CN103975832B - A kind of white wing silvery birch plantlet in vitro outside sprout-cultivating-bottle method - Google Patents
A kind of white wing silvery birch plantlet in vitro outside sprout-cultivating-bottle method Download PDFInfo
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Abstract
The invention discloses a kind of white wing silvery birch plantlet in vitro outside sprout-cultivating-bottle seedling-cultivating method, comprising: cultivating process totally seven steps such as the cultivation of the foundation of aseptic culture body, plantlet in vitro, the domestication of unrooted bottle seedling, transplanting method and the management after transplanting.The inventive method can solve the present situation that in white wing silvery birch bottle, rooting rate is very low, both the link of taking root in bottle can be simplified, turn improve transplanting survival rate, and upgrowth situation is good, thus shorten the tissue culture time, accelerate seedling propagation speed, decrease and cultivate space, simplify production link, save production cost.White wing silvery birch outside sprout-cultivating-bottle technology of the present invention is that factorial praluction provides reliable technical guarantee.
Description
Technical field
The present invention relates to a kind of plantlet in vitro outside sprout-cultivating-bottle method of seeds, be specifically related to a kind of white wing silvery birch plantlet in vitro outside sprout-cultivating-bottle method, belong to ecological management new seeds artificial breeding technique field.
Background technology
Silvery birch originates in Oceania, evergreen, trunk is straight, tree-like attractive in appearance, especially save during the blossom season, serving as a contrast with orange-yellow flower in Wan Lvcong, is landscape tree and street tree, and Chinese Guangdong, Guangxi, each cities and towns, Yunnan are all introduced a fine variety, for one of main green tree species in city, silvery birch kind about 60 genus, 1300 kinds, China introduce a fine variety as seen as Grevillea banksii R. Br, horse felt silvery birch, super silvery birch, Jones silvery birch, Canberra silvery birch, by silvery birch, bronze silvery birch, moonlight silvery birch, spend the kind such as silvery birch, white wing silvery birch in vain.White wing silvery birch originates in Australia, is a kind of shrub, and growth is fast; branches and leaves are luxuriant, crawl in earth's surface, well developed root system; go deep into soil layer, few damage by disease and insect, resistance; resistance to arid necessarily and water wet; its bark and leaf are to flue dust and toxic gas better resistance, stronger than religion to the absorbing capacity of sulphur dioxide and fluoride, strong adaptability; can drought-resistant dead soil, be ecology of mining areas protection, highway two takes up the fine tree species of the slope treatments such as water channel.
Due to white wing silvery birch seed-setting rate and germination rate low, cottage propagation is very difficult, and in tissue culture propagation bottle, rooting rate only has about 20%, and cultivation period is long, and transplanting survival rate is low, and production cost is high, is difficult to the development adapting to a large amount of nursery stock demand.
Application for a patent for invention 101743908A discloses a kind of method of fast breeding Grevillea banksii R. Br, mainly comprise the selection of material and sterilization, callus cultivation, the propagation of callus, the induction of indefinite bud and expand numerous, the culture of rootage of indefinite bud, the step such as transplanting and hardening.This invention utilizes tissue culture rapid propagation technique, and not only breeding cycle is short, can whole year production, obtains a large amount of best buy nursery stocks, and can ensure the superiority of Grevillea banksii R. Br kind.Application for a patent for invention 102301909A discloses the method for the efficient Fast-propagation of a kind of orange silvery birch.It, by selecting suitable cutting, carries out suitable process to cutting, in suitable cottage propagation period, cuttage is in the nutrition cup that Nutrition Soil is housed, be positioned in cuttage cool canopy and carry out breeding child care management, then through suitable cultivation management technology, and obtain the orange silvery birch seedling of high-quality.This invention achieves the efficient Fast-propagation of orange silvery birch, by adopting the method for the efficient Fast-propagation of orange silvery birch of the present invention, orange silvery birch cottage propagation survival rate can reach 80% to 90%, plant height can be produced in 8 to 10 months and reach 100cm to 120cm, leading thread reaches the orange silvery birch seedling of high-quality of 0.6cm to 0.8cm, can apply, for afforestation industry provides the excellent orange silvery birch nursery stock of high-quality on the gardening of gardens.
