CN103636502B - A kind of promote lanzhou lily group train bulb expand grow method - Google Patents
A kind of promote lanzhou lily group train bulb expand grow method Download PDFInfo
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Abstract
The present invention relates to a kind of promote lanzhou lily group train bulb expand grow method, belong to plant seedling and breed field, specifically relate to a kind of method promoting to induce in lanzhou lily tissue cultures bulb development.The present invention is by improving lanzhou lily group culturation rapid propagating technology, i.e. the induction of clump Shoot propagation, clove, the expanding of clove, expanding of bulb to impel its bulb to expand development growth with the formation four-stage of strong sprout and whole plant.Operating process and culture test tube seedling similar, cultivate rear acquisition to have and expand the whole plant of bulb, the breeding starting material that can be used as production directly moves into field planting, increase substantially reproduction coefficient, shorten the growth cycle of lanzhou lily, be conducive to extensive plantation and the popularization of lanzhou lily, there is good economic benefit and social benefit.
Description
Technical field
The invention belongs to field of plant cultivation, is specifically a kind of method promoting to induce in lanzhou lily tissue cultures bulb development.
Background technology
Lanzhou lily belongs to herbaceos perennial, because its underground bulb is stacked obvolvent by tens of lobe scale, has the meaning of hundred synthesis and gains the name.Lanzhou lily is one of national four giant lily strains, is also the sweet edible lily of unique taste, as edible lily simultaneously, its local flavor is sweet and refreshing, without bitter taste, be referred to as " sweet lily ", and its scale inclusion is tight, color and luster is pure white, meat is plump, has very high edible, medication and ornamental value.
Lanzhou lily growth cycle is very long, and in Production of Large Fields, the clove sprouted by scale grows to commodity kind ball, needs for six years.Lily mainly carries out division propagation by clove, and a strain lily can only obtain 1 ~ 3 clove every year, and clove is through naturally developing into complete plant, and the general time needing 3 years, reproduction speed is very limited, and easily causes bulb to degenerate.
Currently report that its common ground is as follows about lanzhou lily tissue culture technique: explant many employings scale, main cultivation stage has Multiplying culture, bulb induce to cultivate and culture of rootage; 6-benzyl aminoadenine (6-BA), methyl α-naphthyl acetate (NAA) growth regulator is added based on basal medium MS in Multiplying culture, bulb induce adds 6-BA, NAA, IBA, KT based on basal medium MS in cultivating, and does not carry out the cultivation of expanding of bulb; The article on " gardens gardening " such as Wang Liyan " lanzhou lily and david lily test-tube plantlet tie bulb research ", reports by filling a prescription as the medium of 1/2MS+0.5mg/L IBA in vitro ties the method that Bulb culture can form the lanzhou lily bulb of diameter 1cm.Current, research lanzhou lily bulb expands the study hotspot budding into ectogenesis starting material, carries out the method for production application, have not been reported at set of applications training bulb as bulb reproduction starting material.
Patent CN03135142.5 discloses a kind of method for quickly breeding of lanzhou lily, concrete preparation method is: choose lanzhou lily scale, the segment of leaf and root, after adopting explant sterilization, access is containing the medium of different plant hormone, induction produces bud, root media is moved into after squamous subculture, be transplanted into plantation when test tube height of seedling is about 3-5 centimetre and practice seedling matrix, the induction of scale indefinite bud and to grow medium be soon MS, BA2mg/L, NAA0.2mg/L, root, leaf induction and propagating culture medium are MS, BA2mg/L, NAA0.4mg/L, rooting of vitro seedling medium is 1/2MS, NAA0.3mg/L, temperature 27 ± 2 DEG C, intensity of illumination 2000LX, light application time 12h/d, pH is 5.8.This invention lanzhou lily scale lining can be made to develop into lily seedling that bulb is 2 ~ 4mm, but bulb is less, directly cannot enter field planting.
