CN107047318B - A kind of external rapid propagation method of grande passerage herb - Google Patents
A kind of external rapid propagation method of grande passerage herb Download PDFInfo
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- CN107047318B CN107047318B CN201710470075.XA CN201710470075A CN107047318B CN 107047318 B CN107047318 B CN 107047318B CN 201710470075 A CN201710470075 A CN 201710470075A CN 107047318 B CN107047318 B CN 107047318B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of external rapid propagation method of grande passerage herb, step are as follows：(1) aseptic explant of grande passerage herb is prepared；(2) aseptic explant is inoculated on fast breeding culture medium, carries out the induction and rapid amplifying breeding of regrowth；(3) regrowth of inductive formation is transferred in MS minimal mediums and carries out rooting induction；(4) the Lepidium apetalum complete regenerated plant after taking root is transplanted into normal growth in cultivation matrix.The present invention is the domestic and international first external fast breeding technique of grande passerage herb, effectively expands the breeding coefficient of grande passerage herb, overcomes grande passerage herb seed and expands numerous territory restriction problem.
Description
Technical field
The present invention relates to field of plant tissue culture technique, more particularly to a kind of external quick breeding of grande passerage herb
Method.
Background technology
Grande passerage herb is herbaceos perennial, south of europe, the Africa for being distributed in 1800~4250 meters of height above sea level is northern,
Western part of Asia and middle part, a variety of degeneration-resistant characters such as have Salt And Alkali Tolerance, drought resisting waterlogging.Grande passerage herb all herbal medicine, the traditional Chinese medical science think
Bacillary dysentery, enteritis can be controlled；Its seed contains sitosterol and cardiac glycoside composition, can strengthen heart contraction, decreased heart rate, to exhaustion
Heart can increase the output quantity of blood, reduce vein pressure.
According to the record of Chinese Plants will, grande passerage herb is divided into the grande passerage herb (former mutation) for originating in Qinghai
(Lepidium latifolium L.var.latifolium) and originate in light fruit grande passerage herb (mutation) north in North China solely
Row dish (Lepidium latifolium L.var.affine).With being continually developed to grande passerage herb edibility, people
Gradually find to originate in grande passerage herb (former mutation) (the Lepidium latifolium in Qinghai
L.var.latifolium it is that a kind of nutritive value is high) rich in vitamin A, the various trace elements such as C, Yi Jitong, calcium, potassium
New type of health vegetables.It can feed pig, sheep as feed, be the more universal forage plant in NORTHWEST CHINA salt-soda soil.In addition, solely
For row colza rich in abundant aliphatic acid, yield is higher by 30% or so than winter rape average level, is a kind of new oil crops.
In view of grande passerage herb strong stress resistance, and have higher application value in edible, medicinal, feeding etc., therefore
With good promotion development prospect.But originate in grande passerage herb (former mutation) (the Lepidium latifolium in Qinghai
L.var.latifolium) be distributed in the Northwest at home, although eastern region can normal growth it is solid, it is tied
Seed majority dysontogenesis is difficult to obtain normal seedling so that the cultivation of grande passerage herb is promoted and is extremely restricted.
Therefore, exploitation is a kind of is directed to grande passerage herb (former mutation) (the Lepidium latifolium for originating in Qinghai
L.var.latifolium external fast breeding technique), the cultivation for grande passerage herb are promoted tool and are of great significance.
The content of the invention
For above-mentioned prior art, it is an object of the invention to provide a kind of external quick breeding side of grande passerage herb
Method.Instant invention overcomes grande passerage herb east China area reproductive efficiency is low the problem of, has greatly facilitated wide leaf and has walked alone
The cultivation of dish is promoted.
To achieve the above object, the present invention adopts the following technical scheme that：
The first aspect of the present invention, there is provided a kind of external rapid propagation method of grande passerage herb, step are as follows：
(1) aseptic explant of grande passerage herb is prepared；
(2) aseptic explant is inoculated on fast breeding culture medium, carries out the induction and rapid amplifying breeding of regrowth；
(3) regrowth of inductive formation is transferred in MS minimal mediums and carries out rooting induction；
(4) the grande passerage herb complete regenerated plant after taking root is transplanted into normal growth in cultivation matrix.
