CN109169273A - A kind of breeding method of bletilla proliferation pseudobulb - Google Patents
A kind of breeding method of bletilla proliferation pseudobulb Download PDFInfo
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- CN109169273A CN109169273A CN201811023533.6A CN201811023533A CN109169273A CN 109169273 A CN109169273 A CN 109169273A CN 201811023533 A CN201811023533 A CN 201811023533A CN 109169273 A CN109169273 A CN 109169273A
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- proliferation
- bletilla
- generation
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- seedling
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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
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of breeding methods of bletilla proliferation pseudobulb, the breeding method of the bletilla proliferation pseudobulb is by sterile seedling squamous subculture, survival rate of the squamous subculture 5 times bletilla proliferation pseudobulbs for starting to obtain bletilla proliferation pseudobulb after proliferation, and obtain in domestication hardening is 99% or more.The bletilla test tube seedling come is turned out through the invention, and than conventional seed asepsis live streaming (although also known as tissue-cultured seedling), no matter speed of growth, Fen Tiller ability or pseudobulb size have significant difference.
Description
Technical field
The invention belongs to technical field of cultivation, and in particular to a kind of breeding method of bletilla proliferation pseudobulb.
Background technique
Bletilla is orchid family bletilla category herbaceos perennial, is used as medicine with stem tuber have astringing to arrest bleeding, the function such as detumescence and promoting granulation
Effect, however the artificial propagation techniques of bletilla are not mature enough, traditional division propagation mode growth cycle is long, and breeding coefficient is small, in addition
Field bletilla pollination rate is low with seed germination rate, so that bletilla market provides a large amount of seedling, and opens for bletilla new
Reproduction technique.So far, the tissue culture quick breeding research report in relation to bletilla is more, but major part is broken up by callus
, the technical problem being proliferated by pseudobulb cannot be fundamentally solved, therefore, developing one kind can solve above-mentioned technology hardly possible
The method of topic is very important.
Summary of the invention
The purpose of the present invention is to provide a kind of breeding methods of bletilla proliferation pseudobulb.
The object of the present invention is achieved like this, and the breeding method of the bletilla proliferation pseudobulb is grown directly from seeds by sterile
Seedling squamous subculture obtains bletilla proliferation pseudobulb after starting proliferation for squamous subculture 5 times, and obtained bletilla proliferation pseudobulb is being tamed and dociled
Changing the survival rate in hardening is 99% or more.
The present invention from pseudobulb occur and be proliferated angle, by sterile seedling several generations switching culture (or
Claim squamous subculture), the seedling differentiated by callus is eliminated, and by changing plant hormone induction and retaining aseptic seedling base
Portion divides Tiller seedling, can be proliferated by pseudobulb in 4 each every generation of switching.Pseudobulb is proliferated this approach previous
Report in seldom exist, even if certain researchers have carried out similar research, but its coefficient is very low；And innovative point of the invention is just
It is this: after sterile seedling derives, carries out 2 generation squamous subcultures；Callus and its generation are eliminated since the 3rd generation
Seedling, be carried out continuously for 2 generations, proliferation at this time relies primarily on a Fen Tiller and carries out；Proliferation is all by pseudobulb side since the 5th generation
Formula carries out, and effective multiplication factor greatly improves, up to 4.0 or more.The method although slightly show too long in time, by taking root
The thick shape of test tube seedling come is turned out, pseudobulb has just started to expand rapidly in bottle and occur with pseudobulb bud；
In following greenhouse domestication hardening survival rate also up to 99% or more, existing document report is greatly exceeded.In addition, passing through this
Invention turn out come bletilla test tube seedling, than conventional seed asepsis live streaming (although also known as tissue-cultured seedling) no matter the speed of growth, point
Tiller ability or pseudobulb size have significant difference.
