CN1528114A - Konjak in vitro rapid breeding method - Google Patents
Konjak in vitro rapid breeding method Download PDFInfo
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- CN1528114A CN1528114A CNA031469825A CN03146982A CN1528114A CN 1528114 A CN1528114 A CN 1528114A CN A031469825 A CNA031469825 A CN A031469825A CN 03146982 A CN03146982 A CN 03146982A CN 1528114 A CN1528114 A CN 1528114A
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Abstract
The present invention discloses a quickly breeding method of jonjak, said method includes the following five steps: (1) introduction; (2). obtaining sterile explant; (3) induction and proliferation of bud; (4) induction and culture of root; and (5) transplantation of test-tube plantlet.
Description
Technical field
The present invention relates to the cultivating method of plant variety, the method for in-vitro rapid propagation of concrete De Shi Amorphophalus rivieri potato.
Background technology
Amorphophalus rivieri potato (Tacca chantrieri Andre) has another name called arrow root potato, LAOHUXU, and black butterflies etc. are the herbaceos perennials of arrow root potato section, are the important representative kinds of China arrow root potato section, are born in the tropical rain forest more.The inflorescence of Amorphophalus rivieri potato is large-scale, flower, inflorescence involucre and the equal atropurpureus of interior bract, and this rarely has in flowers.Bract bar shaped in it, reach tens centimetres, quite unique, beautiful; The plant type of Amorphophalus rivieri potato is tall and straight, and no matter leaf belt leather matter is spent, inflorescence, plant all have very high ornamental value.The strong stem of root is usually used in controlling stomach ulcer, hypertension, hepatitis, external application burn, scald, sore wound, and among the people being used for loses weight, and the strong stem of root has very high medical value.The natural propagation of You Yu Amorphophalus rivieri potato is subjected to the influence of environment bigger; add artificial interference; its ecotope of depending on for existence is subjected to serious destruction day by day; the distribution of Amorphophalus rivieri potato is at present dwindled day by day; plant quantity constantly reduces, and ， Amorphophalus rivieri potato is listed in the three-level protective vulnerable species Zhong " Chinese Plants " announced in 91 years.
The Amorphophalus rivieri potato mainly relies on seminal propagation under field conditions (factors).Broadcast but need promptly adopt promptly, seed nature germination rate is the also available division propagation of Di ， Amorphophalus rivieri potato very, but reproduction coefficient is little.The fruit of Amorphophalus rivieri potato is a berry, but its berry is shrivelled midway or the person of physically well developing hangs over the outer power propagation of the last need of Hua Ting always, but it relies on the ability of external force not strong, thereby influences its propagation and breeding; Though be berry on the one hand, plant has certain toxicity, be difficult to by insect transmission, in addition, the feature that does not have remarkable anemophily to propagate.In addition, You Yong Amorphophalus rivieri potato among the people is made medicinal custom, thereby is subjected to artificial interference big.The Amorphophalus rivieri potato is a underground, and the habitat of the destruction also Shi Amorphophalus rivieri potato of forest environment has been subjected to threat, therefore, and in the minimizing day by day of natural world ， Amorphophalus rivieri potato.
Summary of the invention
The objective of the invention is to shortcoming at the prior art existence; Ti has supplied the method for in-vitro rapid propagation of Yi Zhong Amorphophalus rivieri potato; the population of this class De Amorphophalus rivieri in imminent danger potato is enlarged; carry out its good seed large-scale production, thereby on the basis of protection rare species resource, make it obtain develop and useining.
The method for in-vitro rapid propagation of the Amorphophalus rivieri potato of Ben Faming may further comprise the steps:
The first step, introduce a fine variety
To introduce De Amorphophalus rivieri potato from Yunnan and become strain to cut off leaf to stay root-like stock, root to take back, placing shade density is 80～90% cool canopy.Be planted in pond sludge: peat soil: in the matrix of rough sand=6: 3: 1, the humidity that keeps air is more than 80%.
The acquisition of second step, aseptic explant
The acquisition of the aseptic seedling of seed can obtain by aseptic seeding;
The aseptic seeding of seed has following two kinds of methods:
(1) indoor aseptic seeding
Seed is adopted the back at once with the running water flushing from maternal plant, and 70% alcohol is handled 10～15s, and aseptic water washing 3～5 times again with 0.1% mercuric chloride sterilization 3-5min, is inoculated behind the aseptic water washing 5 times, and dark culturing, temperature remain on 25～30 ℃.
