CN104521756A - Method for producing trichosanthes tissue culture seedlings - Google Patents
Method for producing trichosanthes tissue culture seedlings Download PDFInfo
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- CN104521756A CN104521756A CN201410814578.0A CN201410814578A CN104521756A CN 104521756 A CN104521756 A CN 104521756A CN 201410814578 A CN201410814578 A CN 201410814578A CN 104521756 A CN104521756 A CN 104521756A
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- lfs
- trichosanthes
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- snakegourd
- plantlet
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Abstract
The invention discloses a method for producing trichosanthes tissue culture seedlings. The method comprises the following steps: (1) collecting single-node stems with axillary buds from strong trichosanthes plants, and performing conventional disinfection and sterilization; and (2) cutting the stems into an MS improved solid medium added with LFS for performing trichosanthes tissue culture seedling production, wherein the formula of the MS improved solid medium is 0.1-0.5mg.L<-1>(MS+LFS). The MS improved solid medium can induce explants to directly grow to the trichosanthes tissue culture seedlings with complete into roots and buds. The method disclosed by the invention has the advantages of simple production procedure, high tissue culture seedling quality and low cost.
Description
Technical field
The invention belongs to Plant Biotechnology field of tissue culture, be specifically related to a kind of method of producing snakegourd plantlet in vitro.
Technical background
Snakegourd (Trichosanthes kirilowii Maxim.) is called sky melon, is hung melon, crow melon, Curcurbitaceae snake gourd.Be one of important traditional Chinese medicine material of China, in medical treatment with health care, have very high using value and DEVELOPMENT PROSPECT widely.But, in its plantlet in vitro merchandized handling, almost all select the 6-BA (N in the basic element of cell division (CTKs) so far bar none
6-BA, N
6-benzyladenine) and growth hormone (Auxin) in the growth regulator such as IAA (heteroauxin), NAA (methyl α-naphthyl acetate) or IBA (indolebutyric acid), according to different proportionings, to be re-dubbed needed for " explant startup ", " expanding numerous without offspring subculture " and three production stages such as " Regenerated plant is taken root " three kinds of not identical medium respectively, progressively to complete the production of plantlet in vitro successively.Production routine bothers, and planting percent is low, and cost is high.
Summary of the invention
It is simple that the technical problem to be solved in the present invention is to provide a kind of production routine, and planting percent is high, the method for the production snakegourd plantlet in vitro that cost is low.
The present invention solves the problems of the technologies described above with following technical scheme:
Produce a method for snakegourd plantlet in vitro, comprise the following steps:
(1) gather single stem segment of band axillalry bud from healthy and strong snakegourd plant and carry out routine disinfection sterilizing;
(2) described stem segment cuttage is carried out the production of snakegourd plantlet in vitro in the MS improvement solid culture medium being added with spirit hair element-LFS, the formula that described MS improves solid culture medium is MS+LFS 0.1 ~ 0.5mgL
-1.
The formula that described MS improves solid culture medium is MS+LFS 0.3mgL
-1.
Remarkable advantage of the present invention is:
Using the single hormone culture-medium be mixed with by LFS to complete needs to produce with " explant Primary culture base ", " without offspring subculture medium " and " Regenerated plant root media " three kinds of plantlet in vitro that medium just can complete traditionally, thus greatly simplify snakegourd plantlet in vitro production routine, improve seedling efficiency, reduce production cost.
Embodiment
Following embodiment only for illustration of the present invention, instead of is used for limiting the scope of the invention.When not deviating from the present invention's spirit and essence, the amendment do method of the present invention, step or condition or replacement, all belong to scope of the present invention.
Embodiment 1:
Gather single stem segment of band axillalry bud from healthy and strong snakegourd plant and carry out routine disinfection sterilizing; By above-mentioned stem segment cuttage in using MS+LFS 0.1mgL
-1the solid culture medium of preparation, without the need to adding any other plant hormone and conditioning agent again, cultivating 30d and just directly can induce the normal germination and growth of 60% snakegourd axillalry bud explant, then building up the regeneration plant that root, seedling are complete, i.e. plantlet in vitro.These regeneration plant cauline leaf forms are normal, and average plant height reaches 3.8 ㎝, the average 2.5/strain of adventive root, can as subculture material, continue to expand in same medium numerous, without the need to preparing " subculture medium " in addition again.Also can transplant by bottle outlet, sell, without the need to establishing " program of taking root " and preparation " root media " again.Therefore, MS+LFS 0.1mgL is used
-1the solid culture medium of this single hormone just can substitute the snakegourd plantlet in vitro production overall process that three kinds of conventional medium just can complete.
