CN105993865A - Cultivation method for quercus variabilis aseptic seedling - Google Patents
Cultivation method for quercus variabilis aseptic seedling Download PDFInfo
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- CN105993865A CN105993865A CN201510114224.XA CN201510114224A CN105993865A CN 105993865 A CN105993865 A CN 105993865A CN 201510114224 A CN201510114224 A CN 201510114224A CN 105993865 A CN105993865 A CN 105993865A
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Abstract
The invention belongs to the field of seedling growing, and specifically relates to a cultivation method for quercus variabilis aseptic seedlings, and especially the quercus variabilis aseptic seedling can be used to inoculate truffles. An objective of the invention is to provide the cultivation method for quercus variabilis aseptic seedlings. A safe potassium permanganate solution is used to disinfect, processing process is simple, seedling growing time is saved, and seedling growing effect is substantial. The quercus variabilis aseptic seedlings obtained by the cultivation method are especially suitable to be used to inoculate truffles. The method comprises steps: (1) selecting seeds; (2) disinfecting and treating the seeds; (3) accelerating germination; (4) sowing; (5) performing seedling emergence management. Using the method can effectively improve truffle mycorrhizal seedling formation time, improves quercus variabilis seed emergence rate and planting percent, improves seedling uniformity, and substantially improves seedling quality. The quercus variabilis seedlings cultured by the method can be directly used for inoculation of truffles.
Description
Technical field
The invention belongs to nursery field, being specifically related to is the breeding method of a kind of cork oak aseptic seedling, and especially this cork oak is aseptic
Seedling can be used for inoculating truffles.
Background technology
Cork oak (Quercus variabilis) is that China is distributed one of the widest seeds within the border, is also important cork, tannic extract money
Source seeds, with multiple Applying Ectomycorrhizal Fungi symbiosis.Under natural conditions, in scondary forest is formed, it is topmost one-tenth that stump sprouts Seedling
Seedling mode, seedling complete life cycle by environment and seedling side fibrous root few affected the biggest.Cork oak planting by stock survival rate is only
30～50%.
In existing report, in " a kind of method for cultivating cork oak seedlings " of Application No. CN201210289748.9, exist and adopt
With asking of the cumbersome process such as formalin disinfectant solution, region restriction, seed soaking time oversize (46～52h), repeatedly root-inducing powder process
Topic.In recent years, scholars are more to applied research on afforestation of the seedling of cork oak, are embodied in control root, radicle short more
Cut, on Mycorrhizal Seedling, cultivation of container seedling etc., there are some researches show, such as " impact on cork oak growth of seedling of the Arbuscular Mycorrhizal Fungi bacterium "
[Li Hao etc., Agriculture of Anhui science, 2013 (6): 2536-2538.] report, cork oak Mycorrhizal Seedling, there is seedling percent high, young
Seedling growth is fast, seedling mycorhiza forms fast and that seedling quality is high feature.Truffles Seedling when cultivating more selects Pinus armandi Franch-P. Komavovii Lavl., oriental white oak, Semen Castaneae
Deng seeds, inventor finds that cork oak has clear superiority on environmental suitability, shortening nursery, inoculation time, but in routine
In the nursery of cork oak field, disease, seedling quality are difficult to control to, and cause cork oak transplanting survival rate low, use the present invention to cultivate
Method, it is achieved that cork oak is cultivated overall process Human disturbance and controls, and has cultivated the aseptic strong sprout being suitable to inoculate truffles, has widened
It utilizes field.
Summary of the invention
It is an object of the invention to provide the breeding method of a kind of cork oak aseptic seedling, have employed safer disinfecting solution of potassium permanganate,
Treatment process is simple, save seedling raise period and nursery effect is more notable, and the cork oak aseptic seedling that above-mentioned breeding method obtains is outstanding
It is applicable to inoculate truffles.
