CN104604687A - Method for inducing clustered shoots to rapidly propagate butterfly orchid by utilizing pedicle stem segments after bud cutting - Google Patents
Method for inducing clustered shoots to rapidly propagate butterfly orchid by utilizing pedicle stem segments after bud cutting Download PDFInfo
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Abstract
The invention discloses a method for propagating plants rapidly. The method comprises the following steps that butterfly orchid pedicle auxiliary buds after flowering are induced to form nutritional buds, and then the nutritional buds are cut, and the abandoned pedicle stem segments act as explants, and are directly inoculated into an induction culture medium for inducing clustered buds; the novel induced buds are cut off to be inoculated to the propagation culture medium for subculture propagation culture; finally rooting and strenghening are carried out and tissue culture seedlings are transplanted. By the method, the auxiliary buds on the butterfly orchid pedicles after flowering and abandoned pedicle materials after bud cutting germinate in a manually prepared culture medium for multiple times to generate novel buds. The method has the advantages that not only is the normal flower period of butterfly orchid not affected, but also butterfly orchid mother plants are not hurt, the butterfly orchid pedicle stem segments are recycled for 2-3 times; pedicle materials are used fully, waste is turned into wealth; the propagation velocity and the production of butterfly orchid tissue culture seedlings are improved greatly; the produced seed seedlings are strong and neat, and are convenient to manage in a factory.
Description
Technical field
The invention belongs to Moth orchid propagation technique field, the bennet stem section induced bundle after bud is cut in especially a kind of utilization is sprouted the method for propagating quickly butterfly orchid.
Background technology
Moth orchid (Phalaenopsis sp.) belongs to the orchid family Phalaenopsis plant, found in 1750, great majority originate in moist Asia, and natural distribution is in A Longmu, Burma, each island, the Indian Ocean, the Malay Peninsula, the South Sea Islands, Philippine down to islands, the low latitudes torrid zone such as TaiWan, Chinas.Initial species has kind more than 70, and crossbreed is innumerable especially, and the change of its flower color and decorative pattern emerges in an endless stream, spend in vain be, safflower system, pink system, chrysanthemum system, reticulate pattern system, tiger spot system, kind series such as some line system etc.Its flower pattern is peculiar, and attitude is graceful, and pattern is gorgeous, and the florescence is permanent, and have the good reputation of " orchid queen ", ornamental value is high, is loved by the people, and market demand is increasing.
Phalaenopsis, in single stem aerial orchid, seldom grows side shoot, is difficult to carry out conventional vegetative propagation; Moth orchid can not complete pollination under field conditions (factors) in addition, is difficult to obtain seed, even if carry out artificial pollination to obtain seed, not containing endosperm in its seed, also be difficult under natural conditions sprout, germination rate is extremely low, and therefore Moth orchid is also difficult to utilize seed to carry out sexual propagation.Method for tissue culture is mainly utilized to carry out the Fast-propagation of Moth orchid at present, belong to the vegetative propagation of Moth orchid, also so-called clonal propagation, the tissue cultures of Moth orchid can obtain a large amount of high-quality, neat Moth orchid young plant in a short time, have that the rate of increase is high, speed is subject to seasonal restrictions soon, or not can whole year production and easily remove the advantages such as virus, adapting to the wilderness demand in market, is also the important channel of factorial seedling growth.Moth orchid tissue cultures program is generally: (1) chooses explant blade, root, stem apex, bennet etc.; (2) induced synthesis protocorm; (3) protocorm is bred in a large number; (4) plantlet is differentiated; (5) strong plantlets and rootage; (6) greenhouse is transplanted.Can find out that from said procedure (1) explant that Moth orchid tissue cultures is chosen can be blade, root, stem apex, bennet etc., but above-mentioned explant all there is certain injury to maternal plant or affects the maternal plant normal florescence.What the tissue cultures of current Moth orchid was conventional has two kinds of approach, one is that conventional bennet induction vegetative bud is bred, if the method adopt existing petal but the bennet of not blooming or blooming as explant, Moth orchid normal florescence and ornamental value can be affected, and reproduction coefficient is generally about 2.0-3.0 in every two months, the plantlet in vitro produced in the short time is limited; Two is utilize aseptically sowing seeds method to carry out the production of Moth orchid plantlet in vitro, and the seedling variation that the method breeds is more, and seedling early growth is inconsistent.