Summary of the invention
The object of the invention is the deficiency existed for existing seedling growing process, a kind of seedling-cultivating method of effective white wing silvery birch plantlet in vitro outside sprout-cultivating-bottle is provided, reach that growing-seedling period is short, seedling growth is vigorous, method of operating is simple and feasible, production cost is lower, for large-scale popularization uses these seeds to provide reliable seedling object.
Object of the present invention is achieved by the following technical programs: a kind of seedling-cultivating method of white wing silvery birch plantlet in vitro outside sprout-cultivating-bottle, comprises the following steps:
(1) foundation of aseptic culture body: choose healthy and strong select tree, plucks without damage by disease and insect, the shoot that has no mechanical damage, is trimmed to the stem section with 1-2 axillalry bud, through laboratory cultures, induction explant, forms sterile propagation material, set up aseptic culture body, obtain sterile propagation body;
(2) without the cultivation of offspring: sterile propagation body to be inoculated in proliferated culture medium that to carry out expansion numerous;
(3) preparation before transplanting seedlings: choose that the Multiple Buds expanding numerous rear acquisition carries out inoculating, cultivates, hardening;
(4) prepare seedling medium: be implanted in cool canopy to cultivate in the dish of cave and carry out, first prepare seedling medium, seedling medium is tamped cavities, trickle drenches seedling medium;
(5) transplant: unrooted tissue-culture container seedling is transplanted in seedling medium, completes bud seedling Artificial Control condition of culture from provisions to the process of seedling autotrophy;
(6) management after transplanting: to the management carrying out water, fertilizer and anti-bacteria without offspring in two months that transplant;
(7) seedling culture: carry out seedling culture after the transplanting of bud seedling is taken root to height of seedling 25-35 centimetre transplanting, concrete grammar is two months after the transfer, root growth very flourishing, in the dish of cave, root system keeps together, now seedling carefully to be taken out from cave, do not hurt shoot root, be transplanted to bigger being equipped with in the seedling-growing container of Nutrition Soil to cultivate, for keeping out of the direct sun, palpus 75% shady net shading, carry out full exposure Routine Management after growth is stable, within 4 months, raw height of seedling 30 centimetres can go out garden field planting.
The concrete grammar that the foundation of step (1) described aseptic culture body adopts is: by clean for the stem section running water after pruning, at the mercury chloride sterilization 3 minutes of use for laboratory 0.1%, again with aseptic washing 4 times, one bottle one section is inoculated in MS+6BA(6-benzyl purine, lower same) 2mg/l+NAA(methyl α-naphthyl acetate, down together) on 0.05mg/l medium, cultivate 28-35 days, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium, through squamous subculture, and can growth and breeding continuously, complete the foundation of aseptic culture body.
The numerous concrete grammar of step (2) described expansion is inoculated in the proliferated culture medium of MS+6BA0.5-2mg/l+NAA0.05-0.2mg/l by sterile propagation body, be placed on room temperature 28 ± 2 DEG C, intensity of illumination is cultivated 30 days in the culturing room of 1500lx, and its proliferation times reaches 3.2 times.
Preparation before transplanting seedlings described in step (3) is: cut the bud that in Multiple Buds, high 20 ± 5mm is long and be inoculated in MS medium, be placed on room temperature 28 ± 2 DEG C, intensity of illumination is cultivated 10 days on the culturing rack of 3000lx, be placed into outdoor 75% shading condition lower refining seedling 15-20 days again, when unrooted bottle seedling leaf launches, elongation is healthy and strong, prepares to transplant.
Seedling medium described in step (4) is mixed to get in the ratio of 7:3 by peat soil and coconut palm chaff.
The concrete grammar of step (5) described transplanting is: by carefully taking out without offspring in bottle, clean and stick the medium on seedling, speed is stained with water of taking root, be transplanted in the cave dish filling up seedling medium, a strain bud seedling is planted in each cave, transplants thickness of earth-fill cover and is no more than 1 centimetre, final singling water of drenching after having transplanted, adopt cultivation temperature 25-28 DEG C, cool canopy with 75% shading net shading.
In two months after step (6) described transplanting, water management is: matrix will be kept moistening in two weeks after transplanting, relative air humidity reaches 95%, prevents leaves water loss, progressively controls water supply in media afterwards, and relative moisture controls 60 ± 5%, with hestening rooting.