Usually so that just generation induction, squamous subculture, culture of rootage obtain plantlet in vitro in lanzhou lily tissue culturing system, the plantlet in vitro obtained can be transplanted to grown in field, be difficult to develop into the breeding starting material that can produce, main cause is that plantlet in vitro does not possess bulb or bulb is too small and weak, so in lanzhou lily group training system, the growth of research bulb enlarges into the main focus in order to cultivate kind of ball.
Summary of the invention
Under above-mentioned background, the present invention, by improving lanzhou lily group culturation rapid propagating technology, impels its bulb to expand development growth.Operating process and culture test tube seedling similar, cultivate rear acquisition to have and expand the whole plant of bulb, the breeding starting material that can be used as production directly moves into field planting, present invention reduces the growth cycle of lanzhou lily, be conducive to lanzhou lily extensively plant and promote, there is good economic benefit and social benefit.
The invention provides and a kind ofly promote that lanzhou lily group is trained bulb and expanded the method for growing, operation is comparatively simple, to environment and equipment requirement low, specific implementation method is as follows:
1, obtain lanzhou lily explant: select the lanzhou lily kind ball taken when mid-October, peel off injured scale, select the scale outer, middle level is clean, for subsequent use with drying after tap water.
2, sterilization inoculation: scale step 1 dried is put on superclean bench, used for 75% alcohol immersion 20 ~ 30 second, soak 10 ~ 12 minutes with the mercuric chloride solution of 0.1% again, then use aseptic water washing 3 times, the scale rinsed well is placed on superclean bench and dries.
3, lanzhou lily tissue cultures induction bulb expands growths, and the technology path adopting for four stages cultivated is induced bulb to be formed and grown and expands growth:
1. first stage clump Shoot propagation: scale step 2 dried is inoculated into F1 medium and cultivates, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000 ~ 4000Lux, photoperiod is 9h Light/15h Dark, described F1 medium comprises MS medium, 0.2 ~ 0.4mg/L 6-benzyl aminoadenine, 0.04 ~ 0.06mg/L methyl α-naphthyl acetate, 20 ~ 40g/L sucrose and 4.5g/L agar, and the cultivation of this stage carries out more than 2 times;
2. the induction of second stage clove: by the clump bud seedling cutting removing blade of step 3 the 1. step gained, the sprout of remainder is transferred to F2 medium, with medium F2 for bulb induce Development culture base carries out induced synthesis and the propagation of bulb, condition of culture is: cultivation cycle is 30 ~ 40 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000 ~ 4000Lux, and the photoperiod is 9h Light/15h Dark, and described F2 medium comprises MS medium, 80 ~ 100g/L sucrose and 4.5g/L agar;
3. the expanding of phase III clove: the differentiation bulb of step 3 2. step gained is cut down, and excise blade, be transplanted to F3 medium, the growth of expanding of clove is carried out for bulb expands Development culture base with medium F3, condition of culture is: cultivation cycle is 30 ~ 40 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000 ~ 4000Lux, photoperiod is 9h Light/15h Dark, described F3 medium comprises 1.5 times of MS medium, 0.12 ~ 0.18mg/L 6-benzyl aminoadenine, 0.12 ~ 0.18mg/L gibberellin, 60 ~ 80g/L sucrose and 4.5g/L agar,
4. the formation of expanding with strong sprout and whole plant of fourth stage bulb: the bulb that expands of step 3 3. step gained is cut down, and excise blade, moved to F4 medium, grow and the medium of strong plantlets and rootage for bulb expands further with medium F4, that carries out bulb expands growth and the formation of whole plant, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000 ~ 4000Lux, photoperiod is 9h Light/15h Dark, described F4 medium comprises MS, 0.8 ~ 1.2mg/L paclobutrazol, 80 ~ 100g/L sucrose and 4.5g/L agar.
Embodiment
Embodiment 1
1, obtain lanzhou lily explant: select the lanzhou lily kind ball taken when mid-October, peel off injured scale, select the scale outer, middle level is clean, for subsequent use with drying after tap water.
