CN104542294A - Rooting culture method for tamarix chinensis tissue culture seedlings - Google Patents
Rooting culture method for tamarix chinensis tissue culture seedlings Download PDFInfo
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- CN104542294A CN104542294A CN201510017370.0A CN201510017370A CN104542294A CN 104542294 A CN104542294 A CN 104542294A CN 201510017370 A CN201510017370 A CN 201510017370A CN 104542294 A CN104542294 A CN 104542294A
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- chinese tamarisk
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Abstract
The invention discloses a subculture method of tamarix chinensis tissue culture seedlings. The subculture method is a one-step seedling culturing method. According to the method, an active carbon component is added into a culture medium, the phenomenon that a tamarix chinensis explants is browned is reduced, the culture period is shortened, the consumption of sugar is reduced, the production cost is reduced, the material cost is further reduced, the stability of inheritable characters of the tamarix chinensis are ensured, a large quantity of high-quality tamarix chinensis tissue culture seedlings are obtained in a short period, and an important significance to the industrialization of the tissue culture of the tamarix chinensis is exerted. The method is applicable to rooting culture of the tamarix chinensis tissue culture seedlings and is further applicable to the subculture of seedlings planted in greenhouses or the tissue culture seedlings.
Description
Technical field
The invention belongs to domestication of plants field, relate to the cultural method of a Plants, be specifically related to a kind of method of Chinese tamarisk plantlet in vitro culture of rootage.
Background technology
Chinese tamarisk is the raw woody plant of salt, there is high Salt And Alkali Tolerance, cold-resistant, drought-resistant, the water-fast good characteristics such as wet, vitality is indomitable, can vigorous growth on the saline land of salt content more than 3 ‰, on saline land, not soil removal and replacement just can survive, and is coast protection forest and heavy salinized first-selected seeds of afforesting.China Chinese tamarisk also has powerful salt-secreting capacity and flourishing root system, the salinity absorbed can be excreted by blade, plays to check winds and fix drifting sand, improve the effects such as soil.The ecology improvement of Chinese tamarisk to the construction of alkaline land improving, saline land greening, coast protection forest and saline land with good characteristic has great importance.In addition, Chinese tamarisk or a kind of traditional Chinese medicine, there is dispelling wind, induce sweat, promoting eruption, deciphering effect.
Chinese tamarisk belongs to suspend Tamarix, and about pointing out in the cultured in vitro research of Chinese tamarisk that in Chinese tamarisk tissue culture expanding propagation, optimum minimal medium is MS, conventional Hormone is 6-BA, KT and NAA, and conventional Hormone of taking root is NAA.Chinese tamarisk tissue culture expanding propagation is mainly divided into the acquisition of aseptic seedling, the acclimatization and transplants of the screening of proliferated culture medium, the screening of root media, seedling of taking root.But the content disclosed in prior art is only limitted to laboratory theory research quality, cannot directly put into production.In addition, in aseptic seedling acquisition process, the disinfectant that use mercuric chloride etc. are poisonous, causes Brown phenomenon serious, is difficult to form industrialization and produces.And be mostly using Multiplying culture and culture of rootage as two independently stages in prior art, in actual industrialization process, cost of labor and Material Cost large.
Chinese tamarisk plantlet in vitro has browning in subculture process, and the quinones substance that plant corpus discharges makes the medium around plant become sepia, but plant itself there is no brownization.This phenomenon does not occur in explant Induction Process, but all can occur in subculture process and after root media of transferring, the plantlet in vitro producing browning still can grow and take root, but growth rate and rooting rate are subject to obvious impact, situation seriously also can cause plantlet in vitro base portion gradually brownization and jeopardize its existence.Because browning only appears in minority plantlet in vitro, and have certain relation with the physiological status of explant material, therefore need choose the eugonic explant that children is tender when explant selection as far as possible, need in subculture process to abandon old branch part, only by new first portion subculture, part browning can be reduced like this, but the plantlet in vitro negligible amounts that this operation is turned out, be not suitable for large-scale production.
