CN1640244A

CN1640244A - Tufted bamboo tissue culture rapid breeding rooting method

Info

Publication number: CN1640244A
Application number: CN 200410097715
Authority: CN
Inventors: 杨本鹏; 张树珍; 昝丽梅
Original assignee: State Key Laboratory Of Tropical Bioscience And Biotechnlogy Tropical Agricultural Academe Of China; Institute of Tropical Bioscience and Biotechnology Chinese Academy of Tropical Agricultural Sciences
Current assignee: State Key Laboratory Of Tropical Bioscience And Biotechnlogy Tropical Agricultural Academe Of China; Institute of Tropical Bioscience and Biotechnology Chinese Academy of Tropical Agricultural Sciences
Priority date: 2004-11-24
Filing date: 2004-11-24
Publication date: 2005-07-20
Anticipated expiration: 2024-11-24
Also published as: CN1305371C

Abstract

The present invention discloses a rooting method for tissue culture and quick propagation of bushy bamboo. Said method includes the following steps: using cladium of bamboo as explant, on the bud-inducing culture medium culturing and constructing sterile line, on the enrichment culture medium making sterile seedling undego the bushy bud quick proparation process to form lots of sprouts, making plant division and strengthening seedlings, then successively inoculating them into first rooting culture medium and second rooting culture medium so as to culture and induce out complete root system.

Description

The rooting method of tufted bamboo tissue culture rapid breeding

Technical field

The present invention relates to a kind of rooting method of the bamboo of growing thickly, relate in particular to a kind of rooting method that utilizes tissue culture to breed fast.

Background technology

Bamboo is the important forest resources, regeneration capacity is strong, growth rapidly, the phase of becoming a useful person is short, output is high, has once the characteristics that afforestation can " forever " utilization, is described as " cutting unbeaten industry ".Its bamboo wood has that longitudinal strength is good, elasticity is big, wear-resistant, corrosion-resistant, counter-bending, can be mothproof, can be fire-retardant etc. characteristics, compare advantages such as having hardness is strong, stability is high, the gummed degree is good, beautiful appearance with timber.About more than 1,600 ten thousand tons of bamboo wood is produced in the whole world per year, except that traditional being used to builds a house, makes furniture and handicraft, is mainly used in and makes bamboo fiber board and senior bamboo paper, and its demand grows with each passing day; The young shoot of bamboo---bamboo shoots, fresh and crisp, nutritious, it is rich in 18 seed amino acids and the necessary trace element of multiple human body, be natural health food, developed into series of products such as bright bamboo shoot, dried bamboo shoots, bamboo shoot can, also become a big pillar product of bamboo resource development and use; Bamboo still is the four seasons evergreen species green for a long time, the underground bamboo whip system (scattered bamboo) that it is crisscross or the rhizome system (bamboo grows thickly) of the bamboo root and stem of certain plants and fibrous root prosperity have powerful fixing of soil and conservation of water function, its acrial part is taken out bamboo shoot every year, although cut aged bamboo stalk every year selectively and take bright bamboo shoot, also can keep comparatively stable forest form structure, thus bamboo grove conserve water and soil, water conservation, aspect such as regulate the climate, purify air and beautify the environment also have the unrivaled advantage of general forest.Therefore the bamboo resource that good bamboo kind is bred fast to the development and utilization preciousness has great importance.

Tissue culture about bamboo, at first use India's perverse bamboo seed zygotic embryo evoked callus from nineteen eighty-two Metha etc., formed since plumule and the regeneration plant, some researchs have been carried out both at home and abroad in this respect, mainly be to adopt materials such as seed, embryo, inflorescence, flower pesticide, blade, tender stem apex, young root, ripe bamboo sprout sections as explant, obtain regeneration plant by callus approach and axillalry bud germinating approach, but the rooting efficiency of tissue cultivating seedling is not really desirable.

Dendrocalamus sinicus and imperial bamboo are two kinds of maximum in the world bamboos that grow thickly, its bar Gao Keda 20～30m, thick 15～the 30cm of stem, the wall thickness of bar base portion can reach 3～4cm, long 30～the 40cm of internode, bamboo wood output is 5～8 times of mao bamboon, and its bamboo shoot are fit to the processing dried bamboo shoots again, and they are two rising bamboo kinds.This bamboo kind only has fragmentary distribution in Chinese yunnan province some areas, and its ring seedling cultivation and rooting is very difficult, adopts conventional method can't cultivate seedling in enormous quantities.The method of research and utilization tissue culture obtains the key that a large amount of high quality seedlings is imperial bamboo of development and industry thereof, and the very difficulty but its tissue cultivating seedling is taken root does not reach effect with conventional rooting method for tissue culture and medium, does not satisfy demand.

