CN100355333C - Bulbil konjak sprout breeding method - Google Patents
Bulbil konjak sprout breeding method Download PDFInfo
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- CN100355333C CN100355333C CNB2004100407230A CN200410040723A CN100355333C CN 100355333 C CN100355333 C CN 100355333C CN B2004100407230 A CNB2004100407230 A CN B2004100407230A CN 200410040723 A CN200410040723 A CN 200410040723A CN 100355333 C CN100355333 C CN 100355333C
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Abstract
The present invention relates to a bulbule konjak sprout breeding method. The method for breeding a bulbule konjak by tissue cultivation technology comprises: bulbules or leaves or leaf stalks or tuber sprouts (tender sprouts) or scale leaves or tuber slices of bulbule konjaks are firstly put into MS or 1/2 MS or N6 basal cultivation media, and 0 to 5 mg. L <-1> of plant hormones such as BA or Kt, or the combination of plant hormones such as BA+NAA or BA+IAA or Kt+NAA or Kt+IAA is added to the basal cultivation media, wherein the concentration of sucrose is 3%, a pH value is 5.8, light irradiation time is 12h. d <-1>, illumination intensity is from 1000 to 1500 lx, and a cultivation temperature is 24+/-1 DEG C; callus blocks or blocks of adventitious bud tissue are moved into cultivation media with MS+BA0.5+NAA0.2 to grow continuously, wherein the callus blocks or the blocks of adventitious bud tissue are generated outside all of the explants; after the irregular shapes of the callus blocks or the blocks of adventitious bud tissue are formed, the callus blocks or the blocks of adventitious bud tissue are cut for secondary cultivation; test-tube sprout clusters are then transferred to 1/2 MS cultivation media only containing growth hormones for rootage cultivation; the test-tube sprouts are directly transferred into basal substances without special treatment after 20 days.
Description
Technical field: the invention belongs to biological technical field, particularly, relate to the mating system of a plant species bulbil konjak (Amorphophallus bulbifer) seedling.
Background technology: bulbil konjak (Amorphophallus bulbifer), be commonly called as " red konjaku ", be the perennial perennial root herbaceous plant of Araeceae (Araceae).Its underground loose stem tuber can be used as health care and functional food, and has a wide range of applications at aspects such as industry and medicine.Particularly outstanding is, and its GM (glucomannans) content substantially exceeds white konjaku, and have the output height, disease resistance is strong and advantage such as wide adaptability, is worth focus development.Konjaku cultivation sowing quantity is big, at the bottom of the stem tuber reproduction coefficient; Grain weight is few, and variation is big.Along with the development of konjaku industry, ten minutes active demand high-quality konjaku seedling.Do not see in the prior art bulbil konjak tissue culture and fast numerous successful report are arranged.
Summary of the invention: the mating system that the object of the present invention is to provide a kind of bulbil konjak (Amorphophallusbulbifer) seedling.
The objective of the invention is to realize by following technical scheme:
A kind of bulbil taro seedling breeding method of rubbing is utilized the tissue culture technique breeding bulbil taro that rubs, and getting the bulbil of bulbil konjak or blade or petiole or root or stem tuber bud earlier is tender shoots or scale leaf or stem tuber section, at MS or 1/2MS or N
6Additional 0-5mg.L in the minimal medium
-1Plant hormone BA or Kt or plant hormone combination BA+NAA or BA+IAA or Kt+NAA or Kt+IAA, sucrose concentration 3%, pH value=5.8, light application time 12hd
-1Intensity of illumination 1000-1500lx, cultivation temperature is 24 ± 1 ℃ and cultivates down, continued growth in the medium of callus that above each explant is produced and indefinite bud piece of tissue immigration MS+BA 0.5+NAA0.2 then, cut subculture after forming irregular structure, again test tube bud clump is changed on the 1/2MS medium that only contains growth hormone and carry out culture of rootage, after 20 days, test-tube plantlet directly do not moved in the matrix of specially treated, do not get final product.
Embodiment: followingly further specify flesh and blood of the present invention, but content of the present invention is not exceeded with embodiment with embodiments of the invention.
Embodiment 1
One, experiment material and method
The bulbil of field acquisition bulbil konjak, and plant leaf blade, petiole, root, stem tuber bud (tender shoots), scale leaf and stem tuber section.After cleaning up,, use 0.2%HgCl again with 70% ethanol disinfection several seconds with flowing water
2Sterilize 10-30 minute (the concrete time decides according to the material size).For several times at last, up to the disinfectant residue is removed with sterile water wash.Minimal medium (MS, 1/2MS, N have been tested
6), plant hormone and combination (BA, Kt, BA+NAA, BA+IAA, Kt+NAA and Kt+IAA etc.) and phytohormone concentration (0-5mg.L
-1) wait the influence of explant evoked callus with the differentiation indefinite bud.Sucrose concentration 3%, pH value=5.8, light application time 12hd
-1, intensity of illumination 1000-1500lx, cultivation temperature is 24 ± 1 ℃.
