CN113207689A - Rapid propagation method for directly inducing bulblet by lily stem segment - Google Patents
Rapid propagation method for directly inducing bulblet by lily stem segment Download PDFInfo
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- CN113207689A CN113207689A CN202110559352.0A CN202110559352A CN113207689A CN 113207689 A CN113207689 A CN 113207689A CN 202110559352 A CN202110559352 A CN 202110559352A CN 113207689 A CN113207689 A CN 113207689A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a rapid propagation method for directly inducing bulblets by lily stem segments, which is characterized in that after lily flowers fall, tender stems at the middle upper part of lily are cut, the stems are washed clean by running water, 2/3 leaves are cut off by a double-sided blade, and the stems with 1/3 leaves are put into a wide-mouth bottle. The invention provides a rapid propagation method for directly inducing bulblets by lily stem segments, which has the advantages of early induction starting, obviously shortened cultivation time, 4d of beginning protrusion of axilla of a leaf, cultivation of the emergence of a seed bulb primordium at the axilla of one week of the leaf, cultivation of seed bulbs formed at the axilla of two weeks of the leaf, cultivation for one month, realization of induction and expansion cultivation of the seed bulbs by using a culture medium MS +2mg/L6-BA +90g/L sucrose, simple cultivation process and pollution reduction, wherein the induction starting time of the seed bulbs can reach the bulb specification, namely the diameter reaches 1-2 cm.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a rapid propagation method for directly inducing bulblets by lily stem segments.
Background
Lily is perennial bulb flower of lily of Liliaceae, has various flower types and colors, has high ornamental, edible and medicinal values, and is known as the king of bulbous flower. For a long time, the lily is propagated by the traditional bulb division, scale cuttage and overground bulblet, the problems of low propagation coefficient, virus accumulation, seed nature degradation and the like exist, the yield and the quality of the lily are seriously influenced, and the large-scale production of the lily bulb and cut flowers is restricted.
Plant tissue culture technology is one of the important research techniques and means in life science. By using the tissue culture technology, the propagation speed of the lily can be accelerated, the growth cycle of the lily bulbs can be shortened, and the defects of bulb breeding can be overcome, so that the tissue culture technology is widely applied to the commercialization and industrialization process of the lily. Tissue culture of lily is usually achieved through 5 routes. The first is organotypic, i.e. the process of forming clumpy buds through axillary bud development and the generation of adventitious buds; secondly, organogenesis, namely a mode of differentiating explants to form callus and differentiating organs; thirdly, the embryogenesis type, namely the process that the explant forms embryoid according to the embryogenesis mode or forms callus at first and then forms an embryoid regeneration plant through the callus; the fourth is a bulblet type, namely, the squamae forms a bulblet with roots on the paraxial surface or the edge directly; fifth, sporozoite type, i.e., culture with mature or immature spores. At present, the lily is usually propagated by taking scales as explants and inducing bulb-type formation of seed bulbs through organogenesis and scales.
In addition, the tissue culture propagation can also be carried out through seeds, bulbels, leaves, flower organs and the like, but the problems of long period and low induction rate exist, and the production requirement of the lilies cannot be met. Therefore, a rapid propagation method for directly inducing bulblet by lily stem segments is provided.
Disclosure of Invention
Based on the technical problems in the background art, the invention provides a rapid propagation method for directly inducing bulblets by lily stem segments, so as to solve the problems in the background art.
The invention provides the following technical scheme: a rapid propagation method for directly inducing bulblets by lily stem segments comprises the following steps:
A. obtaining an explant: cutting the tender stem at the middle upper part of the lily after the lily flowers fall, washing the cut stem clean with running water, cutting 2/3 leaves with a double-sided blade, and putting the stem with 1/3 leaves into a wide-mouth bottle;
B. surface sterilization: placing the wide-mouth bottle on a superclean bench for surface sterilization: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, and rinsing with sterile water;
C. seed ball induction and expansion culture: the sterilized stem is cut into single stem nodes with 1cm of leaf stalks and the length of which is 1.5cm, and the single stem nodes are inoculated on a culture medium which is added with 6-benzylamino adenine (6-BA), naphthylacetic acid (NAA) and cane sugar with different concentrations in an MS basic culture medium, and are induced and expanded to culture under the induction conditions of 25 +/-1 ℃, the illumination intensity of 2000lx and the illumination period of 16 hours/day.
