CN103988780A - Method for directly inducing adventitious buds of butterfly orchid - Google Patents
Method for directly inducing adventitious buds of butterfly orchid Download PDFInfo
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- CN103988780A CN103988780A CN201410221460.7A CN201410221460A CN103988780A CN 103988780 A CN103988780 A CN 103988780A CN 201410221460 A CN201410221460 A CN 201410221460A CN 103988780 A CN103988780 A CN 103988780A
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Abstract
After butterfly orchid is treated by an outdoor hormone combination (TDZ+GA3+NAA) and a browning prevention solution (AC+PVP+Vc0.5g/L+glutathione), adventitious buds can be directly induced, the content of explant total phenols and PAL activity can be inhibited, and rapid plant growth is promoted. Due to the outdoor hormone treatment, hormones can be absorbed by the explant in advance, hormonal equilibrium needed by adventitious bud germination is achieved, and the time needed for absorbing nutrition in a bottle is shortened. Browning generated in the explant is greatly inhibited due to the outdoor treatment, so that rapid growth of the adventitious buds and seedlings is promoted. According to the explant, the adventitious buds of the butterfly orchid are directly induced without passing a callus stage, excellent traits of stock plants can be kept, variation generated in the culture process is reduced, and the method has high significance in large-scale production.
Description
Technical field
A kind of method that the present invention relates to direct induction Moth orchid indefinite bud, belongs to biotic induce technical field.
Background technology
Moth orchid (Phalaenopsis spp.) is the orchid family Phalaenopsis flowers.Because its flower type is peculiar, beautiful in colour, the florescence is lasting, have the good reputation of " cattleya queen ".
According on August 15th, 2013 king of Yantai City Inst. of Agricultural Science, Shandong Prov. often rue deliver at " northern gardening " about in " Moth orchid group culturation rapid propagating technology architectural study ", utilizing the tissues such as seed, bennet axillalry bud, stem apex, blade, bennet internode, the tip of a root and organ is explant, induction under certain condition forms protocorm, then breeds by protocorm.January in 2012 Sichuan Province on the 20th Sichuan institute of nationalities environment and life science be Tang Dehua deliver in " Agriculture of Anhui science " about in " research of brownization of Phalaenopsis Explant in Vitro control ", in the method that prevents Brown, adopt and add adsorbent, anti-browning agent, natural organic matter, regulating illumination and cultivation temperature and explant is heat-treated etc., but both there is certain weak point: active carbon can effectively be controlled brown stain, be but unfavorable for differentiation; The membership that adds of antioxidant increases pollution; Thermal treatment generally all needs design temperature interval and time period, and convalescence want 24-48 hour needs to work continuously, program comparision complexity.
Summary of the invention
The present invention has overcome deficiency of the prior art, has filled up in prior art and has lacked the outdoor pretreated blank that shifts to an earlier date, and a kind of method of rapid, high volume breeding butterfly orchid seedling plant is provided.
Technical scheme of the present invention is:
A method for direct induction Moth orchid indefinite bud, is characterized in that, comprises the following steps:
(1) adventitious bud inducing processing, first possess fecundity or perennial healthy and strong Moth orchid plant leaf front sprays exogenous hormone from inside to outside to raw then, in the time that bennet is extracted out, bennet is from bottom to top sprayed to nutritive element, every 25-40 days sprays once, sprays 3-5 time altogether;
(2) anti-browning processing in vegetative period, plant and bennet are sprayed to after the process of exogenous hormone 7 days, the front of blade is sprayed to anti-browning mixed solvent liquid from inside to outside, bennet is from bottom to top sprayed to anti-browning mixed solvent liquid, every 8-15 days sprays once.
Described adventitious bud inducing is processed and anti-browning processing in vegetative period is all carried out outdoor.