Specific embodiment
The breeding method of bletilla proliferation pseudobulb of the present invention is by sterile seedling squamous subculture, squamous subculture
Survival rate of the bletilla proliferation pseudobulbs for starting for 5 times to obtain bletilla proliferation pseudobulb after proliferation, and obtain in domestication hardening be
99% or more.
The described bletilla proliferation pseudobulb breeding method specifically includes the following steps:
1) sterile seedling is accessed in 1 generation proliferated culture medium, obtained for 1 generation in 20~40 DEG C of temperature dark culture 50~70 days
It is proliferated seedling；
2) 1 generation proliferation seedling is accessed in 2 generation proliferated culture mediums, obtained for 2 generations in 20~40 DEG C of temperature dark culture 50~70 days
It is proliferated seedling；
3) 2 generations proliferation seedling is accessed in 3 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination
Culture obtains 3 generations proliferation seedling for 30~50 days；
4) 3 generations proliferation seedling is accessed in 4 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination
Culture obtains 4 generations proliferation seedling for 30~50 days；
5) 4 generations proliferation seedling is accessed in Fen Tiller culture medium, 21~23 DEG C of temperature, illuminance 1500~2000Lx illumination cultivation
Obtain within 20~40 days object bletilla proliferation pseudobulb；
Wherein, 1 generation proliferated culture medium are as follows: MS+ banana puree 40~60g/L+, 10~30g/L of sucrose, pH value 5.0~6.0；
2 generation proliferated culture mediums are as follows: MS+ banana puree 40~60g/L+, 10~30g/L of sucrose, agar 0.3~0.5%, pH value
5.0~6.0；
3 generation proliferated culture mediums are as follows: 0.05~0.5mg/L+ of MS+ banana puree 40~60g/L+, 10~30g/L+2,4-D of sucrose
0.5~1.5mg/L of NAA, agar 0.3~0.5%, pH value 5.0~6.0；
Fen Tiller culture medium are as follows: 0.01~0.05mg/L of 1/2MS+AC 0.5~1.5g/L+, 70~90g/L+6-BA of banana puree
+ NAA 0.5~1.5mg/L+, 10~20g/L of sucrose, pH value 6.5~7.5.
The 1 generation proliferated culture medium are as follows: MS+ banana puree 50g/L+ sucrose 20g/L, pH value 5.8.
The 2 generation proliferated culture mediums are as follows: MS+ banana puree 50g/L+ sucrose 20g/L, agar 0.4%, pH value 5.5.
The 3 generation proliferated culture mediums are as follows: MS+ banana puree 50g/L+ sucrose 20g/L+2,4-D 0.2mg/L+NAA
1.0mg/L, agar 0.4%, pH value 5.8.
The Fen Tiller culture medium are as follows: 1/2MS+AC 1.0g/L+ banana puree 80g/L+6-BA 0.03mg/L+NAA
1.0mg/L+ sucrose 15g/L, pH value 7.0.
Embodiment 1
1) sterile seedling is accessed in 1 generation proliferated culture medium, obtains 1 generation proliferation seedling in 20 DEG C of temperature dark culture 70 days；
2) 1 generation proliferation seedling is accessed in 2 generation proliferated culture mediums, obtains 2 generations proliferation seedling in 20 DEG C of temperature dark culture 70 days；
3) 2 generations proliferation seedling is accessed in 3 generation proliferated culture mediums, 25 DEG C of temperature, illuminance 1500~2000Lx illumination cultivation
Obtain within 30 days 3 generations proliferation seedling；
4) 3 generations proliferation seedling is accessed in 4 generation proliferated culture mediums, 20 DEG C of temperature, illuminance 1500~2000Lx illumination cultivation
Obtain within 50 days 4 generations proliferation seedling；
5) 4 generations proliferation seedling is accessed in Fen Tiller culture medium, 21 DEG C of temperature, 1500~2000Lx of illuminance illumination cultivation 40 days
Obtain object bletilla proliferation pseudobulb；
Wherein, 1 generation proliferated culture medium are as follows: MS+ banana puree 40g/L+ sucrose 10g/L, pH value 5.0；
2 generation proliferated culture mediums are as follows: MS+ banana puree 40g/L+ sucrose 10g/L, agar 0.3%, pH value 5.0；
3 generation proliferated culture mediums are as follows: MS+ banana puree 40g/L+ sucrose 10g/L+2,4-D 0.05mg/L+NAA 0.5mg/L,
Agar 0.3%, pH value 5.0；
Fen Tiller culture medium are as follows: 1/2MS+AC 0.5g/L+ banana puree 70g/L+6-BA 0.01mg/L+NAA 0.5mg/L+ sugarcane
Sugared 10g/L, pH value 6.5.