Preferably, in the step a, the lily can be selected from one of ornamental lily 'Sobang', edible and medicinal lily pellet and iron-baked lily.
Preferably, the step B is performed by rinsing with sterile water for 4-6 times, and each time is 3 min.
Preferably, the bulbs can be produced after one month of culture in the step C, and the diameter of the bulbs is 1-2 cm.
The invention provides a rapid propagation method for directly inducing bulblets by lily stem segments, which has the advantages that the induction start is early, the cultivation time is obviously shortened, 4d leaf axils are cultured to start to bulge, seed sphere primordium appears at one week leaf axils, seed spheres are formed at two weeks leaf axils, the seed spheres can reach the sphere output specification after being cultured for one month, the diameter reaches 1-2cm, compared with the traditional scale propagation, the induction time of the seed spheres can be shortened by one month, the sphere forming process only needs one step, namely, the induction and the expansion culture of the seed spheres can be realized by using a culture medium MS +2mg/L6-BA +90g/L cane sugar, the culture process is simple, the pollution chance is reduced, the ornamental value is not influenced, and the digging damage to the seed spheres in the resource collection process is avoided.
Drawings
FIG. 1 is a schematic diagram of the steps of the present invention;
FIG. 2 is a diagram of the induction seed sphere of the 'Sobang' lily stem section of the present invention;
FIG. 3 is a diagram of the shape of the ball of the Dandelion stem induced seed of the present invention;
FIG. 4 is a diagram of the induction seed sphere of Lilium Candidum of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-4, the present invention provides a technical solution: a rapid propagation method for directly inducing bulblets by lily stem segments comprises the following steps:
A. obtaining an explant: cutting the tender stem at the middle upper part of the lily after the lily flowers fall, washing the cut stem clean with running water, cutting 2/3 leaves with a double-sided blade, and putting the stem with 1/3 leaves into a wide-mouth bottle;
B. surface sterilization: placing the wide-mouth bottle on a superclean bench for surface sterilization: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, and rinsing with sterile water;
C. seed ball induction and expansion culture: the sterilized stem is cut into single stem nodes with 1cm of leaf stalks and the length of which is 1.5cm, and the single stem nodes are inoculated on a culture medium which is added with 6-benzylamino adenine (6-BA), naphthylacetic acid (NAA) and cane sugar with different concentrations in an MS basic culture medium, and are induced and expanded to culture under the induction conditions of 25 +/-1 ℃, the illumination intensity of 2000lx and the illumination period of 16 hours/day.
Example one
Cutting the tender stem at the middle upper part of the lily after the lily flowers fall, washing the cut stem clean with running water, cutting 2/3 leaves with a double-sided blade, and putting the stem with 1/3 leaves into a wide-mouth bottle; placing the wide-mouth bottle on a superclean bench for surface sterilization: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, and rinsing with sterile water; the sterilized stem is cut into single stem nodes with 1cm of leaf stalks and the length of which is 1.5cm, and the single stem nodes are inoculated on a culture medium which is added with 6-benzylamino adenine (6-BA), naphthylacetic acid (NAA) and cane sugar with different concentrations in an MS basic culture medium, and are induced and expanded to culture under the induction conditions of 25 +/-1 ℃, the illumination intensity of 2000lx and the illumination period of 16 hours/day.
Example two
Cutting the tender stem at the middle upper part of the lily after the lily falls behind, washing with running water, cutting 2/3 leaves with a double-sided blade, putting the stem with 1/3 leaves into a wide-mouth bottle, wherein the lily can be one of ornamental lily 'Sobang', edible and medicinal lily pellet and iron-baked lily; placing the wide-mouth bottle on a superclean bench for surface sterilization: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, and rinsing with sterile water; the sterilized stem is cut into single stem nodes with 1cm of leaf stalks and the length of which is 1.5cm, and the single stem nodes are inoculated on a culture medium which is added with 6-benzylamino adenine (6-BA), naphthylacetic acid (NAA) and cane sugar with different concentrations in an MS basic culture medium, and are induced and expanded to culture under the induction conditions of 25 +/-1 ℃, the illumination intensity of 2000lx and the illumination period of 16 hours/day.