Described exogenous hormone is mixture or MS mineral element or the hormonal composition of MS mineral element and hormonal composition.Hormonal composition is by 0.01-0.05mg/L phenyl thiadiazolyl group urea (TDZ), 0.05-0.5mg/L gibberellin (GA
3) and 0.05-0.5mg/L methyl α-naphthyl acetate (NAA) mix, wherein the mass ratio of TDZ, GA3, NAA is 1:10～15:10～15, the mass ratio of MS mineral element and hormonal composition is 20000～25000:1.More preferably: described hormonal composition is by 0.01mg/L phenyl thiadiazolyl group urea (TDZ), 0.1mg/L gibberellin (GA
3) and 0.1mg/L methyl α-naphthyl acetate (NAA) mix, wherein TDZ, GA
3, NAA mass ratio be 1:12:15, the mass ratio of MS mineral element and hormonal composition is 23000:1.
Anti-browning mixed solvent liquid is mixed by active carbon (AC) leaching liquor of 0.3-1.5g/L, the polyvinylpyrrolidone (PVP) of 0.5-2.0g/L, the ascorbic acid (Vc) of 0.2-0.8g/L and the glutathione of 0.2-0.8g/L, and wherein the mass ratio of AC leaching liquor, PVP, Vc, glutathione is 1:1～5:0.3～1:0.3～1.More preferably: anti-browning mixed solvent liquid is mixed by active carbon (AC) leaching liquor of 1.0g/L, the polyvinylpyrrolidone (PVP) of 1.0g/L, the ascorbic acid (Vc) of 0.5g/L and the glutathione of 0.5g/L, and wherein the mass ratio of AC leaching liquor, PVP, Vc, glutathione is 1:3.6:0.45:0.45.
Induction Moth orchid indefinite bud practical technique advantage:
1. the indoor Induction Process of routine will constantly be adjusted hormone prescription, and each adjustment all will repeat many experiments, time and materials is all caused to certain loss, and outdoor processing is simple to operate, not loss material, saved greatly experimental period, induction effect is remarkable simultaneously, can rapid, high volume breeding indefinite bud seedling in the short time.
2. outdoor HORMONE TREATMENT can make explant absorb in advance hormone, reaches and sprouts the required hormonal balance of indefinite bud, has shortened a bottle interior required time of nutrient absorption.
3. outdoor processing has suppressed brownization producing in explant greatly, thereby makes to have promoted indefinite bud and seedling ramp.
4. explant directly induces Moth orchid indefinite bud can keep the merit of maternal plant without the callus stage, generation variation in minimizing incubation, important in inhibiting in large-scale production.
Brief description of the drawings
Fig. 1 is Moth orchid adventitious bud inducing;
Fig. 2 is the increment of Moth orchid indefinite bud;
Fig. 3 is that Moth orchid individual plant is cultivated.
Embodiment
Embodiment 1
Adventitious bud inducing technical finesse:
For promoting smoothly effectively the formation of indefinite bud, first possesses fecundity (being transitioned into reproduction reproductive phase by nourishing and growing) or perennial healthy and strong Moth orchid plant sprays 6.44 × 10 to raw then
6g MS mineral element+hormone combinations combination (NAA of the GA3+150g0.1mg/L of the TDZ+120g0.1mg/L of 10g0.01mg/L), spray MS mineral element, spray hormone combinations, 4 processing of contrast, each pack processing is containing 10 basin plant individualities, and test is established repetition 3 times.Processing time starts from early October, and every 40d carries out once, sprays altogether 3 times.While spraying, adopt the little watering can of 150ml, kettle mouth furnishing goes out vaporific optimum state aligning plant leaf front and evenly sprays from inside to outside at every turn, has sprayed rear water smoke and has been attached to not mobile being advisable of blade surface, and vacuum side of blade does not spray.In the time that bennet at the beginning of 11 months is extracted out, start bennet to spray, while spraying, direction is from top to bottom, has sprayed rear water smoke and has been attached to not mobile being advisable of bennet surface.Treated Moth orchid is placed in green house and cultivates, until gather explant.Inoculate and add up every data, data statistical approach after 30 days: differentiation number: the number of the actual generation of inoculation explant differentiation situation; Bud sum: produce the bud sum that differentiation situation explant breaks up; Differentiation rate: the explant number that produces the bud number/inoculation of differentiation situation; Height of seedling: the average height (measuring height/Bud Differentiation sum of all Bud Differentiations) of the Bud Differentiation producing; Experimental result is in table 1 in detail.