The breeding method of cork oak aseptic seedling of the present invention comprises the steps:
(1) seed selection；
(2) sterilization: rinse seed with water, the potassium permanganate solution immersion 1～4h with concentration is 0.2%～0.6% (mass fraction):
Ambient temperature 25～32 DEG C, potassium permanganate solution floods more than seed 2cm；Then seed airtight 15～30min is encapsulated with sealed bag,
Use aseptic water washing again；
(3) accelerating germination: soak seed 4～12h with sterilized water that initial temperature is 50～60 DEG C, then by concentration 50～120mg/L
Gibberellins solution soaks seed 12～24h, is placed in the incubator of 25～32 DEG C during seed soaking；
(4) sowing: be placed on by seed in the germination box keeping bottom humid, is placed in the incubator of 25～30 DEG C 4～7 days
" show money or valuables one carries unintentionally " to visible seed, can sow in substrate；
Described sowing media is: 0.8～1.5:0.5～1.2:0.7～1.0 mixing by volume of perlite, Vermiculitum, water, takes out after sterilizing
Substrate, is cooled to room temperature, and regulation substrate pH is to 6.5～7.5；
(5) emerge management: sowing is unearthed successively to seedling, move to growth cabinet is cultivated, by following condition of culture circulate into
Row A section and B section:
A section: illuminance 33%～66%, time 8～11 hours, humidity 75%～90%, temperature 25 DEG C～32 DEG C；
B section: illuminance 0%, time 13～16 hours, humidity 75%～95%, temperature 25 DEG C～32 DEG C.
Use breeding method of the present invention, after seedling age reaches March, just can transplant, cultivate gained cork oak Seedling and be particularly suited for and block
Bacterium inoculates.
In technique scheme, step (1) described seed selection, for carrying out grain-by-grain seed selection, rejects the seed that disease pest is damaged and bad colour is normal,
Select color and luster, size uniform and full, without going mouldy, free from insect pests, the most floating seed；Seed source is harvesting ripe voluntarily
Novel species, cleanliness is 90～96%；Or buy to authoritative kind of a unit of selling.
In technique scheme, during step (4) sowing, it is placed in the incubator of 25～30 DEG C 4～7 days and " reveals to visible seed
In vain ", plant 1 time with aseptic washing every day period.This is owing to seed is in germination process, and on the one hand seed can ferment generation ethanol
The material occurred etc. harmful root system, is on the other hand, due to surface, substantial amounts of endophyte and external source bacterium to occur, so needing to clean.
In technique scheme, during step (4) sowing, substrate is laid in seedlings nursing plate, and the seedlings nursing plate used is plastic crate dish,
The fly net of surrounding pad 30～60 mesh, it is deep that seed base-root grows end the most upright sowing 1～2cm, and spacing 0.5～1cm is placed on
In the environment of temperature is more than 20 DEG C.
In technique scheme, during step (4) sowing, the condition of substrate sterilizing is sterilizing 40min～2h under the conditions of 121 DEG C.
In technique scheme, during step (4) sowing, the substrate pH value after the sterilized cooling of the perlite of employing, Vermiculitum is
6.5, according to truffles growth characteristics, quick lime regulation substrate pH can be used to 6.5～7.5；Keeping germination box bottom humid is to use
Filter paper pads is in germination cassette bottom portion, and described filter paper is qualitative filter paper；Germination box is 13cm × 19cm × 12cm high-quality ABS transparent box；
Seedlings nursing plate specification is 32cm × 28cm × 10cm.
Wherein, the reason that step (2) sterilization uses sealed bag encapsulation seed airtight is to reduce disease further, as prevented standing
Rot, canker etc..
