CN106416909A - Method for obtaining explants of Quercus variabilis Bl tissue culture - Google Patents
Method for obtaining explants of Quercus variabilis Bl tissue culture Download PDFInfo
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- CN106416909A CN106416909A CN201610903196.4A CN201610903196A CN106416909A CN 106416909 A CN106416909 A CN 106416909A CN 201610903196 A CN201610903196 A CN 201610903196A CN 106416909 A CN106416909 A CN 106416909A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
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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
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D1/00—Fertilisers containing potassium
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/40—Fertilisers incorporated into a matrix
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Environmental Sciences (AREA)
- Organic Chemistry (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Inorganic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Pest Control & Pesticides (AREA)
- Cultivation Of Plants (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a method for obtaining explants of Quercus variabilis Bl tissue culture. The method comprises the following processes: selecting seeds and sowing, transplanting and managing bud seedlings, planting and managing mother plants and obtaining the explants, planting the mother plants after being subjected to two-round selection of fine provenance and excellent strong seedlings, and cutting suitable sprouted branches from the mother plants to be sterilized and inoculated. According to the invention, the mother plants are managed and sprouted branches are collected both very conveniently, the obtained explants are excellent in quality, low in pollution rate and death rate, high in germination rate of adventitious buds, neat in sprouting and strong in operability, and meets the needs for establishing a rapid propagation system of the Quercus variabilis Bl tissue culture, so that the method for obtaining the explants of the Quercus variabilis Bl tissue culture has stronger economic benefits, social benefits and ecological benefits.
Description
Technical field
The invention belongs to technical field of plant asexual propagation, it is related to a kind of cork oak tissue cultures explant acquisition methods.
Background technology
Cork oak（Quercus variabilis Bl）Also known as cork oak, tertia Qinggang, hemp oak, it is Fagaceae
（Fagaceae）Oak belongs to arbor, and tree crown is extensively avette, and trunk is many, taupe, deep lobe, and phellem layer is special thick.Sprig is filbert, male flower
Sequence is born in annual shoot bottom, the life of female flower list or twin and annual shoot axil.May at florescence；The fruit next year 9-10 month is ripe.In being
The important commerical tree species of state.Cork oak enjoys light, and is often born in mountain region tailo, but treelet is preferably having lateral shelter.To weather, soil
Strong adaptability.It is resistant to -20 DEG C of low temperature, in the acidity of pH4-8, in neutral and calacareous soil, all have a growth, also drought-resistant,
Barren and with deep, fertile, suitably moistening and well-drained loam and sandy loam optimum, intolerant to ponding.Product Liaoning,
Hebei, Shanxi, Shaanxi, Gansu, Shandong, Jiangsu, Anhui, Zhejiang, Jiangxi, Fujian, Taiwan, Henan, Hubei, Hunan, Guangdong, wide
The provinces and regions such as west, Sichuan, Guizhou, Yunnan.The tailo of below 800 meters of height above sea level is generally born in by North China, and southwest is up to height above sea level
2000-3000 rice.
At present, the common planting type of cork oak is afforestation seeding.Although current cork oak has successfully utilized tissue-culturing rapid propagation
Technology has carried out nursery to the superior families selecting and has afforested, but in tissue culture procedures, explant material generally makes
With big tree fringe bar and base portion rudiment etc..But big majority of setting is located at remote forest zone, plucks inconvenience, rudiment is difficult, quantity is few and is difficult to
Sterilizing, therefore big tree coppice shoot is difficult to meet actual production nursery demand.And In vitro Embryo, aseptic seedling, cotyledon and needle etc. are made
For explant material, obtain the speed of adventitious bud and quantity fast not as good as stem section and many it is difficult to become the common used material of tissue cultures.
Content of the invention
The present invention, in order to overcome the deficiencies in the prior art, provides a kind of harvesting conveniently, quality is good, and quantity is many, sterilizing easily, can
Meet the cork oak tissue cultures explant acquisition methods of cork oak tissue-culturing quick-propagation Establishing needs.