Summary of the invention
Bennet stem section induced bundle after the object of the present invention is to provide a kind of utilization to cut bud is sprouted the method for propagating quickly butterfly orchid, solve Moth orchid plant division slower, not easily sprout under seed natural conditions, plant breeding speed is slow, commercial seedling output is low, growth cycle is inconsistent, flowering time is uneven, can not the problem such as blooming control and listing in enormous quantities.
To achieve these goals, the present invention adopts following technical scheme, induced vegetative bud cut the discarded bennet stem section after bud as explant to open the Moth orchid pedicel axillary buds spent, directly be inoculated into the induction carrying out bud in inducing culture, the sprouting induced is cut to be inoculated in proliferated culture medium and carries out shoot proliferation cultivation, finally carry out the transplanting of strong plantlets and rootage and plantlet in vitro.
Concrete step is:
(1) selection of explant: choose the butterfly orchid variety grown fine without damage by disease and insect, treat that its flower withers, the florescence is mistake, is cut by whole piece bennet with scissors, and the pedicel axillary buds stem section (one section of one bud) cutting 3-4cm long is explant material;
(2) preparation of medium:
Bud inducement medium: MS+6-BA2.5mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6:
Shoot propagation medium: MS+6-BA5.0mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6:
Rooting and hardening-off culture base: 1/2MS+NAA1.0mg/L+ banana puree 100g/L, sucrose 20g/L, pH value is 5.6;
(3) explant is disinfected: by above-mentioned explant material, after rinsing 30-60min under running water, in superclean bench, put into the alcoholic solution soaking disinfection 45S of 75%, after this alcohol is blotted with aseptic filter paper, put into the NaClO thimerosal sterilization 15-20min of 7-10%, then on aseptic filter paper, the bract of axillalry bud is carefully peelled off, the NaClO thimerosal putting into 5% is again sterilized 5min, with aseptic water washing 3-5 time after sterilizing, it is for subsequent use that filter paper suck dry moisture is placed in culture dish;
(4) Fiber differentiation and condition of culture: by the band axillalry bud bennet after strict sterilization, two ends stem section contacted with thimerosal with aseptic operation cutter are cut away along 45 DEG C of angles, bennet bud point becomes 45 DEG C of angle oblique cuttings to enter in bud inducement medium upward with medium plane, cultivation temperature 25 ± 2 DEG C, light application time 12-14h/d, intensity of illumination is 1600-2000Lx, and induction time is about 30-45d;
(5) Multiplying culture and condition of culture: cut from bennet by the sprouting that step (4) induces, 2-3 strain is a group, is inoculated in proliferated culture medium and carries out shoot proliferation, condition of culture and the same step of incubation time (4):
(6) by step (5), just in generation, cuts the bennet stem section renewed vaccination after bud in inducing culture, puts culturing rack and cultivates, condition of culture and the same step of incubation time (4);
(7) cut cutting the Multiple Buds that the bennet stem section chain induction after bud goes out from bennet, 2-3 strain is one group, is inoculated in proliferated culture medium and carries out shoot proliferation, condition of culture and the same step of incubation time (4);
(8) step (6) and step (7) 2-3 time is repeated;
(9) strong plantlets and rootage and condition of culture: when breeding the sprout obtained and reaching 2-5cm height, under aseptic condition, Multiple Buds is separated into individual plant, is seeded in Rooting and hardening-off culture base and carries out strong plantlets and rootage, the same step of condition of culture (4), the strong plantlets and rootage time is 2-3 month;
(10) training tissue culture seedling and transplanting: to grow to 3-5cm high when plantlet in vitro, there is 2-5 bar root, 3-6 sheet leaf, during the long 2-3cm of leaf, hardening can be carried out, tissue culture bottle is placed on natural lighting condition lower refining seedling 5-7 days, opens tissue culture bottle lid and continue hardening 2-3 days, to strengthen the adaptive capacity of bottle seedling to outdoor environment; From bottle, carefully take out plantlet in vitro again, clean root medium, gently wrap up in root with sterilized sphagna and plant in 36 hole dishes, keep temperature about 25 DEG C, after air humidity 80-85%, 25-30 days, when having new root young leaves to grow, regularly spray carbendazim 1000 times of liquid.
