Dendrobium Jingjie vitro studies
【Abstract】 Objective To achieve the sterile stem rapid propagation of Dendrobium candidum. Methods Dendrobium plantlets Jingjie inoculated at different concentrations of MS medium supplemented with NAA-induced axillary bud sprouting,
and with different hormone combinations and organic additives on the subculture medium and induction of rooting. Results NAA 0.4 mg / L ,6-BA 5 mg / L when the germination rate of
axillary buds Dendrobium 93.33%; NAA 0.3 mg / L ,6-BA 2 mg / L
when the bundle seedlings rooting rate 96.67%, banana juice and potato juice can significantly promote the proliferation of multiple shoot clumps and strong seedling root. Conclusion Dendrobium stem suitable for in vitro rapid propagation
system.
Key words Dendrobium sterile Jingjie vitro hormone concentrations of organic additives
Abstract: ObjectiveTo develop a rapid propagation of Dendrobium candidum using the aseptic nodal segments of shoots.MethodsAxenic nodal segments were first germinated on
Murashige and Skoog (MS) basal medium supplemented with different concentration NAA.The seedlings were subcultured and rooted on the medium containing different concentration hormones and organic susbstance.Results93.33% of the axenic
nodal segments of Dendrobium candidum germinated on the medium with NAA 0.4 mg / L ,6-BA 5 mg / L and 96.67% of explant
rooted on the medium containing NAA 0.3 mg / L ,6-BA 2 mg /
L.The growth of explant was promoted by the extract of banana and potato.ConclusionThe in vitro propagation of Dendrobium candidum from axenic nodal segment has been established.
Key words: Dendrobium candidum; Axenic nodal segments; In
vitro propagation; Concentration of hormone; Organic susbstance
Dendrobium was first seen in "Shan Hai Jing", medicinal contained only in the "Shen Nong's Herbal Classic", then the ages of various herbs are therefore correspondingly recorded out at home. In the "Compendium of Materia Medica" was listed as top grade. Dendrobium so far is still commonly used in
Chinese medicine Ziyin Qingfei, Shengjinzhike, Yang Wei Chufan to the drug, its scope of application is more tending to widen. Recent studies have found that Dendrobium Dendrobium contains polysaccharide [1], alkaloids [2 ~ 4], drumstick
Philippines, and Erianin [5] and other compounds, as well as seven kinds of essential amino acids [6] and a variety of trace elements. Dendrobium with modern pharmacological showed anti-tumor [7], anti-aging, strengthen immunity [8] and other
functions, cardiovascular, digestive, respiratory and eye diseases have obvious therapeutic effects [9].
Dendrobium Dendrobium candidum is the current version of "Chinese Pharmacopoeia" collection contained in the five kinds of commonly used Dendrobium is one of the top grade of
Dendrobium class medicine. Dendrobium sect, also known as black grass, metal Chlorophytum, iron Piran, belong to Orchidaceae Dendrobium epiphytes. Processed Dendrobium Dendrobium maple is also known as the Big Dipper, the market
is expensive, is an economic value and are high medical value of precious Chinese medicine. However, due to over-harvesting
in recent years, the ecological destruction of the environment and other reasons, increasing Kui Dendrobium wild resources have dried up, while demanding their habitat, natural and low reproduction rate factors further Dendrobium brink of extinction, has now been listed as endangered the protection of TCM varieties [10]. The use of in vitro rapid propagation of Dendrobium Dendrobium to resolve the current contradiction between resources and needs of an important way. At present the study on the Dendrobium focused on the original bulb proliferation, differentiation, etc. [11,12], to Jing Jie as the material, to achieve rapid propagation of Dendrobium
candidum in vitro research is still rare reports. This study was sterile stem segments by in vitro culture, establishment of a suitable system for rapid propagation of Dendrobium candidum.
1 Materials and methods
1.1 Materials
Choices and to handle the growth of our laboratory culture maintained strong, node thick of Dendrobium plantlets as the material, cut a length of 1 ~ 1.5 cm with a 1 ~ 2 axillary buds of a small stem.