Two, experimental result
Inducing and breed of 1 bud
1.1 the bulbil of bulbil bulbil konjak is at suitable culture medium (1/2MS+BA 2mgL
-1(unit down together)+IAA0.2) goes up 9d and begins to sprout (broadcasting 170d in advance than soil).The big first end in contact medium of the bulbil place 3-7 sheet that splits is the cauliflower shape.24d produces the clump bud, and each bulbil has 13 buds at most.The beginning bud is by several pieces scale leaf parcels, and petiole extends then, and solely leaf is carried out, and 98d can obtain the complete test-tube plantlet of high about 10cm.Along with incubation time prolongs, whole bulbil callusization, and have above indefinite bud is distributed in.Can cut the shoot proliferation rate and reach 50d/l: 3.
1.2 stem apex stem tuber bud is placed on the medium of MS+BA 1.0+NAA0.1, its incision 15d produces white yellow warty callus, the little brown point-like of last cloth thing, and 42d produces indefinite bud, about 4 buds/piece.The rate of increase is 50d/l: 2.
1.3 the undercut section is seeded in MS+BA with the undercut section
0.2+ NAA
2Medium on, 39d produces white granular callus group in the stem tuber segment, 69d produces indefinite bud, about 2 bud/sections, the about 75% differentiation clump bud of statistics after 3 months, it is 1: 2 that clone is built up the back 50d rate of increase.
1.4 scale leaf scale leaf (promptly wrapping up the scale of tender shoots) is seeded on the medium of MS+BA1+NAA0.2,24d explant callusization is also grown up, and inductivity reaches 100%, and 47d has 50% differentiation clump bud, and more than every 10 bud, growth rate is 50d/l: 4.
1.5 blade is 0.5-1cm with blade cuts
2Fritter, be seeded on the medium of MS+BA 0.5+NAA0.1, produce callus at nearly vein place about 4d, 49d produces the clump bud, about 6 buds/piece, inductivity reaches 54%, differentiation rate 38.7%, rate of increase 50d/l: 3.
1.6 petiole is cut into petiole on the medium that the segment that is about 0.8cm is seeded in MS+BA0.3, the segment of 18d about 78% produces callus, and generally in morphologic lower end or two head lengths, but the callus that the lower end produces is more flourishing.39d breaks up indefinite bud, and differentiation rate is 100%, and more than every hop count bud to 10 bud, growth rate is 30d/l: 4.
1.7 the stem tuber stripping and slicing is seeded in the stem tuber stripping and slicing on the medium of MS+Kt2+IAA0.2, cardinal extremity is closely cultivated in the stripping and slicing of about 20d stem tuber and periphery grows the yellowish-brown callus, and along with the prolongation of incubation time, the stem tuber stripping and slicing of band callus is grown up, last callusization fully, and differentiate indefinite bud on its surface, and be scattered shape, can cut subculture.
Inducing of 2 similar tuberosity things
More than the callus that produces of each explant and indefinite bud piece of tissue on the medium of MS+BA 0.5+NAA0.2, can continue to grow up, form irregular similar tuberosity structure, the epidermis light green, inner white can be cut subculture, bud can grow tall and take root on it.
3 take root and transplant
Test tube bud clump is cut into the simple bud bar, is placed on the 1/2MS medium that only contains growth hormone, and about 20d, the long root of budling base portion, rooting rate reaches about 95%, has formed complete test-tube plantlet.With with similar tuberosity thing (size is about 1cm
2To the test-tube plantlet of 2.5cm * 1.5cm), without practicing seedling, directly immigration is not in the matrix of specially treated, and survival rate reaches 100%, but low level management behind the 30d.Now successfully shift out 4 batches.
Bulbil konjak is only leaf at occurring in nature, sees the phenomenon of 1 strain, 7 leaves in the group training process.Scale leaf also has 3-4 sheet nearly.Scale leaf is easier to success as explant in the group training.Train the later stage in group, the clump bud produces for a long time, makes the propagation object with scale leaf, seldom produces browning, and the material acquisition easily, and this can be used as a new way of breeding test-tube plantlet.Scale leaf produces callus and becomes graininess, and microscopy is the cells,primordial of meristematic cell tissue, therefore can guarantee its genetic stability.
Bulbil konjak is because of being rich in aldehydes matter, brownization very easily in incubation, external-talent's necrosis.The present invention is combined as BA+NAA or BA+IAA by changing plant hormone, or the consumption of reduction BA+NAA etc., solved this problem.Bulbil konjak tissue culture and fast numerous successful report are not arranged in the prior art.Root segment is cultivated successfully and is not also appeared in the newspapers in the training of Amorphophallus group.The tissue of the same plant species of description of test of the present invention is different with organ position of living in, and it requires also different to condition of culture.
