CN103621408A - Making method for artificial hybrid liriodendron seeds - Google Patents
Making method for artificial hybrid liriodendron seeds Download PDFInfo
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- CN103621408A CN103621408A CN201310681360.8A CN201310681360A CN103621408A CN 103621408 A CN103621408 A CN 103621408A CN 201310681360 A CN201310681360 A CN 201310681360A CN 103621408 A CN103621408 A CN 103621408A
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Abstract
The invention discloses a making method for artificial hybrid liriodendron seeds. The making method comprises the steps of cultivating hybrid liriodendron cotyledon embryos, making hybrid liriodendron cotyledon embryo capsules, preparing artificial hybrid liriodendron seed coats and the like. The artificial seeds made by the making method can replace natural tree seeds, so that the liriodendron foresting period is shortened; furthermore, the making method cannot be limited by seasons and can be applied to large-scale and large-batch high-efficiency production of the artificial hybrid liriodendron seeds; sufficient seedlings are supplied to large-scale planting of hybrid liriodendrons.
Description
Technical field
The invention belongs to biotechnology and agricultural engineering field, be specifically related to a kind of hybridized Chinese tuliptree artificial seed preparation method.
Background technology
Chinese tulip tree, claims again Liriodendron chinense, Ivy Tree Bark, ninebark, maple He Shu etc., is the plant of Magnoliaceae Liriodendron.This genus is only deposited two kinds now, be distributed in China, north subtropical area from Zhejiang of east, the Chinese tulip tree on ,Zhong Yue border, Yunnan that Fujian extends to the west and south and be distributed in the yellow poplar of eastern united states.Liriodendron plant in classification of angiosperm status in more original position.Although area is wide, is microcommunity more at present and distributes.The Chinese tulip tree being wherein distributed in has within Chinese territory been listed in national secondary Precious, Rare, Endangered protected trees.
Liriodendron Chinense form is perfectly straight, tree-like attractive in appearance; Leaf peculiar, blade is roomy, nick, and the unobstructed general layout in both sides has a sliver, and tip cuts shape, leaf just like as Chinese ancient costume " mandarin jacket ", therefore be commonly called as Liriodendron chinense; Faint yellow large-scale flower is opened on branch in spring top, and its shape is if cup is like bowl, and light is bright pleasing, and ornamental value is high, and fruit is conical, also extremely attractive in appearance.These seeds have stronger resistance to sulphur dioxide and chlorine, are widely used in afforestation.In addition, the light red brown of Liriodendron plant timber, texture is perfectly straight, and color and luster is attractive in appearance, structure is fine and closely woven, and material is light and soft, and planing surface is bright and clean, is suitable for as plywood, paper pulp, furniture and indoor decoration material, also can be for building, shipbuilding etc., economic worth is high, is also important commerical tree species.
As the rare famous and precious yellow poplar of viewing and admiring landscape tree in the world, same tree performance is tall and graceful, leaf beauty, and tree-like rectifying, tree order is neat, spends large and beautiful.Aged branch bar is open and flat, or micro-letting droop.At every florescence, tree pressed in fine jade English, and emerald green curtain covers ground, beautiful gracefulness.But two seeds all exist natural ripening rate only to have 1% left and right, the defect of sexual propagation ability, seedling cannot meet the needs of producing with seedling.
With Liriodendron Chinense and yellow poplar, be that parent carries out artificial hybridization, obtain hybrid, be hybridized Chinese tuliptree.Hybridized Chinese tuliptree, has obvious hybrid vigour, not only tree-like more perfectly straight than parent, and floral leaf is more deep and remote beautiful, and many virtues that match in excellence or beauty, overlook many grass; And growth is also rapider.The air pollution that these seeds are dispersed and cause without other seeds fruit hairs simultaneously.
At present, hybridized Chinese tuliptree nursery stock " supply falls short of demand ".Agricultural University Of Nanjing successfully develops hybridized Chinese tuliptree somatic embryo and occurs and plant regeneration technique.But the enforcement of this technology still can't meet the demand to hybridized Chinese tuliptree nursery stock in current production, therefore need to develop hybridized Chinese tuliptree nursery stock production technology more efficiently.The production of artificial seed, will improve the production efficiency of hybridized Chinese tuliptree nursery stock undoubtedly effectively.
