CN106416910A - White tassel stalk tissue culture and explants extraction method - Google Patents

White tassel stalk tissue culture and explants extraction method Download PDF

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Publication number
CN106416910A
CN106416910A CN201610903197.9A CN201610903197A CN106416910A CN 106416910 A CN106416910 A CN 106416910A CN 201610903197 A CN201610903197 A CN 201610903197A CN 106416910 A CN106416910 A CN 106416910A
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sprout
days
acquisition methods
solution
dead ears
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陈杏林
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Liuzhou Agriculture-Countryside-Farmer Science And Technology Ltd
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Liuzhou Agriculture-Countryside-Farmer Science And Technology Ltd
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G17/00Cultivation of hops, vines, fruit trees, or like trees
    • A01G17/005Cultivation methods
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00Growth substrates; Culture media; Apparatus or methods therefor
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05DINORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
    • C05D1/00Fertilisers containing potassium
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05GMIXTURES 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
    • C05G5/00Fertilisers characterised by their form
    • C05G5/40Fertilisers incorporated into a matrix

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Botany (AREA)
  • Organic Chemistry (AREA)
  • Pest Control & Pesticides (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Inorganic Chemistry (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention disclose a white tassel stalk tissue culture and explants extraction method, including the process of selecting seeds and sowing seeds, shoot seedling transplanting and managing, and planting mother plant and managing explants extracting, planting mother plant with the quality seeds and quality strong seedlings after a two round selection, disinfecting and transplanting after suitable coppice shoots are cut from the mother plant. The mother plants are very convenient to manage and the coppice shoots are easy to gather, the explants are high in quality, low in pollution extent and death rate, high in adventitious bud germination rate, the shoots are tidily developed, strongly operable, the method satisfies the requirements of the rapid reproducing system built for white tassel stalk tissue culture, and has comparatively strong economic effect, social effect and ecological effect.

Description

A kind of dead ears Ke's tissue cultures explant acquisition methods
Technical field
The invention belongs to technical field of plant asexual propagation, it is related to a kind of dead ears Ke's tissue cultures explant acquisition methods.
Background technology
Dead ears Ke(Lithocarpus craibianus Barn.), it is Fagaceae(Fagaceae)Lithocarpus (Lithocarpus)Arbor, arbor, up to 20 meters.Annual shoot, blade back and female inflorescence axle all have brown color or canescence wax squama Layer.Leaf keratin, avette or ovate-elliptic, long 12-19 centimetre, sometimes shorter or longer, wide 4-7 centimetre, dilute wider, top is long Point, the short point of base portion to wide wedge shape, full edge, the hypomere of middle arteries is thick on blade face, and vein there are about 8-12 bar, often recessed in splitting channel-shaped on blade face Fall into, offshoot does not show or mays be seen indistinctly, and after tender leaf is done, often there is glossy gloss on blade face, biennial leaf is then grey dark;Petiole is long 1-2.5 li Rice.Tassel locusta armpit is given birth to, dilute for panicle, up to 15 centimetres;The top of female inflorescence often raw minority male flower, up to 30 lis Rice, 3-5-7 collection of female flower generates cluster, style long 1-1.2 millimeter.Acorn-cup spheroidal or slightly flat, top is in often mamillary short projection, Footpath 15-20 millimeter, full bag nut, occasionally there are top cracking warp, therefore nut part exposes, squamella triangle, apex point Shape, fits perfectly in shell wall, and imbricate arranges, clearly, positioned at the slightly long and narrow of acorn-cup top and to curved under acorn-cup oral area, by yellowish-brown, Fine platy, the wax squama being slightly close to;The nearly spheroidal of nut, footpath 13-18 millimeter, by very sparse thin volt hair, the hair at top is closeer, really Navel is raised, accounts for the 1/3 of nut area.The month at florescence 8-9, fruit same period next year is ripe.Produce South-west Sichuan, south of Yunnan and the west and south Various places.It is born in 700 meters of mountain region shaws of 1 500-2, is more common in and hillside fields is relatively dried, Chang Yin is cut down, therefore generate shrub shape, It is usually about 10 meters of dungarunga.Laos, Northern Thailand also have.
At present, dead ears Ke resource is still in wild distribution, but with keeping the demand of Plant Diversity, scale people Work cultivation dead ears Ke's resource is imperative.Although current dead ears Ke successfully using group culturation rapid propagating technology to the excellent family selecting System has carried out nursery and has afforested, but in tissue culture procedures, explant material is usually used big tree fringe bar and base portion rudiment Deng.But big majority of setting is located at remote forest zone, plucks inconvenience, rudiment is difficult, quantity is few and is difficult to sterilize, therefore big coppice shoot of setting is difficult to Meet actual production nursery demand.And In vitro Embryo, aseptic seedling, cotyledon and needle etc. be as explant material, it is indefinite to obtain The speed of 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 dead ears Ke's tissue cultures explant acquisition methods of dead ears Ke's tissue-culturing quick-propagation Establishing needs.
In order to realize foregoing invention purpose, technical scheme is as follows:
A kind of dead ears Ke's tissue cultures explant acquisition methods, main operational steps are as follows:
(1)Seed selection and sowing:Choose no disease and pests harm, dead ears Ke's 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:Yellow soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 3: 2: 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 dead ears Ke, 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 dead ears Ke's 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, dead ears Ke's 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:Yellow soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 3: 2: 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, dead ears Ke's 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:Yellow soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 3: 2: 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, dead ears Ke's 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:Yellow soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 3: 2: 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, dead ears Ke's 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:Yellow soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 3: 2: 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 dead ears Ke's 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, dead ears Ke's 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:Yellow soil: coconut palm chaff of becoming thoroughly decomposed: vermiculite: plant ash is 4: 3: 2: 1;
(4)Explant obtains:Clip rudiment bar, prunes needle, carry out disinfection inoculation.
2. a kind of dead ears Ke's 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 dead ears Ke's 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 dead ears Ke's 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 dead ears Ke's 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 dead ears Ke's 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 dead ears Ke's 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%.
CN201610903197.9A 2016-10-17 2016-10-17 White tassel stalk tissue culture and explants extraction method Pending CN106416910A (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105265058A (en) * 2015-11-04 2016-01-27 张莘蔓 Method for improving germination rate of Lithocarpus craibianus seeds
CN105367270A (en) * 2015-12-02 2016-03-02 罗杰 Special compound fertilizer for lithocarpus craibianus and preparation method
CN105393777A (en) * 2015-12-02 2016-03-16 黄钱英 LithocarpuscraibianusBarn container nursery method
CN105409550A (en) * 2015-12-02 2016-03-23 黄钱英 Method for culturing large-scale Lithocarpus craibianus Barn. seedlings
CN105474999A (en) * 2015-12-02 2016-04-13 罗杰 Asexual rapid reproduction method of lithocarpus craibianus Barn.

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105265058A (en) * 2015-11-04 2016-01-27 张莘蔓 Method for improving germination rate of Lithocarpus craibianus seeds
CN105367270A (en) * 2015-12-02 2016-03-02 罗杰 Special compound fertilizer for lithocarpus craibianus and preparation method
CN105393777A (en) * 2015-12-02 2016-03-16 黄钱英 LithocarpuscraibianusBarn container nursery method
CN105409550A (en) * 2015-12-02 2016-03-23 黄钱英 Method for culturing large-scale Lithocarpus craibianus Barn. seedlings
CN105474999A (en) * 2015-12-02 2016-04-13 罗杰 Asexual rapid reproduction method of lithocarpus craibianus Barn.

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