CN100551232C - The tissue cultivation rapid breeding method of a kind of santal - Google Patents

The tissue cultivation rapid breeding method of a kind of santal Download PDF

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Publication number
CN100551232C
CN100551232C CNB2007100329822A CN200710032982A CN100551232C CN 100551232 C CN100551232 C CN 100551232C CN B2007100329822 A CNB2007100329822 A CN B2007100329822A CN 200710032982 A CN200710032982 A CN 200710032982A CN 100551232 C CN100551232 C CN 100551232C
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santal
stage
root
bud
illumination
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CN101213936A (en
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马国华
胡玉姬
张新华
何长信
周俊芳
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South China Botanical Garden of CAS
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South China Botanical Garden of CAS
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Abstract

The tissue cultivation rapid breeding method of a kind of santal (Santalum album), the present invention is an explant with the almug tender shoots, induction period, clump bud by indefinite bud are bred each cultivation stage such as stage, the stage of taking root and transplanting stage, allotment and preferably induce, breed, take root, the culture medium prescription and the condition of culture of each cultivation stage such as transplanting, realized the tissue-culturing quick-propagation of santal seedling.Can effectively solve the high-quality santal seedling supply problem in the santal plantation.

Description

The tissue cultivation rapid breeding method of a kind of santal
Technical field
The tissue culture that the present invention relates to plant is cultivated propagation technique, specifically, relates to the tissue culture rapid propagation technique of almug.
Background technology
Santal ( SantalumAlbum) be the Santalaceae aiphyllium.Mainly be distributed in Southeast Asia, Australia and the Pacific region.The history in existing thousands of years of human use santal is because of its timber can be used for the technology engraving and refine sandalwood oil especially famous.In the area, Asia, the sandalwood that smoulders becomes the important ceremony of religions such as Buddhism, Hinduism, Islam.Sandalwood is the raw material that extract sandalwood oil, and it contains compositions such as α and β santalol, can prepare perfume, spices, perfumed soap and scent etc.Sandalwood oil has antimicrobial component, and as traditional Chinese medicine, sandalwood oil can be treated some obstinate diseases such as skin disease such as hemorrhoid, acne, dysentery, gonorrhoea; In addition, the still very desirable sedative of sandalwood oil.International trade santal heartwood is per ton at present sells 47,000 dollars, and its price is in the trend of rising always; And sandalwood price per jin at home is in RMB more than 160 yuan---santal is the high plant of unit are income second in the world, has nothing to be ashamed of for recognize the most expensive in the world timber of valency with jin.Santal have the title of " green gold ", and its economic well-being of workers and staff is 5-10 a times of other forests.Owing to be subjected to the driving of santal timber and sandalwood oil great number interests thereof, cause a large amount of minimizings and the disappearance of primary santal trees.Gain the name because of being distributed with the primary tree of a large amount of santals in the past in the Honolulu on island, the Pacific Ocean, but because excessive felling, Honolulu of today has not had the santal natural distributed.Past, India always was main cultivation and the exported country of santal, because unordered felling, its santal resource almost exhausts.The natural distributed of the no santal of China, the past is always from external import, and is bigger always to its finished product of santal or half-finished demand.The santal seed growing success of Indonesia overseas Chinese present is accepted in botanical garden, south China in 1962; Botanical garden, south China in 1980 is directly introduced a fine variety good santal kind from India and is also succeedd.Through the development test of decades, we have screened tens and have satisfied the required santal host of santal normal growth.Development test through domestic a plurality of areas shows that santal is tropical in south, south China, cultivation can be promoted in the subtropical zone.The at present domestic santal plantation upsurge that just starting.Supply falls short of demand for high-quality santal seedling.Yet because santal male and female flowering organ is different ripe, by offspring's proterties high separation of seminal propagation, interindividual variation is very big, and this is starved of the complete tissue culture quick breeding system of a cover of setting up for the needs plantation rare tree species of decades.In the world wide, the research work of much carrying out the santal tissue culture has been arranged at present, the example of several report santal tissue culture has also been arranged at home.But the santal tissue culture is taken root into seedling but success at home, promptly is that the research report of complete santal tissue culture does not appear in the newspapers as yet.