Management fertile in two months after step (6) described transplanting is: the 28-32 days after transplanting, sprays the potassium dihydrogen phosphate of 0.1%, afterwards, sprays the potassium dihydrogen phosphate of a time 0.1% every two weeks.
In two months after step (6) described transplanting, the management of anti-bacteria is: within after transplanting the 3rd day, adopt 0.1% tpn spraying disinfection sterilizing, can effectively prevent germ from growing, 0.1% tpn of spraying every 10-15 days afterwards, effectively can improve survival rate.
Seedling culture described in step (7) Nutrition Soil used is mixed to get in the ratio of 1:1 by yellow soil and coconut palm chaff.
The present invention relative to the beneficial effect of prior art is: the inventive method can solve the present situation that in white wing silvery birch bottle, rooting rate is very low, both the link of taking root in bottle can be simplified, turn improve transplanting survival rate, and upgrowth situation is good, thus shorten the tissue culture time, accelerate seedling propagation speed, decrease and cultivate space, simplify production link, save production cost.White wing silvery birch outside sprout-cultivating-bottle technology of the present invention is that factorial praluction provides reliable technical guarantee.
Embodiment
Be described in further details the present invention below by embodiment, these embodiments are only used for the present invention is described, do not limit the scope of the invention.
Embodiment 1MS culture medium prescription contains the drug variety of following nutritive element and the ratio of quality in every 1 liter, and be dissolved in the water and be mixed with, PH is adjusted to 5.8
Add water to 1000ml, in addition additional 6-benzyladenine 0.5-3.0mg/l, methyl α-naphthyl acetate 0.01-0.2mg/l.
Embodiment 2MS culture medium prescription contains the drug variety of following nutritive element and the ratio of quality in every 1 liter, and be dissolved in the water and be mixed with, PH is adjusted to 5.8
Add water to 1000ml, additional in addition: 6-benzyladenine 0.2-2.0mg/l, methyl α-naphthyl acetate 0.01-0.2mg/l.
Embodiment 3 is carried out according to following steps:
(1) foundation of aseptic culture body: choose healthy and strong select tree, pluck without damage by disease and insect, the shoot that has no mechanical damage, be trimmed to the stem section with 1-2 axillalry bud, clean with running water, at the mercury chloride sterilization 3 minutes of use for laboratory 0.1%, again with aseptic washing 4 times, one bottle one section is inoculated on MS+6BA2mg/l+NAA0.05mg/l medium, cultivate 30 days, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium, through squamous subculture, and can growth and breeding continuously, complete the foundation of aseptic culture body;
(2) without the cultivation of offspring: be inoculated in the proliferated culture medium of MS+6BA0.5mg/l+NAA0.05mg/l by sterile propagation body, be placed on room temperature 28 DEG C, intensity of illumination is cultivated 30 days in the culturing room of 1500lx, and its proliferation times reaches 3.2 times;
(3) preparation before transplanting seedlings: cut the bud that in Multiple Buds, high 20 ± 5mm is long and be inoculated in MS medium, be placed on room temperature 29 DEG C, intensity of illumination is cultivated 10 days on the culturing rack of 3000lx, be placed into outdoor 75% shading condition lower refining seedling 15 days again, when unrooted bottle seedling leaf launches, elongation is healthy and strong, prepares to transplant;
(4) prepare seedling medium: be mixed to get seedling medium with peat soil and coconut palm chaff in the ratio of 7:3, seedling medium is tamped the cavities of cultivating cave and coiling, trickle drenches seedling medium;
(5) transplant: be implanted in in cool canopy and carry out, by carefully taking out without offspring in bottle, clean and stick the medium on seedling, speed is stained with water of taking root, and is transplanted in the cave dish filling up seedling medium, a strain bud seedling is planted in each cave, transplant thickness of earth-fill cover and be no more than 1 centimetre, final singling water of drenching after having transplanted, adopt cultivation temperature 25 DEG C, the cool canopy shading net shading of 75%, completes bud seedling Artificial Control condition of culture from provisions to the process of seedling autotrophy;
(6) management after transplanting: to the management carrying out water, fertilizer and anti-bacteria without offspring in two months that transplant, as follows:
A water management: matrix will be kept moistening in two weeks after transplanting, relative air humidity 95%, progressively controls water supply in media afterwards, and relative moisture controls 60%;
The management of b fertilizer: 28 days after transplanting, spray the potassium dihydrogen phosphate of 0.1%, afterwards, sprays the potassium dihydrogen phosphate of a time 0.1% every two weeks;
The management of c anti-bacteria: adopt 0.1% tpn spraying disinfection sterilizing in after transplanting the 3rd day, afterwards every spraying in 10 days 0.1% tpn;
(7) seedling culture: be two months after the transfer, root growth very flourishing, in the dish of cave, root system keeps together, now seedling carefully to be taken out from cave, not hurt shoot root, be transplanted to bigger being equipped with in the seedling-growing container of Nutrition Soil (being mixed to get in the ratio of 1:1 by yellow soil and coconut palm chaff) and cultivate, keep out of the direct sun, 75% shady net shading, carry out full exposure Routine Management after growth is stable, within 4 months, raw height of seedling 30 centimetres can go out garden field planting.