The advantage of cultural method of the present invention is as follows:
1) use method for culturing seedlings of the present invention, select high-quality kind, suitable concentration disinfecting solution of potassium permanganate, thermophilic to break dormancy, loosen
The substrate that ventilative water retention property is good, finally significantly improves seedling side root generating capacity, seedling quality and germination percentage；
2) use the inventive method nursery respective container Seedling can reduce production cost, and germination percentage is more than 90%；
3) aseptic seedling using the present invention to cultivate can meet the requirement of inoculation truffles, and forming truffle mycorrhiza fungi Seedling can be effectively improved
Seedling replanting survival rate and the time of shortening mycorrhiza fungi seeding cultivating；
4) substrate that method for culturing seedlings of the present invention is selected is loose ventilative, and can do can be wet, can regulate main root length, energy according to stromal thickness
Effectively regulate root/shoot ratio, be easily formed strong sprout；
5) operating procedure is simple.
Generally, cork oak planting by stock survival rate only has 30～50%, by method for culturing seedlings of the present invention, can be effectively improved bolt
Skin oak percentage of seedgermination and planting percent, improve seedling regularity, significantly improves seedling quality, the cork that the application present invention cultivates
Oak Seedling can be directly used for the inoculation of truffles, also improves the time that truffle mycorrhiza fungi Seedling is formed, to China's truffles artificial culture process and
Afforestation industry is respectively provided with outstanding contributions.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is further illustrated, and following each embodiment is merely to illustrate the present invention, to the present invention
Do not limit.
Embodiment 1
The breeding method of a kind of cork oak aseptic seedling for inoculating truffles, comprises the following steps:
(1) novel species of harvesting ripe voluntarily, cleanliness is 94%；
(2) after plucking seed, carry out grain-by-grain seed selection, reject the seed that disease pest is damaged and bad colour is normal, select color and luster, size uniform
And full, without going mouldy, free from insect pests, the most floating seed；
(3) 2h (ambient temperature 30 DEG C, sterilization are processed with the disinfecting solution of potassium permanganate of 0.5% after tap water rinse seed 3 times
Liquid is advisable to flood more than seed 2cm), encapsulate the airtight 30min of seed with sealed bag, then with aseptic water washing 5 times；
(4) after by initial temperature being the aseptic water seed soaking 4h of 55 DEG C, then soak seed 24h by the Gibberellins solution of 100mg/L,
And seed soaking process is placed in the incubator of 30 DEG C；Seed is placed on bottom the most again be lined with in the germination box of moistening filter paper, puts
Put in the incubator of 30 DEG C, through 5 days (every day period plants 1 time with aseptic washing), it is seen that seed starts " showing money or valuables one carries unintentionally ", i.e.
Can sow in substrate；Seedlings nursing plate uses plastic crate dish, the fly net of surrounding pad 60 mesh, and substrate is down to wherein and pave, seed
It is deep that radicle grows end upright sowing 1.5cm downwards, spacing 1cm, be placed on temperature more than 20 DEG C in the environment of；
Described sowing media is: after " perlite+Vermiculitum+water " is according to the proportioning mix homogeneously of volume ratio 1:1:1, be placed in 121 DEG C
Under the conditions of, sterilizing 1h, end subject to sterilization, take out substrate, be cooled to room temperature, such substrate, measure through pH meter and learn pH
It is 6.5, according to truffles growth characteristics, with quick lime regulation substrate pH to 7；Described filter paper is qualitative filter paper；Germination box specification
For 13cm × 19cm × 12cm high-quality ABS transparent box；Seedlings nursing plate uses specification 32cm × 28cm × 10cm；
(5), after sowing 8 days, it is seen that seedling is unearthed successively, then carrying seed tray is circulated by following condition to growth cabinet
Cultivate (first paragraph: temperature 25 DEG C, time 8h, illuminance 33%, humidity 85%；Second segment temperature 28 DEG C, time 16h,
Illuminance 0%, humidity 92%), after seedling age reaches March, randomly select 20 strains in its root situation of body formula Microscopic observation, really
Determine root growth good after, transplant seedling in nutritive cube, just for the inoculation of truffles.