The a small amount of brownization seedling of report is had in prior art, the medium that only more need renew after there is brownization or different parts seedling being moved to same medium, browning is then resolved substantially, also no longer brownization is there is in what brownization was serious after replacing two subculture, but this operation not only increases operating procedure, also add production cost.
Therefore, study a kind of browning that can overcome Chinese tamarisk explant, shorten the forming seedling through one step culture method of cultivation cycle, have great importance.
Summary of the invention
The technical problem to be solved in the present invention, be to provide a kind of method of Chinese tamarisk plantlet in vitro culture of rootage, for forming seedling through one step culture method, with the addition of active carbon composition in the medium, overcome the phenomenon of Chinese tamarisk Brown, shorten cultivation cycle, reduce the consumption of sugar, to realizing, Chinese tamarisk group training industrialization is significant, not only reduces production cost, more reduces Material Cost, ensure that the stable of Chinese tamarisk genetic character simultaneously, realize obtaining a large amount of high-quality Chinese tamarisk plantlet in vitro wood in a short time.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
A method for Chinese tamarisk plantlet in vitro culture of rootage, it carries out according to following steps order:
(1) acquisition of aseptic seedling
With newborn spray or tender shoots for explant, rinse twice with washing powder water soaking, rear clear water rinses 30 ~ 50min; Use 70 ~ 75% alcohol disinfecting 20 ~ 30s again, aseptic water washing 2 times, afterwards with 0.05 ~ 0.1% mercuric chloride 100ml and 0.05ml Tween-20 sterilization 10-15min, finally in superclean bench, use aseptic water washing 6 ~ 8 times, remove the water on explant surface, prune, explant through pruning is forwarded in culture medium A and cultivates, condition of culture is: illumination cultivation 13h, and intensity of illumination is 1500 ~ 3500 luxs, and temperature is 23 ~ 27 DEG C; Cultivate and namely obtain aseptic seedling after 15-20 days;
(2) culture of rootage is bred
Aseptic seedling continued in culture medium A, cultivate 44-49 days, namely light green, the blade of get Ye Se is unfolded, and to take root seedling without vitrifying, the Chinese tamarisk group training of state of growing thickly of falling leaves without yellow leaf; Gained seedling is that plant height is greater than 1.8cm, and root is grown up in 1cm, and radical is greater than 2.
Limit as one of the present invention, described Chinese tamarisk group training seedling of taking root directly can be transplanted and to booth, to be planted or carry out squamous subculture prepare the training of Chinese tamarisk group and to take root seedling.
As the further restriction of above-mentioned restriction, described subculture method is seedling excision root system of the training of Chinese tamarisk group being taken root, each clump of seedling is divided into 7 or 8 little Cong, each little Cong is with 6 ~ 10 sprigs, average height 1-3cm, by every little Cong tissue culture plant inoculation in culture medium A, the inoculation degree of depth is 0.3 ~ 0.5cm, cultivate after 44-49 days, the training of Chinese tamarisk group is taken root seedling; Wherein culture of rootage condition is: illumination cultivation 13h, intensity of illumination 1500-3500 lux, and temperature is 23-27 DEG C.
Limit as another kind of the present invention, described culture medium A is medium based on MS, also be added with the sucrose of 15 ~ 20g/L, the agar of 5.5 ~ 7.0g/L, the 6-BA of 0.05 ~ 0.5mg/L, the KT of 0.1 ~ 0.3mg/L and mass fraction are the active carbon of 0.05-0.15%, adjust ph to 6.0 ~ 6.9.
As the further restriction of above-mentioned restriction, described pH value to 6.5.
As the further restriction of above-mentioned restriction, the consumption of described sucrose is 20g/L.