The grande passerage herb is grande passerage herb (former mutation) (the Lepidium latifolium for originating in Qinghai
L.var.lat ifolium)。
In step (1), the preparation method of aseptic explant is：The Lepidium apetalum seed of mature and plump is selected, is sterilized, Ran Houbo
Kind shreds the whole strain of 10~14 days seedling age Lepidium apetalum seedling after sprouting as the external nothing quickly bred on MS minimal mediums
Bacterium explant.
Preferably, the method for sterilizing is：It is 2% sodium hypochlorite sterilizing 25min with mass concentration, is then rushed with sterilized water
Wash.
In step (2), the fast breeding culture medium is sucrose, the 1mg/L 6- that 30g/L is added in MS minimal mediums
BA, 0.25mg/L NAA, 6~8g/L agar, regulation pH are 5.8.
In step (2), the condition that the induction of regrowth and rapid amplifying breeding use is 23~25 DEG C of culture, and daily 14
Hour illumination, 10 hours dark, 14~21 days subcultures once, coinduction culture 28~42 days.
In step (3), condition that rooting induction uses for：23~25 DEG C of cultures, daily illumination in 14 hours, 10 hours black
Secretly, coinduction culture 20 days.
Beneficial effects of the present invention：
(1) present invention is directed to grande passerage herb (former mutation) (the Lepidium latifolium for originating in Qinghai
L.var.latifolium a kind of rapid propagation method) is provided first, is that grande passerage herb is (particularly magnificent in east China
North, East China) popularization and application provide technical guarantee.
(2) fast breeding technique of the invention is used as explant, structure using the whole strain chopping of seedling is sprouted under condition of tissue culture
Incomplete seedling can also utilize, therefore not influenceed by embryonic development is bad, reduce explant materials difficulty, expand explant
Body carrys out source range.
(3) in the present invention, it is 8.54~11.04 to expand numerous coefficient, i.e. every aseptic seedlings ultimately form the flat of regeneration plant
Equal number.Above steps is an organic whole, conditional, effectively increases the reproductive efficiency of grande passerage herb.
To sum up, the present invention is the first external fast breeding technique of grande passerage herb for originating in Qinghai both at home and abroad, is effectively expanded
The big breeding coefficient of grande passerage herb, overcome the territory restriction problem of grande passerage herb seminal propagation.
Brief description of the drawings
The Figure of description for forming the part of the application is used for providing further understanding of the present application, and the application's shows
Meaning property embodiment and its illustrate be used for explain the application, do not form the improper restriction to the application.
Fig. 1：Pick up from the grande passerage herb aseptic seeding in Qinghai；
Fig. 2：The induction of regrowth and quick breeding；
Fig. 3：Culture of rootage；
Fig. 4：Plant after transplanting.
Embodiment
It is noted that described further below is all exemplary, it is intended to provides further instruction to the application.It is unless another
Indicate, all technologies used herein and scientific terminology are with usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe embodiment, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form, additionally, it should be understood that, when in this manual using term "comprising" and/or " bag
Include " when, it indicates existing characteristics, step, operation, device, component and/or combinations thereof.
As background technology is introduced, cultivation of the external fast breeding technique of grande passerage herb for grande passerage herb is developed
Tool is promoted to be of great significance.Based on this, present applicant proposes a kind of external rapid propagation method of grande passerage herb.
In a kind of embodiment of the application, there is provided a kind of external rapid propagation method of grande passerage herb, step
It is as follows：
(1) preparation of aseptic explant：The Lepidium apetalum seed of mature and plump is selected, is 2% sodium hypochlorite with mass concentration
Sterilize 25min, after aseptic water washing 4 times, is seeded on MS minimal mediums, by 10~14 days seedling age Lepidium apetalum seedling after sprouting
Whole strain chopping is as the external aseptic explant quickly bred.
(2) aseptic explant is inoculated on fast breeding culture medium, carries out the induction and rapid amplifying breeding of regrowth；It hurry up
The condition that speed amplification breeding uses is 23~25 DEG C of cultures, daily illumination in 14 hours, 10 hours dark, 14~21 days subculture one
It is secondary, coinduction culture 28~42 days.