(6) truffle mycorrhiza fungi inoculation；The method using Publication No. CN103070013A to disclose carries out truffle mycorrhiza fungi inoculation.
(7) mycorhiza detection；Take out nursery stock after inoculating 5 months, typical exotrophic mycorrhiza is taken off with aseptic pincet, at body
Brush off earth (or with ultrasonic waves for cleaning) with hairbrush under visor, clean up with distilled water, root system is placed directly in 0.7～11.5 times
The formation state of stereomicroscopy Microscopic observation mycorhiza and feature, and add up mycorhiza infection rate, infection rate=infection Seedling number/inoculation Seedling
Number × 100%.Mycorhiza infection intensity uses the method for randomization to add up, infection intensity=infection root segment number/investigation root segment number
× 100%.
(8) result；Related data of the present invention is as shown in table 1.
Table 1 relevant experimental data of the present invention
Note:
A representative employing " Wang Shiguo, Wang Shibao. cork oak seedling-raising technique and seedling growth research. Ningxia agriculture and forestry science and technology, 2012,53 (02): 20-21. " method
The truffles inoculation that the seedling cultivated is carried out.
B represents the truffles inoculation that the seedling that patent CN201210289748.9 " a kind of method for cultivating cork oak seedlings " cultivated is carried out.Infection rate and
Infection intensity is to detect data after truffles inoculation May, seedling data before remaining inoculation.
From table 1, breeding method of the present invention occupies certain on raising percentage of seedgermination, side radical, side root length, root/shoot ratio
Advantage, and widened its application, cork oak seedling inoculation truffles infection rate, the mycorhiza cultivated by the inventive method are infected
Index occupies advantage than other method.
Embodiment described above is only to be described embodiment of the present invention, does not enter the spirit and scope of the present invention
Row limits；The content that the present invention is claimed all is recorded in detail in the claims.
Claims (9)
1. the breeding method of cork oak aseptic seedling, it is characterised in that: comprise the steps:
(1) seed selection；
(2) sterilization: rinse seed with water, by concentration be 0.2～the potassium permanganate solution of 0.6% (mass fraction) soaks 1～4
H: ambient temperature 25～32 DEG C, potassium permanganate solution floods more than seed 2cm；Then with sealed bag encapsulation seed airtight 15～
30min, then use aseptic water washing；
(3) accelerating germination: soak seed 4～12h with sterilized water that initial temperature is 50～60 DEG C, then by concentration 50～120mg/L
Gibberellins solution soak seed 12～24h, be placed on during seed soaking in the incubator of 25～32 DEG C；
(4) sowing: be placed on by seed in the germination box keeping bottom humid, is placed in the incubator of 25～30 DEG C 4～7
It " shows money or valuables one carries unintentionally " to visible seed, can sow in substrate；
Described sowing media is: 0.8～1.5:0.5～1.2:0.7～1.0 mixing by volume of perlite, Vermiculitum, water, takes out after sterilizing
Substrate, is cooled to room temperature, and regulation substrate pH is to 6.5～7.5；
(5) emerge management: sowing is unearthed successively to seedling, move to growth cabinet is cultivated, by following condition of culture circulate into
Row A section and B section:
A section: illuminance 33%～66%, time 8～11 hours, humidity 75%～90%, temperature 25 DEG C～32 DEG C；
B section: illuminance 0%, time 13～16 hours, humidity 75%～95%, temperature 25 DEG C～32 DEG C.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: step (1) described seed selection
For carrying out grain-by-grain seed selection, rejecting the seed that disease pest is damaged and bad colour is normal, selection color and luster, size are uniform and full, nothing is gone mouldy, nothing
Damage by worms, the most floating seed.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: during step (4) sowing,
Being placed in the incubator of 25～30 DEG C 4～7 days and " show money or valuables one carries unintentionally " to visible seed, every day period plants 1 time with aseptic washing.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: during step (4) sowing,
Substrate is laid in seedlings nursing plate, and the seedlings nursing plate used is plastic crate dish, the fly net of surrounding pad 30～60 mesh, seed embryo root length
Go out end upright sowing 1～2cm downwards deep, spacing 0.5～1cm, be placed on temperature more than 20 DEG C in the environment of.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: during step (4) sowing,
The condition of substrate sterilizing is sterilizing 40min～2h under the conditions of 121 DEG C.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: during step (4) sowing,
Use quick lime regulation substrate pH to 6.5～7.5.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: during step (4) sowing,
Keeping germination box bottom humid is to use filter paper pads in germination cassette bottom portion, and described filter paper is qualitative filter paper.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: during step (4) sowing,
Germination box is 13cm × 19cm × 12cm high-quality ABS transparent box.