Owing to have employed above-mentioned technical scheme, compared with prior art, acquired technological progress is in the present invention:
The invention provides the method for Chinese tamarisk plantlet in vitro culture of rootage, for forming seedling through one step culture method, with the addition of active carbon composition in the medium, decrease the phenomenon of Chinese tamarisk Brown, shorten cultivation cycle, reducing the consumption of sugar, not only reduce production cost, more reduce Material Cost, ensure that the stable of Chinese tamarisk genetic character simultaneously, realizing obtaining a large amount of high-quality Chinese tamarisk plantlet in vitro wood in a short time significant to realizing Chinese tamarisk group training industrialization.
The present invention is applicable to the culture of rootage of Chinese tamarisk plantlet in vitro, is further used for the squamous subculture of plantation or plantlet in vitro in booth.
The present invention is described in further detail below in conjunction with specific embodiment.
Embodiment
The method of embodiment 1 one kinds of Chinese tamarisk plantlet in vitro culture of rootage
A method for Chinese tamarisk plantlet in vitro culture of rootage, it carries out according to following steps order:
(1) acquisition of aseptic seedling
With newborn spray or tender shoots for explant, rinse twice with washing powder water soaking, rear clear water rinses 30min; Use 70% alcohol disinfecting 20s again, aseptic water washing 2 times, afterwards with 0.1% mercuric chloride 100ml and 0.05ml Tween-20 sterilization 10min, finally in superclean bench, use aseptic water washing 6 times, remove the water on explant surface, prune, explant through pruning is forwarded in culture medium A and cultivates, condition of culture is: illumination cultivation 13h, and intensity of illumination is 2000 luxs, and temperature is 25 DEG C; Cultivate and namely obtain aseptic seedling after 20 days;
(2) culture of rootage is bred
Aseptic seedling continued in culture medium A, cultivate 45 days, namely light green, the blade of get Ye Se is unfolded, and to take root seedling without vitrifying, the Chinese tamarisk group training of state of growing thickly of falling leaves without yellow leaf;
Wherein, culture medium A is medium based on MS, is also added with the sucrose of 20g/L, the agar of 6.0g/L, and the KT of the 6-BA of 0.25mg/L, 0.2mg/L and mass fraction are the active carbon of 0.1%, adjust ph to 6.5.
Gained seedling is plant height is 1.8cm, and root is long is 1cm, and radical is 2.The Chinese tamarisk group training turned out seedling of taking root directly can be transplanted to booth plantation or carry out squamous subculture and prepared the training of Chinese tamarisk group and to take root seedling.
Carry out directly transplanting when planting to booth, method is: seedling of the training of Chinese tamarisk group being taken root, after eccysis base portion medium, by its direct plantation on the seedbed being paved with sandy soil, during transplanting, root system is buried in the earth, carry out conventional booth maintenance subsequently, after 30 days, booth transplanting survival rate reaches 85%, the seedling growth potential neat and consistent and the training of Chinese tamarisk group is taken root.And the general survival rate of Chinese tamarisk cuttage is 80%, and cuttage seeding growing way differs, and height is uneven.
When carrying out squamous subculture, cultural method is: seedling of the training of Chinese tamarisk group being taken root excision root system, and each clump of seedling is divided into 7 little Cong, and each little Cong is with 8 sprigs, average height 2cm, by every little Cong tissue culture plant inoculation in culture medium A, the inoculation degree of depth is 0.3cm, and condition of culture is: illumination cultivation 13h, intensity of illumination 2000 lux, temperature is 25 DEG C, cultivates after 45 days, the training of Chinese tamarisk group is taken root seedling.This Chinese tamarisk group training seedling of taking root can proceed squamous subculture circulation, to take root seedling, again carry out planting in squamous subculture and booth to obtain the training of Chinese tamarisk group.