The present invention compared to the prior art its advantage is:
The present invention disclose a kind of utilization cut bud after bennet stem section induced bundle to sprout the method for propagating quickly butterfly orchid, sleeping bud on the faded Moth orchid bennet of flower and the bennet stem section after cutting bud thereof are repeatedly sprouted in the medium of artificial preparation, then carry out breeding and culture of rootage, the Phalaenopsis plants that final acquisition is complete.The conventional bennet that utilizes induces vegetative bud to breed, growth coefficient in two months is about 2.0, and adopts as above technical scheme, makes the growth coefficient of each Moth orchid pedicel axillary buds bring up to about 14.0-26.0, be equivalent to efficiency and improve 20 times, save production cost greatly.The advantage of this method is that neither affecting the Moth orchid normal florescence need not injure Moth orchid maternal plant again, Moth orchid bennet stem section can also be recycled 2-3 time, bennet material can be made full use of, turn waste into wealth, substantially increase proliferative speed and the output of Moth orchid plantlet in vitro, the seedling produced is healthy and strong, neat, is convenient to seedling factory management.
Embodiment
Below in conjunction with specific embodiment 1, the present invention will be further described.
Embodiment 1:
(1) selection of explant: choose the black Jack plant of butterfly orchid variety without damage by disease and insect, treat that flower withers, the florescence is mistake, is cut by whole piece bennet with scissors, the pedicel axillary buds stem section (one section of one bud) cutting 3-4cm long is explant material;
(2) preparation of medium:
Bud inducement medium: MS+6-BA2.5mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6:
Shoot propagation medium: MS+6-BA5.0mg/L+NAA0.5mg/L, sucrose 20g/L, pH value is 5.6:
Rooting and hardening-off culture base: 1/2MS+NAA1.0mg/L+ banana puree 100g/L, sucrose 20g/L, pH value is 5.6;
(3) explant is disinfected: by above-mentioned explant material, after rinsing 30-60min under running water, in superclean bench, put into the alcoholic solution soaking disinfection 45S of 75%, after this alcohol is blotted with aseptic filter paper, put into the NaClO thimerosal sterilization 12-15min of 7-10%, aging material sterilizing 15-20min, then on aseptic filter paper, the bract of axillalry bud is carefully peelled off, the NaClO thimerosal putting into 5% is again sterilized 5min, with aseptic water washing 3-5 time after sterilizing, it is for subsequent use that filter paper suck dry moisture is placed in culture dish;
(4) Fiber differentiation and condition of culture: by the band axillalry bud bennet after strict sterilization, two ends stem section contacted with thimerosal with aseptic operation cutter are cut away along 45 DEG C of angles, bennet bud point becomes 45 DEG C of angle oblique cuttings to enter in bud inducement medium upward with medium plane, cultivation temperature 25 ± 2 DEG C, light application time 12-14h/d, intensity of illumination is 1600-2000Lx, and induction time is about 30-45d;
(5) Multiplying culture and condition of culture: cut from bennet by the sprouting that step (4) induces, 2-3 strain is a group, is inoculated in proliferated culture medium and carries out shoot proliferation, culturing room's condition and the same step of incubation time (4);
(6) by step (5), just in generation, cuts the bennet stem section renewed vaccination after bud in inducing culture, puts culturing rack and cultivates, condition of culture and the same step of incubation time (4);
(7) cut cutting the Multiple Buds that the bennet stem section chain induction after bud goes out from bennet, 2-3 strain is one group, is inoculated in proliferated culture medium and carries out shoot proliferation, condition of culture and the same step of incubation time (4);
(8) step (6) and step (7) 2-3 time is repeated;
(9) strong plantlets and rootage and condition of culture: when breeding the sprout obtained and reaching 2-5cm height, sterile working, is separated into individual plant by Multiple Buds, is seeded in Rooting and hardening-off culture base and carries out strong plantlets and rootage, culturing room's conditional synchronization rapid (4), the strong plantlets and rootage time is 2-3 month;
(10) training tissue culture seedling and transplanting: when plantlet in vitro grows to 3-5cm height, there is 2-5 bar root, 3-6 sheet leaf, during the long 2-3cm of leaf, hardening can be carried out, tissue culture bottle is placed on natural lighting condition lower refining seedling 5-7 days, opens tissue culture bottle lid hardening 2-3 days, to strengthen the adaptive capacity of bottle seedling to outdoor environment; From bottle, carefully take out plantlet in vitro again, clean root medium, gently wrap up in root with sterilized sphagna and plant in 36 hole dishes, keep temperature about 25 DEG C, air humidity 80-85%, after 30 days, when having new root young leaves to grow, spray carbendazim 1000 times of liquid, regularly spray, survival rate is to 95%.
