CN109793010A - A kind of liquid composite bacteria agent and its application method promoting Growth of Dendrobium candidum - Google Patents
A kind of liquid composite bacteria agent and its application method promoting Growth of Dendrobium candidum Download PDFInfo
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- CN109793010A CN109793010A CN201910193312.1A CN201910193312A CN109793010A CN 109793010 A CN109793010 A CN 109793010A CN 201910193312 A CN201910193312 A CN 201910193312A CN 109793010 A CN109793010 A CN 109793010A
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Abstract
The invention discloses a kind of liquid composite bacteria agents and its application method for promoting Growth of Dendrobium candidum, the liquid composite bacteria agent includes mycorrhizal fungi dendrobium mixture (Mycena dendrobii) and lanthanum nitrate, it can also include auxin, seedling root, lanthanum nitrate and auxin are inoculated with dendrobium mixture, and the dendrobe tissue culture seedling aerial part for being transplanted to greenhouse after domestication and growing is handled with spray pattern.The liquid composite bacteria agent formed using both dendrobium mixture and lanthanum nitrate, it significantly improves dendrobium nobile seedling percent and promotes growth, including index characters such as plant height, single plant weight, root, stem, leaves, and the composite bacteria agent of dendrobium mixture, lanthanum nitrate and auxin three composition is used to significantly improve the active constituent contents such as dendrobium nobile polysaccharide from medicinal materials and alkaloid.This planting technology method is simple, is easy to be widely applied, small investment, and the quality for the dendrobium nobile medicinal material produced meets Chinese Pharmacopoeia requirement.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to it is a kind of promote Growth of Dendrobium candidum liquid composite bacteria agent and its
Application method.
Background technique
Dendrobium candidum (Dendrobium officinale) is Chinese tradition rare traditional Chinese medicine, has very high medicinal valence
Value and economic value, are listed in country-level wild protection plant, Perenniporia martius has widely distributed in China.However,
Due to the artificial destruction excavated without restraint with primitive environment, leads to the serious imbalance of relation between supply and demand, pushed dendrobium candidum people
The development of work cultivation and its industrialization.The setting percentage of wild Dendrodium is very low under natural conditions, in addition seed does not have endosperm directly to mention
For nutrition, early period, seedling was difficult to sprout, and its transplanting survival rate is very low.Dendrobium candidum artificial growth is all made of tissue-cultured seedling at present,
The problems such as there is at high cost, transplanting survival rate is low, slow growth, low output.
Plant growth-promoting microbial inoculum has become the hot spot of current research, and plant mycorrhizal fungi and interior life with growth-promoting functions are thin
Bacterium is due to having the nutriments such as enough carbon sources, nitrogen source, and the protection by plant tissue or root, it is easier to play a role.Bacterium
Mycorrhiza fungi, which is mainly reflected in the facilitation of host plant, can preferably absorb nutriment, be conducive to secondary metabolite
Accumulation etc., while absorption of the host plant to mineral elements such as nitrogen, phosphorus can be promoted, to promote the life of host plant
It is long.Currently, microbial bacterial agent is mostly single bacterium, the report of mix bacterium agent is few.Single pure culture inoculum lacks dendrobium candidum
Nutrient required for growing, nursery initial performances are bad, it is difficult to promote.
Therefore, the correlation and liquid composite bacteria agent of dendrobium nobile mycorrhizal fungi and host plant are studied to dendrobium candidum group
The growth-promoting functions of training seedling are of great significance.
Summary of the invention
It is an object of the invention in place of overcome the deficiencies in the prior art, provide a kind of liquid for promoting Growth of Dendrobium candidum
Bluk recombination microbial inoculum and its application method cause to grow slow, medicinal plant money for solving dendrobium candidum due to lacking mycorrhizal fungi
The insufficient problem in source, using two kinds of liquid composite bacteria agents, can greatly promote dendrobium nobile Seedling Height, stem length, stem thickness, root long,
Root thickness, radical, section length, joint number, the number of blade, leaf length, leaf width etc., additionally it is possible to which significantly improving dendrobium nobile polysaccharide from medicinal materials and alkaloid etc. has
Effective component content；This planting technology method is simple, is easy to be widely applied, small investment, the quality for the dendrobium nobile medicinal material produced
Meet Chinese Pharmacopoeia requirement.