In order to realize foregoing invention purpose, technical scheme is as follows：
A kind of cork oak tissue cultures explant acquisition methods, main operational steps are as follows：
（1）Seed selection and sowing：Choose no disease and pests harm, the cork oak superior families seed of full grains is sowed after vernalization process；
（2）Sprout transplanting and management：Sprout is transplanted to specification after being unearthed be in 15cm × 15cm non-woven fabrics Seedling bag, optionally drenches
Water, spray, fertilising；
（3）Maternal plant plantation and management：Work as step（2）Transplanting nursery stock is taken root after stablizing, and being transplanted to specification is 30cm × 30cm container
Cup in, build cool canopy, select strong sprout as maternal plant be transplanted to equipped with the container cup of matrix in carry out maternal plant plantation, regularly topdress, plus
The strong prevention and control of plant diseases, pest control；The component of described matrix and weight ratio are：Red heart soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 4: 1: 1；
（4）Explant obtains：Clip rudiment bar, prunes needle, carry out disinfection inoculation.
Above-described type of seeding refers to for the seed after routine disinfection and hot-water soak to be placed in 100～140 DEG C
Dry in baking oven, after naturally cooling to room temperature, seed is seeded in and to be crossed using the disinfecting solution of potassium permanganate of volumetric concentration 0.3%
In sand bed.
Above-described sprout transplanting is to be transplanted in non-woven fabrics Seedling bag as high 6～8 cm of sprout with management method,
Apply mass concentration 0.5% composite fertilizer's solution every 7～10 days, after 40 days, sprout is taken root and stablized.
Above step（3）Choose height of seedling 40～60 cm, ground diameter 0.3～0.6 cm, trunk leads to straight, the few container seedling of side shoot
As maternal plant.
Above-described maternal plant management method is to build the shade of high 1.8～2.5 m with the shade net that shade density is 40～60%
Canopy, is 0.5～1% composite fertilizer's solution every volumetric concentration of fertilising in 7～15 days, every 20 days dense to foliage-spray one inferior quality
Spend the potassium dihydrogen phosphate for 0.2%～0.5%；After maternal plant is planted 30 days, in blocking trunk at the cm of ground 30.
Above-described explant acquisition methods are clip length 6～9cm, and the rudiment bar of diameter 0.2～0.4 cm uses volume
The alcoholic solution of concentration 75% is carried out after spraying disinfection to rudiment bar, wipes out terminal bud at the cm of rudiment bar top 0.5～1.0, remaining
Lower part is from top to bottom trimmed to the stem section of 3.0～4.0 cm length as explant, and every rudiment bar takes 1～2 stem section.
Active ingredient >=48% of the NPK in above-described composite fertilizer solution.
In contrast to prior art, the present invention has the advantage that and has the beneficial effect that：
1st, the present invention is to carry out maternal plant plantation in nursery, facilitates maternal plant management and coppice shoot collection；Pass through when maternal plant selects
The two-wheeled of fine provenance and excellent strong sprout selects, and can ensure that the fine quality of acquired explant well, and in pipe
Under reason, coppice shoot can constantly be sprouted, and growth is fast, can meet the needs of large-scale production.
2nd, the present invention carries out cutting strunk in time and promotees to sprout to maternal plant, both ensure that enough sprout growth spaces, and preferably
Maintain the equilibrium of nutrition；Choose the rudiment bar being suitable for as explant material, its degree of lignification and physiological ageing degree can
Preferably meet tissue culture needs.
4th, the maternal plant matrix that the present invention prepares according to the physiological property of cork oak, material is easy to get, balanced in nutrition, decreases life
Produce cost, improve the survival rate of maternal plant.
5th, the present invention, after sterilization rudiment bar, wipes out terminal bud in its top, can remove the tender terminal bud of children being injured by thimerosal,
Ensure that the postvaccinal survival rate of explant；Every coppice shoot from top to bottom 1-2 stem section of clip it is ensured that to take stem section to be respectively provided with stronger
Power of regeneration it is ensured that during adventitious bud inducing adventitious bud concentrate and sprout, rudiment is neat.
6th, the maternal plant management of the present invention and coppice shoot collection are quite convenient to, and the explant being obtained is best in quality, pollution rate,
The death rate is low, and adventitious bud germination rate is high, and rudiment is neat, workable, meets cork oak tissue-culturing quick-propagation system
The demand set up, has stronger economic benefit, social benefit and ecological benefits.
Specific embodiment
With reference to specific embodiment, the invention will be further described.