Artificial seed its essence is that it is that somatic embryo or embryoid, indefinite bud, terminal bud and axillalry bud etc. are partly wrapped in the capsule that nutrient can be provided that tissue is cultivated to the bathmic explant of biologically active that has producing; and there is defencive function and the shell that prevents mechanical damage at its additional one deck of wrapping up in; form a kind of structure that is similar to seed, there is the function that natural seed sprouting emerges.Compare natural seed, artificial seed can not be subject to the restriction of season, environment each side natural endowment to carry out extensive in enormous quantities efficient making to produce, and while making the selecting of agricultural production, sowing becomes possibility.Than test-tube plantlet, artificial seed volume is little, and transportation and preservation are convenient, with low cost, have reduced transplanting domestication process.Therefore, develop the manufacturing technology of hybridized Chinese tuliptree artificial seed, will greatly improve the planting scale of hybridized Chinese tuliptree.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of hybridized Chinese tuliptree artificial seed, to improve hybridized Chinese tuliptree seedling production efficiency, meet the growing demand to hybridized Chinese tuliptree nursery stock in current production.
In order to solve above technical problem, the concrete technical scheme that the present invention adopts is as follows:
A preparation method for hybridized Chinese tuliptree artificial seed, is characterized in that comprising the following steps:
Step 1, cultivates hybridized Chinese tuliptree cotyledonary embryos
When hybridized Chinese tuliptree somatic embryo grows to cotyledonary embryos, from medium, take out cotyledonary embryos, aseptic inoculation is cultivated on cotyledonary embryos growth medium; The pH of described cotyledonary embryos growth medium is 5.8, and composition is MS+IAA0.5mg/L+ZT2mg/L+ glutamic acid 400mg/L+3.0% sucrose+0.7% agar; Condition of culture is for take LED ruddiness as light source, at light intensity 35 μ molm
-2s
-1under condition, illumination is 14 hours;
Step 2, makes hybridized Chinese tuliptree cotyledonary embryos capsule
Preparing hybrid Chinese tulip tree artificial endosperm, and carry out sterilization treatment; By aseptic artificial endosperm and CaCl
2solution is placed in respectively different beakers; Under gnotobasis, by described cotyledonary embryos grown cultures basal growth the cotyledonary embryos after 7 days put into described artificial endosperm, in every milliliter of artificial endosperm, contain 1 cotyledonary embryos; Then with the suction pipe of internal diameter 4mm, a cotyledonary embryos is sucked together with artificial endosperm, drop to CaCl
2in solution, every drop volume 0.8ml, solidifies after 10min, forms hybridized Chinese tuliptree cotyledonary embryos capsule;
Described artificial endosperm is the sodium alginate hydrosol, and composition is 4.0% sodium alginate+MS+NAA0.4mg/L+2,4-D0.2mg/L+3.0% sucrose;
Step 3, prepares hybridized Chinese tuliptree Artificial Seed Coats
By sodium alginate and gelatin, according to mass ratio, be that 4:1 mixes that to be made into mass concentration be 5% sodium alginate/glutin Artificial Seed Coats embedding liquid, and through sterilization treatment; By aseptic Artificial Seed Coats embedding liquid and 2.0%CaCl
2solution is placed in respectively different beakers; Take out described hybridized Chinese tuliptree cotyledonary embryos capsule, clean by sterile water wash, put into described Artificial Seed Coats embedding liquid, after one minute, take out, then be put into 2.0%CaCl
2in solution, complexing is solidified 10 minutes, forms artificial seed capsule, is hybridized Chinese tuliptree artificial seed, takes out described hybridized Chinese tuliptree artificial seed, with filter paper, blots liquid on it, stores and waits to sprout.
Described MS is Murashige and Skooge medium, and IAA is heteroauxin, and ZT is zeatin; NAA is α-naphthaleneacetic acid; 2,4-D is 2,4-dichlorphenoxyacetic acid.
The composition of described MS is:
Macroelement, KNO
31900mg/L, NH
4nO
31650mg/L, MgSO
47H
2o370mg/L,, KH
2pO
4170mg/L, CaCl
22H
2o440mg/L;
Trace element, MnSO
44H
2o22.3mg/L, ZnSO
47H
2o8.6mg/L, H
3bO
3,6.2mg/L, KI0.83mg/L, NaMoO
44H
2o0.25mg/L, CuSO
45H
2o0.025mg/L, CoCl
26H
2o0.025mg/L;
Iron salt solutions, FeSO
44H
2o27.8mg/L, Na
2-EDTA37.3mg/L;
Organic principle, glycine 2.0mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride, 0.5mg/L, nicotinic acid 0.5mg/L, inositol 100mg/L.
Tool beneficial effect of the present invention.The present invention replaces natural trees seed with artificial seed, can save a large amount of seeds, shortens Chinese tulip tree and grows into forest the cycle; The present invention can not be subject to seasonal restrictions, and extensive in enormous quantities High-efficient Production hybridized Chinese tuliptree artificial seed, for the implant mass of hybridized Chinese tuliptree provides sufficient seedling; Material source of the present invention is easy, operates simple and easy; The present invention compares with test-tube plantlet, and volume is little, and transportation and preservation are convenient, with low cost, reduce rooting culture process.