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (2)
1. the bennet stem section induced bundle after bud is cut in a utilization is sprouted the method for propagating quickly butterfly orchid, it is characterized in that: induced vegetative bud cut the discarded bennet stem section after bud as explant with opening the Moth orchid pedicel axillary buds spent, directly be inoculated into the induction carrying out Multiple Buds in inducing culture, the sprouting induced is cut to be inoculated in proliferated culture medium and carries out shoot proliferation cultivation, finally carry out the transplanting of strong plantlets and rootage and plantlet in vitro.
2. the bennet stem section induced bundle after bud is cut in utilization according to claim 1 is sprouted the method for propagating quickly butterfly orchid, it is characterized in that:
Concrete step is:
(1) selection of explant: choose the butterfly orchid variety grown fine without damage by disease and insect, treat that its flower withers, the florescence is mistake, is cut by whole piece bennet with scissors, and the pedicel axillary buds stem section (one section of one bud) cutting 3-4cm long is explant material;
(2) preparation of medium:
Bud inducement medium: MS+6-BA2.5mg/L+NAA0.5 mg/L, sucrose 20 g/L, pH value is 5.6;
Shoot propagation medium: MS+6-BA5.0mg/L+NAA0.5mg/L, sucrose 20 g/L, pH value is 5.6;
Rooting and hardening-off culture base: 1/2MS+NAA1.0mg/L+ banana puree 100g/L, sucrose 20 g/L, pH value is 5.6;
(3) explant is disinfected: by above-mentioned explant material, after rinsing 30-60min under running water, in superclean bench, put into the alcoholic solution soaking disinfection 45S of 75%, after this alcohol is blotted with aseptic filter paper, put into the NaClO thimerosal sterilization 15-20min of 7-10%, then on aseptic filter paper, the bract of axillalry bud is carefully peelled off, the NaClO thimerosal putting into 5% is again sterilized 5min, with aseptic water washing 3-5 time after sterilizing, it is for subsequent use that filter paper suck dry moisture is placed in culture dish;
(4) Fiber differentiation and condition of culture: by the band axillalry bud bennet after strict sterilization, two ends stem section contacted with thimerosal with aseptic operation cutter are cut away along 45 DEG C of angles, bennet bud point becomes 45 DEG C of angle oblique cuttings to enter in bud inducement medium upward with medium plane, cultivation temperature 25 ± 2 DEG C, light application time 12-14h/d, intensity of illumination is 1600-2000Lx, and induction time is about 30-45d;
(5) Multiplying culture and condition of culture: cut from bennet by the sprouting that step (4) induces, 2-3 strain is a group, is inoculated in proliferated culture medium and carries out shoot proliferation, condition of culture and the same step of incubation time (4);
(6) by step (5), just in generation, cuts the bennet stem section renewed vaccination after bud in inducing culture, puts culturing rack and cultivates, condition of culture and the same step of incubation time (4);
(7) cut cutting the Multiple Buds that the bennet stem section chain induction after bud goes out from bennet, 2-3 strain is one group, is inoculated in proliferated culture medium and carries out shoot proliferation, condition of culture and the same step of incubation time (4);
(8) step (6) and step (7) 2-3 time is repeated;
(9) strong plantlets and rootage and condition of culture: when breeding the sprout obtained and reaching 2-5cm height, under aseptic condition, Multiple Buds is separated into individual plant, is seeded in Rooting and hardening-off culture base and carries out strong plantlets and rootage, the same step of condition of culture (4), the strong plantlets and rootage time is 2-3 month;
(10) training tissue culture seedling and transplanting: to grow to 3-5cm high when plantlet in vitro, there is 2-5 bar root, 3-6 sheet leaf, during the long 2-3cm of leaf, hardening can be carried out, tissue culture bottle is placed on natural lighting condition lower refining seedling 5-7 days, opens tissue culture bottle lid and continue hardening 2-3 days, to strengthen the adaptive capacity of bottle seedling to outdoor environment; From bottle, carefully take out plantlet in vitro again, clean root medium, gently wrap up in root with sterilized sphagna and plant in 36 hole dishes, keep temperature about 25 DEG C, after air humidity 80-85%, 25-30 days, when having new root young leaves to grow, regularly spray carbendazim 1000 times of liquid.