The method of embodiment 2-6 Chinese tamarisk plantlet in vitro culture of rootage
Embodiment 2-6 is respectively a kind of method of Chinese tamarisk plantlet in vitro culture of rootage, similar to the cultural method of embodiment 1, and difference is only the difference of the index of wherein involved technical parameter and obtained Chinese tamarisk plantlet in vitro, shown in table specific as follows:
The technical parameter of embodiment 2-6 squamous subculture, as shown in the table:
The screening technique of embodiment 7 culture medium A
The screening of condition has been carried out to culture medium A, at medium based on MS, sucrose 20g/L, agar 5.8g/L, when pH is adjusted to 6.5, adds different types of nutrient or additive wherein, upgrowth situation for Chinese tamarisk plantlet in vitro has obvious difference, especially, while minimizing brown material, ensure the basic demand (quantity etc. as plant height, root length and root) of plantlet in vitro, detailed process is as shown in the table:
As seen from the above table, after with the addition of active carbon composition, observe the upgrowth situation of Chinese tamarisk plantlet in vitro, greatly reduce the appearance of brown material, and the condition of rooting of plantlet in vitro is good, meets the requirement proceeding squamous subculture and directly carry out greenhouse gardening.
Carried out corresponding experiment to the addition of active carbon again, find, and the concentration of charcoal non activated has been higher, its browning reduces the also proportional minimizing of degree, and the upgrowth situation of Chinese tamarisk plantlet in vitro also has a declining tendency, and specific experiment result is as follows:
As seen from the above table, during active carbon employing variable concentrations, the upgrowth situation of the Chinese tamarisk plantlet in vitro of turning out is different, when concentration of activated carbon is 0.05-0.15%, the upgrowth situation of Chinese tamarisk plantlet in vitro is good, state of growing thickly, the average 5-6 of growth coefficient, average plant height 1.8cm, plantlet in vitro is taken root for about 15 days, cultivates 44-49 days, the long > 1cm of root, radical >=2, occur without brown material; And when active carbon concentration lower than 0.05% time, still have brown material occur; When active carbon concentration higher than 0.15% time, although brown material does not occur again, the upgrowth situation of Chinese tamarisk plantlet in vitro declines, value-added coefficient and average plant height reduce, reason is, concentration of activated carbon increase can adsorb the nutriment in medium, and the upgrowth situation of Chinese tamarisk plantlet in vitro is declined.
The above is only preferred embodiment of the present invention, is not restriction the present invention being made to other form, and any those skilled in the art may utilize above-mentioned technology contents to be changed or be modified as the Equivalent embodiments of equivalent variations as enlightenment.But everyly do not depart from technical solution of the present invention content, according to technical spirit of the present invention to the simple modification done by above embodiment, equivalent variations and remodeling, still belong to the protection domain of the claims in the present invention.
Claims (6)
1. a method for Chinese tamarisk plantlet in vitro culture of rootage, is characterized in that it carries out according to following steps order:
(1) acquisition of aseptic seedling
With newborn spray or tender shoots for explant, rinse twice with washing powder water soaking, rear clear water rinses 30 ~ 50min; Use 70 ~ 75% alcohol disinfecting 20 ~ 30s again, aseptic water washing 2 times, afterwards with 0.05 ~ 0.1% mercuric chloride 100ml and 0.05ml Tween-20 sterilization 10-15min, finally in superclean bench, use aseptic water washing 6 ~ 8 times, remove the water on explant surface, prune, explant through pruning is forwarded in culture medium A and cultivates, condition of culture is: illumination cultivation 13h, and intensity of illumination is 1500 ~ 3500 luxs, and temperature is 23 ~ 27 DEG C; Cultivate and namely obtain aseptic seedling after 15-20 days;
(2) culture of rootage is bred
Aseptic seedling continued in culture medium A, cultivate 44-49 days, namely light green, the blade of get Ye Se is unfolded, and to take root seedling without vitrifying, the Chinese tamarisk group training of state of growing thickly of falling leaves without yellow leaf; Gained seedling is that plant height is greater than 1.8cm, and root is grown up in 1cm, and radical is greater than 2.
