CN107155896B - A method of promote African Chrysanthemum tissue culture to transplant seedling rooting - Google Patents
A method of promote African Chrysanthemum tissue culture to transplant seedling rooting Download PDFInfo
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- CN107155896B CN107155896B CN201710567470.XA CN201710567470A CN107155896B CN 107155896 B CN107155896 B CN 107155896B CN 201710567470 A CN201710567470 A CN 201710567470A CN 107155896 B CN107155896 B CN 107155896B
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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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- 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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Abstract
The present invention provides a kind of method of promotion African Chrysanthemum tissue culture transplanting seedling rooting, by pouring India's pyriform spore, promote African Chrysanthemum tissue culture transplanting seedling rooting using India's pyriform spore, improve root system and then promote plant strain growth, furthermore the present invention can also the problems such as regeneration plant root system caused by preferably multiple subculture is weak, slow growth, weak seedling, be of great significance to the promotion of African Chrysanthemum plant commodity value.Step of the present invention is simple, strong operability, significant effect.
Description
Technical field
The present invention relates to African Chrysanthemum seedling-raising technique field more particularly to a kind of sides for promoting African Chrysanthemum tissue culture transplanting seedling rooting
Method.
Background technique
African Chrysanthemum is composite family herbaceos perennial, is one of big cut-flower in the world five.African Chrysanthemum is not pollinated from flower, and tradition is non-
Continent chrysanthemum produces to be bred frequently with the method for division propagation, but division propagation breeding coefficient is lower, easily accumulate disease, plant division after
Generation growth is uneven, seedling is weak, kind is easily degenerated.Tissue-cultured seedling has stabilization characteristics of genetics, growing way uniformity, fast, the degeneration-resistant energy of growth
The strong advantage of power, therefore African Chrysanthemum production both at home and abroad mostly uses tissue-cultured seedling at present.It, will be tight if tissue culture transplanted seedling root system development is bad
Ghost image rings plant strain growth, reduces its commodity value, the tissue culture transplanted seedling of multiple subculture be even more will appear weak root system, slow growth,
The problems such as growing way is weak, therefore need to induce group that is primary rather than using multiple subculture again at regular intervals mostly in production
Train seedling.Therefore certain means is taken to promote African Chrysanthemum tissue culture transplanting seedling rooting that there is certain meaning to African Chrysanthemum the factorial production
Justice.
India's pyriform spore is a kind of class arbuscular mycorrhizal fungi that can manually cultivate, numerous research shows that it is to root system of plant
Formation, plant strain growth, plant resistance capacity have apparent facilitation.But it there is no to apply in African Chrysanthemum production at present
Report.African Chrysanthemum tissue culture transplanting seedling rooting is promoted once to be proved using India's pyriform spore feasible, it will there is biggish life
Produce application value and economic value.The present invention provides it is a kind of using India's pyriform spore promote African Chrysanthemum tissue culture transplanting seedling rooting and
Restore the method for the African Chrysanthemum tissue culture transplanted seedling root activity of multiple subculture.
Summary of the invention
The present invention provides a kind of methods of promotion African Chrysanthemum tissue culture transplanting seedling rooting, can efficiently solve African Chrysanthemum group
Training transplanted seedling root system development is poor and has a series of problems of its generation.
Technical solution of the present invention:
A method of promote African Chrysanthemum tissue culture to transplant seedling rooting, preparation, African Chrysanthemum including India's pyriform spore fermentation liquid
The transplanting of tissue-cultured seedling, India's pyriform spore fermentation liquid pour, and verify the effect of India's pyriform spore fermentation liquid hestening rooting later.Pass through
India's pyriform spore is poured, promotes African Chrysanthemum tissue culture transplanting seedling rooting using India's pyriform spore, improve root system and then promotes plant raw
Long, furthermore the present invention can also regeneration plant root system be weak, slow growth, seedling is weak, survival rate is low caused by preferably multiple subculture etc.
Problem is of great significance to the promotion of African Chrysanthemum plant commodity value.Step of the present invention is simple, strong operability, effect are aobvious
It writes.