Summary of the invention

The rooting method that the purpose of this invention is to provide a kind of tufted bamboo tissue culture rapid breeding.

The rooting method of the tufted bamboo tissue culture rapid breeding that the present invention is designed comprises the process of setting up, shoot proliferation incubation, the root induction process of no bacterial strain system, the transplanting process of test-tube plantlet.The process of setting up of described no bacterial strain system is that sprout sections with bamboo is as explant, earlier carry out surface sterilization with ethanol, and then sterilize in the immersion mercuric chloride solution, then take out and clean with sterile water, and carry out lucifuge on the bud inducing culture and cultivate with being inoculated in behind the aseptic filter paper suck dry moisture, change over to again under the natural daylight and cultivate, to induce the sprouting of sprout axillalry bud.Described shoot proliferation incubation is being inoculated on the proliferated culture medium after the formed bud cutting of inducing culture stage, and under illumination condition, cultivate, form the bud of growing thickly to induce, the bud of growing thickly that then will form is divided into the bud clump again and is inoculated in and carries out shoot proliferation on the identical medium.Described root induction process be with the bud of growing thickly cut into several buds be Dan Cong be inoculated in first root media cultivate after, be transferred to again in second root media and cultivate, grown the root system of adorning to promote it.The transplanting process of described test-tube plantlet is that the bottle seedling of taking root is moved on to the natural room temperature lower refining seedling, take out and transplant on " coconut palm chaff+river sand+a little earth " matrix after seedling cleans medium, and keep moistening, when treating seedling length, can move into the secondary nursery and cultivate into commercial seedling to certain altitude.More than said each medium all be to be basal medium with the MS medium, and be added with other components, to satisfy the needs of each process.

The rooting method of tufted bamboo tissue culture rapid breeding provided by the present invention has solved the difficult problem that the bamboo tissue cultivating seedling is difficult to take root, for the quick breeding of good bamboo kind provides technical guarantee, to play positive impetus to the development and utilization of bamboo resource, have important economic and social benefit.

Embodiment

The present invention is described in detail below in conjunction with embodiment.

The rooting method of the tufted bamboo tissue culture rapid breeding that the present invention is designed comprises the process of setting up, shoot proliferation incubation, the root induction process of no bacterial strain system, the transplanting process of test-tube plantlet.

1, the process of setting up of no bacterial strain system was learnt from else's experience bamboo sprout bar selected and was removed blade carefully, after the liquid detergent solution soaking, rinse well with running water, branch is cut into the segment of band joint, ethanol with 70%-80% under aseptic condition carries out surface sterilization, again sections is taken out and drop into disinfection in the mercuric chloride solution, constantly shake during this time, make the ring section be immersed in the mercuric chloride solution all the time; Take out the sections aseptic water washing, then use the aseptic filter paper suck dry moisture; Forward is inoculated into sections on the ready inducing culture, and first lucifuge was cultivated 6-8 days, changes under the natural daylight again and cultivates, to induce the sprouting of sprout axillalry bud.Described inducing culture is to be basal medium with the MS medium, is added with 6BA 1.8～2.2mg.L then in proportion ^-1, NAA0.1～0.3mg.L ^-1, PVP200～280mg.L ^-1, sucrose 20～40g.L ^-1, carragheen 6～8g.L ^-1

2, the shoot proliferation incubation is being inoculated on the proliferated culture medium after the formed bud cutting of inducing culture stage, cultivated 18-20 days down or under the illumination condition of 800～1200Lx prior to the natural daylight scattered light, form the bud of growing thickly to induce, the bud of growing thickly of formation is divided into bud clump with 2～3 buds again and is inoculated in and carries out shoot proliferation on the identical proliferated culture medium.Described proliferated culture medium is to be basal medium with the MS medium, is added with 6BA 1.8～2.2mg.L then in proportion ^-1, KT0.3～0.6mg.L ^-1, Sucus Cocois 80～150ml.L ^-1, sucrose 20～40g.L ^-1, carragheen 6～8g. ^-1