Claims (1)
1, bulbil konjak seedling breeding method, utilize method for tissue culture breeding bulbil konjak, it is characterized in that getting bulbil, blade, petiole, root, stem tuber bud, scale leaf or the stem tuber of bulbil konjak, section is cultivated in following medium as explant: additional BA 2mg.L in the 1/2MS minimal medium
-1+ IAA 0.2mg.L
-1, or the MS minimal medium in additional Kt 2mg.L
-1+ IAA0.2mg.L
-1, or the MS minimal medium in additional BA0.2-1mg.L
-1+ NAA 0.1-2mg.L
-1, sucrose concentration 3% in above-mentioned each medium, pH value=5.8, light application time 12hd
-1, intensity of illumination 1000-15001x, cultivation temperature is 24+1 ℃ cultivates down, and above each explant callus that produces and the indefinite bud piece of tissue that each explant produces are moved into MS+BA 0.5mg.L
-1+ NAA0.2mg.L
-1Medium in continued growth, after forming irregular stem tuber shape structure, the cutting subculture changes the test tube bud clump that produces on the 1/2MS medium that only contains growth hormone over to then and carries out culture of rootage, after 20 days, the direct immigration of test-tube plantlet that obtains is not passed through in the matrix of specially treated.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2004100407230A CN100355333C (en) | 2004-09-17 | 2004-09-17 | Bulbil konjak sprout breeding method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100407230A CN100355333C (en) | 2004-09-17 | 2004-09-17 | Bulbil konjak sprout breeding method |
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| CN1596618A CN1596618A (en) | 2005-03-23 |
| CN100355333C true CN100355333C (en) | 2007-12-19 |
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| CNB2004100407230A Expired - Fee Related CN100355333C (en) | 2004-09-17 | 2004-09-17 | Bulbil konjak sprout breeding method |
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100421547C (en) * | 2005-11-21 | 2008-10-01 | 云南耕耘魔芋资源开发有限责任公司 | Bulbil konjak artificial induced flowering asexual breeding method |
| CN102498880A (en) * | 2011-11-04 | 2012-06-20 | 云南农业大学 | Cutting seedling raising method of rodgersflower rhizome stem tubers |
| CN102763597B (en) * | 2012-08-15 | 2013-11-20 | 云南省农业科学院生物技术与种质资源研究所 | Two-generation in one year reproduction method for konjac cross breeding |
| CN102939902A (en) * | 2012-11-21 | 2013-02-27 | 昆明学院 | Rapid propagation method of amorphophallus bulbifer through tissue culture |
| CN106417034A (en) * | 2016-11-14 | 2017-02-22 | 昆明学院 | Reproduction method for tissue culture of amorphophallus bulbifer |
| CN107211891B (en) * | 2017-05-27 | 2020-09-04 | 安康市农业科学研究所 | Konjak callus direct seeding and rapid propagation technology |
| CN111602596B (en) * | 2020-07-17 | 2021-10-01 | 中国热带农业科学院橡胶研究所 | Method for obtaining regeneration plant by tissue culture of bulbil konjak flower device |
| CN112106665A (en) * | 2020-10-28 | 2020-12-22 | 中国科学院昆明植物研究所 | Culture medium for one-step seedling formation of amorphophallus rivieri and application and tissue culture method |
| CN113678653B (en) * | 2021-09-24 | 2022-12-23 | 云南省热带作物科学研究所 | Konjac seedling propagation method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1295790A (en) * | 1999-11-14 | 2001-05-23 | 云南省农业科学院生物技术研究所 | Amorphophallus rivieri group seedling-culturing batch production and cultivation technology |
| CN1493186A (en) * | 2003-08-09 | 2004-05-05 | 云南省农业科学院生物技术研究所 | Breeding technology of konjak testtube taro |
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2004
- 2004-09-17 CN CNB2004100407230A patent/CN100355333C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1295790A (en) * | 1999-11-14 | 2001-05-23 | 云南省农业科学院生物技术研究所 | Amorphophallus rivieri group seedling-culturing batch production and cultivation technology |
| CN1493186A (en) * | 2003-08-09 | 2004-05-05 | 云南省农业科学院生物技术研究所 | Breeding technology of konjak testtube taro |
Non-Patent Citations (4)
| Title |
|---|
| 影响魔芋愈伤组织形成的几个因素 马林，张玲，李山峰.广西植物，第23卷第6期 2003 * |
| 魔芋( Amor phophallus)离体繁殖研究 柳，俊，谢从华，余展深，刘，勇.华中农业大学学报，第20卷第3期 2001 * |
| 魔芋组织培养及快繁技术研究 吕世安，沈艳芬，黄元勋，陈永波，柳文录，盛德贤，于斌武.湖北民族学院学报(自然科学版)，第21卷第2期 2003 * |
| 魔芋组织培养技术研究进展 刘贵周，谢庆华，赵庆云，谢世清.中国农学通报，第19卷第4期 2003 * |
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