Method particularly includes:
(1) preparation of India's pyriform spore fermentation liquid
India's pyriform spore bacterial strain is activated, then the aerial hyphae of picking colony marginal portion is inoculated in PDA solid medium
On；It is placed in 28 DEG C of dark culturing case and cultivates 5 d, picking mycelia is in the PDA liquid medium equipped with 100 ml sterilizing
It is 28 DEG C in temperature in 250ml triangular flask, revolving speed is culture 3 days in the shaking table of 120 r/min, is received after mono layer gauze filters
Collect filtrate, being diluted with water to spore concentration is 2 × 107A/ml obtains India's pyriform spore fermentation liquid.
(2) transplanting of African Chrysanthemum tissue-cultured seedling
The 30 days height of having cultivated in root media for choosing continuous subculture 5-20 generation are the African Chrysanthemum tissue culture of 3 ~ 4cm
Seedling；It is transplanted after hardening of uncapping 2 days into the flowerpot of the humus equipped with basin body product 3/4, one plant of every basin；Flowerpot diameter used is 10
It cm, is highly 10 cm.
(3) India's pyriform spore fermentation liquid pours
50 ml India pyriform spore fermentation liquids are nearby poured into African Chrysanthemum tissue culture transplanted seedling root system.It is placed in 25 ± 2 DEG C of constant temperature
It is cultivated in culturing room, culturing room's intensity of illumination is 1300 ± 200 lx, and light application time is 12 h/d.
The invention has the following advantages that
1, India's pyriform spore used in the present invention is a kind of class mycorrhizal fungi that can manually cultivate, and expands complicated and simple list.
2, India's pyriform spore fermentation liquid preparing process is simple, and strong operability can obtain bulk fermentation within a short period of time
Liquid.
3, India's pyriform spore fermentation liquid pours easy to operate.
4, the present invention can promote African Chrysanthemum tissue culture transplanted seedling root growth, and the quality and commodity valence of African Chrysanthemum can be improved
Value.
5, the present invention can restore the African Chrysanthemum tissue culture transplanted seedling root activity of multiple subculture, can effectively solve due to multiple
Regeneration plant root system caused by subculture is weak, slow growth, the problems such as seedling is weak, survival rate is low, has biggish production practices application
Value and economic value.
Detailed description of the invention
The African Chrysanthemum plant in 5 generation of subculture of 30 days (A) and 90 days (B) after Fig. 1 transplanting, from left to right successively are as follows: India's pears
Shape spore fermentation liquor treatment group, India's pyriform spore fermentation liquor treatment group of sterilizing and PDA liquid medium control group.
The African Chrysanthemum plant in 10 generation of subculture of 30 days (A) and 90 days (B) after Fig. 2 transplanting, from left to right successively are as follows: India
Pyriform spore fermentation liquor treatment group, India's pyriform spore fermentation liquor treatment group of sterilizing and PDA liquid medium control group.
The African Chrysanthemum plant in 15 generation of subculture of 30 days (A) and 90 days (B) after Fig. 3 transplanting, from left to right successively are as follows: India
Pyriform spore fermentation liquor treatment group, India's pyriform spore fermentation liquor treatment group of sterilizing and PDA liquid medium control group.
The African Chrysanthemum plant in 20 generation of subculture of 30 days (A) and 90 days (B) after Fig. 4 transplanting, from left to right successively are as follows: India
Pyriform spore fermentation liquor treatment group, India's pyriform spore fermentation liquor treatment group of sterilizing and PDA liquid medium control group.
The African Chrysanthemum plant (A) handled without India's pyriform spore in 20 generation of Fig. 5 subculture and the Africa for thering is India's pyriform spore to handle
Chrysanthemum plant survives situation (B).
Specific embodiment
The following further describes the technical solution of the present invention with reference to specific embodiments, but the present invention is not limited in
This.
Embodiment 1
1, the preparation of India's pyriform spore fermentation liquid
India's pyriform spore bacterial strain is activated, then the aerial hyphae of picking colony marginal portion is inoculated in new PDA solid training
It supports on base.It is placed in 28 DEG C of dark culturing case and cultivates 5 d, picking mycelia is in the PDA Liquid Culture equipped with 100 ml sterilizing
It is 28 DEG C in temperature in the 250ml triangular flask of base, revolving speed is culture 3 days in the shaking table of 120 r/min, is filtered through mono layer gauze
After collect filtrate, be diluted with water to spore concentration be 2 × 107A/ml obtains India's pyriform spore fermentation liquid.