(3) regrowth of inductive formation is transferred in MS minimal mediums and carries out rooting induction；The bar that rooting induction uses
Part is：23~25 DEG C of cultures, daily illumination in 14 hours, 10 hours dark, coinduction culture 20 days.
(4) the Lepidium apetalum complete regenerated plant after taking root is transplanted into normal growth in cultivation matrix.
In order that the technical scheme of the application can clearly be understood by obtaining those skilled in the art, below with reference to tool
The embodiment of body describes the technical scheme of the application in detail.
Test material used is the conventional test material in this area in the embodiment of the present invention, can pass through commercial channel
It is commercially available.
Culture medium used, specific as follows in the embodiment of the present invention：
MS culture mediums：Commercially available prod.
Fast breeding culture medium：30g/L sucrose, 1mg/L 6-BA, 0.25mg/L NA are added in MS minimal mediums
A, 6g/L agar, regulation pH are 5.8；With 121 DEG C of high pressure moist heat sterilization 20min after preparation.
Cultivation matrix is cultivation matrix conventional in the prior art.
Embodiment 1：The grande passerage herb normal seedling harvested in Qinghai is numerous soon
Specific method is as follows：
(1) acquisition of aseptic explant：Mature and plump person is chosen in the grande passerage herb seed of Qinghai Area from harvesting,
Sterilized 25min with 2% sodium hypochlorite, after aseptic water washing 4 times, be seeded on MS minimal mediums, 10 after aseptic seedling sprouting
My god, whole strain seedling is uniformly cut into 10 pieces as the aseptic explant quickly bred in vitro.
(2) aseptic explant is inoculated on fast breeding culture medium, condition of culture is 23~25 DEG C and cultivated, the daily 14 small time
According to, 10 hours dark, once, numerous in vitro seedling of regeneration grande passerage herb is expanded in coinduction culture 28 days, acquisition to 14 days subcultures in vitro,
The average number that every piece of explant forms Regeneration in Vitro seedling in this step is 1.104 plants；
(3) the Regeneration in Vitro seedling of above-mentioned inductive formation is transferred in MS minimal mediums, continued with the condition in step (2)
Carry out rooting induction, rooting rate 100%.
(4) the Lepidium apetalum complete regenerated plant after taking root is transplanted into cultivation matrix, makes it according to conventional cultivation mode
Into normal growth, this step transplanting survival rate is 100%.
To sum up, the average plant number that every aseptic seedlings of grande passerage herb in Qinghai ultimately form regeneration plant is harvested
Mesh, that is, expand numerous coefficient as 11.04.
Grande passerage herb aseptic seeding, regrowth induction and quick breeding, culture of rootage and cultivation after plant point
Not as Figure 1-4.
Embodiment 2：The grande passerage herb radicle missing seedling harvested in Shandong is numerous soon
Specific method is as follows：
(1) acquisition of aseptic explant：Mature and plump person is chosen in the grande passerage herb seed of Shandong District from harvesting,
Sterilized 25min with 2% sodium hypochlorite, after aseptic water washing 4 times, be seeded on MS minimal mediums, 14 after aseptic seedling sprouting
My god, the whole strain seedling for lacking radicle is uniformly cut into 10 pieces as the aseptic explant quickly bred in vitro.
(2) aseptic explant is inoculated on fast breeding culture medium, condition of culture is 23~25 DEG C and cultivated, the daily 14 small time
According to, 10 hours dark, once, numerous in vitro seedling of regeneration grande passerage herb is expanded in coinduction culture 42 days, acquisition to 21 days subcultures in vitro,
The average number that every piece of explant forms Regeneration in Vitro seedling in this step is 0.854 plant；
(3) the Regeneration in Vitro seedling of above-mentioned inductive formation is transferred in MS minimal mediums, continued with the condition in step (2)
Carry out rooting induction, rooting rate 100%.
(4) the Lepidium apetalum complete regenerated plant after taking root is transplanted into cultivation matrix, makes it according to conventional cultivation mode
Into normal growth, this step transplanting survival rate is 100%.