Embodiment 4 is carried out according to following steps:
(1) foundation of aseptic culture body: choose healthy and strong select tree, pluck without damage by disease and insect, the shoot had no mechanical damage, be trimmed to the stem section with 1-2 axillalry bud, clean with running water, at the mercury chloride sterilization 3 minutes of use for laboratory 0.1%, again with aseptic washing 4 times, one bottle one section is inoculated in MS+6BA(6-benzyl purine, lower same) 2mg/l+NAA(methyl α-naphthyl acetate, down together) on 0.05mg/l medium, cultivate 28-35 days, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium, through squamous subculture, and can growth and breeding continuously, complete the foundation of aseptic culture body,
(2) without the cultivation of offspring: be inoculated in the proliferated culture medium of MS+6BA2mg/l+NAA0.1mg/l by sterile propagation body, be placed on room temperature 30 DEG C, intensity of illumination is cultivated 30 days in the culturing room of 1500lx, and its proliferation times reaches 3.2 times;
(3) preparation before transplanting seedlings: cut the bud that in Multiple Buds, high 20 ± 5mm is long and be inoculated in MS medium, be placed on room temperature 31 DEG C, intensity of illumination is cultivated 10 days on the culturing rack of 3000lx, be placed into outdoor 75% shading condition lower refining seedling 20 days again, when unrooted bottle seedling leaf launches, elongation is healthy and strong, prepares to transplant;
(4) prepare seedling medium: be mixed to get seedling medium with peat soil and coconut palm chaff in the ratio of 7:3, seedling medium is tamped the cavities of cultivating cave and coiling, trickle drenches seedling medium;
(5) transplant: be implanted in in cool canopy and carry out, by carefully taking out without offspring in bottle, clean and stick the medium on seedling, speed is stained with water of taking root, and is transplanted in the cave dish filling up seedling medium, a strain bud seedling is planted in each cave, transplant thickness of earth-fill cover and be no more than 1 centimetre, final singling water of drenching after having transplanted, adopt cultivation temperature 28 DEG C, the cool canopy shading net shading of 75%, completes bud seedling Artificial Control condition of culture from provisions to the process of seedling autotrophy;
(6) management after transplanting: to the management carrying out water, fertilizer and anti-bacteria without offspring in two months that transplant, as follows:
A water management: matrix will be kept moistening in two weeks after transplanting, relative air humidity 95%, progressively controls water supply in media afterwards, and relative moisture controls 62%;
The management of b fertilizer: 30 days after transplanting, spray the potassium dihydrogen phosphate of 0.1%, afterwards, sprays the potassium dihydrogen phosphate of a time 0.1% every two weeks;
The management of c anti-bacteria: adopt 0.1% tpn spraying disinfection sterilizing in after transplanting the 3rd day, afterwards every spraying in 15 days 0.1% tpn;
(7) seedling culture: be two months after the transfer, root growth very flourishing, in the dish of cave, root system keeps together, now seedling carefully to be taken out from cave, not hurt shoot root, be transplanted to bigger being equipped with in the seedling-growing container of Nutrition Soil (being mixed to get in the ratio of 1:1 by yellow soil and coconut palm chaff) and cultivate, keep out of the direct sun, 75% shady net shading, carry out full exposure Routine Management after growth is stable, within 4 months, raw height of seedling 30 centimetres can go out garden field planting.
