CN104737911A - Quick cultivation method for rhizoma bletillae tissue culture seedlings - Google Patents

Quick cultivation method for rhizoma bletillae tissue culture seedlings Download PDF

Info

Publication number
CN104737911A
CN104737911A CN201510143716.1A CN201510143716A CN104737911A CN 104737911 A CN104737911 A CN 104737911A CN 201510143716 A CN201510143716 A CN 201510143716A CN 104737911 A CN104737911 A CN 104737911A
Authority
CN
China
Prior art keywords
bletilla
condition
medium
breeding method
fast breeding
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201510143716.1A
Other languages
Chinese (zh)
Inventor
李宏哲
蔡忠虎
熊浩宇
张超
彭安顺
杨林芳
夏光惠
姚雪娇
岳蕾
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Yunnan University of Traditional Chinese Medicine TCM
Original Assignee
Yunnan University of Traditional Chinese Medicine TCM
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Yunnan University of Traditional Chinese Medicine TCM filed Critical Yunnan University of Traditional Chinese Medicine TCM
Priority to CN201510143716.1A priority Critical patent/CN104737911A/en
Publication of CN104737911A publication Critical patent/CN104737911A/en
Pending legal-status Critical Current

Links

Landscapes

  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention discloses a quick cultivation method for rhizoma bletillae tissue culture seedlings. The method comprises the following steps: selecting intact non-cracked rhizoma bletillae capsules and disinfecting the capsules by 75% alcohol for 30 seconds; then soaking and disinfecting the capsules by using 0.2% mercury bichloride for 10 minutes; washing 6 times by sterile water; fully and thoroughly washing, airing and uniformly spreading on a culture medium to cultivate; and when the seeds germinate to obtain seedlings which are about 2cm long, inoculating 3-4 seedlings as a cluster to a strong sprout culture medium to be cultured. Compared with the conventional tissue culture seedlings obtained by sterile germination of rhizoma bletillae seeds, the method just needs subculture once, so that the time consumed is short and at least three months are saved.