To sum up, the average plant number that every aseptic seedlings of grande passerage herb in Shandong ultimately form regeneration plant is harvested
Mesh, that is, expand numerous coefficient as 8.54.
Comparative example 1：
The spire and tender stem of the grande passerage herb in Qinghai are originated in collection, disinfect, the wide leaf after disinfecting is walked alone
Dish spire is cut into 0.5cm2The fragment of size, tender stem are cut into 0.5-1.0cm segment, and the explant as callus induction connects
Kind in filling in the triangular flask of aseptic culture medium, the composition of culture medium is：1.5mg/L2,4-D+1.0mg/ is added in MS culture mediums
L IAA+0.5mg/L 6-BA。
The triangular flask for being inoculated with explant is placed in 25 degrees Celsius, cultivated under 1800-2000lx intensities of illumination.Will be raw
Lepidium apetalum complete regenerated plant after root is transplanted into cultivation matrix, and normal growth is made it into according to conventional cultivation mode.
In this comparative example, the survival rate of transplanting is 82%, expands numerous coefficient as 5.02.
Comparative example 2：
Material used is replaced with to the spire and tender stem for the grande passerage herb for picking up from Tangshan Area, remaining processing method is same
Comparative example 1.
In this comparative example, the survival rate of transplanting is 86%, expands numerous coefficient as 6.13.
There to be different genetic backgrounds (to pick up from Qinghai, Shandong and Tang respectively it can be seen from above-described embodiment and comparative example
The grande passerage herb on mountain) material carry out tissue cultures, tissue culture system needed for it is different, does not have comparativity in tissue culture system
With referring to property.In addition, for the grande passerage herb for originating from Qinghai, the selection of its tissue culture system has for expanding numerous effect
There is very crucial influence, experimental research find that, the external rapid propagation method of grande passerage herb of the invention, pass through regeneration
The step such as the induction of seedling and rapid amplifying breeding, rooting induction preferably with combination, it is extraordinary to realize to being collected in green grass or young crops
The quick breeding of the grande passerage herb in sea.
The preferred embodiment of the application is the foregoing is only, is not limited to the application, for the skill of this area
For art personnel, the application can have various modifications and variations.It is all within spirit herein and principle, made any repair
Change, equivalent substitution, improvement etc., should be included within the protection domain of the application.
Claims (1)
1. the external rapid propagation method of a kind of grande passerage herb, it is characterised in that step is as follows：
（1）Prepare the aseptic explant of grande passerage herb：The Lepidium apetalum seed of mature and plump is selected, is 2% chlorine with mass concentration
Sour sodium sterilizes 25min, with aseptic water washing 4 times, is then seeded on MS minimal mediums, by 10 ~ 14 days seedling ages after sprouting
The whole strain chopping of Lepidium apetalum seedling is as the external aseptic explant quickly bred；
（2）Aseptic explant is inoculated on fast breeding culture medium, carries out the induction and rapid amplifying breeding of regrowth；Regrowth
Induction and the rapid amplifying condition that uses of breeding cultivated for 23 ~ 25 DEG C, daily illumination in 14 hours, 10 hours dark, 14 ~
21 days subcultures once, coinduction culture 28 ~ 42 days；
（3）The regrowth of inductive formation is transferred in MS minimal mediums and carries out rooting induction；The condition that rooting induction uses
For：23 ~ 25 DEG C of cultures, daily illumination in 14 hours, 10 hours dark, coinduction culture 20 days；
（4）Lepidium apetalum complete regenerated plant after taking root is transplanted into normal growth in cultivation matrix；
The grande passerage herb is the grande passerage herb for originating in Qinghai（Lepidium latifolium L. var. latifolium）；
Step（2）In, the fast breeding culture medium be added in MS minimal mediums 30g/L sucrose, 1mg/L 6-BA,
0.25mg/L NAA, 6 ~ 8g/L agar, regulation pH are 5.8.
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| CN102511396B (en) * | 2011-10-14 | 2013-07-17 | 新疆维吾尔自治区中药民族药研究所 | In vitro rapid breeding method of medicinal plant Lipidium meyenii |
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