1.2 axillary buds sprouting would be sterile stem
segments were inoculated on to add 30 g * L-1 sucrose, 8.0 g *
L-1 agar, pH 5.6 in MS medium. One 6-BA 5 mg * L-1, and
different concentrations of NAA (0 ~ 0.8 mg * L-1). Culture
temperature is 25 ?, light intensity of 2 000 lux,
photoperiod of 12 h * d-1. On a regular basis the effects of different concentrations of growth hormone on the germination of Dendrobium candidum role of axillary buds.
1.3 propagation of adventitious buds sprouting of axillary buds will have been inoculated in the MS 30 g * L-1
sucrose NAA 0.4 mg * L-1 6-BA 5 mg * L-1, and add 200 g * L-1
of the bananas, potatoes, watermelon , bean sprouts proliferation medium and in medium without natural additives as the control, pH were 5.6, The effects of different extracts on the proliferation of multiple shoot clumps. Ibid culture conditions.
1.4 strong seedling roots to grow robust, the plant height of 2 cm or so plagued seedlings were inoculated on MS 30 g * L-1 sucrose 8.0 g * L-1 agar, hormone concentrations
were NAA 0.1,0.3,0.5 mg * L-1 and 6-BA 2,4 mg * L-1, or MS 30
g * L-1 sucrose NAA 0.3 mg * L-1 6-BA 2 mg * L-1 8.0 g * L-1
agar, add 200 g * L -1 different extracts of the rooting
medium on rooting and strong seedling trials conducted. Ibid culture conditions.
1.5 Statistics seedling growth conditions in vitro cultured plantlets inoculated 50 d after the induction rate were observed statistics bud, bud proliferation and number, plant height, root induction rate, root number, root length, plant robust degree and extent of browning.
2 Results
2.1 of different concentrations of auxin induction of axillary bud on the stem segment role of 6-BA for the 5 mg *
L-1, different concentrations of NAA-induced germination of
stem axillary buds if the results shown in Table 1. In Table 1
we can see from the NAA axillary buds of Dendrobium Stem with the concentration of the role of germination. When adding the concentration is low, NAA can significantly promote the germination of axillary buds, NAA concentration of 0.4 mg * L-
1, the axillary buds germination rate, multiplication to achieve the highest, 93.33% and 1.97, respectively, and the leaves are dark green, thick stems, the formation of a large number of multiple shoot clumps. The NAA concentrations continue to rise, the germination rate and multiplication of axillary buds decreased significantly, but also part of the material due to necrosis, resulting in experiments reduce the number of total shoots. Experiment also found that when the NAA concentration is too high, some of the axillary buds after germination occurs distortion, differentiation of flower buds, and withered gradually die. The results showed that low concentrations of NAA can promote the germination of Dendrobium axillary buds, induced by bud differentiation,
plant growth robust. But the NAA concentration is too high, it will inhibit the germination and plant growth of axillary buds and to promote flower bud formation, more suitable for the cultivation of Dendrobium watch. According to the experimental
results to NAA 0.4,6-BA 5 mg * L-1 as a subculture medium, the
test-tube seedlings subculture, the results of the growth of good, robust plants, the proliferation of multiple high (up to 3.2 times). Table 1 NAA on axillary bud germination of Dendrobium (omitted) reposted elsewhere in the paper for free download http://
2.2 The different extracts on shoot proliferation of different extracts on the proliferation of the role of Dendrobium buds was markedly different. Bananas, potatoes, watermelon can significantly improve the shoot multiplication, in which banana juice most conducive to the proliferation of Dendrobium buds, reaching 4.98 times, followed by potatoes, to 4.90-fold. Banana juice and potato juice can not only promote the differentiation of bud, but also conducive to the growth of shoots an average of 1.30 cm, respectively, and Ya Zhang 1.39 cm, plants are very robust. Bean sprout juice on shoot proliferation and growth does not promote the role of plant robust levels were generally not suitable for the cultivation
of Dendrobium candidum. In addition, the control group in the training process, banana group, watermelon group suffered from varying degrees of medium browning, materials, phenomena such as the roots of black, but the potato group does not appear in
any medium Browning. The results in Table 2. Table 2 Effects of different extracts on the proliferation of Dendrobium buds (omitted)