2. the method for Chinese tamarisk plantlet in vitro culture of rootage according to claim 1, is characterized in that: described Chinese tamarisk group training seedling of taking root directly can be transplanted to booth plantation or carry out squamous subculture and prepared the training of Chinese tamarisk group and to take root seedling.
3. the method for Chinese tamarisk plantlet in vitro culture of rootage according to claim 2, it is characterized in that: described subculture method is seedling excision root system of the training of Chinese tamarisk group being taken root, each clump of seedling is divided into 7 or 8 little Cong, each little Cong is with 6 ~ 10 sprigs, average height 1-3cm, by every little Cong tissue culture plant inoculation in culture medium A, the inoculation degree of depth is 0.3 ~ 0.5cm, cultivate after 44-49 days, the training of Chinese tamarisk group is taken root seedling; Wherein culture of rootage condition is: illumination cultivation 13h, intensity of illumination 1500-3500 lux, and temperature is 23-27 DEG C.
4. the method for the Chinese tamarisk plantlet in vitro culture of rootage according to any one of claim 1-3, it is characterized in that described culture medium A is medium based on MS, also be added with the sucrose of 15 ~ 20g/L, the agar of 5.5 ~ 7.0g/L, the 6-BA of 0.05 ~ 0.5mg/L, the KT of 0.1 ~ 0.3mg/L and mass fraction are the active carbon of 0.05-0.15%, adjust ph to 6.0 ~ 6.9.
5. the method for Chinese tamarisk plantlet in vitro culture of rootage according to claim 4, is characterized in that: described pH value to 6.5.
6. the method for Chinese tamarisk plantlet in vitro culture of rootage according to claim 4, is characterized in that: the consumption of described sucrose is 20g/L.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105052738A (en) * | 2015-07-31 | 2015-11-18 | 青岛正杰实业有限公司 | Tamarix chinensis tissue culture and rapid propagation method |
| CN106508678A (en) * | 2016-10-27 | 2017-03-22 | 甘肃省治沙研究所 | Tamarix taklamakanensis tissue culture and rooting method |
| CN106550872A (en) * | 2016-11-02 | 2017-04-05 | 甘肃省治沙研究所 | A kind of Ramulus et Folium Tamariciss Rapid Rooting culture medium and Ramulus et Folium Tamariciss method for tissue culture |
| CN109548649A (en) * | 2017-11-01 | 2019-04-02 | 岭南生态文旅股份有限公司 | A kind of M. laxiflora tissue cultures inducer substance and its application method |
| CN116034873A (en) * | 2022-12-02 | 2023-05-02 | 武威市林业科学研究院 | Tamarix chinensis tissue rapid propagation method |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105052738A (en) * | 2015-07-31 | 2015-11-18 | 青岛正杰实业有限公司 | Tamarix chinensis tissue culture and rapid propagation method |
| CN106508678A (en) * | 2016-10-27 | 2017-03-22 | 甘肃省治沙研究所 | Tamarix taklamakanensis tissue culture and rooting method |
| CN106550872A (en) * | 2016-11-02 | 2017-04-05 | 甘肃省治沙研究所 | A kind of Ramulus et Folium Tamariciss Rapid Rooting culture medium and Ramulus et Folium Tamariciss method for tissue culture |
| CN109548649A (en) * | 2017-11-01 | 2019-04-02 | 岭南生态文旅股份有限公司 | A kind of M. laxiflora tissue cultures inducer substance and its application method |
| CN116034873A (en) * | 2022-12-02 | 2023-05-02 | 武威市林业科学研究院 | Tamarix chinensis tissue rapid propagation method |
| CN116034873B (en) * | 2022-12-02 | 2023-11-17 | 武威市林业科学研究院 | Tamarix chinensis tissue rapid propagation method |
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