Embodiment 1：
Choose no disease and pests harm, the cork oak superior families seed of full grains, after routine disinfection and hot-water soak, seed is put
Dry in 100～140 DEG C of baking oven, after naturally cooling to room temperature, seed is seeded in the permanganic acid using volumetric concentration 0.3%
In the sand bed that potassium solution was sterilized.
After sprout is unearthed, being transplanted to specification as high 6～8 cm of sprout is in 15cm × 15cm non-woven fabrics Seedling bag, depending on feelings
Condition water drenching, spray, and applied mass concentration 0.5% composite fertilizer's solution every 7～10 days, the wherein NPK in composite fertilizer's solution
Active ingredient >=48%, after 40 days, sprout is taken root and is stablized.
After transplanting nursery stock is taken root and stablized, choose height of seedling 40～60 cm, ground diameter 0.3～0.6 cm, trunk leads to straight, side shoot
As maternal plant, be transplanted to specification is 30cm × 30cm and equipped with the container cup of matrix to few container seedling, the group of described matrix
Divide and weight ratio is：Red heart soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 4: 1: 1.Taken with the shade net that shade density is 40～60%
Build the cool canopy of high 1.8～2.5 m, be 0.5～1% composite fertilizer's solution every volumetric concentration of fertilising in 7～15 days, composite fertilizer is molten
Active ingredient >=48% of the NPK in liquid；The potassium dihydrogen phosphate that every 20 days is 0.2～0.5% to mass concentration of foliage-spray
Solution；After maternal plant is planted 30 days, in blocking trunk at the cm of ground 30.
Clip length 6～9cm, the rudiment bar of diameter 0.2～0.4 cm, with the alcoholic solution of volumetric concentration 75% to rudiment bar
After carrying out spraying disinfection, at the cm of rudiment bar top 0.5～1.0, wipe out terminal bud, remaining part is from top to bottom trimmed to 3.0～
The stem section of 4.0 cm length takes 1～2 stem section as explant, every rudiment bar.
Embodiment 2：
Choose no disease and pests harm, the cork oak superior families seed of full grains, after routine disinfection and hot-water soak, seed is put
Dry in 110 DEG C of baking oven, after naturally cooling to room temperature, seed is seeded in potassium permanganate using volumetric concentration 0.3% molten
In the sand bed that liquid disinfectant is crossed.
After sprout is unearthed, being transplanted to specification as high 6～8 cm of sprout is in 15cm × 15cm non-woven fabrics Seedling bag, depending on feelings
Condition water drenching, spray, and mass concentration 0.5% composite fertilizer's solution was applied every 8 days, wherein the NPK's in composite fertilizer's solution is effective
Composition >=48%, after 40 days, sprout is taken root and is stablized.
After transplanting nursery stock is taken root and stablized, choose height of seedling 45～50 cm, ground diameter 0.3～0.4 cm, trunk leads to straight, side shoot
As maternal plant, be transplanted to specification is 30cm × 30cm and equipped with the container cup of matrix to few container seedling, the group of described matrix
Divide and weight ratio is：Red heart soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 4: 1: 1.Build height with the shade net that shade density is 50%
The cool canopy of 2.0 m, is 0.5% composite fertilizer's solution every volumetric concentration of fertilising in 8 days, effective one-tenth of the NPK in composite fertilizer's solution
Divide >=48%；The potassium dihydrogen phosphate that every 20 days is 0.2～0.3% to mass concentration of foliage-spray；When maternal plant plants 30
After it, in blocking trunk at the cm of ground 30.
Clip length 6～8cm, the rudiment bar of diameter 0.2～0.3 cm, with the alcoholic solution of volumetric concentration 75% to rudiment bar
After carrying out spraying disinfection, wipe out terminal bud at the cm of rudiment bar top 0.5, remaining part is from top to bottom trimmed to the stem of 3.0cm length
Duan Zuowei explant, every rudiment bar takes 1～2 stem section.
Embodiment 3：
Choose no disease and pests harm, the cork oak superior families seed of full grains, after routine disinfection and hot-water soak, seed is put
Dry in 120 DEG C of baking oven, after naturally cooling to room temperature, seed is seeded in potassium permanganate using volumetric concentration 0.3% molten
In the sand bed that liquid disinfectant is crossed.