EXAMPLE III
Cutting the tender stem at the middle upper part of the lily after the lily falls behind, washing with running water, cutting 2/3 leaves with a double-sided blade, putting the stem with 1/3 leaves into a wide-mouth bottle, wherein the lily can be one of ornamental lily 'Sobang', edible and medicinal lily pellet and iron-baked lily; placing the wide-mouth bottle on a superclean bench for surface sterilization: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, rinsing with sterile water for 3min each time for 4-6 times; the sterilized stem is cut into single stem nodes with 1cm of leaf stalks and the length of which is 1.5cm, and the single stem nodes are inoculated on a culture medium which is added with 6-benzylamino adenine (6-BA), naphthylacetic acid (NAA) and cane sugar with different concentrations in an MS basic culture medium, and are induced and expanded to culture under the induction conditions of 25 +/-1 ℃, the illumination intensity of 2000lx and the illumination period of 16 hours/day.
Example four
Cutting the tender stem at the middle upper part of the lily after the lily falls behind, washing with running water, cutting 2/3 leaves with a double-sided blade, putting the stem with 1/3 leaves into a wide-mouth bottle, wherein the lily can be one of ornamental lily 'Sobang', edible and medicinal lily pellet and iron-baked lily; placing the wide-mouth bottle on a superclean bench for surface sterilization: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, rinsing with sterile water for 3min each time for 4-6 times; cutting the sterilized stem into 1.5cm single stem nodes with 1cm petiole, inoculating on MS basic culture medium supplemented with 6-benzylamino adenine (6-BA), naphthylacetic acid (NAA) and sucrose at different concentrations, inducing and culturing under the induction conditions of 25 + -1 deg.C, illumination intensity of 2000lx and illumination period of 16 hr/day, and culturing for one month to obtain the bulb with diameter of 1-2 cm.
EXAMPLE five
After the ornamental lily 'Sobang' blossoms, cut the tender stem (1/4 stems from the top downwards) at the upper part and wash clean with running water, cut off the leaf of 2/3 with a double-sided blade, put the stem with 1/3 leaves into a wide-mouth bottle, and carry out surface sterilization on a super clean bench: treating with 75% alcohol for 30s and 10% NaClO for 10min, and washing with sterile water for 3min for 4-6 times; cutting the killed stem into single stem sections with the length of 1.5cm and the leaf stalks of 1cm, inoculating the single stem sections on an MS +6-BA2.0mg/L + sucrose 90g/L culture medium, carrying out induction culture under the conditions of 25 +/-1 ℃, illumination intensity of 2000lx and light cycle of 16 h/day, culturing 4d of leaf axilla to start to protrude, culturing one week of leaf axilla to generate a seed bulb primordium, culturing two weeks of leaf axilla to form seed bulbs, and culturing for one month, wherein the seed bulbs can reach the bulb-out specification, namely the diameter reaches 1-2 cm.
EXAMPLE six
Cutting the tender stem at the upper part (1/3 stems from the top downwards) of the edible and medicinal lily root and the ironmaking firecracker after the lily flowers fall, cleaning the stem by running water, cutting off 2/3 leaves by using a double-sided blade, putting the stem with 1/3 leaves into a wide-mouth bottle, and performing surface sterilization on a superclean bench: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, rinsing with sterile water for 4-6 times (each for 3 min), cutting the sterilized stem into 1.5cm long single stem with 1cm petiole, inoculating on MS +6-BA2.0mg/L + sucrose 90g/L culture medium, performing induction culture at 25 + -1 deg.C, illumination intensity of 2000lx, and light cycle of 16 h/day for one month, and culturing the seed ball with diameter of 1-2 cm.