Summary of the invention
The objective of the invention is to propose the tissue cultivation rapid breeding method of a kind of santal, the inventor is in the research of carrying out the santal tissue culture, basic process from the santal tissue culture, comprise the inducing of bud, breed, take root, stage of transplanting etc., screening optimum medium and condition of culture, make santal can realize its effective breeding, thereby realize purpose of the present invention.
The tissue cultivation rapid breeding method of santal proposed by the invention comprises following cultivation stage:
A) induction period of indefinite bud: get the high-quality almug tender shoots that grows up more than 10 years, after sterilization, be transferred on the inducing culture and cultivate illumination every day 50-100 μ mol m -2s -15-14 hour, until inducing the formation indefinite bud; Described inducing culture adds agar 0.6g L with the MS minimal medium -1, coconut milk 50-100ml L -1, sucrose 30g L -1, pH value 5.4-5.8 is in addition with addition of NAA0.1-1.0mg L -1+ BAP 0.5-2.0mg L -1
B) the clump bud breeding stage: indefinite bud is transferred on the propagating culture medium to set up clump bud breeding system, illumination every day 80-120 μ mol m -2s -110-14 hour, one month to the one and a half months subculture once, described propagating culture medium is with the MS minimal medium, additional agar 0.6g L -1, sucrose 30g L -1, pH value 5.4-5.8, other adds BAP 0.5-2.0mg L -1+ NAA 0.1-0.5mg L -1+ IBA0.5-2.0mg L -1
C) take root the stage: will grow to cut and be transferred on the root media illumination and cultivate root induction, illumination every day 100-200 μ mol m to the clump bud more than the 4-5cm -2s -110-14 hour, cultivate 1-2 month root induction, behind the rooting of vitro seedling blake bottle forwarded to and cultivate 1-2 month in the greenhouse with strong sprout, described root media is with the MS minimal medium, additional culture matrix vermiculite 30-40g L -1(being beneficial to ventilation takes root), sucrose 20-30g L -1, pH value 5.4-5.8, other adds IBA 0.3-0.5mg L -1
D) the transplanting stage: test-tube seedling transplanting that will long root is to outdoor and suitable host plant co-incubation.
Through test, use the present invention to carry out the tissue-culturing quick-propagation of almug, the test-tube plantlet survival rate can reach 95%, can effectively solve the high-quality santal seedling supply problem in the santal plantation.
Embodiment
Embodiment one
1, explant
The young tender axillalry bud of the adult high-quality santal plant of gathering from santal garden, South China Botanical Garden;
2, incubation
A) induction period: santal tender shoots explant is through 75% alcohol disinfecting after 10 seconds, after aseptic washing 2 times, changes in 0.1% mercuric chloride 10 minutes over to, after anhydrously forward on the inducing culture illumination after bud being cut after washing 3 times to and cultivate.Inducing culture adopts the MS minimal medium, additional agar 0.6g L -1, coconut milk 100ml L -1, sucrose 30g L -1, pH value 5.5; Other adds NAA 0.5mg L -1+ BAP 1.0mg L -1, illumination every day 50-100 μ mol m -2s -110 hours.
B) the breeding stage: indefinite bud is transferred on the propagating culture medium illumination every day 120 μ mol m -2s -114 hours, propagating culture medium adopted MS minimal medium, agar 0.6g L -1, sucrose 30g L -1, pH value 5.5; Propagating culture medium has comprised BAP0.8mg L -1+ NAA 0.3mg L -1+ IBA0.8mg L -1, a month subculture is once bred frequency and is reached 5-8;
C) take root the stage: the clump bud that will grow is cut and is transferred on the root media, and root media adopts the MS minimal medium, additional vermiculite 30g L -1, sucrose 30g L -1, pH value 5.5; Other adds IBA 0.3mg L -1, illumination every day 200 μ mol m -2s -114 hours, get the above clump bud of 4-5cm and cut, be placed on this root media and cultivate, can take root in 2 months (can either guarantee to take root, also can make test-tube plantlet can not form clump bud or lateral bud); Behind the rooting of vitro seedling blake bottle forwarded in the greenhouse and took exercise one month, promote the strongr of santal test-tube plantlet.