After table 1 inoculation, 30d is at different disposal Moth orchid differentiation adventitious buds and growing state
Anti-browning processing in vegetative period:
The pretreated research of anti-browning in vegetative period is specially 30d before collection explant leaf and bennet is sprayed to anti-browning mixed solvent liquid (glutathione of the Vc+4.5g0.5g/L of the PVP+4.5g0.5g/L of AC leaching liquor+36g1g/L of 10g1g/L) and contrast (not spraying anti-brown liquid) 2 processing.Anti-browning is processed and is sprayed hormone at plant and bennet and carry out after 7 days, every 30d once, totally 3 times.The method that anti-browning mixed solvent liquid sprays is identical with the HORMONE TREATMENT of 1.1 Leafs and bennet spray method.Explant is different exogenous hormones bennet axillalry buds after treatment, adds up every data after 40 days.It is 15.83% that the explant that result of the test shows to spray anti-browning liquid produces brown rate, and the brown rate of generation that does not spray anti-browning liquid is 81.68%.Experimental result is in table 2 in detail.
Table 2 different explants is processed the brownization degree table of comparisons
Embodiment 2
Adventitious bud inducing technical finesse:
For promoting smoothly effectively the formation of indefinite bud, first possesses fecundity (being transitioned into reproduction reproductive phase by nourishing and growing) or perennial healthy and strong Moth orchid plant sprays 5.2 × 10 to raw then
6g MS mineral element+hormone combinations combination (NAA of the GA3+140g0.4mg/L of the TDZ+110g0.3mg/L of 10g0.05mg/L), spray MS mineral element, spray hormone combinations, 4 processing of contrast, each pack processing is containing 10 basin plant individualities, and test is established repetition 3 times.Processing time starts from early October, and every 40d carries out once, sprays altogether 3 times.While spraying, adopt the little watering can of 150ml, kettle mouth furnishing goes out vaporific optimum state aligning plant leaf front and evenly sprays from inside to outside at every turn, has sprayed rear water smoke and has been attached to not mobile being advisable of blade surface, and vacuum side of blade does not spray.In the time that bennet at the beginning of 11 months is extracted out, start bennet to spray, while spraying, direction is from top to bottom, has sprayed rear water smoke and has been attached to not mobile being advisable of bennet surface.Treated Moth orchid is placed in green house and cultivates, until gather explant.Inoculate and add up every data, data statistical approach after 30 days: differentiation number: the number of the actual generation of inoculation explant differentiation situation; Bud sum: produce the bud sum that differentiation situation explant breaks up; Differentiation rate: the explant number that produces the bud number/inoculation of differentiation situation; Height of seedling: the average height (measuring height/Bud Differentiation sum of all Bud Differentiations) of the Bud Differentiation producing; Experimental result is in table 3 in detail.
After table 3 inoculation, 30d is at different disposal Moth orchid differentiation adventitious buds and growing state
Anti-browning processing in vegetative period:
The pretreated research of anti-browning in vegetative period is specially 30d before collection explant leaf and bennet is sprayed to anti-browning mixed solvent liquid (glutathione of the Vc+8.3g0.33g/L of the PVP+6.2g0.25g/L of AC leaching liquor+20g0.68g/L of 10g0.38g/L) and contrast (not spraying anti-brown liquid) 2 processing.Anti-browning is processed and is sprayed hormone at plant and bennet and carry out after 7 days, every 30d once, totally 3 times.The method that anti-browning mixed solvent liquid sprays is identical with the HORMONE TREATMENT of 1.1 Leafs and bennet spray method.Explant is different exogenous hormones bennet axillalry buds after treatment, adds up every data after 40 days.It is 17.1% that the explant that result of the test shows to spray anti-browning liquid produces brown rate, and the brown rate of generation that does not spray anti-browning liquid is 86.7%.Experimental result is in table 4 in detail.