The technical solution adopted by the present invention to solve the technical problems first is that:
A kind of liquid composite bacteria agent promoting Growth of Dendrobium candidum, including 3~5g/L dendrobium mixture (Mycena
Dendrobii), 4~6mg/L lanthanum nitrate.
In one embodiment: further including 190~210mg/L auxin.
In one embodiment: after the dendrobium mixture preparation method includes: dendrobium mixture actication of culture, switching is cultivated in PDA
In base, in 22~25 DEG C constant temperature stationary culture 14~20 days, then access in PDA culture medium and carry out fermented and cultured 7~14 days, i.e.,
?.
The technical solution adopted by the present invention to solve the technical problems second is that:
A kind of application method for the liquid composite bacteria agent promoting Growth of Dendrobium candidum, in the liquid composite bacteria agent, dendrobium nobile
Small mushroom is inoculated in dendrobium candidum seedling root, and dendrobium nobile mycorrhizal fungi and host plant is made full use of to form Mycorrhizal Symbiosiss relationship, nitre
Sour lanthanum, auxin are sprayed on dendrobium candidum seedling aerial part.
In one embodiment: in the liquid composite bacteria agent, dendrobium mixture was every 28~32 days inoculation dendrobium candidum seedling roots
Portion is primary, and lanthanum nitrate, auxin are primary every 9~11 days sprinkling dendrobium candidum seedling aerial parts.
In one embodiment: the inoculum concentration of the dendrobium mixture is 4~6mL.
In one embodiment: the dendrobium candidum seedling is the dendrobe tissue culture seedling that greenhouse growth is transplanted to after domestication.
The technical program compared with the background art, it has the following advantages:
In the liquid composite bacteria agent of promotion Growth of Dendrobium candidum of the invention, dendrobium mixture mycorrhizal fungi can improve tissue-cultured seedling
Plant height and stem thickness, avoided after liquid treating agent is made be inoculated with by fungi when bacteria containing amount it is excessively high and to lose symbiosis flat
The problem of weighing apparatus, will not because of culture medium residual and pollute tissue-cultured seedling；The lanthanum nitrate of suitable concentration is in dendrobium candidum mycorhiza shape
At when with certain facilitation, (with the increase of inoculation nitric acid la concn, the fungal infection rate of dendrobium candidum also can be therewith
Increase)；Not only dendrobium candidum seedling amount is big after addition auxin, and growing-seedling period is short, and operation is convenient, and mutation phenomenon does not occur.It adopts
After liquid composite bacteria agent culture of the invention, dendrobium nobile Seedling Height, training branch amount, stem length, stem thickness, section length, joint number, root
Length, root thickness, radical, the number of blade, leaf length, leaf width, dendrobium polysaccharide, dendrobium nobile alkaloid etc. obviously increase, to dendrobium candidum children
Seedling growth has good facilitation.Meanwhile liquid treating agent does not influence its infection ability after composite bacteria agent is made.
Detailed description of the invention
Present invention will be further explained below with reference to the attached drawings and examples.
Fig. 1 is different liquids composite bacteria agent treated dendrobium nobile seedling percent schematic diagram.
Fig. 2 is different liquids composite bacteria agent treated stem of Dendrobium appearance index schematic diagram.
Fig. 3 is different liquids composite bacteria agent treated dendrobium nobile root appearance index schematic diagram.
Fig. 4 is different liquids composite bacteria agent treated dendrobium nobile blade appearance index schematic diagram.
Fig. 5 is different liquids composite bacteria agent treated the whole strain appearance index schematic diagram of dendrobium nobile.
Fig. 6 is different liquids composite bacteria agent treated stem of Dendrobium fresh weight dry weight contrast schematic diagram.
Fig. 7 is different liquids composite bacteria agent treated dendrobium polysaccharide content schematic diagram (note: between different letter expressions processing
Significant difference；" * * " indicates extremely significant with control group otherness).
Fig. 8 is different liquids composite bacteria agent treated dendrobium nobile alkaloid schematic diagram (note: different letter expressions processing
Between significant difference；" * * " indicates extremely significant with control group otherness).
Specific embodiment
The contents of the present invention are illustrated below by embodiment:
Present embodiments provide a kind of liquid composite bacteria agent for promoting Growth of Dendrobium candidum, comprising: 4g/L dendrobium mixture
(Mycena dendrobii), 5mg/L lanthanum nitrate.