(2) outdoor sowing
Seed is adopted the back at once with the running water flushing from maternal plant, broadcasts in the peat soil with the mode of program request, puts degree of shading and is in 80～90% the cool canopy, and the maintenance peat soil is moistening, and the relative moisture of air remains on more than 80%.
The inducing and breed of the 3rd step, bud
Inducing and breeding mainly from aseptic seedling of bud when aseptic seedling grows tall 1.5～2.0cm, downcut stem apex, is inoculated in medium (3), (4) and cultivates.Beginning first and second is commissioned to train when supporting, the stem apex base portion expands, color begins to transfer to faint yellow, and grows white to flaxen budlet, from the third generation, begin to extract out green blade, second or three generations after blade is extracted out, the spire induced strong goes out the white budlet of the about 0.1～0.2cm of diameter, and these budlets are received in medium (3), (4), about 30～35 days, these budlets all grew up to the about 0.3～0.5cm of diameter, high about 1.5cm, with the suitable strong bud of taking root of 1～2 leaf.
Medium: minimal medium is MS, 6-BA, KT, NAA, the IBA of additional variable concentrations, and main culture medium is:
(1)1/2MS，
(2)MS；
(3) MS+6-BA1.0～0.5+NAA0.3～0.05mg/L (following unit together);
(4)MS+6-BA2.0～0.5+KT2.0～1.0+NAA0.3～0.05；
(5)MS+6-BA0.5+IBA0.3
(6)1/2MS+IBA0.5；
Above medium adds agar 7g/L, pH5.4～5.6, and the consumption of medium (1), (2), (6) sugar is 20g/L, and the sugared consumption of medium (3)～(5) is 30g/L, and cultivation temperature is 25～30 ℃.Seed is cultivated with dark before rudiment, and all the other cultivate intensity of illumination is 1500～2000lx, illumination every day 10～12h;
The 4th step, the inducing and cultivating of root
The diameter that obtains in the step 3 clump bud greater than 0.3mm is inoculated among the 1/2MS that contains NAA, IBA, receives in the medium (6) again and cultivate, the plant base portion promptly can grow sturdy new after 30～35 days; The diameter that obtains in the step 3 can directly be inoculated in the medium (6) and carries out culture of rootage greater than the clump bud of 0.3mm.
The transplanting of the 5th step, test-tube plantlet
Hardening is 10～15 days in the cool canopy that places the test-tube plantlet that is about to transplant outside the culturing room or be about to transplant, the medium that clean test-tube plantlet adheres to, plant river sand: in the matrix of peat soil=6: 4, the seedbed need shade 90%, keep air humidity more than 80%, temperature remains on 22～30 ℃, transplants the back and sprays the 1/2MS nutrient solution in per 3 days.
The present invention compared with prior art has following advantage:
1, Zai Zi Ran circle Amorphophalus rivieri potato is with seed or division propagation, and the speed of breeding is slow, and this is one of its reason in imminent danger.The rapid propagation in vitro technology of this Xiang Amorphophalus rivieri potato is not at home and abroad appeared in the newspapers.Utilize this method can advise mould greatly and give birth to the good seedling of organizing of product Amorphophalus rivieri potato, thereby carry the population number of high Amorphophalus rivieri potato greatly, protected this endangered species.
2, the asexual quick-breeding method of this method employing has overcome the variation that produces in the seminal propagation, is selecting on the excellent basis, on a large scale Ti Gong Amorphophalus rivieri potato good seed and aseptic tissue cultivating seedling.
3, no matter be seminal propagation or division propagation You Yu Amorphophalus rivieri potato is low in natural world natural propagation rate, population is rare, it further develops and is restricted, and this method can be advised the living good seedling of organizing of Amorphophalus rivieri potato that produces of mould greatly, and this provides wide prospect for further Kai Fali with the Amorphophalus rivieri potato.
Embodiment
1, introduces a fine variety
Introduce the Amorphophalus rivieri potato originate in Yunnan in December, 1996 becomes strain 5 strains to cut off leaf to stay root-like stock, root to take back from tropical plants garden, Xishuangbanna, Yunnan, put degree of shading and be in 85% the cool canopy, be planted in pond sludge: peat soil: in the matrix of rough sand=6: 3: 1, keeping the relative moisture of air is 90%.
2, the acquisition of aseptic explant
The acquisition of the aseptic seedling of seed can obtain by indoor, outdoor aseptic seeding:
Collected seed respectively from 1997～2000 years and to carry out germination test, (withered on Hua Ting with it is standard) is adopted and is broadcast after the August in every year berry maturation, broadcasts back 60 days statistics germination rates.