After sprout is unearthed, being transplanted to specification as high 6～8 cm of sprout is in 15cm × 15cm non-woven fabrics Seedling bag, depending on feelings
Condition water drenching, spray, and mass concentration 0.5% composite fertilizer's solution was applied every 9 days, wherein the NPK's in composite fertilizer's solution is effective
Composition >=48%, after 40 days, sprout is taken root and is stablized.
After transplanting nursery stock is taken root and stablized, choose height of seedling 50～55 cm, ground diameter 0.4～0.5 cm, trunk leads to straight, side shoot
As maternal plant, be transplanted to specification is 30cm × 30cm and equipped with the container cup of matrix to few container seedling, the group of described matrix
Divide and weight ratio is：Red heart soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 4: 1: 1.Taken with the shade net that shade density is 40～60%
Build the cool canopy of high 2.2 m, be 1% composite fertilizer's solution every volumetric concentration of fertilising in 10 days, the NPK in composite fertilizer's solution has
Effect composition >=48%；The potassium dihydrogen phosphate that every 20 days is 0.3～0.4% to mass concentration of foliage-spray；When maternal plant kind
After planting 30 days, in blocking trunk at the cm of ground 30.
Clip length 7～8cm, the rudiment bar of diameter 0.2～0.3cm, with the alcoholic solution of volumetric concentration 75%, rudiment bar is entered
After row spraying disinfection, wipe out terminal bud at the cm of rudiment bar top 0.5～1.0, remaining part is from top to bottom trimmed to 3.0～4.0
The stem section of cm length takes 1～2 stem section as explant, every rudiment bar.
Embodiment 4：
Choose no disease and pests harm, the cork oak superior families seed of full grains, after routine disinfection and hot-water soak, seed is put
Dry in 140 DEG C of baking oven, after naturally cooling to room temperature, seed is seeded in potassium permanganate using volumetric concentration 0.3% molten
In the sand bed that liquid disinfectant is crossed.
After sprout is unearthed, being transplanted to specification as high 6～8 cm of sprout is in 15cm × 15cm non-woven fabrics Seedling bag, depending on feelings
Condition water drenching, spray, and mass concentration 0.5% composite fertilizer's solution was applied every 10 days, wherein the NPK's in composite fertilizer's solution has
Effect composition >=48%, after 40 days, sprout is taken root and is stablized.
After transplanting nursery stock is taken root and stablized, choose height of seedling 55～60 cm, ground diameter 0.5～0.6 cm, trunk leads to straight, side shoot
As maternal plant, be transplanted to specification is 30cm × 30cm and equipped with the container cup of matrix to few container seedling, the group of described matrix
Divide and weight ratio is：Red heart soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 4: 1: 1.Build height with the shade net that shade density is 60%
The cool canopy of 2.5 m, is 1% composite fertilizer's solution every volumetric concentration of fertilising in 15 days, effective one-tenth of the NPK in composite fertilizer's solution
Divide >=48%；The potassium dihydrogen phosphate that every 20 days is 0.4～0.5% to mass concentration of foliage-spray；When maternal plant plants 30
After it, in blocking trunk at the cm of ground 30.
Clip length 8～9cm, the rudiment bar of diameter 0.3～0.4 cm, with the alcoholic solution of volumetric concentration 75% to rudiment bar
After carrying out spraying disinfection, wipe out terminal bud at the cm of rudiment bar top 0.5～1.0, remaining part is from top to bottom trimmed to 4.0 cm
As explant, every rudiment bar takes 1～2 stem section to long stem section.
Claims (7)
1. a kind of cork oak tissue cultures explant acquisition methods it is characterised in that：Main operational steps are as follows：
（1）Seed selection and sowing：Choose no disease and pests harm, the cork oak superior families seed of full grains is sowed after vernalization process；
（2）Sprout transplanting and management：Sprout is transplanted to specification after being unearthed be in 15cm × 15cm non-woven fabrics Seedling bag, optionally drenches
Water, spray, fertilising；
（3）Maternal plant plantation and management：Work as step（2）Transplanting nursery stock is taken root after stablizing, and being transplanted to specification is 30cm × 30cm container
Cup in, build cool canopy, select strong sprout as maternal plant be transplanted to equipped with the container cup of matrix in carry out maternal plant plantation, regularly topdress, plus
The strong prevention and control of plant diseases, pest control；The component of described matrix and weight ratio are：Red heart soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 4: 1: 1；
（4）Explant obtains：Clip rudiment bar, prunes needle, carry out disinfection inoculation.
