CN103548686B - Flower of Japanese Camellia tissue cultured seedling root media - Google Patents
Flower of Japanese Camellia tissue cultured seedling root media Download PDFInfo
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- CN103548686B CN103548686B CN201310532889.3A CN201310532889A CN103548686B CN 103548686 B CN103548686 B CN 103548686B CN 201310532889 A CN201310532889 A CN 201310532889A CN 103548686 B CN103548686 B CN 103548686B
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Abstract
The invention discloses a kind of Flower of Japanese Camellia tissue cultured seedling root media, be made up of the component of following concentration: saltpetre 80mg/L, four water-calcium nitrate 150mg/L, magnesium sulfate heptahydrate 350mg/L, potassium primary phosphate 100mg/L, Repone K 65mg/L, sodium sulfate 200mg/L, white sugar 20g/L, carrageenin 6.0g/L, gac 100mg/L, indole-3-butyric acid 2.0mg/L, indolylacetic acid 0.1mg/L.The substratum that the most applicable Flower of Japanese Camellia tissue cultured seedling that the present invention finally obtains is taken root is MV+IBA2.0mg/L+IAA0.1mg/L+ gac 100mg/L, and rooting rate reaches as high as 73.3%, mean elements 2.2, average root is long is 2.4cm, and plant is taken out high fast, leaf leaf look all normal, and root system is more flourishing.
Description
Technical field
The invention belongs to field of plant tissue culture technique, relate to a kind of Flower of Japanese Camellia tissue cultured seedling root media.
Background technology
Flower of Japanese Camellia (Camellia japonica L.) calls camellia, camellia, thorn apple tree etc., and being Angiospermae Dicotyledoneae Theales Theaceae Camellia (Camelliao L.) plant, is one of China's tradition famous flower, kind nearly over one hundred kind.Because of evergreen all the year round, that the flowers are in blossom is gorgeous, fragrant, flower appearance and color charming, usually used as a kind of perennial woody ornamental flower of elegance, be suitable for the plantation of the aspect such as family potted plant, afforestation, there is very large ornamental value, practical value and afforestation and be worth.In addition, Flower of Japanese Camellia also has a lot of pharmaceutical use, and root, the Hua Junke of Flower of Japanese Camellia are used as medicine, and has effect of the cool blood of convergence, hemostasis.It is reported, Flower of Japanese Camellia plant has very strong absorption CO
2ability, to SO
2, H
2s, Cl
2, HF, the obnoxious flavour such as flue dust have extremely strong resistance, thus play an important role in environment protection, purifying air.
The modes of reproduction of Flower of Japanese Camellia is mainly divided into sexual propagation and vegetative propagation.
Sexual propagation general seed is bred, and seed has very strong variation ability through the new plant after planting become, and thus becomes the important way that Flower of Japanese Camellia is bred.But chronic required for this modes of reproduction, and a lot of famous and precious self fertility of rare kind is poor, not easily sets seeds.Thus, sexual propagation Flower of Japanese Camellia can not reach the object of fast breeding offspring, at this moment in order to obtain the camellia flower seedling of more multiple characters improved seeds fast, just must carry out artificial asexual multiplication.
Vegetative propagation is also known as vegetative reproduction.Vegetative propagation has the characteristic keeping maternal good character, this offspring's vitality is vigorous, bloom, result comparatively early, breeding potential is very high, and the breeding cycle shortens greatly relative to sexual propagation.Conventional method has cuttage, grafting, press strip and tissue culture propagation.
Cottage propagation: with mid-June and best by the end of August.The selection of cutting directly affects its surviving rate, all must have very high requirement during cuttage to matrix, temperature, humidity.Due to the impact by reproductive material in season, the breeding coefficient of cottage propagation is low, and not easily takes root, and is not suitable for scale operation.
Propagation by grafiting: the highest with graft survival rate in 5 ~ June, after taking over a job, rudiment takes out the tip soon, within general about 60 days, rudiment can take out the tip.But the operation easier of grafting is comparatively large, not easily grasps.Required cost of labor is larger.