2, the transplanting of African Chrysanthemum tissue-cultured seedling
The 30 days height of having cultivated in root media for choosing 5 generation of continuous subculture are the African Chrysanthemum tissue-cultured seedling of 3 ~ 4cm.
It transplants after hardening of uncapping 2 days into the flowerpot equipped with 3/4 humus, one plant of every basin.Flowerpot diameter used is 10 cm, is highly
10 cm。
3, India's pyriform spore fermentation liquid pours
50 ml India pyriform spore fermentation liquids are nearby poured into African Chrysanthemum tissue culture transplanted seedling root system.Sterilizing India is set simultaneously
Pyriform spore fermentation liquor treatment group and PDA liquid medium control group.It is placed in 25 ± 2 DEG C of constant temperature incubation room and cultivates, culturing room
Intensity of illumination is 1300 ± 200lx, and light application time is 12 h/d.The three groups of African Chrysanthemums of observation in 30 days and 90 days are planted after transplanting respectively
Strain root growth situation is simultaneously taken pictures.
The African Chrysanthemum tissue culture transplanted seedling in 5 generations of continuous subculture is after transplanting 30 days (Figure 1A): at India's pyriform spore fermentation liquid
Reason group African Chrysanthemum transplanting seedling rooting situation is best, and fibrous root is more.India's pyriform spore fermentation liquor treatment group of sterilizing is taken second place, PDA liquid
Body medium controls root growth situation is worst.90 days (Figure 1B) after transplanting, India's pyriform spore fermentation liquor treatment group and goes out
India's pyriform spore fermentation liquor treatment group African Chrysanthemum root system fibrous root number of bacterium is significantly more than PDA liquid medium.
Embodiment 2
1, the preparation of India's pyriform spore fermentation liquid
India's pyriform spore bacterial strain is activated, then the aerial hyphae of picking colony marginal portion is inoculated in new PDA solid training
It supports on base.It is placed in 28 DEG C of dark culturing case and cultivates 5 d, picking mycelia is in the PDA Liquid Culture equipped with 100 ml sterilizing
It is 28 DEG C in temperature in the 250ml triangular flask of base, revolving speed is culture 3 days in the shaking table of 120 r/min, is filtered through mono layer gauze
After collect filtrate, be diluted with water to spore concentration be 2 × 107A/ml obtains India's pyriform spore fermentation liquid.
2, the transplanting of African Chrysanthemum tissue-cultured seedling
The 30 days height of having cultivated in root media for choosing 10 generation of continuous subculture are the African Chrysanthemum tissue culture of 3 ~ 4 cm
Seedling.It transplants after hardening of uncapping 2 days into the flowerpot equipped with 3/4 humus, one plant of every basin.Flowerpot diameter used is 10 cm, height
For 10 cm.
3, India's pyriform spore fermentation liquid pours
50 ml India pyriform spore fermentation liquids are nearby poured into the African Chrysanthemum tissue culture transplanted seedling root system in 10 generations of continuous subculture.
Sterilizing India's pyriform spore fermentation liquor treatment group and PDA liquid medium control group are set simultaneously.It is placed in 25 ± 2 DEG C of constant temperature incubation
Indoor culture, culturing room's intensity of illumination are 1300 ± 200 lx, and light application time is 12 h/d.Respectively 30 days and 90 days after transplanting
It observes three groups of African Chrysanthemum plant root growing states and takes pictures.
The African Chrysanthemum tissue culture transplanted seedling in 10 generations of continuous subculture is after transplanting 30 days (Fig. 2A), at India's pyriform spore fermentation liquid
Reason group and sterilizing India's pyriform spore fermentation liquor treatment group African Chrysanthemum plant take root situation be slightly better than PDA liquid medium control group
Most preferably, wherein India's pyriform spore fermentation liquor treatment group plant root is best.(Fig. 2 B), PDA liquid medium 90 days after transplanting
Control group plant root is poorer than the African Chrysanthemum tissue culture transplanted seedling in 5 generations of continuous subculture.It India's pyriform spore fermentation liquor treatment group and goes out
India's pyriform spore fermentation liquor treatment group African Chrysanthemum root system fibrous root number of bacterium is significantly more than PDA liquid medium control group, wherein
India's pyriform spore fermentation liquor treatment group plant root is best.