Embodiment 2:
Gather single stem segment of band axillalry bud from healthy and strong snakegourd plant and carry out routine disinfection sterilizing; By above-mentioned stem segment cuttage in using MS+LFS 0.5mgL
-1the solid culture medium of preparation, without the need to adding any other plant hormone and conditioning agent again, cultivating 25d and just directly can induce the normal germination and growth of 100% snakegourd axillalry bud explant, then building up the regeneration plant that root, seedling are complete, i.e. plantlet in vitro.These regeneration plant cauline leaf forms are normal, and average plant height reaches 5.2 ㎝, the average 3.5/strain of adventive root.These regeneration plants can, as subculture material, expand numerous in same medium, without the need to preparing " subculture medium " in addition again.Also can transplant by bottle outlet, sell, without the need to establishing " program of taking root " and preparation " root media " again.Therefore, MS+LFS 0.5mgL is used
-1the solid culture medium of this single hormone just can substitute the snakegourd plantlet in vitro production overall process that three kinds of conventional medium just can complete.
Embodiment 3:
Below by the inventive method, through preferred formula MS+LFS 0.3mgL
-1when the single hormone solid culture medium of preparation and routine three kinds of medium containing hormon used carry out the production of snakegourd plantlet in vitro, the comparison at " startup ", " subculture " and " taking root " three critical stages:
1. " startup of the axillalry bud explant " stage:
Explant: gather single stem segment of band axillalry bud from healthy and strong plant and carry out routine disinfection sterilizing.Often organize medium 30 bottles, every bottle graft kind 1 explant.Eliminate pollution bottle after cultivating 7d, often organize and get 5 bottles at random, observe statistics: planting percent=seedling explant number/inoculation explant number × 100%, average plant height (terminal bud is to joint of taking root), average indefinite radical (bar/strain).
A-1 group: solid culture medium of the present invention: MS+LFS 0.3mgL
-1;
B-1 group: traditional Primary culture base MS+6-BA 0.3mgL
-1+ NAA0.06mgL
-1;
Result:
A-1 group: cultivate 20d, planting percent reaches 80%, 25d and reaches 100%, cauline leaf form is normal, and average plant height reaches 5.4 ㎝.
Be cultured to 15d, start have adventive root to occur, the average 3.2/strain of 25d adventive root, builds up complete regenerated plant.
B-1 group: cultivate 20d, planting percent only 20%, 25d nearly 30%, average plant height 2.2 ㎝, do not have adventive root to occur completely, blade is obviously less.
2. " subculture the expands numerous " stage:
Subculture material: the normal sterile seedling that above-mentioned A-1 group obtains is cut two ends on clean work station, gets+2 ~+4 joints, cuts into single-unit stem with bud, excise blade simultaneously, reserve part petiole protection axillalry bud.Cuttage is in following two groups of medium respectively, often organizes 12 bottles, every bottle of 5 stem sections.Eliminate pollution bottle after cultivating 7d, often organize and get 5 bottles at random, statistics morphological indexes is the same.
A-2 group: solid culture medium of the present invention: MS+LFS 0.3mgL
-1;
B-2 group: traditional subculture expands breeding culture medium: MS+6-BA 0.2mgL
-1+ NAA0.1mgL
-1;
Result:
A-2 group: cultivate 15d Rooting percent average out to 50%, 25d and reach 100%.Average every strain has root 3.5, average root long 4.3 ㎝, average plant height 4.8 ㎝.Cauline leaf growth is normal, builds up the regeneration plant of complete stalwartness.These regeneration plants can directly be transplanted to nutrition cup and carry out nursery sale by bottle outlet, also can continue shoot proliferation.Anatomical proof, the adventive root of these regeneration plants occurs from marrow meristematic tissue, difficult drop-off when washing seedling, transplanting survival rate >=95%.
B-2 group: cultivate 25d and still occur without adventive root, average plant height 3.2cm.Cauline leaf form is normal.
3. " propagation seedling rooting " stage:
To take root material: with above-mentioned B-2 group gained without offspring, cuttage is in following 2 groups of medium respectively.Often organize 10 bottles, every bottle of 5 stem sections.Statistics: Rooting percent=root of hair strain number/inoculation strain number × 100%; Average indefinite radical (bar); Average adventive root long (cm).