The induction result of the seedball is shown in the table, wherein the induction rate of the seedball of MS +2mg/L6-BA +90g/L cane sugar is the highest, and the seedball can be induced by 100%.
| Numbering | 6-BA(mg/L) | NAA(mg/L) | Sucrose (g/L) | Inductivity (%) |
| 1 | 0 | 0 | 30 | 4.76 |
| 2 | 0 | 0.05 | 60 | 38.46 |
| 3 | 0 | 0.1 | 90 | 40.41 |
| 4 | 1 | 0 | 60 | 86.67 |
| 5 | 1 | 0.05 | 90 | 65.75 |
| 6 | 1 | 0.1 | 30 | 52.82 |
| 7 | 2 | 0 | 90 | 100 |
| 8 | 2 | 0.05 | 30 | 58.42 |
| 9 | 2 | 0.1 | 60 | 78.95 |
In the invention, the induction is started early, the cultivation time is shortened remarkably, the axilla of the 4d leaves are cultured to begin to bulge, the axilla of one week is cultured to generate a seed bulb primordium, the axilla of two weeks is cultured to form seed bulbs, the seed bulbs can reach the bulb-out specification after being cultured for one month, namely the diameter reaches 1-2cm, compared with the traditional scale propagation, the induction time of the seed bulbs can be shortened for one month, the bulb-forming process only needs one step, namely the induction and the expansion cultivation of the seed bulbs can be realized by using one culture medium MS +2mg/L6-BA +90g/L cane sugar, the cultivation process is simple, and the pollution chance is reduced.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (4)
1. A rapid propagation method for directly inducing bulblet by lily stem segments is characterized in that: the method comprises the following steps:
A. obtaining an explant: cutting the tender stem at the middle upper part of the lily after the lily flowers fall, washing the cut stem clean with running water, cutting 2/3 leaves with a double-sided blade, and putting the stem with 1/3 leaves into a wide-mouth bottle;
B. surface sterilization: placing the wide-mouth bottle on a superclean bench for surface sterilization: treating with 75% alcohol for 30s, treating with 10% NaClO for 10min, and rinsing with sterile water;
C. seed ball induction and expansion culture: the sterilized stem is cut into single stem nodes with 1cm of leaf stalks and the length of which is 1.5cm, and the single stem nodes are inoculated on a culture medium which is added with 6-benzylamino adenine (6-BA), naphthylacetic acid (NAA) and cane sugar with different concentrations in an MS basic culture medium, and are induced and expanded to culture under the induction conditions of 25 +/-1 ℃, the illumination intensity of 2000lx and the illumination period of 16 hours/day.
2. The rapid propagation method for directly inducing bulblets by lily stem segments according to claim 1, which is characterized in that: in the step A, the lily can be one of ornamental lily 'Sobang', edible and medicinal lily pellet and irongun lily.
3. The rapid propagation method for directly inducing bulblets by lily stem segments according to claim 1, which is characterized in that: and B, rinsing with sterile water for 4-6 times in each step for 3 min.
4. The rapid propagation method for directly inducing bulblets by lily stem segments according to claim 1, which is characterized in that: and C, the bulbs can be produced after the cultivation in the step C is carried out for one month, and the diameter of the bulbs is 1-2 cm.
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Citations (6)
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| US5058318A (en) * | 1990-07-17 | 1991-10-22 | Tammen James F | Process for the production of lilium species |
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| CN109315288A (en) * | 2017-07-31 | 2019-02-12 | 赵义平 | A kind of tissue culture and rapid propagation method of lily |
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| CN110810240A (en) * | 2019-11-08 | 2020-02-21 | 甘肃爽口源生态科技股份有限公司 | Stem tissue culture and rapid propagation method for Lanzhou lily plants |
-
2021
- 2021-05-21 CN CN202110559352.0A patent/CN113207689B/en active Active
Patent Citations (6)
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| US5058318A (en) * | 1990-07-17 | 1991-10-22 | Tammen James F | Process for the production of lilium species |
| CN106386070A (en) * | 2016-08-31 | 2017-02-15 | 连云港西诺花卉种业有限公司 | Method for breeding bulblets with Oriental Hybrids plant stem tissue |
| CN109315288A (en) * | 2017-07-31 | 2019-02-12 | 赵义平 | A kind of tissue culture and rapid propagation method of lily |
| CN110268980A (en) * | 2019-07-01 | 2019-09-24 | 甘肃爽口源生态科技股份有限公司 | A kind of lanzhou lily stem apex detoxification and tissue culture and rapid propagation method |
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