Blackwater Valerian tissue culture and rapid propagation of
【Abstract】 Objective To Blackwater Valerian callus
induction medium and culture conditions to study the growth, build Blackwater Valerian rapid propagation system. Methods
filter explants, adding different types and concentrations of growth regulators on the Blackwater Valerian for callus induction, shoot proliferation and rooting culture cultivation. The results suitable for callus induction medium: MS 0.5 mg / LNAA 5.0 mg/L6 BA. Bud proliferation of
cultured to MS 0.2 mg / LNAA 10.0 mg/L6 BA is appropriate.
Rooting culture with MS 0.1mg/LNAA better. Conclusion can be achieved using tissue culture techniques Blackwater Valerian multiply quickly.
Key words Blackwater Tissue Culture and Rapid Propagation of Valeriana
AbstractObjectiveTo provide some new evidences for rapid propagation of Valeriana amurensis Smir. Ex Kom through studying on medium with callus tissue culture and the condition of growth.MethodsPrimary culture of different
explants, culture of cluster buds and their rooting culture were conducted on medium of treatment combinations of adding different hormones. ResultsThe appropriate medium for different culture stages were MS 0.5 mg / LNAA 5.0 mg/L6 BA
in callus culture induction, MS 0.2 mg / LNAA 10.0 mg/L6 BA
in differentiation and subculture of cluster buds, MS 0.1 mg / LNAA in rooting culture.ConclusionRapid propagation can be achieved with plant tissue culture in Valeriana amurensis Smir. ex Kom.
Key wordsValeriana amurensis Smir. Ex Kom; Tissue culture; Rapid propagation
Blackwater Valerian (Valeriana amurensis Smir. Ex Kom) for the Valerianaceae (Valerianaceae) Valeriana (Valeriana Linn.) Perennial herbs. I have made in the Northeast,
concentrating in the Changbai Mountain and its Yumo area. Medicine part of the root and rhizome. Blackwater contains valerian main iridoid, sesquiterpenes, alkaloids and flavonoids four types of chemical composition [1], have a calming, antispasmodic, anti-tumor activity, such as the role
of antidepressant activity, in particular its a-half times
terpenoid and ester compounds Valerian has great potential for use is expected to seek a new type of antidepressants and sedatives [2]. Present Valerian extract and its preparations
are very popular in the international sales of the top 10 listed herbal [3].
Our multi-use of wild resources, Valerian is not
conducive to mass production. In recent years, tourism development and the natural environment due to the destruction
and uncontrolled excavation, wild valerian state resources have declined sharply. Also, because Valeriana plants perennial herb, natural growth is very slow and tends to be more depletion of wild resources. Carry out Blackwater
Valerian tissue culture and rapid propagation of the research, has become more pressing and important. In regard to the genus Valeriana in tissue culture of a small number of reports, Jian-Rong Chen et al [4] on the induction of callus was Valerian preliminary study. After checking the literature, on the Blackwater Valerian tissue culture have not yet see the relevant reports. This study was designed by Blackwater Valerian tissue culture studies for the establishment to provide the basis of its rapid propagation system.
1 Materials and methods
1.1 Materials used in this study were collected from Blackwater Valerian Yichun in Heilongjiang Green Lake area, through the Pharmacy Teaching and Research Professor Liu Juan Jiamusi university students identified as Blackwater Valerian Valeriana amurensis Smir. Ex Kom. Experiment were analytically pure reagents.
1.2 Instrument SA-1600-2JZ Multi Clean console (Shanghai Jia Feng Horticultural Products Co., Ltd.); ZPQ-280D
intelligent climate incubator (Heilongjiang East Extension
Instrument Manufacturing Co., Ltd.); YXQ-LS-75SLL Vertical
Pressure Steam eliminate bacteria device (Boxun Industrial Co., Ltd. Shanghai Medical Equipment Factory); electronic balance FA2004 (Shanghai Kohei Technology Co., Ltd.).
1.3 Medium and culture conditions of use of MS basic medium, adding 0.8% agar, 3% sucrose and different concentrations of plant growth regulator, adjust pH value to 5.8, 121 ? autoclave 30min, standby. All training were carried out under the following conditions: temperature (25 ?
1) ?, illumination time 12h / d, light intensity 1500 ~ 2000 lx.
1.4 means the material by running water 30 min later, placed in sterile ultra-clean bench vaccination. 70% alcohol before rinsing 30s, then 0.1% HgCl2 disinfection 8 min,
finally washed with sterile water, 7 to 8 times, will be disinfected after the shoot tip, stem segments and petioles about 1 ~ 1.5 cm cut into small bits, leaves cut into 0.5 cm2 size, was inoculated into a pre-preparation of a good culture
medium for callus induction.