Description

A kind of fast breeding method of bletilla plantlet in vitro
Technical field
The invention belongs to traditional Chinese medicine planting technology field, be specifically related to a kind of fast breeding method of bletilla plantlet in vitro.
Background technology
Bletilla is perennial herb.Pseudobulb block root shape, white, plump, there is finger-like difference.Spend beautiful greatly, aubergine, can view and admire for cultivation.Pseudobulb is containing gum bletilla, starch, volatilization wet goods composition, and have the effects such as convergence, tonifying lung hemostasis, detumescence, oral administration external application, is applied to the diseases such as hemoptysis haematemesis, traumatism and bleeding, sore swollen toxin, chapped skin, pulmonary tuberculosis hemoptysis, ulcer haemorrhage.Bletilla is a kind of wild plant day by day in imminent danger, and the condition of seed germination is harsh, needs and mycosymbiosis, adds that Seedling Stage is longer, to environment sensitive, is therefore difficult to effectively be bred by seed under natural environment situation.Along with the exhaustion of wild resource, the rising all the way of the Bletilla striata medicinal materials market price, has broken through 400 yuan/kg high point.Under stimulation so at high price, finally remaining wild bletilla resource is estimated to exhaust in the near future.Under the ordering about of interests, plantation family start to seek bletilla artificial cultivation, but due to seedling very deficient, be difficult to scale development, even can not start to walk.Therefore, the technical problem of seedling breeding must be solved, just can break the restriction of bletilla seedling bottleneck.
Summary of the invention
The object of the present invention is to provide a kind of fast breeding method of bletilla plantlet in vitro.The method is simple to operate, and the seed germination time is short, the cost-saving and time.
The present invention realizes especially by following technical scheme:
A fast breeding method for bletilla plantlet in vitro, comprises the following steps:
1) select intact bletilla capsule with 75% alcohol-pickled sterilization 30s;
2) with 0.2% mercuric chloride by capsule soaking disinfection 10min, aseptic water washing 6 times, fully rinses well;
3) will disinfect and the capsule dried cut, seed is sown on medium equably, cultivates according to condition of culture;
4) seedling after seed germination having been grown to 2 centimetres is forwarded in strong seedling culture base by 3 ~ 4 one clump and cultivates.
Further, step 3) described in medium prepare by the following method: adopt C medium, add each mother liquor successively, NAA 1.0mg/L, potato 200g/L (stripping and slicing liquor 0.5h, only get juice), banana 40g/L pulls an oar, sucrose 30g/L, agar 4g/L, constant volume, surveys pH value to 6.0.
Further, step 3) described in condition of culture be: 25 ± 2 DEG C, illuminance be in the culturing room of 1500lux, 12h illumination, 12h dark cultivate three months.
Further, step 4) described in strong seedling culture base prepare by the following method: adopt 1/2MS medium, add each mother liquor successively, NAA 0.5mg/L, 6-BA 0.2mg/L, potato tubers making beating directly adds 10g/L, banana making beating 75g/L, sucrose 20g/L, agar 4g/L, active carbon 0.4g/L, constant volume, surveys pH value to 6.0.
Further, step 4) in condition of culture be: 25 ± 2 DEG C, illuminance is 1500lux, 12h illumination, cultivate in the culturing room of 12h dark, pseudobulb to be grown and root can carry out hardening.
Beneficial effect of the present invention is: adopt seed group training, the seedling pollution rate obtained is low, differentiation rate is high, stabilization characteristics of genetics, active principle content is high, compared with the method obtains plantlet in vitro with traditional bletilla seed asepsis sprouting simultaneously, only needs subculture once, used time is short, has saved at least trimestral time.
Embodiment
Below in conjunction with embodiment, the present invention is described further, the following stated, only to preferred embodiment of the present invention, not do other forms of restriction to the present invention, any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed to the Equivalent embodiments of equal change.Everyly do not depart from the present invention program's content, any simple modification done following examples according to technical spirit of the present invention or equivalent variations, all drop in protection scope of the present invention.
Embodiment 1
One, bletilla seed asepsis sprouting
1, the preparation of medium
Adopt C medium, add each mother liquor successively, NAA 1.0mg/L, potato 200g/L (stripping and slicing liquor 0.5h, only gets juice), banana 40g/L pulls an oar, sucrose 30g/L, agar 4g/L.Constant volume, about surveying pH value to 6.0.
2, bletilla capsule/seed disinfection
This operates on superclean bench and carries out.If bletilla capsule is intact not splitting, the thorough wiping bletilla of the alcohol with 75% capsule, or soak 30s, pour out alcohol; If capsule ftractures, the pouch parcel seed made with highdensity silk stocking, sterilization method is the same.
Use the mercuric chloride of 0.2% by above-mentioned capsule/seed disinfection 10min again, sterile water fully rinses 6 times.Capsule/seed is dried in the air to parching completely on superclean bench.
3, bletilla seed inoculation
Will disinfect and the capsule dried incision, or directly the seed disinfected is sown on medium equably, density appropriateness.
4, cultivate
The seed inoculated is placed on 25 ± 2 DEG C, illuminance be in the culturing room of 1500lux (12 h light, 12 h dark) cultivate three months.
Two, the strong plantlets and rootage of bletilla plantlet in vitro
1, the preparation of Rooting and hardening-off culture base
Adopt 1/2MS medium, add each mother liquor successively, NAA 0.5mg/L, 6-BA 0.2mg/L, potato tubers making beating directly adds 10g/L, banana making beating 75g/L, sucrose 20g/L, agar 4g/L, active carbon 0.4g/L.Constant volume, about surveying pH value to 6.0.
2, inoculate
Seedling seed germination having been grown to 2 cm is forwarded in strong seedling culture base by 3 ~ 4 one clump.
3, cultivate
The seedling inoculated is placed on 25 ± 2 DEG C, illuminance is cultivate in the culturing room of 1500lux (12 h light, 12 h dark).The bletilla having grown pseudobulb and root after three months can carry out hardening.
Embodiment 2
On December 18th, 2013, the experiment of bletilla seed asepsis sprouting is carried out in laboratory, Yunnan University of Traditional Chinese Medicine Yi Jianji building 6301 ~ 6303.Select 10 capsules, sow 400 bottles altogether.Because bletilla seed is tiny, and maturity between capsule is inconsistent, concrete emergence rate can only rough estimate for being greater than 80%.
After cultivating three wheat harvesting periods, on March 25th, 2014, the bletilla seedling growing to 2 ~ 3 centimetres high is gone to squamous subculture in strong seedling culture base by 3 ~ 4 one clump.
After cultivating three wheat harvesting periods, on July 15th, 2014, the bletilla seedling growing pseudobulb and root is delivered to little Shao planting base, Kunming.After the hardening of ten days, plant in ground, cover pine needle and take shade net.After two months, most seedling grows fine, and statistics survival rate is to 92.36%.In January, 2015, dig out bletilla pseudobulb statistics size after seedling, mean size is 1.78 centimetres.
Facts have proved that the enforcement of this method can shorten the cultivation cycle of bletilla, and good result can be reached.