Embodiment
Below in conjunction with specific embodiment, technical scheme of the present invention is described in further details.
This enforcement is just made hybridized Chinese tuliptree artificial seed according to following process.
1. the cultivation of hybridized Chinese tuliptree cotyledonary embryos
When hybridized Chinese tuliptree somatic embryo grows to cotyledonary embryos, from medium, choose cotyledonary embryos, aseptic inoculation is cultivated on cotyledonary embryos growth medium.The composition of cotyledonary embryos growth medium is: MS+IAA0.5mg/L+ZT2mg/L+ glutamic acid 400mg/L+3.0% sucrose+0.7% agar.MS is Murashige and Skooge medium, and IAA is heteroauxin, and ZT is zeatin.Condition of culture: illumination 14 hours, light intensity 35 μ molm
-2s
-1.Light source is LED ruddiness.The pH5.8 of medium.
2. the making of hybridized Chinese tuliptree cotyledonary embryos capsule
Preparing hybrid Chinese tulip tree artificial endosperm, sterilizing.By aseptic artificial endosperm and CaCl
2solution is placed in respectively different beakers.Under gnotobasis, by cotyledonary embryos grown cultures basal growth the cotyledonary embryos after 7 days put into artificial endosperm, 1 cotyledonary embryos of every ml artificial endosperm, sucks a cotyledonary embryos with the suction pipe of internal diameter 4mm together with artificial endosperm, drop to CaCl
2in solution, every drop volume 0.8ml left and right, solidifies after 10min, forms hybridized Chinese tuliptree cotyledonary embryos capsule.The artificial endosperm is here actually the sodium alginate hydrosol, and its composition is: 4.0% sodium alginate+MS+NAA0.4mg/L+2,4-D0.2mg/L+3.0% sucrose.Here MS is Murashige and Skooge medium, and NAA is α-naphthaleneacetic acid, and 2,4-D is 2,4-dichlorphenoxyacetic acid.
3. the preparation of hybridized Chinese tuliptree Artificial Seed Coats
Sodium alginate is mixed and is made into the Artificial Seed Coats embedding liquid that mass concentration is 5% sodium alginate/glutin according to mass ratio 4:1 with gelatin, carry out sterilizing.By aseptic Artificial Seed Coats embedding liquid and 2.0%CaCl
2solution is placed in respectively different beakers.By described hybridized Chinese tuliptree cotyledonary embryos capsule, take out totally by sterile water wash, put into described Artificial Seed Coats embedding liquid, after one minute, take out, be put into 2.0%CaCl
2in solution, complexing is solidified 10 minutes, forms artificial seed capsule, is hybridized Chinese tuliptree artificial seed, takes out hybridized Chinese tuliptree artificial seed, with filter paper, blots liquid on it, stores and waits to sprout.
MS composition is:
Macroelement: KNO
31900mg/L, NH
4nO
31650mg/L, MgSO
47H
2o370mg/L,, KH
2pO
4170mg/L, CaCl
22H
2o440mg/L;
Trace element: MnSO
44H
2o22.3mg/L, ZnSO
47H
2o8.6mg/L, H
3bO
3,6.2mg/L, KI0.83mg/L, NaMoO
44H
2o0.25mg/L, CuSO
45H
2o0.025mg/L, CoCl
26H
2o0.025mg/L;
Iron salt solutions: FeSO
44H
2o27.8mg/L, Na
2-EDTA37.3mg/L;
Organic principle: glycine 2.0mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride, 0.5mg/L, nicotinic acid 0.5mg/L, inositol 100mg/L.
Implementation result experiment contrast
For implementation result of the present invention is described, proportioning to capsulating material in Artificial Seed Coats embedding liquid is tested, sodium alginate is mixed with three kinds of colloidal substances of gelatin, shitosan, tracasol of the conventional capsulating material of different proportion respectively, being made into mass concentration is the hydrosol of 5% sodium alginate/conventional capsulating material, also be Artificial Seed Coats embedding liquid, according to the step described in summary of the invention, make the sub-artificial seed of hybridized Chinese tuliptree, the sprouting situation of observing the sub-artificial seed of hybridized Chinese tuliptree, result is as shown in table 1.
Coated hybridized Chinese tuliptree of Artificial Seed Coats material of the colloidal substance mixed preparing of table 1. sodium alginate and different proportion
The germination rate of artificial seed
As can be seen from Table 1, although make Artificial Seed Coats material with sodium alginate separately, the germination rate of artificial seed can reach 75%, sodium alginate/glutin 4:1 combination matching better effects if, the germination rate of artificial seed can reach 81%, so the present invention selects this combination, also illustrate that the sub-artificial seed germination rate of hybridized Chinese tuliptree of made of the present invention is higher, be greater than significantly the planting percent of test-tube seedling transplanting, simultaneously, save incubation time, reduced rooting culture process, improved productivity ratio.