3, the root induction process bud of will growing thickly cuts into 2～3 buds and is 1 clump and is inoculated into and cultivated in first root media 15～20 days, is transferred to cultivate in second root media just can grow complete root system in 20～25 days again, and rooting rate reaches 95%.Described first root media is to be basal medium with the 1/2MS medium, is added with 2 in proportion then, 4-D 0.1～2.0mg.L ^-1+ NAA1.8～2.2mg.L ^-1+ IAA1.8～2.2mg.L ^-1, sucrose 20～40g.L ^-1, carragheen 6～8g.L ^-1Described second root media is to be basal medium with the 1/2MS medium, is added with NAA1.8～2.2mg.L then in proportion ^-1+ IAA1.8～2.2mg.L ^-1, sucrose 20～40g. ^-1, carragheen 6～8g.L ^-1

4, the transplanting process of test-tube plantlet moved on to the natural room temperature lower refining seedling to the bottle seedling of taking root 5～7 days, 70% thiophanate methyl or 1000 times of carbendazols with 1000 times after taking out seedling and cleaning medium soak, transplant again on " coconut palm chaff+river sand+a little earth " matrix about the about 70-90% of obscurity, and keeping moistening, transplanting survival rate reaches more than 98%.When treating seedling length, can move into the secondary nursery and cultivate into commercial seedling to 16-25cm.

Claims

1, a kind of rooting method of tufted bamboo tissue culture rapid breeding, it comprises the process of setting up, shoot proliferation incubation, the root induction process of no bacterial strain system, the transplanting process of test-tube plantlet, it is characterized in that:

(1), the process of setting up of no bacterial strain system was learnt from else's experience bamboo sprout bar selected and was removed blade carefully, after the liquid detergent solution soaking, rinse well with running water, branch is cut into the segment of band joint, ethanol with 70%-80% under aseptic condition carries out surface sterilization, again sections is taken out and drop into disinfection in the mercuric chloride solution, constantly shake during this time, make the ring section be immersed in the mercuric chloride solution all the time; Take out the sections aseptic water washing, then use the aseptic filter paper suck dry moisture; Forward is inoculated into sections on the ready inducing culture, and first lucifuge was cultivated 6-8 days, changes under the natural daylight again and cultivates, to induce the sprouting of sprout axillalry bud.Described inducing culture is to be basal medium with the MS medium, is added with 6BA 1.8～2.2mg.L then in proportion ^-1, NAA0.1～0.3mg.L ^-1, PVP200～280mg.L ^-1, sucrose 20～40g.L ^-1, carragheen 6～8g.L ^-1

(2), the shoot proliferation incubation is being inoculated on the proliferated culture medium after the formed bud cutting of inducing culture stage, cultivated 18-20 days down or under the illumination condition of 800～1200Lx prior to the natural daylight scattered light, form the bud of growing thickly to induce, the bud of growing thickly of formation is divided into bud clump with 2～3 buds again and is inoculated in and carries out shoot proliferation on the identical proliferated culture medium.Described proliferated culture medium is to be basal medium with the MS medium, is added with 6BA1.8～2.2mg.L then in proportion ^-1, KT 0.3～0.6mg.L ^-1, Sucus Cocois 80～150ml.L ^-1, sucrose 20～40g.L ^-1, carragheen 6～8g.L ^-1

(3), the root induction process bud of will growing thickly cuts into 2～3 buds and is 1 clump and is inoculated into and cultivated in first root media 15～20 days, is transferred to cultivate in second root media just can grow complete root system in 20～25 days again, rooting rate reaches 95%.Described first root media is to be basal medium with the 1/2MS medium, is added with 2 in proportion then, 4-D 0.1～2.0mg.L ^-1+ NAA1.8～2.2mg.L ^-1+ IAA1.8～2.2mg.L ^-1, sucrose 20～40g.L ^-1, carragheen 6～8g.L ^-1Described second root media is to be basal medium with the 1/2MS medium, is added with NAA1.8～2.2mg.L then in proportion ^-1+ IAA1.8～2.2mg.L ^-1, sucrose 20～40g.L ^-1, carragheen 6～8g.L ^-1

(4), the transplanting process of test-tube plantlet moved on to the natural room temperature lower refining seedling to the bottle seedling of taking root 5～7 days, 70% thiophanate methyl or 1000 times of carbendazols with 1000 times after taking out seedling and cleaning medium soak, transplant again on " coconut palm chaff+river sand+a little earth " matrix about the about 70-90% of obscurity, and keeping moistening, transplanting survival rate reaches more than 98%.When treating seedling length, can move into the secondary nursery and cultivate into commercial seedling to 16-25cm.