2.3 of different hormone combinations on rooting of Dendrobium seedlings inoculated in the profusion of different
hormone combinations of the rooting medium, 15 d can be observed after a few bottles of root formation, to 20 d, the first two groups rooting rate reached 58%, 40 d, when reached 96.67%, followed by rooting rate is no longer growing. Proportion of different hormone-induced Dendrobium plagued
rooting ability of significantly different. Lower concentrations of 6-BA in favor of Dendrobium roots; and NAA at low concentrations, its ability to promote rooting and the concentration was positively correlated, when the
concentration is too high (0.5 mg * L-1), its reduced capacity
to promote rooting. When the NAA 0.3 mg * L-1 ,6-BA 2 mg * L-
1, the highest rate of rooting of Dendrobium, reaching 96.67% at this time plant height, root number, root length and the
degree of all able-bodied best, but the bud lower
multiplication. Therefore, in order to induce rooting for the purpose, the optimum hormonal ratio of NAA 0.3 mg * L-1 ,6-BA
2 mg * L-1. The results in Table 3. Table 3 different hormone combinations on rooting of Dendrobium (omitted)
2.4 Different extracts of roots of Dendrobium candidum
Different extracts of Dendrobium candidum in various aspects of culture have a significant effect strong seedling roots. Banana juice most beneficial Dendrobium strong sprout
roots, whether it is the proliferation of multiple shoots, plant height, rooting rate, root number, root length, add the banana juice were significantly better than the control group, root thick, clean, dark green leaf color, stem Thick short.
Potato juice can promote the proliferation and growth of seedlings have arisen, there is conducive to obtaining healthy stout test-tube seedlings, no browning phenomenon, but
inhibited rooting. The role of watermelon juice and potato juice the contrary, it can significantly improve the rooting rate and root number, but the weak plants, stems Zoysia less
root part of the leaves turn yellow wilt, roots thin and short, partial loss of geotropic. The results in Table 4. Table 4 different extracts of the roots of Dendrobium
(omitted)
3 Conclusions and discussion
The experimental results show that in suitable culture conditions, Dendrobium sterile stem segments can be highly effective germination. In the MS NAA 0.4 mg * L-1 6-BA 5 mg *
L-1 sucrose for 50 d after germination rate of axillary buds, multiplication highest, 93.33% and 1.97, respectively, plant growth and robust; to the medium to subculture, its multiplication to 3.2 times. In order to stem as the material, to achieve rapid propagation of Dendrobium candidum, you can broaden the sources of tissue culture materials, speed up breeding, promoting large-scale production of genetic
stability of plantlets shape.
Commonly used in tissue culture of organic additives, such as banana juice, watermelon juice and potato juices are rich in amino acids, hormones and vitamins and other organic and natural ingredients are more complex compounds, their cell and tissue proliferation and differentiation of a clear catalytic role [13,14]. This study shows that sprouts juice
right Dendrobium does not promote the role of propagation of adventitious buds is not suitable as the proliferation of cultured Dendrobium candidum organic additives. Banana, watermelon, potatoes on the propagation of adventitious buds
have a facilitating role. One banana juice on the propagation of adventitious buds, roots occurred in varying degrees to promote, help develop strong sprout. Potato juice can significantly improve the shoot multiplication, but inhibit the formation of roots. In plant propagation, the medium cost is an important part of production costs [13], using potatoes instead of the banana can significantly reduce the Dendrobium factory costs of production. Watermelon juice is able to promote root formation, increase the number of roots, but roots thin and short, partial loss of gravitropism, vertical upward growth, weak plant growth. QIN Gai-Hua et al [15] found
that adding organic matter alone, the banana juice most conducive to the proliferation of Steller's blue of the
original bulb, while adding banana juice and potato juice, the effect is more than banana juice alone. Whether the use of
different organic compounds with a similar effect can be achieved still needs further study.
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