Embodiment 5 is carried out according to following steps:
(1) foundation of aseptic culture body: choose healthy and strong select tree, pluck without damage by disease and insect, the shoot that has no mechanical damage, be trimmed to the stem section with 1-2 axillalry bud, clean with running water, at the mercury chloride sterilization 3 minutes of use for laboratory 0.1%, again with aseptic washing 4 times, one bottle one section is inoculated on MS+6BA2mg/l+NAA0.05mg/l medium, cultivate 35 days, sprout green bud from axil, the green bud of clip is inoculated on proliferated culture medium, through squamous subculture, and can growth and breeding continuously, complete the foundation of aseptic culture body;
(2) without the cultivation of offspring: be inoculated in the proliferated culture medium of MS+6BA1mg/l+NAA0.2mg/l by sterile propagation body, be placed on room temperature 30 DEG C, intensity of illumination is cultivated 30 days in the culturing room of 1500lx, and its proliferation times reaches 3.2 times;
(3) preparation before transplanting seedlings: cut the bud that in Multiple Buds, high 20 ± 5mm is long and be inoculated in MS medium, be placed on room temperature 26 DEG C, intensity of illumination is cultivated 10 days on the culturing rack of 3000lx, be placed into outdoor 75% shading condition lower refining seedling 18 days again, when unrooted bottle seedling leaf launches, elongation is healthy and strong, prepares to transplant;
(4) prepare seedling medium: be mixed to get seedling medium with peat soil and coconut palm chaff in the ratio of 7:3, seedling medium is tamped the cavities of cultivating cave and coiling, trickle drenches seedling medium;
(5) transplant: be implanted in in cool canopy and carry out, by carefully taking out without offspring in bottle, clean and stick the medium on seedling, speed is stained with water of taking root, and is transplanted in the cave dish filling up seedling medium, a strain bud seedling is planted in each cave, transplant thickness of earth-fill cover and be no more than 1 centimetre, final singling water of drenching after having transplanted, adopt cultivation temperature 26 DEG C, the cool canopy shading net shading of 75%, completes bud seedling Artificial Control condition of culture from provisions to the process of seedling autotrophy;
(6) management after transplanting: to the management carrying out water, fertilizer and anti-bacteria without offspring in two months that transplant, as follows:
A water management: matrix will be kept moistening in two weeks after transplanting, relative air humidity 95%, progressively controls water supply in media afterwards, and relative moisture controls 65%;
The management of b fertilizer: 32 days after transplanting, spray the potassium dihydrogen phosphate of 0.1%, afterwards, sprays the potassium dihydrogen phosphate of a time 0.1% every two weeks;
The management of c anti-bacteria: adopt 0.1% tpn spraying disinfection sterilizing in after transplanting the 3rd day, afterwards every spraying in 12 days 0.1% tpn;
(7) seedling culture: be two months after the transfer, root growth very flourishing, in the dish of cave, root system keeps together, now seedling carefully to be taken out from cave, not hurt shoot root, be transplanted to bigger being equipped with in the seedling-growing container of Nutrition Soil (being mixed to get in the ratio of 1:1 by yellow soil and coconut palm chaff) and cultivate, keep out of the direct sun, 75% shady net shading, carry out full exposure Routine Management after growth is stable, within 4 months, raw height of seedling 30 centimetres can go out garden field planting.