2 Results
2.1 Different Explant on Blackwater Valerian callus induction Blackwater Valerian order to determine the best callus induction site, the shoot tip, stem, petiole base, petioles and leaves were inoculated in the middle containing the MS 0.2 mg / LNAA (NAA) 5.0 mg/L6 BA (6 amino benzyl
adenine) of the medium. 20 d after inoculation, statistics callus induction rate (see Table 1). The results showed that containing MS 0.2 mg / LNAA 5.0mg/L6 BA to MS medium, to
the base of shoot tips and petioles as explants induced callus is more appropriate, with inter-section of stem is more
appropriate for inducing multiple shoot clumps . Reposted elsewhere in the paper for free download http://
Table 1 Effect of different explants on callus induction (omitted)
2.2 The different plant growth regulators on Blackwater Valerian callus induction and growth of different concentrations investigated ratio of NAA and 6-BA on callus
induction and organizational growth. Table 2.
Table 2 NAA and 6-BA of different concentrations of
combinations (omitted)
Multiple types of initial callus culture generated after subculture of callus regeneration capacity of different types of forms in different. Callus produced by the initial
training, there are loose texture, hard, color white, yellow, light green, dark green. Where C Series media are growing more satisfactory (see Table 3). Callus medium No. C4 compact structure, smooth surface, after subculture of callus to
maintain its robust growth, cultured 25 d after the formation of visible white roots. , No. C1 and normal growth medium after subculture, callus texture, moderate, and long-term
culture no root differentiation, which features a stronger ability of callus meristem can be used as cell culture.
Table 3 C Series formula of shoot-tip callus growth
effects (abbreviated)
2.3 arisen between the stem bud induction and
proliferation of cultured seedlings with different concentrations of the ratio of 6-BA and NAA on the Blackwater
between valerian with node-induced stem sections. The results
showed that after 30d of culture, part of the section of the medium between the parts of a long shoot. Among them, No. B5 medium to long shoots shorter periods of time, and grow
faster. Stem segments in the training begins when you start swelling, and occur more frequently in the base of the green dots, with the growth and differentiation of plants out of a small bud, bud gradually increasing the number of buds formed
plexus, the profusion of cut shoots transferred to different concentrations of ratio of 6-BA and NAA medium, which is still
No. B5 medium bud proliferation effect is better.
2.4 rooting culture rooted as long as no more than 1 cm when transferred to MS NAA (0.05,0.10,0.20,0.40 mg / L) in the rooting medium. One MS 0.10 mg / LNAA medium to form the root of a white thick and fast growth. 3 weeks time, will be able to produce good root system.
3 Discussion
3.1 explant on callus induction effect in this
experiment, the Blackwater shoot tip culture Valerian callus after the first, and then cut end of petiole and stem callus, and finally the callus of leaf breakage. This indicates that the site easy to split the exuberant callus-based, easy to cut
the damaged area of callus. The reason for this difference may be due to strong growth period of the materials with higher levels of endogenous hormones, and then divide easier. Therefore, before callus induction, the selection of appropriate organizational part of it is very necessary, should try to choose the undifferentiated or differentiated to a lesser extent the use of organizations as explants.
3.2 Plant Growth Regulators on Callus Induction Blackwater valerian is generally believed that the impact of
the key to success lies in callus culture conditions. Among them, plant growth regulators is extremely important. In this study, the use of 6-BA and NAA in different concentrations to induce callus ratio can be seen when higher concentrations of
NAA and 6-BA concentration is relatively low, the explants formed callus is relatively much more desirable. Thus, for callus induction, they should join the amount of growth hormone and less accession to cytokinin. To be able to form a callus on media comparison of several groups found that, in the medium containing 6-BA in the callus can be a long-term
subculture and does not wither. The reason may be 6-BA can
delay tissue senescence, and thus slow down or inhibit the browning [5].
3.3 Plant Growth Regulators on Blackwater Valerian proliferation and growth of seedlings in this experiment the use of 6-BA and NAA concentrations in different ratio to carry out the proliferation of cultured seedlings was found to increase with the 6-BA concentration of the faster growth of seedlings, but the 6-BA concentration of not more than 10.0 mg / L. NAA in the process of adventitious bud differentiation plays an important role, but the concentration of not more than 1.0 mg / L. Thus, for the proliferation of cultured
shoots, should be added to moderate cytokinin, plus a small amount of auxin. ZHANG Lan-ying, etc. This leaves a wide
valerian results are consistent with the reports.
3.4 Plant Growth Regulators on Blackwater inducing effects of valerian roots in this experiment using 4 different
concentrations of NAA rooting medium and found that relatively low concentrations of NAA to take root more easily. This may be due to the concentration of auxin on rooting have an impact. Its concentration is low, the promotion of rooting. This also demonstrates that NAA is more appropriate in Blackwater valerian root.
In summary, the experiments were determined callus induction, clusters of buds develop and the best rooting culture medium formula. Blackwater used valerian-induced
petiole and stem produce callus method for a test, resulting in a relatively short period, a large number of multiple shoot clumps, and then rooting, thus providing a scientific basis
for its rapid propagation. For the stem and petiole callus induction, its effective medicinal ingredients and changes in the content, but also need to be further in-depth study.
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