2, sterilization inoculation: scale step 1 dried is put on superclean bench, used for 75% alcohol immersion 20 second, then soaks 10 minutes with the mercuric chloride solution of 0.1%, then use aseptic water washing 3 times, be placed on superclean bench by the scale rinsed well and dry.
3, the scale that step 2 is dried is inoculated into the F1 medium be made up of MS, 0.3mg/ L 6-benzyl aminopurine, 0.05 mg/L methyl α-naphthyl acetate, 30g/L sucrose and 4.5g/L agar to cultivate, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3000Lux, and the photoperiod is 9h Light/15h Dark.
This stage cultivates 3 times.
The lanzhou lily bulb cultivated is 2 ~ 4mm, directly cannot be placed in field planting.
Embodiment 2
1, obtain lanzhou lily explant: select the lanzhou lily kind ball taken when mid-October, peel off injured scale, select the scale outer, middle level is clean, for subsequent use with drying after tap water.
2, sterilization inoculation: scale step 1 dried is put on superclean bench, used for 75% alcohol immersion 20 second, then soaks 10 minutes with the mercuric chloride solution of 0.1%, then use aseptic water washing 3 times, be placed on superclean bench by the scale rinsed well and dry.
3, lanzhou lily tissue cultures induction bulb expands growths, and the technology path adopting for four stages cultivated is induced bulb to be formed and grown and expands growth:
1. first stage clump Shoot propagation: scale step 2 dried is inoculated into the F1 medium be made up of MS, 0.25mg/ L 6-benzyl aminoadenine, 0.04 mg/L methyl α-naphthyl acetate, 30g/L sucrose and 4.5g/L agar and cultivates, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000Lux, photoperiod is 9h Light/15h Dark, and this stage cultivates 2 times;
2. the induction of second stage clove: by the clump bud seedling cutting removing blade of step 3 the 1. step gained, the sprout of remainder is transferred to the F2 medium be made up of MS, 80g/L sucrose and 4.5g/L agar, with medium F2 for bulb induce Development culture base carries out induced synthesis and the propagation of bulb, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000Lux, and the photoperiod is 9h Light/15h Dark;
3. the expanding of phase III clove: the differentiation bulb of step 3 2. step gained is cut down, and excise blade, be transplanted to the F3 medium be made up of 1.5 times of MS, 0.13mg/ L 6-benzyl aminoadenines, 0.12 mg/L gibberellin, 60g/L sucrose and 4.5g/L agar, the growth of expanding of clove is carried out for bulb expands Development culture base with medium F3, condition of culture is: cultivation cycle is 35 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000Lux, and the photoperiod is 9h Light/15h Dark;
4. the formation of expanding with strong sprout and whole plant of fourth stage bulb: the bulb that expands of step 3 3. step gained is cut down, and excise blade, moved to the F4 medium be made up of MS, 0.9mg/L paclobutrazol, 85g/L sucrose and 4.5g/L agar, grow and the medium of strong plantlets and rootage for bulb expands further with medium F4, that carries out bulb expands growth and the formation of whole plant, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000Lux, and the photoperiod is 9h Light/15h Dark.
After cultivation by four stages, the lanzhou lily bulb parameter of acquisition is as follows:
| To take root quantity | Root length cm | Enclose stem cm | Diameter cm | Bulb weight g |
| 5.26±0.39 | 1.27±0.25 | 3.73±0.41 | 0.85±0.26 | 1.07±0.21 |
The lanzhou lily bulb cultivated through four stages directly can carry out production application as bulb reproduction starting material.
Embodiment 3
1, obtain lanzhou lily explant: select the lanzhou lily kind ball taken when mid-October, peel off injured scale, select the scale outer, middle level is clean, for subsequent use with drying after tap water.
2, sterilization inoculation: scale step 1 dried is put on superclean bench, used for 75% alcohol immersion 20 second, then soaks 10 minutes with the mercuric chloride solution of 0.1%, then use aseptic water washing 3 times, be placed on superclean bench by the scale rinsed well and dry.