Claims (1)
1. a seedling-cultivating method for white wing silvery birch plantlet in vitro outside sprout-cultivating-bottle, is characterized in that: be made up of following steps:
(1) foundation of aseptic culture body: choose healthy and strong select tree, plucks without damage by disease and insect, the shoot that has no mechanical damage, is trimmed to the stem section with 1-2 axillalry bud, through laboratory cultures, induction explant, forms sterile propagation material, set up aseptic culture body, obtain sterile propagation body; The concrete grammar adopted is: by clean for the stem section running water after pruning, at the mercury chloride sterilization 3 minutes of use for laboratory 0.1%, again with aseptic washing 4 times, one bottle one section is inoculated on MS+6BA2mg/l+NAA0.05mg/l medium, cultivates 28-35 days, sprouts green bud from axil, the green bud of clip is inoculated on proliferated culture medium, through squamous subculture, and can growth and breeding continuously, complete the foundation of aseptic culture body;
(2) without the cultivation of offspring: sterile propagation body to be inoculated in proliferated culture medium that to carry out expansion numerous; Expanding numerous concrete grammar is be inoculated in the proliferated culture medium of MS+6BA0.5-2mg/l+NAA0.05-0.2mg/l by sterile propagation body, is placed on room temperature 28 ± 2 DEG C, and intensity of illumination is cultivated 30 days in the culturing room of 1500lx, and its proliferation times reaches 3.2 times;
(3) preparation before transplanting seedlings: cut the bud that in Multiple Buds, high 20 ± 5mm is long and be inoculated in MS medium, be placed on room temperature 28 ± 2 DEG C, intensity of illumination is cultivated 10 days on the culturing rack of 3000lx, be placed into outdoor 75% shading condition lower refining seedling 15-20 days again, when unrooted bottle seedling leaf launches, elongation is healthy and strong, prepares to transplant;
(4) prepare seedling medium: be implanted in cool canopy to cultivate in the dish of cave and carry out, first prepare seedling medium, seedling medium is tamped cavities, trickle drenches seedling medium; Described seedling medium is mixed to get in the ratio of 7:3 by peat soil and coconut palm chaff;
(5) transplant: unrooted tissue-culture container seedling is transplanted in seedling medium, completes bud seedling Artificial Control condition of culture from provisions to the process of seedling autotrophy; The concrete grammar transplanted is: by carefully taking out without offspring in bottle, clean and stick the medium on seedling, speed is stained with water of taking root, be transplanted in the cave dish filling up seedling medium, a strain bud seedling is planted in each cave, transplants thickness of earth-fill cover and is no more than 1 centimetre, final singling water of drenching after having transplanted, adopt cultivation temperature 25-28 ° of C, cool canopy with 75% shading net shading;
(6) management after transplanting: to the management carrying out water, fertilizer and anti-bacteria without offspring in two months that transplant; In two months after transplanting, water management is: matrix will be kept moistening in two weeks after transplanting, relative air humidity reaches 95%, prevents leaves water loss, progressively controls water supply in media afterwards, and relative moisture controls 60 ± 5%; Management fertile in two months after transplanting is: the 28-32 days after transplanting, sprays the potassium dihydrogen phosphate of 0.1%, afterwards, sprays once every two weeks; In two months after transplanting, the management of anti-bacteria is: within after transplanting the 3rd day, adopt 0.1% tpn spraying disinfection sterilizing, afterwards every spraying in 10-15 days once;
(7) seedling culture: carry out seedling culture after the transplanting of bud seedling is taken root to height of seedling 25-35 centimetre transplanting, concrete grammar is two months after the transfer, root growth very flourishing, in the dish of cave, root system keeps together, now seedling carefully to be taken out from cave, do not hurt shoot root, be transplanted to bigger being equipped with in the seedling-growing container of Nutrition Soil to cultivate, for keeping out of the direct sun, palpus 75% shady net shading, carry out full exposure Routine Management after growth is stable, within 4 months, raw height of seedling 30 centimetres goes out garden field planting; Described seedling culture Nutrition Soil used is mixed to get in the ratio of 1:1 by yellow soil and coconut palm chaff.
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| CN104145821B (en) * | 2014-08-25 | 2017-03-29 | 深圳市公园管理中心 | The isolated culture regeneration plant method of yellow fruit silvery birch |
| CN108575359A (en) * | 2018-04-27 | 2018-09-28 | 北京农学院 | A kind of processing of Malus admiring fruit tree training tissue culture seedling and tissue-cultured seedling rooting method |
| CN109479728B (en) * | 2019-01-28 | 2021-08-13 | 胡相伟 | Method for rooting tissue culture seedlings of populus deltoids outside test tubes |
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| 白翅银桦组培快繁技术研究;赵平丽 等;《江西林业科技》;20110630;第12-15页 * |
| 试管苗瓶外生根技术;黄卓忠 等;《广西农业科学》;20070330;第124-126页 * |
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