Table 4 different explants is processed the brownization degree table of comparisons
Embodiment 3
Adventitious bud inducing technical finesse:
For promoting smoothly effectively the formation of indefinite bud, first possesses fecundity (being transitioned into reproduction reproductive phase by nourishing and growing) or perennial healthy and strong Moth orchid plant sprays 5.25 × 10 to raw then
6g MS mineral element+hormone combinations combination (NAA of the GA3+100g0.45mg/L of the TDZ+100g0.5mg/L of 10g0.05mg/L), spray MS mineral element, spray hormone combinations, 4 processing of contrast, each pack processing is containing 10 basin plant individualities, and test is established repetition 3 times.Processing time starts from early October, and every 40d carries out once, sprays altogether 3 times.While spraying, adopt the little watering can of 150ml, kettle mouth furnishing goes out vaporific optimum state aligning plant leaf front and evenly sprays from inside to outside at every turn, has sprayed rear water smoke and has been attached to not mobile being advisable of blade surface, and vacuum side of blade does not spray.In the time that bennet at the beginning of 11 months is extracted out, start bennet to spray, while spraying, direction is from top to bottom, has sprayed rear water smoke and has been attached to not mobile being advisable of bennet surface.Treated Moth orchid is placed in green house and cultivates, until gather explant.Inoculate and add up every data, data statistical approach after 30 days: differentiation number: the number of the actual generation of inoculation explant differentiation situation; Bud sum: produce the bud sum that differentiation situation explant breaks up; Differentiation rate: the explant number that produces the bud number/inoculation of differentiation situation; Height of seedling: the average height (measuring height/Bud Differentiation sum of all Bud Differentiations) of the Bud Differentiation producing; Experimental result is in table 5 in detail.
After table 5 inoculation, 30d is at different disposal Moth orchid differentiation adventitious buds and growing state
Anti-browning processing in vegetative period:
Anti-browning mixed solvent liquid is mixed by active carbon (AC) leaching liquor of 0.3-1.5g/L, the polyvinylpyrrolidone (PVP) of 0.5-2.0g/L, the ascorbic acid (Vc) of 0.2-0.8g/L and the glutathione of 0.2-0.8g/L, and wherein the mass ratio of AC leaching liquor, PVP, Vc, glutathione is 1:1～5:0.3～1:0.3～1.
The pretreated research of anti-browning in vegetative period is specially 30d before collection explant leaf and bennet is sprayed to anti-browning mixed solvent liquid (glutathione of the Vc+10g0.8g/L of the PVP+9.5g0.8g/L of AC leaching liquor+50g1.85g/L of 10g1.5g/L) and contrast (not spraying anti-brown liquid) 2 processing.Anti-browning is processed and is sprayed hormone at plant and bennet and carry out after 7 days, every 30d once, totally 3 times.The method that anti-browning mixed solvent liquid sprays is identical with the HORMONE TREATMENT of 1.1 Leafs and bennet spray method.Explant is different exogenous hormones bennet axillalry buds after treatment, adds up every data after 40 days.It is 15.43% that the explant that result of the test shows to spray anti-browning liquid produces brown rate, and the brown rate of generation that does not spray anti-browning liquid is 85.45%.Experimental result is in table 6 in detail.
Table 6 different explants is processed the brownization degree table of comparisons
Claims (7)
1. a method of directly inducing Moth orchid indefinite bud, is characterized in that, comprises the following steps:
1) adventitious bud inducing processing, first possess fecundity or perennial healthy and strong Moth orchid plant leaf front sprays exogenous hormone from inside to outside to raw then, in the time that bennet is extracted out, bennet is from bottom to top sprayed to nutritive element, every 25-40 days sprays once, sprays 3-5 time altogether;
2) anti-browning processing in vegetative period, plant and bennet are sprayed to after the process of exogenous hormone 7 days, the front of blade is sprayed to anti-browning mixed solvent liquid from inside to outside, in the time that plant bennet is extracted out, bennet is from bottom to top sprayed to anti-browning mixed solvent liquid, every 8-15 days sprays once, obtains the Moth orchid of outdoor induction.