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Cited By (13)
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| CN107278901A (en) * | 2017-07-31 | 2017-10-24 | 王生旭 | A kind of sterilization method of iris bennet bud |
| CN108419676A (en) * | 2018-03-30 | 2018-08-21 | 内蒙古自治区生物技术研究院 | Iris tissue culture medium (TCM) and preparation method thereof |
| CN108651444A (en) * | 2018-05-22 | 2018-10-16 | 山东省农业科学院蔬菜花卉研究所 | A kind of iris pollen preserves and breeding method |
| CN109349105A (en) * | 2018-10-10 | 2019-02-19 | 广西生态工程职业技术学院 | A kind of iris tissue-cultured seedling mating system |
| CN109601386A (en) * | 2019-01-24 | 2019-04-12 | 北京农业职业学院 | The sterilization method of bud explant is cut in a kind of state orchid stem tip tissue culture |
| CN110896856A (en) * | 2019-11-07 | 2020-03-24 | 郑州师范学院 | Method for inducing complete inflorescence in vitro by axillary bud of inflorescence shaft of phalaenopsis |
| CN111264394A (en) * | 2020-04-07 | 2020-06-12 | 无锡向山兰园科技有限公司 | Tissue culture method of cymbidium hybridum |
| CN111758567A (en) * | 2020-05-29 | 2020-10-13 | 福建农林大学 | Phalaenopsis single-bud propagation method adopting symbiotic bacteria |
| CN113197098A (en) * | 2021-06-07 | 2021-08-03 | 芜湖东源新农村开发股份有限公司 | Inoculation method for shortening rapid propagation growth cycle of phalaenopsis |
| CN113575421A (en) * | 2021-08-27 | 2021-11-02 | 中德润萌生态科技(青岛)有限公司 | Aseptic processing method for tissue culture material of orchidaceae plant |
| CN113973713A (en) * | 2021-10-28 | 2022-01-28 | 北京市昌平职业学校 | Butterfly orchid tissue culture method |
| CN115251057A (en) * | 2022-08-26 | 2022-11-01 | 郑州师范学院 | Method for inducing germination of phalaenopsis seedlings by utilizing phytohormone composition |
| CN116686715A (en) * | 2023-07-25 | 2023-09-05 | 云南省农业科学院花卉研究所 | Butterfly orchid tissue culture seedling cultivation method without carrying cymMV and ORSV |
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107278901A (en) * | 2017-07-31 | 2017-10-24 | 王生旭 | A kind of sterilization method of iris bennet bud |
| CN108419676A (en) * | 2018-03-30 | 2018-08-21 | 内蒙古自治区生物技术研究院 | Iris tissue culture medium (TCM) and preparation method thereof |
| CN108651444B (en) * | 2018-05-22 | 2020-11-27 | 山东省农业科学院蔬菜花卉研究所 | Preservation and breeding method for phalaenopsis pollen |
| CN108651444A (en) * | 2018-05-22 | 2018-10-16 | 山东省农业科学院蔬菜花卉研究所 | A kind of iris pollen preserves and breeding method |
| CN109349105A (en) * | 2018-10-10 | 2019-02-19 | 广西生态工程职业技术学院 | A kind of iris tissue-cultured seedling mating system |
| CN109601386A (en) * | 2019-01-24 | 2019-04-12 | 北京农业职业学院 | The sterilization method of bud explant is cut in a kind of state orchid stem tip tissue culture |
| CN110896856A (en) * | 2019-11-07 | 2020-03-24 | 郑州师范学院 | Method for inducing complete inflorescence in vitro by axillary bud of inflorescence shaft of phalaenopsis |
| CN111264394A (en) * | 2020-04-07 | 2020-06-12 | 无锡向山兰园科技有限公司 | Tissue culture method of cymbidium hybridum |
| CN111758567A (en) * | 2020-05-29 | 2020-10-13 | 福建农林大学 | Phalaenopsis single-bud propagation method adopting symbiotic bacteria |
| CN113197098A (en) * | 2021-06-07 | 2021-08-03 | 芜湖东源新农村开发股份有限公司 | Inoculation method for shortening rapid propagation growth cycle of phalaenopsis |
| CN113575421A (en) * | 2021-08-27 | 2021-11-02 | 中德润萌生态科技(青岛)有限公司 | Aseptic processing method for tissue culture material of orchidaceae plant |
| CN113973713A (en) * | 2021-10-28 | 2022-01-28 | 北京市昌平职业学校 | Butterfly orchid tissue culture method |
| CN115251057A (en) * | 2022-08-26 | 2022-11-01 | 郑州师范学院 | Method for inducing germination of phalaenopsis seedlings by utilizing phytohormone composition |
| CN116686715A (en) * | 2023-07-25 | 2023-09-05 | 云南省农业科学院花卉研究所 | Butterfly orchid tissue culture seedling cultivation method without carrying cymMV and ORSV |
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