KR20150001056A - Method of propagating Paulownia coreana using root cutting - Google Patents

Abstract

The present invention relates to a propagation method of Paulownia coreana by root cuttings, comprising the steps of taking the root of an example of Paulownia coreana in mid-October and wet-storing the root in a refrigerator whose temperature is between 2 and 4 degree Celsius; and producing young Paulownia coreana by transplanting the stored root in the bed soil with sand in the spring of next year.

Description

[0001] The present invention relates to a method for propagating Paulownia coreana using root cutting,

The present invention relates to a propagation method of paulownia seedlings using root cutting, and more particularly to a vegetative propagation method for regenerating a plant by a root cutting method using a root of a dormant root.

The results of the microbial propagation of pheasant, which was published at home and abroad, are as follows.

So Woong (1976) showed that the root of the paulownia can be divided into three kinds of lengths and treated with two types of BA, Observing the formation process of stem through adventitia, we observed that cell division occurs mainly in the phloem parenchyma, and that malformed stem develops in its cleavage tissue.

Jagannathan L. and Marcotrigiano M (1986) is Paulownia The hypocotyl of tomentosa was cut into sections and the plants were regenerated. The morphological variation and ploidy level of the regenerated plants were examined and found to be early flowering in some individuals.

Rao et al. (1993) Paulownia In the fortunei , multi - stem was induced with the petiole and leaves of the meal in vitro. Optimal conditions were 4NAA and 20BA in MS medium. Seven days after cultivation, adventitia occurred mainly at the cut end of the petiole, and buds were induced in more than 80% of the cuts. Every 2 weeks, subculture with new medium promoted stem growth. 43 stems per section were obtained and successfully rooted and successfully transplanted into peat mixed soil.

Bergmann and Moon (1997) examined the regeneration of plant through adventitious induction of three kinds of Tungus species as petals and leaves. 10 clones. The optimal medium was the MS medium supplemented with 0.2 ~ 0. mg / L NAA and 5.0 ~ 7.0 mg / L BA.

Ipekci and Gozukirmizi (2004) is Paulownia In elongata , various types of growth embryogenesis were examined by cutting leaf and interlobular tissue. In MS medium, 54.1% calli were induced by TDZ and KN treatment, and 50.7 somatic embryos were averaged in 100mg embryogenic tissue. Somatic embryos were 80% germinated in MS basal medium and developed into plants. Plants were transferred to a mixture of peat and perlite in a 3: 1 mixture and survived 70-80%.

Corredoira et al. (2008) Paulownia Using two types of sections of a mature tomentosa list of materials were tested for plant regeneration through adventitious buds induced by TDZ and IAA treatment. The frequency of regeneration was 85 ~ 87% and the number of stem was 17.6 per section. There was a reaction difference according to the genotype, and the stem was able to root at 90% by IBA treatment for one week, and was successfully activated after 8 weeks in the greenhouse.

As shown in the above example, the tung trees show that effective breeding is possible through in-flight culture. However, Korean paulownia ( Paulownia coreana ) and especially the silkworm breeding method using the dormant root has not yet been developed. Among the micro-breeding techniques, the technique using the bud tissue such as the axillary bud can be an effective means of mass propagation because the mutation of the regenerated plant is small, and the tree with the preserved value Which is advantageous for application to the proliferation preservation of the plant.

[references]

All rights reserved. 1978. A Study on the Origin of the Differentiation of Pseudomonas spp. Korean Journal of Plant Tissue Culture 5 (1): 25-30

Soo Woong Young. 1976. A Study on the Differentiation and Root Propagation from Tungus persicae. Korean Journal of Plant Tissue Culture 4 (1): 4-8

Bergmann BA and Moon HK. 1997. In vitro adventitious shoot production in Paulownia . Plant Cell Rep. 16: 315-319

Corredoira E., Ballester A., Vieitez AM. 2008. Thidiazuron-induced high frequency plant regeneration from leaf explants of Paulownia tomentosa mature trees. Plant Cell Tiss Org Cult. 95: 197-208

Jagannathan L., Marcotrigiano M. 1986. Phenotype and ploidy status of Paulownia tomentosa trees regenerated from cultured hypocotyls. Plant Cell Tiss Org Cult 7: 227-236

Ipekcie and Gozukirmizi N. 2004. Indirect somatic embryogenesis and plant regeneration from leaf and internode ezplants of Paulownia elongata Plant Cell Tiss Org Cult. 79: 341-345

Lloyd G and McCown BH. 1980. Commercially feasible micropropagation of mountain laurel, Kalmia latifolia by use of shoot type culture. Proc Intl Plant Pro Soc 30: 421-427

Murashige T and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-497

Rao CD, Goh CJ and Kumar PP. 1993. High frequency plant regeneration from excised leaves of Paulownia Fortunei . In Vitro Cell Dev Biol 29P: 72-76

Until now, there is no previous research on microbial propagation and propagation technique of 'Seongamdong pongo tree'. In the present invention, conditions for optimizing culture conditions such as selection of medium for efficient micro breeding were investigated and the possibility of root cutting was investigated. Therefore, we studied the surface sterilization method for microbial culture, selection of medium for induction and proliferation of stem from embryo, growth condition control, rooting induction and soil transplantation. The following are the issues to be solved.