Claims (3)
1. a preparation method for hybridized Chinese tuliptree artificial seed, is characterized in that comprising the following steps:
Step 1, cultivates hybridized Chinese tuliptree cotyledonary embryos
When hybridized Chinese tuliptree somatic embryo grows to cotyledonary embryos, from medium, take out cotyledonary embryos, aseptic inoculation is cultivated on cotyledonary embryos growth medium; The pH of described cotyledonary embryos growth medium is 5.8, and composition is MS+IAA 0.5mg/L+ ZT 2mg/L+ glutamic acid 400 mg/L+3.0% sucrose+0.7% agar; Condition of culture is for take LED ruddiness as light source, at light intensity 35 μ molm
-2s
-1under condition, illumination is 14 hours;
Step 2, makes hybridized Chinese tuliptree cotyledonary embryos capsule
Preparing hybrid Chinese tulip tree artificial endosperm, and carry out sterilization treatment; By aseptic artificial endosperm and CaCl
2solution is placed in respectively different beakers; Under gnotobasis, by described cotyledonary embryos grown cultures basal growth the cotyledonary embryos after 7 days put into described artificial endosperm, in every milliliter of artificial endosperm, contain 1 cotyledonary embryos; Then with the suction pipe of internal diameter 4mm, a cotyledonary embryos is sucked together with artificial endosperm, drop to CaCl
2in solution, every drop volume 0.8ml, solidifies after 10 min, forms hybridized Chinese tuliptree cotyledonary embryos capsule;
Described artificial endosperm is the sodium alginate hydrosol, and composition is 4.0% sodium alginate+MS+NAA0.4mg/L+2,4-D 0.2 mg/L+3.0% sucrose;
Step 3, prepares hybridized Chinese tuliptree Artificial Seed Coats
By sodium alginate and gelatin, according to mass ratio, be that 4:1 mixes that to be made into mass concentration be 5% sodium alginate/glutin Artificial Seed Coats embedding liquid, and through sterilization treatment; By aseptic Artificial Seed Coats embedding liquid and 2.0%CaCl
2solution is placed in respectively different beakers; Take out described hybridized Chinese tuliptree cotyledonary embryos capsule, clean by sterile water wash, put into described Artificial Seed Coats embedding liquid, after one minute, take out, then be put into 2.0% CaCl
2in solution, complexing is solidified 10 minutes, forms artificial seed capsule, is hybridized Chinese tuliptree artificial seed, takes out described hybridized Chinese tuliptree artificial seed, with filter paper, blots liquid on it, stores and waits to sprout.
2. the preparation method of a kind of hybridized Chinese tuliptree artificial seed according to claim 1, is characterized in that: described MS is Murashige and Skooge medium, and IAA is heteroauxin, and ZT is zeatin; NAA is α-naphthaleneacetic acid; 2,4-D is 2,4-dichlorphenoxyacetic acid.
3. the preparation method of a kind of hybridized Chinese tuliptree artificial seed according to claim 1, is characterized in that the composition of described MS is:
Macroelement, KNO
31900mg/L, NH
4nO
31650mg/L, MgSO
47H
2o 370mg/L,, KH
2pO
4170mg/L, CaCl
22H
2o 440mg/L;
Trace element, MnSO
44H
2o 22.3mg/L, ZnSO
47H
2o 8.6mg/L, H
3bO
3,6.2mg/L, KI 0.83mg/L, NaMoO
44H
2o 0.25mg/L, CuSO
45H
2o 0.025mg/L, CoCl
26H
2o 0.025mg/L;
Iron salt solutions, FeSO
44H
2o 27.8mg/L, Na
2-EDTA 37.3mg/L;
Organic principle, glycine 2.0mg/L, thiamine hydrochloride 0.1mg/L, puridoxine hydrochloride, 0.5mg/L, nicotinic acid 0.5mg/L, inositol 100mg/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104982336A (en) * | 2015-07-28 | 2015-10-21 | 江苏大学 | Manufacturing method for artificial atractylis lancea seed |
| CN107079626A (en) * | 2017-06-15 | 2017-08-22 | 湖南农业大学 | It is a kind of by plant three-coloured amaranth reduce soil heavy metal cadmium, lead contamination method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104982336A (en) * | 2015-07-28 | 2015-10-21 | 江苏大学 | Manufacturing method for artificial atractylis lancea seed |
| CN107079626A (en) * | 2017-06-15 | 2017-08-22 | 湖南农业大学 | It is a kind of by plant three-coloured amaranth reduce soil heavy metal cadmium, lead contamination method |
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|---|---|
| CN103621408B (en) | 2015-04-22 |
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Granted publication date: 20150422 Termination date: 20181216 |