Embodiment 3
1, the preparation of India's pyriform spore fermentation liquid
India's pyriform spore bacterial strain is activated, then the aerial hyphae of picking colony marginal portion is inoculated in new PDA solid training
It supports on base.It is placed in 28 DEG C of dark culturing case and cultivates 5 d, picking mycelia is in the PDA Liquid Culture equipped with 100 ml sterilizing
It is 28 DEG C in temperature in the 250ml triangular flask of base, revolving speed is culture 3 days in the shaking table of 120 r/min, is filtered through mono layer gauze
After collect filtrate, obtain India's pyriform spore fermentation liquid.
2, the transplanting of African Chrysanthemum tissue-cultured seedling
The 30 days height of having cultivated in root media for choosing 15 generation of continuous subculture are the African Chrysanthemum tissue culture of 3 ~ 4cm
Seedling.It transplants after hardening of uncapping 2 days into the flowerpot equipped with 3/4 humus, one plant of every basin.Flowerpot diameter used is 10 cm, height
For 10 cm.
3, India's pyriform spore fermentation liquid pours
50 ml India pyriform spore fermentation liquids are nearby poured into the African Chrysanthemum tissue culture transplanted seedling root system in 15 generations of continuous subculture.
Sterilizing India's pyriform spore fermentation liquor treatment group and PDA liquid medium control group are set simultaneously.It is placed in 25 ± 2 DEG C of constant temperature incubation
Indoor culture, culturing room's intensity of illumination are 1300 ± 200 lx, and light application time is 12 h/d.Respectively 30 days and 90 days after transplanting
It observes three groups of African Chrysanthemum plant root growing states and takes pictures.
The African Chrysanthemum tissue culture transplanted seedling in 15 generations of continuous subculture is after transplanting 30 days (Fig. 3 A), at India's pyriform spore fermentation liquid
Reason group and sterilizing India's pyriform spore fermentation liquor treatment group African Chrysanthemum plant take root, and slightly better than PDA liquid medium compares situation
Group.90 days after transplanting (Fig. 3 B), Africa of the PDA liquid medium control group plant root than continuous subculture 5 generations and 10 generations
Chrysanthemum plant is poor.India's pyriform spore fermentation liquor treatment group African Chrysanthemum root system fibrous root number of India's pyriform spore fermentation liquor treatment group and sterilizing
It is significantly more than PDA liquid medium control group, wherein India's pyriform spore fermentation liquor treatment group plant root is best.
Embodiment 4
1, the preparation of India's pyriform spore fermentation liquid
India's pyriform spore bacterial strain is activated, then the aerial hyphae of picking colony marginal portion is inoculated in new PDA solid training
It supports on base.It is placed in 28 DEG C of dark culturing case and cultivates 5 d, picking mycelia is in the PDA Liquid Culture equipped with 100 ml sterilizing
It is 28 DEG C in temperature in the 250ml triangular flask of base, revolving speed is culture 3 days in the shaking table of 120 r/min, is filtered through mono layer gauze
After collect filtrate, be diluted with water to spore concentration be 2 × 107A/ml obtains India's pyriform spore fermentation liquid.
2, the transplanting of African Chrysanthemum tissue-cultured seedling
The 30 days height of having cultivated in root media for choosing 20 generation of continuous subculture are the African Chrysanthemum tissue culture of 3 ~ 4cm
Seedling.It transplants after hardening of uncapping 2 days into the flowerpot equipped with 3/4 humus, one plant of every basin.Flowerpot diameter used is 10 cm, height
For 10 cm.
3, India's pyriform spore fermentation liquid pours
50 ml India pyriform spore fermentation liquids are nearby poured into the African Chrysanthemum tissue culture transplanted seedling root system in 20 generations of continuous subculture.
Sterilizing India's pyriform spore fermentation liquor treatment group and PDA liquid medium control group are set simultaneously.It is placed in 25 ± 2 DEG C of constant temperature incubation
Indoor culture, culturing room's intensity of illumination are 1300 ± 200 lx, and light application time is 12 h/d.Respectively 30 days and 90 days after transplanting
It observes three groups of African Chrysanthemum plant root growing states and takes pictures.