Embodiment 2
1) sterile seedling is accessed in 1 generation proliferated culture medium, obtains 1 generation proliferation in 20~30 DEG C of temperature dark culture 50 days
Seedling；
2) 1 generation proliferation seedling is accessed in 2 generation proliferated culture mediums, obtains 2 generations proliferation in 30~40 DEG C of temperature dark culture 50 days
Seedling；
3) 2 generations proliferation seedling is accessed in 3 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination
Culture obtains 3 generations proliferation seedling for 30 days；
4) 3 generations proliferation seedling is accessed in 4 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination
Culture obtains 4 generations proliferation seedling for 50 days；
5) 4 generations proliferation seedling is accessed in Fen Tiller culture medium, 21~23 DEG C of temperature, illuminance 1500~2000Lx illumination cultivation
Obtain within 40 days object bletilla proliferation pseudobulb；
Wherein, 1 generation proliferated culture medium are as follows: MS+ banana puree 60g/L+ sucrose 30g/L, pH value 6.0；
2 generation proliferated culture mediums are as follows: MS+ banana puree 60g/L+ sucrose 30g/L, agar 0.5%, pH value 6.0；
3 generation proliferated culture mediums are as follows: MS+ banana puree 60g/L+ sucrose 30g/L+2,4-D 0.5mg/L+NAA 1.5mg/L, fine jade
Rouge~0.5%, pH value 6.0；
Fen Tiller culture medium are as follows: 1/2MS+AC 1.5g/L+ banana puree 90g/L+6-BA 0.05mg/L+NAA 1.5mg/L+ sugarcane
Sugared 20g/L, pH value 7.5.
Embodiment 3
1) sterile seedling is accessed in 1 generation proliferated culture medium, obtains 1 generation proliferation in 20~40 DEG C of temperature dark culture 60 days
Seedling；
2) 1 generation proliferation seedling is accessed in 2 generation proliferated culture mediums, obtains 2 generations proliferation in 20~40 DEG C of temperature dark culture 60 days
Seedling；
3) 2 generations proliferation seedling is accessed in 3 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination
Culture obtains 3 generations proliferation seedling for 30~50 days；
4) 3 generations proliferation seedling is accessed in 4 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination
Culture obtains 4 generations proliferation seedling for 40 days；
5) 4 generations proliferation seedling is accessed in Fen Tiller culture medium, 21~23 DEG C of temperature, illuminance 1500~2000Lx illumination cultivation
Obtain within 30 days object bletilla proliferation pseudobulb；
Wherein, 1 generation proliferated culture medium are as follows: MS+ banana puree 50g/L+ sucrose 20g/L, pH value 5.8；
2 generation proliferated culture mediums are as follows: MS+ banana puree 50g/L+ sucrose 20g/L, agar 0.4%, pH value 5.5；
3 generation proliferated culture mediums are as follows: MS+ banana puree 50g/L+ sucrose 20g/L+2,4-D 0.3mg/L+NAA 1.0mg/L, fine jade
Rouge 0.4%, pH value 5.5；
Fen Tiller culture medium are as follows: 1/2MS+AC 1.0g/L+ banana puree 80g/L+6-BA 0.03mg/L+NAA 1.0mg/L+ sugarcane
Sugared 15g/L, pH value 7.0.