In another embodiment of the invention, further include 190~210mg/L auxin, that is, promote Growth of Dendrobium candidum
Liquid composite bacteria agent includes: 4g/L dendrobium mixture (Mycena dendrobii), 5mg/L lanthanum nitrate, 200mg/L auxin.
Wherein, the dendrobium mixture in liquid composite bacteria agent is the preparation method is as follows: mycorrhizal fungi dendrobium mixture comes from iron sheet stone
The root of dry measure used in former times, comes from China Forest Microbiological Culture Collection administrative center, and mycorrhizal fungi dendrobium mixture is from the inclined-plane of low-temperature preservation
After test tube strains activation, transferred respectively in the examination for containing PDA culture medium (solid medium contains glucose, potato, agar etc.)
Pipe or culture dish in, in 22~25 DEG C constant temperature stationary culture 14~20 days, taken when mycelia largely or entirely covers with culture medium
It is spare out, it punches respectively in colony edge into bacterium piece, and to be sent out in fritter access PDA potato dextrose agar
Ferment culture 7~14 days.
The step of being cultivated using liquid composite bacteria agent dendrobium candidum include:
By dendrobium nobile sprigging to the mixing for containing 300mL bark, gravel pebbles and raw humus soil 3:1:1 (v/v/v)
In the hole tray of object (hole tray is rectangle, and 5 × 10 grids, each lattice is 4cm diameter).
The dendrobium nobile seedling is placed under greenhouse and is cultivated, temperature is 23~25 DEG C and is managed in conventional manner, comprising: moves
Root water is poured after cultivation, matrix of being subject to irrigates, and is periodically sprayed 4~5 times when maintenance daily, humidity is maintained at 80% or so, to new
Root long gradually reduces spray timings after going out.
Further, every 30 days, with 7~14 days mycorhiza of fermented and cultured in PDA potato dextrose agar
It is primary that fungi dendrobium mixture (4g/L) is inoculated with seedling root.It when inoculation, gently pushs aside around seedling root, is drawn about with suction pipe
5mL dendrobium mixture is seeded in seedling root, then the gently upper original matrix of covering.
Further, it every 10 days, is mixed to get with 5mg/L lanthanum nitrate, the 200mg/L auxin in composite fluid microbial inoculum
Liquid treating agent dendrobium nobile seedling aerial part is sprayed respectively it is primary.It is wet to notice that dendrobium nobile seedling is afraid of, moistens, it should not be too
It is more.
According to the influence of the correlation and different composite liquid agent dendrobium candidum of dendrobium nobile mycorrhizal fungi and host plant,
It is provided with 10 groups of processing, every group of 10 hole trays altogether, each hole tray plants three plants of seedling, and following table 1 is specific processing scheme:
The processing group and concentration of 1 different liquids composite bacteria agent of table
It is cultivated through processing in 6 months, 10 groups of seedling percents are as shown in Figure 1.Experimental result shows, the 4th group of processing mode:
It is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L lanthanum nitrate every 10 days sprinkling seedling aerial parts
The dendrobium nobile seedling highest of single treatment, survival rate 83.0% improve 31.1% than control group survival rate (63.3%).
Seedling is taken out and measures seedling stem length, stem thickness, section length, joint number appearance index respectively, is averaged concrete outcome such as
Shown in Fig. 2.Experimental result shows that different composite liquid handling group dendrobium nobile seedling stem length, stem thickness, section is long, joint number appearance index is equal
It is apparently higher than control group, wherein the 4th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with
5mg/L lanthanum nitrate behaves oneself best every the dendrobium nobile growth of seedling of sprinkling seedling aerial part single treatment in 10 days, stem length 9.63cm,
Stem thickness 4.95mm, the long 1.04cm of section, stipes 9.7, improve 4.1 times, 1.3 times, 1.7 times and 2.3 times than control respectively.
Seedling root long, root thickness, radical appearance index are measured respectively, and it is as shown in Figure 3 to be averaged concrete outcome.Experimental result
It has been shown that, different composite liquid handling group dendrobium nobile seedling root long, root thickness, radical appearance index are obviously higher than control group, wherein the
Four groups of processing modes: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, it was sprayed with 5mg/L lanthanum nitrate every 10 days
The dendrobium nobile growth of seedling of seedling aerial part single treatment behaves oneself best, root long 11.24cm, root thickness 1.23mm, root 10.7, point
1.7 times, 1.6 times, 1.5 times Wei not compareed.