Claims (5)

1. a fast breeding method for bletilla plantlet in vitro, is characterized in that: comprise the following steps:
1) select intact bletilla capsule with 75% alcohol-pickled sterilization 30s;
2) with 0.2% mercuric chloride by capsule soaking disinfection 10min, aseptic water washing 6 times, fully rinses well;
3) will disinfect and the capsule dried cut, seed is sown on medium equably, cultivates according to condition of culture;
4) seedling after seed germination having been grown to 2 centimetres is forwarded in strong seedling culture base by 3 ~ 4 one clump and cultivates.
2. the fast breeding method of a kind of bletilla plantlet in vitro according to claim 1, it is characterized in that: step 3) described in medium prepare by the following method: adopt C medium, add each mother liquor successively, NAA 1.0mg/L, potato juice 200g/L, banana 40g/L pull an oar, sucrose 30g/L, agar 4g/L, constant volume, surveys pH value to 6.0.
3. the fast breeding method of a kind of bletilla plantlet in vitro according to claim 1, it is characterized in that: step 3) described in condition of culture be: 25 ± 2 DEG C, under illuminance is 1500lux condition, cultivate three months in the culturing room of 12h illumination, 12h dark.
4. the fast breeding method of a kind of bletilla plantlet in vitro according to claim 1, it is characterized in that: step 4) described in strong seedling culture base prepare by the following method: adopt 1/2MS medium, add each mother liquor successively, NAA 0.5mg/L, 6-BA 0.2mg/L, potato tubers making beating directly adds 10g/L, banana making beating 75g/L, sucrose 20g/L, agar 4g/L, active carbon 0.4g/L, constant volume, surveys pH value to 6.0.
5. the fast breeding method of a kind of bletilla plantlet in vitro according to claim 1, it is characterized in that: step 4) in condition of culture be: 25 ± 2 DEG C, under illuminance is 1500lux condition, cultivate in the culturing room of 12h illumination, 12h dark, pseudobulb to be grown and root can carry out hardening.
CN201510143716.1A 2015-03-30 2015-03-30 Quick cultivation method for rhizoma bletillae tissue culture seedlings Pending CN104737911A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510143716.1A CN104737911A (en) 2015-03-30 2015-03-30 Quick cultivation method for rhizoma bletillae tissue culture seedlings

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510143716.1A CN104737911A (en) 2015-03-30 2015-03-30 Quick cultivation method for rhizoma bletillae tissue culture seedlings

Publications (1)

Publication Number Publication Date
CN104737911A true CN104737911A (en) 2015-07-01

Family

ID=53578636

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510143716.1A Pending CN104737911A (en) 2015-03-30 2015-03-30 Quick cultivation method for rhizoma bletillae tissue culture seedlings

Country Status (1)

Country Link
CN (1) CN104737911A (en)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105145352A (en) * 2015-08-03 2015-12-16 河南科技大学 Efficient tissue culture and rapid propagation technology for seedlings of bletilla striata
CN105145361A (en) * 2015-09-17 2015-12-16 康美药业(文山)药材种植管理有限公司 Method for efficiently breeding tissue culture seedlings of bletilla striata and planting method of bletilla striata
CN105165368A (en) * 2015-10-15 2015-12-23 向君民 Method for rapidly breeding seedlings by using bletilla striata tubers
CN105210875A (en) * 2015-10-15 2016-01-06 铜仁市金农绿色农业科技有限公司 A kind of bletilla striata quick propagation method
CN106416972A (en) * 2016-08-29 2017-02-22 浙江金石生物科技有限公司 Method for quickly cultivating bletilla striata seedlings
CN107047310A (en) * 2017-05-10 2017-08-18 文山学院 A kind of cultural method of bletilla striata seeds culture seedling
CN109169273A (en) * 2018-09-03 2019-01-11 云南中医学院 A kind of breeding method of bletilla proliferation pseudobulb
CN110679421A (en) * 2019-11-06 2020-01-14 安徽东方金桥农林科技股份有限公司 Bletilla striata seedling cultivation method
CN116649216A (en) * 2023-06-25 2023-08-29 江苏护理职业学院 Pure bletilla striata three-fork tissue culture domesticated seedling raising method

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20030027161A (en) * 2001-09-13 2003-04-07 충청북도 (관리부서:충청북도 농업기술원) New cultivar of Bletilla striata 'CHUNG BUKJARAN'
CN101940158A (en) * 2009-07-09 2011-01-12 正安县绿野科技产业有限责任公司 Method for rapidly propagating plantlets by utilizing bletilla striata seeds
CN103814821A (en) * 2014-01-03 2014-05-28 中华全国供销合作总社南京野生植物综合利用研究所 Method for efficiently and quickly producing bletilla striata seedling
CN104137778A (en) * 2014-07-30 2014-11-12 中国科学院南京分院东台滩涂研究院 Method for culturing bletilla striata protocorm by means of bletilla striata seeds according to fluid suspension culture method