A-3 group: solid culture medium of the present invention: MS+LFS 0.3mgL
-1;
B-3 group: traditional root media MS+NAA 0.3mgL
-1;
Result:
A-3 group: cultivation 15d Rooting percent 30%, 20d reaches 80%, 25d and reaches 100%.Mean elements 3.6/strain, long 4.5 ㎝ of average root.Conventional transplanting survival rate >=95%.
B-3 group: cultivate 15d Rooting percent 15%, 25d 70%, Extending culture just can reach 80% to 35d.Growing fine of seedling.But anatomical proof, this group plant adventive root more than 2/3 occurs from callus, as easy as rolling off a logly when washing seedling comes off, and transplanting survival rate only about 80%, is starkly lower than A-3 group.
Claims (2)
1. produce a method for snakegourd plantlet in vitro, it is characterized in that, comprise the following steps:
(1) gather single stem segment of band axillalry bud from healthy and strong snakegourd plant and carry out routine disinfection sterilizing;
(2) described stem segment cuttage is carried out the production of snakegourd plantlet in vitro in the MS improvement solid culture medium being added with spirit hair element-LFS, the formula that described MS improves solid culture medium is MS+LFS 0.1 ~ 0.5mgL
-1.
2. a kind of method of producing snakegourd plantlet in vitro according to claim 1, is characterized in that, the formula that described MS improves solid culture medium is MS+LFS 0.3mgL
-1.
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| CN201410814578.0A CN104521756B (en) | 2014-12-24 | 2014-12-24 | A kind of method producing Fructus Trichosanthis tissue cultured seedling |
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| CN201410814578.0A CN104521756B (en) | 2014-12-24 | 2014-12-24 | A kind of method producing Fructus Trichosanthis tissue cultured seedling |
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| CN104521756B CN104521756B (en) | 2016-08-17 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105191797A (en) * | 2015-09-29 | 2015-12-30 | 潜山县传文瓜子有限公司 | Snakegourd fruit tissues culture method |
| CN106472311A (en) * | 2016-10-14 | 2017-03-08 | 广西大学 | Method with spirit hair element evoking tobacco calluss |
| CN107360843A (en) * | 2017-08-25 | 2017-11-21 | 无为县翔博生态农业开发有限公司 | A kind of propagation method of Snakegourd Fruit cuttage |
| CN108812311A (en) * | 2018-06-06 | 2018-11-16 | 周庆友 | A kind of method of snakegourd quick reproduction technique |
| CN109089884A (en) * | 2018-08-31 | 2018-12-28 | 湖州德清玖沐农业科技有限公司 | A kind of quick-breeding method of snakegourd seedling |
| CN111213585A (en) * | 2020-02-18 | 2020-06-02 | 鲁东大学 | One-step tissue culture and rapid propagation cultivation process for catalpa bungei |
| CN115735766A (en) * | 2022-11-01 | 2023-03-07 | 颐正源(天津)生态农业科技有限公司 | Snakegourd fruit tissue culture method |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105191797A (en) * | 2015-09-29 | 2015-12-30 | 潜山县传文瓜子有限公司 | Snakegourd fruit tissues culture method |
| CN106472311A (en) * | 2016-10-14 | 2017-03-08 | 广西大学 | Method with spirit hair element evoking tobacco calluss |
| CN106472311B (en) * | 2016-10-14 | 2018-10-19 | 广西大学 | With the method for spirit hair element evoking tobacco callus |
| CN107360843A (en) * | 2017-08-25 | 2017-11-21 | 无为县翔博生态农业开发有限公司 | A kind of propagation method of Snakegourd Fruit cuttage |
| CN108812311A (en) * | 2018-06-06 | 2018-11-16 | 周庆友 | A kind of method of snakegourd quick reproduction technique |
| CN109089884A (en) * | 2018-08-31 | 2018-12-28 | 湖州德清玖沐农业科技有限公司 | A kind of quick-breeding method of snakegourd seedling |
| CN111213585A (en) * | 2020-02-18 | 2020-06-02 | 鲁东大学 | One-step tissue culture and rapid propagation cultivation process for catalpa bungei |
| CN115735766A (en) * | 2022-11-01 | 2023-03-07 | 颐正源(天津)生态农业科技有限公司 | Snakegourd fruit tissue culture method |
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