The breeding method of cork oak aseptic seedling the most according to claim 1, it is characterised in that: during step (4) sowing,
Seedlings nursing plate specification is 32cm × 28cm × 10cm.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106416909A (en) * | 2016-10-17 | 2017-02-22 | 柳州三农科技有限公司 | Method for obtaining explants of Quercus variabilis Bl tissue culture |
| CN106888902A (en) * | 2017-02-08 | 2017-06-27 | 江苏恒诺园林建设有限公司 | It is a kind of to improve the method that cork oak seed sows germination percentage |
| CN108142266A (en) * | 2018-01-15 | 2018-06-12 | 浙江省林业科学研究院 | A kind of Ostrya rehderiana one year old seedling breeding method |
| CN108651250A (en) * | 2018-03-30 | 2018-10-16 | 浙江申诚环境工程有限公司 | A kind of ornamental plant seedling cultivation method |
| CN108934768A (en) * | 2018-07-25 | 2018-12-07 | 四川省农业科学院土壤肥料研究所 | The breeding method of black spore truffle inoculation microbial inoculum and its processing method and mycorhiza |
| CN109220448A (en) * | 2018-08-30 | 2019-01-18 | 中国林业科学研究院林业研究所 | The direct seeding forestation method of cork oak |
| CN109892161A (en) * | 2019-04-15 | 2019-06-18 | 中国科学院昆明植物研究所 | Rare or endangered species version receives green plum seedling from seed artificial breeding method |
| CN110603926A (en) * | 2019-10-16 | 2019-12-24 | 龙橡金林业科技(扬州)有限公司 | Method for improving germination rate of quercus acutissima seeds |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106416909A (en) * | 2016-10-17 | 2017-02-22 | 柳州三农科技有限公司 | Method for obtaining explants of Quercus variabilis Bl tissue culture |
| CN106888902A (en) * | 2017-02-08 | 2017-06-27 | 江苏恒诺园林建设有限公司 | It is a kind of to improve the method that cork oak seed sows germination percentage |
| CN108142266A (en) * | 2018-01-15 | 2018-06-12 | 浙江省林业科学研究院 | A kind of Ostrya rehderiana one year old seedling breeding method |
| CN108651250A (en) * | 2018-03-30 | 2018-10-16 | 浙江申诚环境工程有限公司 | A kind of ornamental plant seedling cultivation method |
| CN108934768A (en) * | 2018-07-25 | 2018-12-07 | 四川省农业科学院土壤肥料研究所 | The breeding method of black spore truffle inoculation microbial inoculum and its processing method and mycorhiza |
| CN109220448A (en) * | 2018-08-30 | 2019-01-18 | 中国林业科学研究院林业研究所 | The direct seeding forestation method of cork oak |
| CN109892161A (en) * | 2019-04-15 | 2019-06-18 | 中国科学院昆明植物研究所 | Rare or endangered species version receives green plum seedling from seed artificial breeding method |
| CN110603926A (en) * | 2019-10-16 | 2019-12-24 | 龙橡金林业科技(扬州)有限公司 | Method for improving germination rate of quercus acutissima seeds |
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