The method of indoor seeding: seed is adopted the back at once with the running water flushing from maternal plant, and 70% alcohol is handled 12s, and aseptic water washing 5 times again with 0.1% mercuric chloride sterilization 3min, is inoculated behind the aseptic water washing 5 times, and dark culturing, temperature remain on 25 ℃.The seed that receive 1 year (1997) only is seeded in 1/2MS and the MS medium, and the result is not all germinateed, for this reason, 1998,1999 with the direct aseptic seeding of said method, obtain 235 of aseptic seeds, but all seeds do not germinate all.2000, divide two groups, with the direct aseptic seeding of said method, but therein one group add antiseptic gauze, seed is placed on the gauze, found that two seed sproutings are arranged having on one group on the gauze, germination rate is 0.8% (seeing Table 1).
Outdoor sowing: seed is adopted the back at once with the running water flushing from maternal plant, broadcasts in the peat soil with the method for program request, puts degree of shading and is in 85% the cool canopy, and the maintenance peat soil is moistening, and the relative moisture of air remains on more than 80%.
3, bud induces and breeds
Inducing and breeding mainly from aseptic seedling of bud when aseptic seedling grows tall 1.8cm, downcut stem apex, is inoculated in medium (3), (4) and cultivates.Beginning first and second is commissioned to train when supporting, the stem apex base portion expands, color begins to transfer to faint yellow, and grows white to flaxen budlet, from the third generation, begin to extract out green blade, second or three generations after blade is extracted out, the spire induced strong goes out the white budlet of the about 0.1cm of diameter, and these budlets are received in medium (3), (4), spend 32 days, these budlets all grow up to diameter 0.35cm, high 1.5cm, with the suitable strong bud of taking root of 1 leaf.
Medium is MS+6-BA1.0～0.5+NAA0.3～0.05mg/L.
Cultivation temperature is 30 ℃.Seed is cultivated with dark before rudiment, and all the other cultivate intensity of illumination is 15001x, illumination every day 12h.
4, root inducing and cultivating
The diameter that obtains in the step 3 is inoculated in the medium (6) greater than the clump bud of 0.3mm cultivates, the plant base portion promptly can grow sturdy new after 33 days.(table Er Wei Amorphophalus rivieri potato rooting rate statistical form).
5, the transplanting of test-tube plantlet
Hardening is 15 days in the cool canopy that places the test-tube plantlet that is about to transplant outside the culturing room or be about to transplant, the medium that clean test-tube plantlet adheres to, plant river sand: in the matrix of peat soil=6: 4, the seedbed need shade 90%, keep air humidity 85%, temperature remains on 25 ℃, transplants the back and sprays the 1/2MS nutrient solution in per 3 days.(the planting percent statistical form of table San Wei Amorphophalus rivieri potato).
Biao Yi Amorphophalus rivieri potato seed germination rate
| Experimental period | Experimental technique | The experiment seed number | Germinative number | Add up to | Germination percentage | ||
| In August, 1997 | Outdoor | 10 | 0 | 30 | 0 | ||
| Indoor | Method one | 10 | 0 | 0 | |||
| Method two | 10 | 0 | 0 | ||||
| In August, 1998 | Outdoor | 50 | 3 | 110 | 6％ | ||
| Indoor | Method one | 30 | 0 | 0 | |||
| Method two | 30 | 0 | 0 | ||||
| In August, 1999 | Outdoor | 50 | 5 | 300 | 10％ | ||
| Indoor | Method one | 150 | 0 | 0 | |||
| Method two | 100 | 0 | 0 | ||||
| In August, 2000 | First group | Outdoor | 100 | 6 | 450 | 6％ | |
| Indoor | Method one | 50 | 0 | 0 | |||
| Method two | 50 | 0 | |||||
| Second group | Method one | 150 | 1 | 0.8％ | |||
| Method two | 100 | 1 | |||||
| Average germination percentage | Indoor: 0.8%, outdoor: 5.5% | ||||||
Biao Er Amorphophalus rivieri potato rooting rate statistical form
| The inoculation date | Plant quantity (strain) | Every strain has root more than two (strain) | Every strain has a root (strain) | Unrooted (strain) | Rooting rate (%) |
| 2001/8/20 | ?100 | ?80 | ?8 | ?12 | ?80 |
| 2001/10/16 | ?100 | ?90 | ?2 | ?8 | ?90 |
| 2002/3/15 | ?100 | ?90 | ?4 | ?6 | ?90 |
| 2002/4/17 | ?100 | ?89 | ?2 | ?9 | ?89 |
| Average rooting rate | 87.25％ | ||||
The planting percent statistical form of Biao San Amorphophalus rivieri potato