D) the test-tube seedling transplanting stage: when high, test-tube plantlet is taken out from bottle to 8-10cm when test-tube plantlet is long in bottle, after cleaning test-tube plantlet moved in the plastic sack that host plant (false wormwood artemisia, catharanthus roseus etc.) are arranged and cultivate.Be mixed with pond sludge, sand, special microbial bacterial agent and root-growing agent (IBA+ABT) etc. in the plastic sack, cultivate 3-4 after individual month, move on to field production.
Embodiment two
1, explant
The young tender axillalry bud of the adult high-quality santal plant of gathering from Long Zhudao santal study base, Guangdong;
2, incubation
A) induction period: santal tender shoots explant is through 75% alcohol disinfecting after 10 seconds, after aseptic washing 2 times, changes in 0.1% mercuric chloride 10 minutes over to, after anhydrously forward on the inducing culture illumination after bud being cut after washing 3 times to and cultivate.Inducing culture adopts the MS minimal medium, additional agar 0.6g L -1, coconut milk 60ml L -1, sucrose 30g L -1, pH value 5.6; Other adds NAA0.2mg L -1+ BAP 0.5mg L -1, illumination every day 50-100 μ mol m -2s -110 hours.
B) the breeding stage: indefinite bud is transferred on the propagating culture medium illumination every day 120 μ mol m -2s -114 hours, propagating culture medium adopted the MS minimal medium, additional agar 0.6g L -1, sucrose 30g L -1, pH value 5.6; Other adds BAP 0.5mgL -1+ NAA 0.1mg L -1+ IBA0.5mg L -1, the one and a half months subculture is once bred frequency and is reached 5-8;
C) take root the stage: the clump bud that will grow is cut and is transferred on the root media, and root media adopts the MS minimal medium, additional vermiculite 35g L -1, be beneficial to ventilation and take root.Sucrose 30g L -1, pH value 5.6; Other adds IBA 0.4mg L -1, illumination every day 200 μ mol m -2s -114 hours, get the above clump bud of 4-5cm and cut, be placed on this root media and cultivated 2 months; Behind the rooting of vitro seedling blake bottle forwarded in the greenhouse and took exercise one month, promote the strongr of santal test-tube plantlet.
D) the test-tube seedling transplanting stage: when high, test-tube plantlet is taken out from bottle to 8-10cm when test-tube plantlet is long in bottle, after cleaning test-tube plantlet moved in the plastic sack that host plant (false wormwood artemisia, catharanthus roseus etc.) are arranged and cultivate.Be mixed with pond sludge, sand, special microbial bacterial agent and root-growing agent (IBA+ABT) etc. in the plastic sack, cultivate 3-4 after individual month, move on to field production.
Embodiment three
1, explant
The young tender axillalry bud of the adult high-quality santal plant of gathering from santal garden, South China Botanical Garden;
2, incubation
A) induction period: santal tender shoots explant is through 75% alcohol disinfecting after 10 seconds, after aseptic washing 2 times, changes in 0.1% mercuric chloride 10 minutes over to, after anhydrously forward on the inducing culture illumination after bud being cut after washing 3 times to and cultivate.Inducing culture adopts the MS minimal medium, additional agar 0.6g L -1, coconut milk 100ml L -1, sucrose 30g L -1, pH value 5.8; Other adds NAA 1.0mg L -1+ BAP 2.0mg L -1, illumination every day 50-100 μ mol m -2s -110 hours.
B) the breeding stage: indefinite bud is transferred on the propagating culture medium illumination every day 120 μ mol m -2s -114 hours, propagating culture medium adopted the MS minimal medium, additional agar 0.6g L -1, sucrose 30g L -1, pH value 5.8; Other adds BAP 1.0mg L -1+ NAA 0.5mg L -1+ IBA1.0mg L -1, one month to the one and a half months subculture once, the breeding frequency reach 5-8;
C) take root the stage: the clump bud that will grow is cut and is transferred on the root media, and root media adopts the MS minimal medium, additional culture matrix vermiculite 40g L -1, sucrose 30g L -1, pH value 5.8; Other adds IBA 0.5mg L -1, illumination every day 200 μ mol m -2s -114 hours, get the above clump bud of 4-5cm and cut, be placed on this root media and cultivated 2 months; Behind the rooting of vitro seedling blake bottle forwarded in the greenhouse and took exercise one month, promote the strongr of santal test-tube plantlet.
D) the test-tube seedling transplanting stage: when high, test-tube plantlet is taken out from bottle to 8-10cm when test-tube plantlet is long in bottle, after cleaning test-tube plantlet moved in the plastic sack that host plant (false wormwood artemisia, catharanthus roseus etc.) are arranged and cultivate.Be mixed with pond sludge, sand, special microbial bacterial agent and root-growing agent (IBA+ABT) etc. in the plastic sack, cultivate 3-4 after individual month, move on to field production.