Mound layering: general at plum rain season press strip, take root after about 60 days, directly potted plant, surviving rate is high.
Cuttage, press strip, propagation by grafiting mode all will be subject to the constraint of natural condition and material self, and rooting rate and surviving rate are not high, can not be used for large-scale production, the development of serious restriction Flower of Japanese Camellia.
Tissue culture propagation: select explant, is cut into 1cm length and is seeded on substratum, carry out shoot proliferation, then transfer on new root media, can take root, then transplant through about 4 weeks after sterilization.
Plant tissue culture is the technology being most widely used, producing larger economic benefit on producing, and has the features such as breeding coefficient is large, reproduction speed fast, the cycle is shorter, nursery stock neat and consistent.Therefore, current garden crop and commodity trees part or major part all adopt rapid propagation in vitro to provide nursery stock, industrialization can produce, meet the demand of market to nursery stock.
Flower of Japanese Camellia is as xylophyta, and tissue culture difficulty is large, and especially take root the stage, this is because in Flower of Japanese Camellia tissue, aldehydes matter content is higher, easily causes the generation of brown stain.Domestic and international report major part adopts the root induction of filter paper bridge liquid nutrient medium, and to plant-growth regulator concentration and type requirements very high, major part adopts NAA to induce taking root of Camellia Plants.Therefore, the rooting propagation system of Flower of Japanese Camellia is studied, selects the conditions such as suitable substratum, sucrose concentration, hormone levels to alleviate the brown stain of Flower of Japanese Camellia tissue cultured seedling, improve rooting rate, find out the culture medium prescription that the most applicable Flower of Japanese Camellia tissue cultured seedling is taken root, significant.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of Flower of Japanese Camellia tissue cultured seedling root media, select best plant growth conditioning agent and substratum to promote that Flower of Japanese Camellia is taken root, improve rooting rate.
For achieving the above object, the invention provides following technical scheme:
Flower of Japanese Camellia tissue cultured seedling root media of the present invention, is made up of the component of following concentration:
Further, the pH value of described root media is 5.8.
Beneficial effect of the present invention is: the present invention has broken the method that necessary employing filter paper bridge liquid nutrient medium is taken root, the MV substratum that optimized choice inorganic salt content is lower is the minimum medium of Flower of Japanese Camellia tissue cultured seedling root induction, and have also been changed the traditional concept adopting NAA root induction in growth hormone kind and concentration, the best plant growth conditioning agent that optimized choice induction Flower of Japanese Camellia tissue cultured seedling is taken root is combined as IBA2.0mg/L+IAA0.1mg/L+ gac 100mg/L; The substratum that the most applicable Flower of Japanese Camellia tissue cultured seedling that the present invention finally obtains is taken root is MV+IBA2.0mg/L+IAA0.1mg/L+ gac 100mg/L, and rooting rate reaches as high as 73.3%, mean elements 2.2, average root is long is 2.4cm, and plant is taken out high fast, leaf leaf look all normal, and root system is more flourishing.
Accompanying drawing explanation
In order to make object of the present invention, technical scheme and beneficial effect clearly, the invention provides following accompanying drawing and being described:
Fig. 1 is that Flower of Japanese Camellia is taken root seedling.
Embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.
The Flower of Japanese Camellia tissue cultured seedling that embodiment of the present invention test materials used provides for landscape flower engineering center of Institute Of Unity and Coherence In Writing Of Chongqing.