| Production time | Produce number (strain) | Field planting number (strain) | Indoor percent of loss (%) | Go out garden number (strain) | The survival rate of transplanting seedlings (%) |
| 2001/8/20-24 | ?1085 | ?1035 | ?4.6 | ?725 | ?70 |
| 2001/10/16 | ?1000 | ?980 | ?2.0 | ?735 | ?75 |
| 2002/1/18-25 | ?2350 | ?2325 | ?1.1 | ?1651 | ?71 |
| 2002/3/15 | ?685 | ?675 | ?1.5 | ?540 | ?80 |
| 2002/4/15 | ?1970 | ?1900 | ?3.6 | ?1520 | ?80 |
| Add up to | ?7090 | ?695 | ?5171 | ||
| Mean (%) | ?2.6 | ?75.2 |
Claims (1)
1, the method for quickly breeding of Yi Zhong Amorphophalus rivieri potato, form by following step successively:
The first step, introduce a fine variety
To introduce De Amorphophalus rivieri potato from Yunnan and become strain to cut off leaf to stay root-like stock, root to take back, placing shade density is 80～90% cool canopy, is planted in pond sludge: peat soil: in the matrix of rough sand=6: 3: 1, the humidity that keeps air is more than 80%;
The acquisition of second step, aseptic explant
The acquisition of the aseptic seedling of seed can obtain by aseptic seeding;
The aseptic seeding of seed has following two kinds of methods:
(1) indoor aseptic seeding
Seed is adopted the back at once with the running water flushing from maternal plant, and 70% alcohol is handled 10～15s, and aseptic water washing 3～5 times again with 0.1% mercuric chloride sterilization 3-5min, is inoculated behind the aseptic water washing 5 times, and dark culturing, temperature remain on 25～30 ℃;
(2) outdoor sowing
Seed is adopted the back at once with the running water flushing from maternal plant, broadcasts in the peat soil with the mode of program request, puts degree of shading and is in 80～90% the cool canopy, and the maintenance peat soil is moistening, and the relative moisture of air remains on more than 80%;
The inducing and breed of the 3rd step, bud
Inducing and breed of bud mainly from aseptic seedling, when aseptic seedling grows tall 1.5～2.0cm, stem apex is downcut, be inoculated in the medium and cultivate, beginning first, two are commissioned to train when supporting, the stem apex base portion expands, color begins to transfer to faint yellow, and grows white to flaxen budlet, from the third generation, begin to extract out green blade, second or three generations after blade is extracted out, the spire induced strong goes out the white budlet of the about 0.1～0.2cm of diameter, and these budlets are received in the medium, about 30～35 days, these budlets all grew up to the about 0.3～0.5cm of diameter, high about 1.5cm, suitable strong bud of taking root with 1～2 leaf;
The 4th step, the inducing and cultivating of root
The diameter that obtains in the step 3 clump bud greater than 0.3mm is inoculated among the 1/2MS that contains NAA, IBA, receives in the medium and cultivate, the plant base portion promptly can grow sturdy new after 30～35 days;
The transplanting of the 5th step, test-tube plantlet
Hardening is 10～15 days in the cool canopy that places the test-tube plantlet that is about to transplant outside the culturing room or be about to transplant, the medium that clean test-tube plantlet adheres to, plant river sand: in the matrix of peat soil=6: 4, the seedbed need shade 90%, keep air humidity more than 80%, temperature remains on 22～30 ℃, transplants the back and sprays the 1/2MS nutrient solution in per 3 days.
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|---|---|---|---|
| CN 03146982 CN1265696C (en) | 2003-09-28 | 2003-09-28 | Konjak in vitro rapid breeding method |
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| CN 03146982 CN1265696C (en) | 2003-09-28 | 2003-09-28 | Konjak in vitro rapid breeding method |
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| CN1265696C CN1265696C (en) | 2006-07-26 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107318305A (en) * | 2017-08-18 | 2017-11-07 | 安康学院 | A kind of konjaku true seed fast breeding method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107318305A (en) * | 2017-08-18 | 2017-11-07 | 安康学院 | A kind of konjaku true seed fast breeding method |
| CN107318305B (en) * | 2017-08-18 | 2020-12-25 | 安康学院 | Rapid breeding method for konjac seedling seeds |
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Granted publication date: 20060726 Termination date: 20130928 |