3, lanzhou lily tissue cultures induction bulb expands growths, and the technology path adopting for four stages cultivated is induced bulb to be formed and grown and expands growth:
1. first stage clump Shoot propagation: scale step 2 dried is inoculated into the F1 medium be made up of MS, 0.3mg/ L 6-benzyl aminopurine, 0.05 mg/L methyl α-naphthyl acetate, 30g/L sucrose and 4.5g/L agar and cultivates, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3500Lux, and the photoperiod is cultivate in this stage of 9h Light/15h Dark to carry out 3 times;
2. the induction of second stage clove: by the clump bud seedling cutting removing blade of step 3 the 1. step gained, the sprout of remainder is transferred to the F2 medium be made up of MS, 95g/L sucrose and 4.5g/L agar, with medium F2 for bulb induce Development culture base carries out induced synthesis and the propagation of bulb, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3500Lux, and the photoperiod is 9h Light/15h Dark;
3. the expanding of phase III clove: the differentiation bulb of step 3 2. step gained is cut down, and excise blade, be transplanted to the F3 medium be made up of 1.5 times of MS, 0.17mg/L 6-benzyl aminopurines, 0.18mg/L gibberellin, 80g/L sucrose and 4.5g/L agar, the growth of expanding of clove is carried out for bulb expands Development culture base with medium F3, condition of culture is: cultivation cycle is 40 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3500Lux, and the photoperiod is 9h Light/15h Dark;
4. the formation of expanding with strong sprout and whole plant of fourth stage bulb: the bulb that expands of step 3 3. step gained is cut down, and excise blade, moved to the F4 medium be made up of MS, 1.1mg/L paclobutrazol, 95g/L sucrose and 4.5g/L agar, grow and the medium of strong plantlets and rootage for bulb expands further with medium F4, that carries out bulb expands growth and the formation of whole plant, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3500Lux, and the photoperiod is 9h Light/15h Dark.
After cultivation by four stages, the lanzhou lily bulb parameter of acquisition is as follows:
| To take root quantity | Root length cm | Enclose stem cm | Diameter cm | Bulb weight g |
| 6.03±0.54 | 1.76±0.25 | 4.07±0.33 | 1.05±0.19 | 1.28±0.27 |
The lanzhou lily bulb cultivated through four stages directly can carry out production application as bulb reproduction starting material.
Embodiment 4
1, obtain lanzhou lily explant: select the lanzhou lily kind ball taken when mid-October, peel off injured scale, select the scale outer, middle level is clean, for subsequent use with drying after tap water.
2, sterilization inoculation: scale step 1 dried is put on superclean bench, used for 75% alcohol immersion 20 second, then soaks 10 minutes with the mercuric chloride solution of 0.1%, then use aseptic water washing 3 times, be placed on superclean bench by the scale rinsed well and dry.
3, lanzhou lily tissue cultures induction bulb expands growths, and the technology path adopting for four stages cultivated is induced bulb to be formed and grown and expands growth:
1. first stage clump Shoot propagation: scale step 2 dried is inoculated into the F1 medium be made up of MS, 0.3mg/L 6-benzyl aminopurine, 0.05mg/L methyl α-naphthyl acetate, 30g/L sucrose and 4.5g/L agar and cultivates, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3000Lux, photoperiod is 9h Light/15h Dark, and this stage cultivates 3 times;
2. the induction of second stage clove: by the clump bud seedling cutting removing blade of step 3 the 1. step gained, the sprout of remainder is transferred to the F2 medium be made up of MS, 100g/L sucrose and 4.5g/L agar, with medium F2 for bulb induce Development culture base carries out induced synthesis and the propagation of bulb, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3000Lux, and the photoperiod is 9h Light/15h Dark;
3. the expanding of phase III clove: the differentiation bulb of step 3 2. step gained is cut down, and excise blade, be transplanted to the F3 medium be made up of 1.5 times of MS, 0.15mg/L 6-benzyl aminopurines, 0.15mg/L gibberellin, 75g/L sucrose and 4.5g/L agar, the growth of expanding of clove is carried out for bulb expands Development culture base with medium F3, condition of culture is: cultivation cycle is 40 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3000Lux, and the photoperiod is 9h Light/15h Dark;
4. the formation of expanding with strong sprout and whole plant of fourth stage bulb: the bulb that expands of step 3 3. step gained is cut down, and excise blade, moved to the F4 medium be made up of MS, 1.0 mg/L paclobutrazols, 80g/L sucrose and 4.5g/L agar, grow and the medium of strong plantlets and rootage for bulb expands further with medium F4, that carries out bulb expands growth and the formation of whole plant, condition of culture is: cultivation cycle is 30 days, cultivation temperature is 25 DEG C, cultivation intensity of illumination is 3000Lux, and the photoperiod is 9h Light/15h Dark.