Claims (6)
1. a kind of breeding method of bletilla proliferation pseudobulb, it is characterized in that: the breeding method of the bletilla proliferation pseudobulb is
By sterile seedling squamous subculture, bletilla proliferation pseudobulb is obtained after starting proliferation for squamous subculture 5 times, and obtained bletilla increases
Growing survival rate of the pseudobulb in domestication hardening is 99% or more.
2. the breeding method of bletilla proliferation pseudobulb according to claim 1, it is characterized in that: the bletilla is proliferated false squama
Stem breeding method specifically includes the following steps:
1) sterile seedling is accessed in 1 generation proliferated culture medium, obtains 1 generation proliferation in 20~40 DEG C of temperature dark culture 50~70 days
Seedling；
2) 1 generation proliferation seedling is accessed in 2 generation proliferated culture mediums, obtains 2 generations proliferation in 20~40 DEG C of temperature dark culture 50~70 days
Seedling；
3) 2 generations proliferation seedling is accessed in 3 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination cultivation
Obtain within 30~50 days 3 generations proliferation seedling；
4) 3 generations proliferation seedling is accessed in 4 generation proliferated culture mediums, 20~25 DEG C of temperature, illuminance 1500~2000Lx illumination cultivation
Obtain within 30~50 days 4 generations proliferation seedling；
5) 4 generations proliferation seedling is accessed in Fen Tiller culture medium, 21~23 DEG C of temperature, 1500~2000Lx of illuminance illumination cultivation 20~
Obtain within 40 days object bletilla proliferation pseudobulb；
Wherein, 1 generation proliferated culture medium are as follows: MS+ banana puree 40~60g/L+, 10~30g/L of sucrose, pH value 5.0~6.0；
2 generation proliferated culture mediums are as follows: MS+ banana puree 40~60g/L+, 10~30g/L of sucrose, agar 0.3~0.5%, pH value 5.0~
6.0；
3 generation proliferated culture mediums are as follows: 0.05~0.5mg/L+NAA of MS+ banana puree 40~60g/L+, 10~30g/L+2,4-D of sucrose
0.5~1.5mg/L, agar 0.3~0.5%, pH value 5.0~6.0；
Fen Tiller culture medium are as follows: 0.01~0.05mg/L+NAA of 1/2MS+AC 0.5~1.5g/L+, 70~90g/L+6-BA of banana puree
0.5~1.5mg/L+, 10~20g/L of sucrose, pH value 6.5~7.5.
3. the breeding method of bletilla proliferation pseudobulb according to claim 2, it is characterized in that: the 1 generation Multiplying culture
Base are as follows: MS+ banana puree 50g/L+ sucrose 20g/L, pH value 5.8.
4. the breeding method of bletilla proliferation pseudobulb according to claim 2, it is characterized in that: the 2 generation Multiplying cultures
Base are as follows: MS+ banana puree 50g/L+ sucrose 20g/L, agar 0.4%, pH value 5.5.
5. the breeding method of bletilla proliferation pseudobulb according to claim 2, it is characterized in that: the 3 generation Multiplying cultures
Base are as follows: MS+ banana puree 50g/L+ sucrose 20g/L+2,4-D 0.2mg/L+NAA 1.0mg/L, agar 0.4%, pH value 5.8.
6. the breeding method of bletilla proliferation pseudobulb according to claim 2, it is characterized in that: the Fen Tiller culture medium
Are as follows: 1/2MS+AC 1.0g/L+ banana puree 80g/L+6-BA 0.03mg/L+NAA 1.0mg/L+ sucrose 15g/L, pH value 7.0.
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Cited By (1)
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| CN109964797A (en) * | 2019-03-22 | 2019-07-05 | 浙江大学 | The method for promoting bletilla pseudobulb to be proliferated/expand |
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Application publication date: 20190111 |