2. the method for direct induction Moth orchid indefinite bud according to claim 1, is characterized in that: adventitious bud inducing is processed and anti-browning processing in vegetative period is all carried out outdoor.
3. the method for direct induction Moth orchid indefinite bud according to claim 1, is characterized in that: described exogenous hormone is mixture or MS mineral element or the hormonal composition of MS mineral element and hormonal composition.
4. the method for direct induction Moth orchid indefinite bud according to claim 3, is characterized in that: described hormonal composition is by 0.01-0.05mg/L phenyl thiadiazolyl group urea (TDZ), 0.05-0.5mg/L gibberellin (GA
3) and 0.05-0.5mg/L methyl α-naphthyl acetate (NAA) mix, wherein TDZ, GA
3, NAA mass ratio be 1:10 ~ 15:10 ~ 15, the mass ratio of MS mineral element and hormonal composition is 20000 ~ 25000:1.
5. the method for direct induction Moth orchid indefinite bud according to claim 4, is characterized in that: described hormonal composition is by 0.01mg/L phenyl thiadiazolyl group urea (TDZ), 0.1mg/L gibberellin (GA
3) and 0.1mg/L methyl α-naphthyl acetate (NAA) mix, wherein TDZ, GA
3, NAA mass ratio be 1:12:15, the mass ratio of MS mineral element and hormonal composition is 23000:1.
6. the method for direct induction Moth orchid indefinite bud according to claim 1, it is characterized in that: anti-browning mixed solvent liquid is mixed by active carbon (AC) leaching liquor of 0.3-1.5g/L, the polyvinylpyrrolidone (PVP) of 0.5-2.0g/L, the ascorbic acid (Vc) of 0.2-0.8g/L and the glutathione of 0.2-0.8g/L, and wherein the mass ratio of AC leaching liquor, PVP, Vc, glutathione is 1:1 ~ 5:0.3 ~ 1:0.3 ~ 1.
7. the method for direct induction Moth orchid indefinite bud according to claim 6, it is characterized in that: described anti-browning mixed solvent liquid is mixed by active carbon (AC) leaching liquor of 1.0g/L, the polyvinylpyrrolidone (PVP) of 1.0g/L, the ascorbic acid (Vc) of 0.5g/L and the glutathione of 0.5g/L, and wherein the mass ratio of AC leaching liquor, PVP, Vc, glutathione is 1:3.6:0.45:0.45.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103875698A (en) * | 2014-04-17 | 2014-06-25 | 四川国光农化股份有限公司 | Plant growth regulating composition and preparation thereof |
| CN105941161A (en) * | 2016-06-25 | 2016-09-21 | 合肥隆扬农业科技有限公司 | Tissue culture method of phalaenopsis |
| CN108184675A (en) * | 2018-03-15 | 2018-06-22 | 江苏恒诺园林建设有限公司 | Rapid germination culture medium for primary culture of sequoia zhongshanensis |
| CN109392649A (en) * | 2018-12-17 | 2019-03-01 | 英德君泓兰花股份有限公司 | A method of promoting the formation of hybrid orchid vegetative bud |
| 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 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103875698A (en) * | 2014-04-17 | 2014-06-25 | 四川国光农化股份有限公司 | Plant growth regulating composition and preparation thereof |
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| CN105941161A (en) * | 2016-06-25 | 2016-09-21 | 合肥隆扬农业科技有限公司 | Tissue culture method of phalaenopsis |
| CN108184675A (en) * | 2018-03-15 | 2018-06-22 | 江苏恒诺园林建设有限公司 | Rapid germination culture medium for primary culture of sequoia zhongshanensis |
| CN108184675B (en) * | 2018-03-15 | 2021-09-07 | 江苏恒诺园林建设有限公司 | Rapid germination culture medium for primary culture of sequoia zhongshanensis |
| CN109392649A (en) * | 2018-12-17 | 2019-03-01 | 英德君泓兰花股份有限公司 | A method of promoting the formation of hybrid orchid vegetative bud |
| 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 |
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| CN103988780B (en) | 2016-04-27 |
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