 23 ℃, 20 ℃ or 25 ℃, and described light intensity is specially 2500lux or 3000lux;
The described 1/2MS medium that adds α-Nai Yisuan of giving birth to is made up of 1/2MS medium and α-Nai Yisuan, and the proportioning of 1/2MS medium and α-Nai Yisuan is 1L: (0.05mg-0.15mg), be specially 1L: 0.1mg, 1L: 0.05mg or 1L: 0.15mg.
4. according to claim 1,2 or 3 described methods, it is characterized in that: described sterilization comprises the steps: described artichoke hybrid seeds peeling, place the mixed solution of washing agent and water to soak 1-2h, flowing water flushing 1-3h, use the ethanol water of 75% (volumn concentration) to soak 10-30s then, aseptic water washing, be 5% aqueous sodium hypochlorite solution sterilization 3-6min with available chlorine content again, aseptic water washing, use 0.1% (quality percentage composition) mercuric chloride solution sterilization 1-4min again, aseptic water washing; The volume ratio of washing agent and water is (0.5-4) in the mixed solution of described washing agent and water: 100;
Described sterilization is specially following 1), 2) or 3) shown in:
1) with described artichoke hybrid seeds peeling, place the mixed solution of washing agent and water to soak 1.5h, flowing water flushing 3h, use the ethanol water of 75% (volumn concentration) to soak 20s then, aseptic water washing is 5% aqueous sodium hypochlorite solution sterilization 4min with available chlorine content again, aseptic water washing, use 0.1% (quality percentage composition) mercuric chloride solution sterilization 2min again, aseptic water washing; The volume ratio of washing agent and water is 2: 100 in the mixed solution of described washing agent and water;
2) with described artichoke hybrid seeds peeling, place the mixed solution of washing agent and water to soak 1h, flowing water flushing 1h, use the ethanol water of 75% (volumn concentration) to soak 10s then, aseptic water washing is 5% aqueous sodium hypochlorite solution sterilization 3min with available chlorine content again, aseptic water washing, use 0.1% (quality percentage composition) mercuric chloride solution sterilization 1min again, aseptic water washing; The volume ratio of washing agent and water is 0.5: 100 in the mixed solution of described washing agent and water;
3) with described artichoke hybrid seeds peeling, place the mixed solution of washing agent and water to soak 2h, flowing water flushing 3h, use the ethanol water of 75% (volumn concentration) to soak 30s then, aseptic water washing is 5% aqueous sodium hypochlorite solution sterilization 6min with available chlorine content again, aseptic water washing, use 0.1% (quality percentage composition) mercuric chloride solution sterilization 4min again, aseptic water washing; The volume ratio of washing agent and water is 4: 100 in the mixed solution of described washing agent and water.
5. according to arbitrary described method among the claim 1-4, it is characterized in that: the constituent of described 1/2MS medium is identical with the MS medium, wherein, the concentration of macroelement in the 1/2MS medium be its in the MS medium concentration 1/2, the concentration of trace element in the 1/2MS medium be its in the MS medium concentration 1/2, the concentration of all the other constituents in the 1/2MS medium is identical with its concentration in the MS medium.
6. cultivate the method that artichoke hybrid seeds obtains the jerusalem artichoke plant for one kind, comprise the steps:
A) cultivate artichoke hybrid seeds according to arbitrary described method among the claim 1-5, obtain the jerusalem artichoke seedling;
B) described jerusalem artichoke seedling is carried out hardening and transplanting, obtain jerusalem artichoke crossbreed plant.
7. method according to claim 6 is characterized in that: in the described method, after described step b), comprise the step of described jerusalem artichoke crossbreed plant being carried out field cultivation.
8. cultivate the method that artichoke hybrid seeds obtains jerusalem artichoke crossbreed stem tuber for one kind, comprise the steps: to obtain jerusalem artichoke crossbreed plant, continue field cultivation to gathering in the crops the jerusalem artichoke stem tuber according to claim 6 or 7 described methods.
9. the application of arbitrary described method in the species of jeruselem artichoke seed selection among the claim 1-8.
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| CN104081999A (en) * | 2014-07-09 | 2014-10-08 | 苏州市美人半岛齐力生态农产品专业合作社 | Jerusalem artichoke planting method |
| CN108633665A (en) * | 2018-05-14 | 2018-10-12 | 廊坊市思科农业技术有限公司 | Selection, implantation methods and the harvesting method of forage type species of jeruselem artichoke |
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| CN101433182A (en) * | 2007-11-16 | 2009-05-20 | 天津市农业生物技术研究中心 | Tissue culture, rapid propagation and transplantation method of American artichoke |
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| CN101433182A (en) * | 2007-11-16 | 2009-05-20 | 天津市农业生物技术研究中心 | Tissue culture, rapid propagation and transplantation method of American artichoke |
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| 《南方农业》 20090331 闫海霞等 菊芋的组织培养与快繁技术研究 58-61 1-9 , 第3期 2 * |
| 《植物生理学通讯》 19971231 王利琳等 菊芋的组织培养及植株再生 358 1-9 , 第5期 2 * |
| 《种子科技》 20081231 申青岭等 菊芋的特征特性及其高产栽培技术 63-64 1-9 , 第3期 2 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104081999A (en) * | 2014-07-09 | 2014-10-08 | 苏州市美人半岛齐力生态农产品专业合作社 | Jerusalem artichoke planting method |
| CN108633665A (en) * | 2018-05-14 | 2018-10-12 | 廊坊市思科农业技术有限公司 | Selection, implantation methods and the harvesting method of forage type species of jeruselem artichoke |
| CN108633665B (en) * | 2018-05-14 | 2020-04-14 | 廊坊市思科农业技术有限公司 | Breeding method, planting method and harvesting method of pasture type jerusalem artichoke variety |
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