1. the screening of minimum medium
Taking root of Flower of Japanese Camellia is very difficult, and this is the general character of xylophyta.In order to find out the high and good substratum of taking root of Flower of Japanese Camellia rooting rate, in the Flower of Japanese Camellia tissue cultured seedling of succeeding transfer culture 30d, select robust growth, consistent Flower of Japanese Camellia tissue cultured seedling as test materials, the budling of being cut into 2 ~ 3cm is inoculated on different root medias, different root medias is respectively with the 1/2MS substratum that inorganic salt content high-micro-element is complete, the 1/2SH that inorganic salt content is medium and the lower MV substratum of inorganic salt content are minimum medium (formula of minimum medium is as shown in table 1), in addition, white sugar 20g/L is all added with in first group of substratum (C1 ~ C3), carrageenin 6.0g/L, gac 100mg/L and indole-3-butyric acid (IBA) 2.0mg/L, white sugar 20g/L is all added with in second group of substratum (C4 ~ C6), carrageenin 6.0g/L, gac 100mg/L, indole-3-butyric acid (IBA) 2.0mg/L and indolylacetic acid (IAA) 0.1mg/L.
Table 1 minimum medium formula
Culture condition is: intensity of illumination 800-1000Lx, culture temperature 26-27 DEG C, lighting delay number 10h/d.Record after 30d after inoculation and manage growing state, root of hair position and number of taking root everywhere, each index and calculation formula as follows:
Namely rooting rate=strain number of taking root/inoculation strain number × 100%(root breakthrough epidermis is counted and is taken root)
Mean elements=total radical/strain number of taking root
Average root is long=overall length/total radical of all
The rooting efficiency of different root media is as shown in table 2, result shows: in the different culture media adding identical plant-growth regulator, the rooting efficiency comparing minimum medium is known, and MV substratum root induction rate is the highest, in first group of process, rooting rate reaches 63.3%, in second group of process, reach 73.3%, and number of on average taking root is up to 2.2, root length is the longest is 2.4, the plant be seeded on this minimum medium is taken out high fast, and leaf leaf look all normal, and root system is more flourishing; And 1/2MS substratum is least ideal culture medium, take root number and rooting rate are all minimum, and the highest rooting rate is only 43.3%, number of on average taking root is up to 0.8, root long the longest be 1.3cm, the plant strain growth be seeded on this kind of minimum medium is slow, plant agingly compares other process and wants soon.
Duncan's new multiple range method is adopted to carry out significance of difference test to three kinds of substratum root induction effects, the results are shown in Table rooting rate one row in 2, it can thus be appreciated that: the Rooting effect of dissimilar minimum medium to Flower of Japanese Camellia tissue cultured seedling is larger, inoculate number identical identical with concentration with the kind of plant-growth regulator time, the rooting rate of three kinds of minimum mediums when first group of process has notable difference, significant difference (P<0.05) between process C2 and C3, between process C1 and C3, difference is not remarkable.
The Rooting effect of table 2 different minimum medium induction Flower of Japanese Camellia tissue cultured seedling
Note: show significant difference (P<0.05) with lowercase alphabet different after column data
Therefore, for Flower of Japanese Camellia tissue cultured seedling root media, the minimum medium that the present invention selects is MV substratum.
2. the screening of plant hormone combination
Plant hormone conventional in plant tissue culture has tethelin and mitogen, and the plant hormone wherein for taking root has common IAA, IBA, NAA tri-kinds, and their kind, concentration and ratio are taken root to tissue cultured seedling and played important regulating effect.Using Flower of Japanese Camellia tissue cultured seedling as test materials, the budling of being cut into 2 ~ 3cm is inoculated into root induction in different root medias; Different root medias all with MV substratum for minimum medium, and be added with white sugar 20g/L, carrageenin 6.0g/L, gac 100mg/L, then add IBA, IAA and NAA of different concns and proportioning.Because of tethelin kind, concentration difference, the rooting rate of different root medias is also variant, the results are shown in Table 3.
The Rooting effect of table 3 different plant-growth regulator induction Flower of Japanese Camellia tissue cultured seedling
Note: show significant difference (P<0.05) with lowercase alphabet different after column data
As shown in Table 3, IBA and NAA process can improve the rooting rate of Flower of Japanese Camellia tissue cultured seedling significantly, and when being used alone, 2.0mg/L IBA induces the rooting rate of Flower of Japanese Camellia to reach 63.3%, the rooting rate that 2.0mg/L NAA induces is then 33.3%, therefore when hormone is used alone, IBA treatment effect is all better than the treatment effect of NAA.But add the IAA of 0.1mg/L on the basis of NAA and IBA after, its rooting rate is all significantly improved, and reaches 46.7% and 70% respectively.