Claims (2)

1. the tissue cultivation rapid breeding method of a santal is characterized in that comprising following cultivation stage:
A) induction period of indefinite bud: get the high-quality almug tender shoots that grows up more than 10 years, after sterilization, be transferred on the inducing culture and cultivate illumination every day 50-100 μ mol m -2s -15-14 hour, until inducing the formation indefinite bud; Described inducing culture adds agar 0.6g L with the MS minimal medium -1, coconut milk 50-100ml L -1, sucrose 30g L -1, pH value 5.4-5.8 is in addition with addition of NAA 0.1-1.0mg L -1+ BAP 0.5-2.0mg L -1
B) the clump bud breeding stage: indefinite bud is transferred on the propagating culture medium to set up clump bud breeding system, illumination every day 80-120 μ mol m -2s -110-14 hour, one month to the one and a half months subculture once, described propagating culture medium is with the MS minimal medium, additional agar 0.6g L -1, sucrose 30g L -1, pH value 5.4-5.8, other adds BAP 0.5-2.0mg L -1+ NAA 0.1-0.5mg L -1+ IBA0.5-2.0mg L -1
C) take root the stage: will grow to cut and be transferred on the root media illumination and cultivate root induction, illumination every day 100-200 μ mol m to the clump bud more than the 4-5cm -2s -110-14 hour, cultivate 1-2 month root induction, behind the rooting of vitro seedling blake bottle forwarded to and cultivate 1-2 month in the greenhouse with strong sprout, described root media is with the MS minimal medium, additional culture matrix vermiculite 30-40g L -1, sucrose 20-30g L -1, pH value 5.4-5.8, other adds IBA0.3-0.5mg L -1
D) the transplanting stage: test-tube seedling transplanting that will long root is to outdoor and suitable host plant co-incubation.
2. santal tissue cultivation rapid breeding method according to claim 1 is characterized in that described host plant is false wormwood artemisia or catharanthus roseus.
CNB2007100329822A 2007-12-29 2007-12-29 The tissue cultivation rapid breeding method of a kind of santal Expired - Fee Related CN100551232C (en)

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CN104585038B (en) * 2015-02-04 2016-09-14 中国科学院华南植物园 Induction method of sandalwood triploid plant
CN105580737B (en) * 2016-03-21 2018-05-01 贵州师范学院 A kind of method for culturing seedlings of Thesium chinese tissue-culture container seedling
CN109744142A (en) * 2017-08-21 2019-05-14 中国林业科学研究院热带林业研究所 A kind of santal tissue-culturing quick-propagation method for culturing seedlings
CN108184666B (en) * 2017-12-29 2019-10-08 中国科学院华南植物园 A kind of method for inspiring xylophyta bearing tree and sprouting the method and the quick breeding seedling based on juvenile form bud of juvenile form bud
CN116868894A (en) * 2023-08-08 2023-10-13 中国林业科学研究院热带林业研究所 Tissue rapid propagation method of santalum album
CN117770139A (en) * 2024-02-26 2024-03-29 东北林业大学 Culture medium for inducing China rose plant to regenerate in vitro and application thereof

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