Root system difference is not after transplanting 30 days (Fig. 4 A), between three groups for the African Chrysanthemum tissue culture transplanted seedling in 20 generations of continuous subculture
Greatly, root system development is weaker.90 days after transplanting (Fig. 4 B), PDA liquid medium control group plant root is than continuous subculture 5
The PDA liquid medium control group African Chrysanthemum plant in generation, 10 generations and 15 generations is poor.India's pyriform spore fermentation liquor treatment group and sterilizing
India's pyriform spore fermentation liquor treatment group African Chrysanthemum root system fibrous root number is significantly more than PDA liquid medium control group, wherein India
Pyriform spore fermentation liquor treatment group plant root is best.Transplanted seedling (Fig. 5 A) survival rate without India's pyriform spore processing group is on 86% left side
The right side, and transplanted seedling (Fig. 5 B) survival rate of India's pyriform spore processing group then can reach 93% or more.
It can be seen that: the transplanting of African Chrysanthemum tissue culture can be remarkably promoted by pouring African Chrysanthemum using India's pyriform spore fermentation liquid
The tissue culture transplanting shoot survival percent that seedling rooting can simultaneously improve the African Chrysanthemum tissue culture transplanted seedling root system of multiple subculture, improve multiple subculture.
The present invention pours African Chrysanthemum tissue culture transplanted seedling using India's pyriform spore fermentation liquid, significantly promotes the shifting of African Chrysanthemum tissue culture
Plant seedling rooting, can preferably improve the tissue culture transplanted seedling root system of multiple subculture, for African Chrysanthemum produce it is significant, have compared with
High application and economic value.
Claims (1)
1. a kind of method for promoting African Chrysanthemum tissue culture transplanting seedling rooting, it is characterised in that: the system including India's pyriform spore fermentation liquid
Standby, African Chrysanthemum tissue-cultured seedling transplanting, India's pyriform spore fermentation liquid pour；
Specifically comprise the following steps:
(1) preparation of India's pyriform spore fermentation liquid
India's pyriform spore bacterial strain is activated, then the aerial hyphae of picking colony marginal portion is inoculated on PDA solid medium；It puts
It is placed in 28 DEG C of dark culturing case and cultivates 5 d, picking mycelia is in the 250ml of the PDA liquid medium equipped with 100 ml sterilizing
It is 28 DEG C in temperature in triangular flask, revolving speed is culture 3 days in the shaking table of 120 r/min, and filter is collected after mono layer gauze filters
Liquid, being diluted with water to spore concentration is 2 × 107A/ml obtains India's pyriform spore fermentation liquid；
(2) transplanting of African Chrysanthemum tissue-cultured seedling
The 30 days height of having cultivated in root media for choosing continuous subculture 5-20 generation are the African Chrysanthemum tissue-cultured seedling of 3 ~ 4 cm；
It is transplanted after hardening of uncapping 2 days into the flowerpot of the humus equipped with basin body product 3/4, one plant of every basin；Flowerpot diameter used is 10
It cm, is highly 10 cm；
(3) India's pyriform spore fermentation liquid pours
50 ml India pyriform spore fermentation liquids are nearby poured into African Chrysanthemum tissue culture transplanted seedling root system；It is placed in the constant temperature training of (25 ± 2) DEG C
Interior culture is supported, culturing room's intensity of illumination is (1300 ± 200) lx, and light application time is 12 h/d.
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| CN108668896B (en) * | 2018-04-24 | 2021-08-31 | 福建农林大学 | Method for improving gerbera jamesonii root rot resistance |
| CN109122095A (en) * | 2018-08-10 | 2019-01-04 | 福建农林大学 | A method of it cultivating high-quality passionflower seedling and improves plant resistance |
| CN110122304A (en) * | 2019-06-14 | 2019-08-16 | 福建农林大学 | A kind of method of banana water planting hardening |
| CN110235665A (en) * | 2019-07-12 | 2019-09-17 | 三明市农业科学研究院 | A kind of method of biological control Africa Black Ostrich |
| CN116138164A (en) * | 2022-07-06 | 2023-05-23 | 三明市农业科学研究院 | Gerbera strong seedling cultivation method |
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