In the process adding NAA, Flower of Japanese Camellia tissue cultured seedling base portion easily produces callus, and callus block is comparatively large, and root mainly has the callus induction of plant base portion to produce, and rooting rate is up to 60%, and root is short and thick, has fibrous root.And the plant base portion of IBA process not easily produces callus, root is mainly directly taken root by the base portion of tissue cultured seedling, and root is elongated need not root, rooting rate reaches 70%.When IBA and NAA concentration is 1.0mg/L, after adding 0.1mg/L IAA, rooting rate reaches 60%, therefore adds a small amount of IAA in each process and be conducive to the induction that Flower of Japanese Camellia tissue cultured seedling takes root, and its rooting rate significantly improves.
The change of each plant growth hormones concentration has a significant effect to Flower of Japanese Camellia tissue cultured seedling rooting efficiency tool, and when IBA is reduced to 1.0mg/L by concentration 2.0mg/L, rooting rate reduces; And the concentration of NAA is when being reduced to 1.0mg/L by 2.0mg/L, rooting rate rises on the contrary to some extent, but the growing state of rooting rate and root is widely different, visible according to the significance of difference analysis of rooting rate after statistics, significant difference (P<0.05) between process C8 and C9, and between other process, difference is not remarkable.
Therefore, the best plant growth conditioning agent that Flower of Japanese Camellia tissue cultured seedling is taken root is induced to be combined as IBA2.0mg/L+IAA0.1mg/L+ gac 100mg/L.
In sum, the substratum that the most applicable Flower of Japanese Camellia tissue cultured seedling that the present invention finally obtains is taken root is MV+IBA2.0mg/L+IAA0.1mg/L+ gac 100mg/L, rooting rate reaches as high as 73.3%, mean elements 2.2, average root is long is 2.4cm, and plant is taken out high fast, and leaf leaf look all normal, root system is more flourishing, as shown in Figure 1.
What finally illustrate is, above preferred embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although by above preferred embodiment to invention has been detailed description, but those skilled in the art are to be understood that, various change can be made to it in the form and details, and not depart from claims of the present invention limited range.
Claims (2)
1. Flower of Japanese Camellia tissue cultured seedling root media, is characterized in that: be made up of the component of following concentration:
2. Flower of Japanese Camellia tissue cultured seedling root media according to claim 1, is characterized in that: the pH value of described root media is 5.8.
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| CN105941159A (en) * | 2016-06-17 | 2016-09-21 | 句容市下蜀窑业茶场 | Seedling strengthening and rooting culture medium for tea tree tissue culture seedlings |
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| CN107135948B (en) * | 2017-06-02 | 2019-07-12 | 广西壮族自治区南宁良凤江国家森林公园 | A kind of method that Camellia nitidissima spray cultivates tissue-cultured seedling under sunlight conditions |
| CN108464243B (en) * | 2018-06-29 | 2021-06-22 | 钦州市农业科学研究所 | Culture medium for tissue culture and rapid propagation of curcuma zedoary |
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| JP4661178B2 (en) * | 2004-11-17 | 2011-03-30 | 日本製紙株式会社 | Plant production method |
| CN100482067C (en) * | 2006-12-18 | 2009-04-29 | 江苏阳光生态农林开发股份有限公司 | Fast tissue culture propagation process for azalea and camellia |
| CN101558742B (en) * | 2009-05-12 | 2012-11-21 | 中国林业科学研究院亚热带林业研究所 | Method for regenerating plant from camellia callus |
| CN103081807B (en) * | 2013-02-01 | 2014-08-20 | 中国林业科学研究院亚热带林业研究所 | Method for regenerating plant by use of callus of camellia japonica |
| CN103329799B (en) * | 2013-06-08 | 2015-01-07 | 上海植物园 | Rapid propagation and seedling growing method for Shuhua camellia tissue culture |
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