Seedling leaves number, leaf length, leaf width appearance index are measured respectively, and it is as shown in Figure 4 to be averaged concrete outcome.Experiment knot
Fruit shows, different composite liquid handling group dendrobium nobile seedling leaves quantity, leaf be long, leaf width appearance index is obviously higher than control group,
Wherein, the 4th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L lanthanum nitrate every
10 days sprinkling the single treatment of seedling aerial part dendrobium nobile growth of seedling behave oneself best, blade quantity 12.8, the long 3.44cm of leaf,
Leaf width 0.93cm, 1.8 times respectively compareed, 2.0 times and 1.3 times.
Seedling Height, whole strain branch amount appearance index are measured respectively, and it is as shown in Figure 5 to be averaged concrete outcome.Experiment knot
Fruit shows that different composite liquid handling group dendrobium nobile Seedling Height appearance index is obviously higher than control group, wherein
A) the 4th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L lanthanum nitrate
It is 22.33cm every the height highest of the whole strain of dendrobium nobile seedling of sprinkling seedling aerial part single treatment in 10 days, is control
2.2 again.
B) second group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, it is red mould with 20mg/L
Element is 3.6 every the whole strain branch quantity highest of dendrobium nobile seedling of sprinkling seedling aerial part single treatment in 10 days, is control
1.1 again.
Root leaf is removed, stem is cut into the section of 0.7~1cm, is cleaned with deionized water, segment is cut into, is weighed, 80 DEG C are dried to
Constant weight is mixed and is crushed, and it is spare to cross 40 meshes.It is as shown in Figure 6 to measure each group stem fresh weight, dry weight result.Experimental result shows, the 4th
Group processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L lanthanum nitrate every the children of sprinkling in 10 days
The stem of Dendrobium fresh weight and dry weight highest of seedling aerial part single treatment, respectively 21.05g and 2.32g, the two are control
4.8 again.
Using 2015 editions pharmacopeia to polysaccharide content of dendrobium candidum measuring method, each group dendrobium polysaccharide content results are measured as schemed
Shown in 7.Experimental result shows that different composite liquid handling group dendrobium nobile seedling polyoses content is all remarkably higher than control group, wherein
A) the 4th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L lanthanum nitrate
It is 36.51% every the dendrobium nobile seedling polyoses content highest of sprinkling seedling aerial part single treatment in 10 days, is control
(22.20%) 1.6 times.
B) the 5th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L nitric acid
Lanthanum, 200mg/L auxin take second place every the dendrobium nobile seedling polyoses content of sprinkling seedling aerial part single treatment in 10 days, are
33.68%, it is 1.5 times for compareing (22.20%).
Using Wan withAcid-dye Colorimetry total alkaloid, it is as shown in Figure 8 to measure each group dendrobium nobile alkaloid result.It is real
It tests the results show that first group and the 5th group of processing mode dendrobium nobile seedling alkaloid are significantly higher than control group, wherein
A) the 5th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L nitric acid
Lanthanum, 200mg/L auxin are every the dendrobium nobile seedling dendrobium nobile alkali content highest of sprinkling seedling aerial part single treatment in 10 days
0.1963%, it is 1.2 times of control (0.1589%).
B) the 4th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L lanthanum nitrate
It is lower every dendrobium nobile seedling dendrobium nobile alkali content of 10 days sprinkling seedling aerial parts, it is 0.0790%, only compares
(0.1589%) 49.7%.
In conclusion survival rate, growth traits and the quality content of different composite liquid handling group dendrobium nobile seedling have significantly
Influence, wherein
A) the 4th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L lanthanum nitrate
Growth and development index of the dendrobium nobile seedling to dendrobium nobile seedling percent sum of seedling aerial part single treatment was sprayed every 10 days
Shape promotes maximum, and general performance is apparently higher than control and other processing groups.
B) the 5th group of processing mode: it is primary every 30 days inoculation seedling roots with 4g/L dendrobium mixture, with 5mg/L nitric acid
Lanthanum, 200mg/L auxin every 10 days sprinkling seedling aerial parts can once promote significantly dendrobium nobile seedling polyoses content and
Alkaloid improves.In officinal dendrobium stem plantation practice,
If preferentially selecting the 4th group of processing, i.e., with 4g/L dendrobium mixture every 30 days (a) using yield as primary goal
It is primary to be inoculated with seedling root, with 5mg/L lanthanum nitrate every 10 days sprinkling seedling dendrobium nobile seedling of aerial part.