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20030027161A (en) * 2001-09-13 2003-04-07 충청북도 (관리부서:충청북도 농업기술원) New cultivar of Bletilla striata 'CHUNG BUKJARAN'
CN101940158A (en) * 2009-07-09 2011-01-12 正安县绿野科技产业有限责任公司 Method for rapidly propagating plantlets by utilizing bletilla striata seeds
CN103814821A (en) * 2014-01-03 2014-05-28 中华全国供销合作总社南京野生植物综合利用研究所 Method for efficiently and quickly producing bletilla striata seedling
CN104137778A (en) * 2014-07-30 2014-11-12 中国科学院南京分院东台滩涂研究院 Method for culturing bletilla striata protocorm by means of bletilla striata seeds according to fluid suspension culture method

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
SYOICHI ICHIHASHI ET.AL.,: "breeding of bletilla hybrids and haploid induction via reduced female gamete parthenogenesis by NAA-Na treatment to establish seedling F1 cultivars in orchid", 《说明书第7段)》 *
和寿星等: "白芨无菌播种育苗技术", 《云南农业科技》 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105145352A (en) * 2015-08-03 2015-12-16 河南科技大学 Efficient tissue culture and rapid propagation technology for seedlings of bletilla striata
CN105145352B (en) * 2015-08-03 2017-06-06 河南科技大学 A kind of bletilla seed height of seedling imitates group culturation rapid propagating technology
CN105145361A (en) * 2015-09-17 2015-12-16 康美药业(文山)药材种植管理有限公司 Method for efficiently breeding tissue culture seedlings of bletilla striata and planting method of bletilla striata
CN105165368A (en) * 2015-10-15 2015-12-23 向君民 Method for rapidly breeding seedlings by using bletilla striata tubers
CN105210875A (en) * 2015-10-15 2016-01-06 铜仁市金农绿色农业科技有限公司 A kind of bletilla striata quick propagation method
CN106416972A (en) * 2016-08-29 2017-02-22 浙江金石生物科技有限公司 Method for quickly cultivating bletilla striata seedlings
CN107047310A (en) * 2017-05-10 2017-08-18 文山学院 A kind of cultural method of bletilla striata seeds culture seedling
CN109169273A (en) * 2018-09-03 2019-01-11 云南中医学院 A kind of breeding method of bletilla proliferation pseudobulb
CN110679421A (en) * 2019-11-06 2020-01-14 安徽东方金桥农林科技股份有限公司 Bletilla striata seedling cultivation method
CN116649216A (en) * 2023-06-25 2023-08-29 江苏护理职业学院 Pure bletilla striata three-fork tissue culture domesticated seedling raising method

Similar Documents

Publication Publication Date Title
CN104737911A (en) Quick cultivation method for rhizoma bletillae tissue culture seedlings
CN104719152B (en) Rhizoma bletillae industrialized seedling method
CN103314861B (en) A kind of dendrobium in vitro cross breeding method
CN104145816B (en) Bletilla striata tissue culture method
CN105145352B (en) A kind of bletilla seed height of seedling imitates group culturation rapid propagating technology
CN105766645B (en) Bletilla tissue-cultured seedling efficient propagation method
CN103583365B (en) Production method of bletilla seed stems
CN104429965B (en) The method that high eyebrow Herba Anoectochili roxburghii seed is bred without hormone quickly tissue culture
CN104663200B (en) A kind of pteridophyte large-scaled propugation method
Kalimuthu et al. In vitro micropropagation of Musa sapientum L.(Cavendish Dwarf)
CN104041412A (en) Rapid propagation method for tissue culture of Guizhou hemiboea cavaleriei
CN105230484A (en) Rapid breeding method of rhizoma polygonati
CN105993964A (en) Technology for carrying out tissue culture breeding by adopting common bletilla pseudobulb seeds
CN105075858B (en) A kind of liquid rapid propagation method of bletilla seed
CN106538251A (en) It is a kind of reduce large leaf Camellia sinensis wither bud disease fast seedling-cultivating method
CN106922536A (en) A kind of method that fast energy-saving cultivates bletilla seedling
CN105191803B (en) A kind of candidum tissue culturing bag seedling production method
CN105706872A (en) Bletilla striata seed direct seeding natural reproduction seedling method
CN106106138B (en) A kind of red palm crossbreeding and rapid propagation method
CN114532225B (en) Tissue culture rapid propagation and cultivation method for paphiopedilum delbrueckii
CN109964815A (en) A kind of bletilla striata aseptic seedling rapid induction Multiple Buds and fast numerous method of taking root
CN108719067A (en) A kind of tissue culture and rapid propagation method of paris polyphylla
CN107821169A (en) A kind of tissue culture method of P. kingianum seedling
CN105230488B (en) A kind of Cymbidium lancifolium leaf tissue culture method for quickly breeding
CN103155858B (en) Rapid lotus corniculatus l. breeding method

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20150701