2. a kind of cork oak tissue cultures explant acquisition methods according to claim 1 it is characterised in that：Described broadcasts
The method of kind refers to dry in the baking oven be placed in the seed after routine disinfection and hot-water soak 100～140 DEG C, naturally cools to
After room temperature, seed is seeded in the sand bed crossed using the disinfecting solution of potassium permanganate of volumetric concentration 0.3%.
3. a kind of cork oak tissue cultures explant acquisition methods according to claim 1 it is characterised in that：Described bud
Seedling transplanting is to be transplanted in non-woven fabrics Seedling bag as high 6～8 cm of sprout with management method, applies an inferior quality every 7～10 days
Concentration 0.5% composite fertilizer's solution, after 40 days, sprout is taken root and is stablized.
4. a kind of cork oak tissue cultures explant acquisition methods according to claim 1 it is characterised in that：Step（3）
Choose height of seedling 40～60 cm, ground diameter 0.3～0.6 cm, trunk leads to directly, and the few container seedling of side shoot is as maternal plant.
5. a kind of cork oak tissue cultures explant acquisition methods according to claim 1 it is characterised in that：Described mother
Strain management method was to build the cool canopy of high 1.8～2.5 m with the shade net that shade density is 40～60%, every fertilising one in 7～15 days
Secondary volumetric concentration is 0.5～1% composite fertilizer's solution, the phosphoric acid that every 20 days is 0.2%～0.5% to mass concentration of foliage-spray
Dihydro potassium solution；After maternal plant is planted 30 days, in blocking trunk at the cm of ground 30.
6. a kind of cork oak tissue cultures explant acquisition methods according to claim 1 it is characterised in that：Described is outer
Implant acquisition methods are clip length 6～9cm, the rudiment bar of diameter 0.2～0.4 cm, with the alcoholic solution pair of volumetric concentration 75%
After rudiment bar carries out spraying disinfection, wipe out terminal bud at the cm of rudiment bar top 0.5～1.0, remaining part is from top to bottom trimmed to
The stem section of 3.0～4.0 cm length takes 1～2 stem section as explant, every rudiment bar.
7. a kind of cork oak tissue cultures explant acquisition methods according to claim 3 or 6 it is characterised in that：Described
Composite fertilizer's solution in NPK active ingredient >=48%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610903196.4A CN106416909A (en) | 2016-10-17 | 2016-10-17 | Method for obtaining explants of Quercus variabilis Bl tissue culture |
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| CN201610903196.4A CN106416909A (en) | 2016-10-17 | 2016-10-17 | Method for obtaining explants of Quercus variabilis Bl tissue culture |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771297A (en) * | 2012-08-15 | 2012-11-14 | 西北农林科技大学 | Method for cultivating cork oak seedlings |
| CN105284235A (en) * | 2015-11-04 | 2016-02-03 | 张莘蔓 | Method for promoting germination of oriental oak seeds |
| CN105393882A (en) * | 2015-12-02 | 2016-03-16 | 黄钱英 | QuercusvariabilisBl container nursery method |
| CN105409687A (en) * | 2015-12-02 | 2016-03-23 | 黄钱英 | Method for culturing large-scale Quercus variabilis Bl seedlings |
| CN105993865A (en) * | 2015-03-16 | 2016-10-12 | 四川省农业科学院土壤肥料研究所 | Cultivation method for quercus variabilis aseptic seedling |
-
2016
- 2016-10-17 CN CN201610903196.4A patent/CN106416909A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771297A (en) * | 2012-08-15 | 2012-11-14 | 西北农林科技大学 | Method for cultivating cork oak seedlings |
| CN105993865A (en) * | 2015-03-16 | 2016-10-12 | 四川省农业科学院土壤肥料研究所 | Cultivation method for quercus variabilis aseptic seedling |
| CN105284235A (en) * | 2015-11-04 | 2016-02-03 | 张莘蔓 | Method for promoting germination of oriental oak seeds |
| CN105393882A (en) * | 2015-12-02 | 2016-03-16 | 黄钱英 | QuercusvariabilisBl container nursery method |
| CN105409687A (en) * | 2015-12-02 | 2016-03-23 | 黄钱英 | Method for culturing large-scale Quercus variabilis Bl seedlings |
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Application publication date: 20170222 |