After cultivation by four stages, the lanzhou lily bulb parameter of acquisition is as follows:
| To take root quantity | Root length cm | Enclose stem cm | Diameter cm | Bulb weight g |
| 5.92±0.47 | 1.63±0.21 | 4.27±0.47 | 1.33±0.22 | 1.43±0.32 |
The lanzhou lily bulb cultivated through four stages directly can carry out production application as bulb reproduction starting material.
Claims (1)
1. the technology path induction lanzhou lily bulb adopting for four stages cultivated expands a method of growing, and it is characterized in that: comprise following four stages:
1. first stage clump Shoot propagation: peel off injured scale, select the scale outer, middle level is clean, after drying with tap water, use 75% alcohol immersion to sterilize for 20 ~ 30 seconds, then soak 10 ~ 12 minutes with the mercuric chloride solution of 0.1%, then dry with after aseptic water washing 3 times, the above-mentioned scale dried is inoculated into F1 medium cultivate, condition of culture is: cultivation cycle is 30 days, and cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000 ~ 4000Lux, and the photoperiod is 9h Light/15h Dark;
2. the induction of second stage clove: by the clump bud seedling of first stage gained cutting removing blade, the sprout of remainder is transferred to F2 medium, with medium F2 for bulb induce Development culture base, carry out induced synthesis and the propagation of bulb, condition of culture is: cultivation cycle is 30 ~ 40 days, cultivation temperature is 25 DEG C, and cultivation intensity of illumination is 2000 ~ 4000Lux, and the photoperiod is 9h Light/15h Dark;
3. the expanding of phase III clove: the differentiation bulb of second stage gained is cut down, and excise blade, be transplanted to F3 medium, with medium F3 for bulb expands Development culture base, carry out the growth of expanding of clove, condition of culture is: cultivation cycle is 30 ~ 40 days, and cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000 ~ 4000Lux, and the photoperiod is 9h Light/15h Dark;
4. the formation of expanding with strong sprout and whole plant of fourth stage bulb: the bulb that expands of phase III gained is cut down, and excise blade, moved to F4 medium, grow and Rooting and hardening-off culture base for bulb expands further with medium F4, that carries out bulb expands growth and the formation of whole plant, and condition of culture is: cultivation cycle is 30 days, and cultivation temperature is 25 DEG C, cultivation intensity of illumination is 2000 ~ 4000Lux, and the photoperiod is 9h Light/15h Dark;
In the described first stage, F1 culture medium prescription is made up of MS medium, 0.3mg/L 6-benzyl aminopurine, 0.05mg/L methyl α-naphthyl acetate, 30g/L sucrose and 4.5g/L agar;
In described second stage, F2 culture medium prescription is made up of MS medium, 100g/L sucrose and 4.5g/L agar;
In the described phase III, F3 culture medium prescription is made up of 1.5 times of MS medium, 0.15mg/L 6-benzyl aminopurine, 0.15mg/L gibberellin, 75g/L sucrose and 4.5g/L agar;
In described fourth stage, F4 culture medium prescription is made up of MS, 1.0mg/L paclobutrazol, 80g/L sucrose and 4.5g/L agar.
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| CN113841613B (en) * | 2021-10-26 | 2022-07-15 | 甘肃省农业科学院生物技术研究所 | Efficient breeding method for lilium davidii germchit |
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