If preferentially selecting the 5th group of processing (b) using quality as primary goal, being connect with 4g/L dendrobium mixture every 30 days
Kind seedling root is primary, primary every 10 days sprinkling seedling aerial parts with 5mg/L lanthanum nitrate, 200mg/L auxin.
If (c) yield and quality is taken into account, the 4th group and the 5th group should be used in combination, the should be used in tissue culture transplantation of seedlings
Four groups of processing then use the 5th group of processing mode after dendrobium nobile adapts to ambient enviroment, i.e., in the 4th group of base to improve its survival rate
200mg/L auxin was increased every 10 days on plinth.
The above is only the preferred embodiment of the present invention, the range implemented of the present invention that therefore, it cannot be limited according to, i.e., according to
Equivalent changes and modifications made by the invention patent range and description, should still be within the scope of the present invention.
Claims (7)
1. a kind of liquid composite bacteria agent for promoting Growth of Dendrobium candidum, it is characterised in that: including 3~5g/L dendrobium mixture
(Mycena dendrobii), 4~6mg/L lanthanum nitrate.
2. the liquid composite bacteria agent according to claim 1 for promoting Growth of Dendrobium candidum, it is characterised in that: further include 190
~210mg/L auxin.
3. the liquid composite bacteria agent according to claim 1 or 2 for promoting Growth of Dendrobium candidum, it is characterised in that: the stone
It after the small mushroom preparation method of dry measure used in former times includes: dendrobium mixture actication of culture, transfers in PDA culture medium, stands training in 22~25 DEG C of constant temperature
Support 14~20 days, then access in PDA culture medium carry out fermented and cultured 7~14 days to get.
4. promote the application method of the liquid composite bacteria agent of Growth of Dendrobium candidum described in a kind of any one of claims 1 to 3,
It is characterized by: dendrobium mixture is inoculated in dendrobium candidum seedling root, lanthanum nitrate, auxin spray in the liquid composite bacteria agent
It spills in dendrobium candidum seedling aerial part.
5. application method according to claim 4, it is characterised in that: in the liquid composite bacteria agent, dendrobium mixture every
Inoculation dendrobium candidum seedling root is primary within 28~32 days, and lanthanum nitrate, auxin were every 9~11 days sprinkling dendrobium candidum seedling grounds
Part is primary.
6. application method according to claim 4 or 5, it is characterised in that: the inoculum concentration of the dendrobium mixture is 4~6mL.
7. application method according to claim 4 or 5, it is characterised in that: the dendrobium candidum seedling is after domestication
It is transplanted to the dendrobe tissue culture seedling of greenhouse growth.
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| CN111727881A (en) * | 2020-06-17 | 2020-10-02 | 安徽皖斛堂生物科技有限公司 | Culture medium and culture method for promoting sterile germination of dendrobium officinale seeds |
| CN112980701A (en) * | 2021-04-17 | 2021-06-18 | 浙江理工大学 | Composite microbial inoculum for promoting germination and growth of dendrobium officinale seeds |
| CN114766341A (en) * | 2022-06-07 | 2022-07-22 | 浙江省亚热带作物研究所(浙南林业科学研究院) | Application of mycorrhizal fungi in cultivation of dendrobium officinale tissue culture seedlings |
| CN114830969A (en) * | 2022-06-07 | 2022-08-02 | 云南菌视界生物科技有限公司 | Organic compound capable of shortening cultivation period of tremella aurantialba and application thereof |
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| CN111727881A (en) * | 2020-06-17 | 2020-10-02 | 安徽皖斛堂生物科技有限公司 | Culture medium and culture method for promoting sterile germination of dendrobium officinale seeds |
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| CN114766341A (en) * | 2022-06-07 | 2022-07-22 | 浙江省亚热带作物研究所(浙南林业科学研究院) | Application of mycorrhizal fungi in cultivation of dendrobium officinale tissue culture seedlings |
| CN114830969A (en) * | 2022-06-07 | 2022-08-02 | 云南菌视界生物科技有限公司 | Organic compound capable of shortening cultivation period of tremella aurantialba and application thereof |
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Application publication date: 20190524 |