① Surface sterilization condition

② Stem induction and proliferation

③ Root induction

④ Purification of soil transplantation

⑤ Root close-up plant induction

In order to solve the above-mentioned problems, the present invention provides a method for sterilizing a stem of a paulownia coreana ; Culturing the surface-sterilized slice to induce stem; Growing said stem; Inducing rooting in the proliferated stem; And a step of purifying the root-inducing individual by transplanting the plant into the culture soil.

The present invention also relates to a method for the production of paulownia coreana ) in the middle of October and stored in a refrigerator maintained at 2-4 ° C; And a step of cutting the stored roots in the sand in the spring of the following year to produce seedlings ( Paulownia < RTI ID = 0.0 > coreana ) breeding methods.

The propagation method according to the present invention can be applied to the propagation of various kinds of hardwood species in a variety of species, and in particular, the young tissue of the aged tree is propagated through a rejuvenation process There are advantages to be able to.

In addition, seedlings such as paulownia trees in the seongnam-myeon can be planted by cutting the roots in a short-cut manner by using the root-cutting method, which is advantageous for the preservation and restoration of the native species with historical characteristics.

Figure 1 is a stem derived from an embryo via primary culture. After the primary culture, uncontaminated sections were used for proliferation.
Fig. 2 shows growth of the multipotent stem derived from the growth medium. It was possible to multiply 3 to 6 times per month under appropriate conditions.
Fig. 3 is an in-flight seedlings undergoing mass propagation in a glass culture bottle. In-flight propagation of paulownia seedlings was effective in the culture bottle of the present invention.
Fig. 4 is a root shape of an individual rooted in a cabin. Within two weeks after rooting, the plants were transferred to the artificial soil to purify the plants.
Fig. 5 is a seedlings planted in an artificial soil and purified. Maintaining high air humidity for the first week after transplantation was a significant factor in survival.
Fig. 6 is a seedlings grown for 3 weeks after purification. The purified saplings showed that the leaves grew rapidly and rapidly.
Fig. 7 is a seedlings growing in a large flower pot. It grows more than 30cm in one month.
Fig. 8 is a stem formed by roots close-fitting in sand. A number of stems were produced in one root, and after 4 to 6 weeks, seedlings were grown through abandonment.

The present invention relates to (a) Paulownia steps to sterilize the surface intercept of stem aekah coreana); (b) primary culturing said surface-sterilized section to induce stem; (c) propagating the induced stem; (d) inducing rooting in the proliferated stem; And (e) transplanting the root-inducing individual into a culture soil to purify the pungent tree.

In the present invention, the surface disinfection is performed by washing the sections with tap water, adding a few drops of Tween 20, bubbling and washing, treating with 70% ethanol (EtOH) for 30 seconds to 1 minute on a clean bench, After treatment with sodium hypochlorite (NaClO) for 8 minutes to 15 minutes, it is washed with sterilized distilled water four times or more, and then, it is wound on a medium.

In the present invention, in the step (b), the surface-sterilized slice is cut with a dissecting knife so as to adhere the grains one by one, and then the WPM base medium is coated with 2.0% sucrose and 0.2% gellan gum in a solid medium Thereby inducing the stem.

In the present invention, step (c) is characterized in that the stem is proliferated in a medium supplemented with 0.2-1.0 mg / L of benzyladenine (BA), 2.0% sucrose and 0.2% gellan gum in a WPM basic medium.

In the present invention, step (d) is characterized by inducing rooting by treating 0.2 to 0.5 mg / L indolebutyric acid (IBA) in a 1 / 2WPM basic medium or a 1 / 2WPM basic medium with a stem grown over 2 cm .

In the present invention, the step (e) is characterized in that the rooted young plants are transplanted into artificial soil for horticulture, and the soil is activated by keeping the air humidity at 90% or more for 1 to 2 weeks.

In the present invention, the artificial plant is characterized in that the volume ratio of peatmos, perlite, and vermiculite is 1: 1: 1.

In the present invention, the paulownia tree may be Sung Sam Mun Paulownia coreana .

The present invention also relates to a method for the production of paulownia coreana ) in the middle of October and stored in a refrigerator maintained at 2-4 ° C; And a step of cutting the stored roots in the sand in the spring of the following year to produce seedlings ( Paulownia < RTI ID = 0.0 > coreana ) .

According to the present invention, the seedlings are produced by cultivating roots sculpted in sandy soil in a purifying greenhouse maintained at 20 to 25 ° C.

In the present invention, the paulownia tree may be Sung Sam Mun Paulownia coreana .

Hereinafter, the present invention will be described in detail.

As an example of the present invention, the optimum conditions for producing the seedlings using the seedling cultivation method and root cutting method are as follows.

① Surface disinfection

Add a few drops of tween 20 to the explants, wash with tap water, sterilize in 70% ethanol (EtOH) for 30 seconds to 1 minute, add 2% sodium hypochlorite (NaClO) for 8-15 minutes It is possible to remove more than 90% of contamination after washing with sterilized distilled water more than 4 times.

② Primary culture Stem induction

The surface-sterilized slices were cut with a dissecting knife so that each of the slices was attached one by one, and then streaked on a WPM (Llyod and McCown 1981) medium in a medium solidified with 2.0% sucrose and 0.2% Occurrence occurs.

③ shoot multiplication

When multiparous stems were induced with 0.2 ~ 1.0 ㎎ / L BA, 2.0% sucrose and 0.2% gellan gum on the WPM medium using the stem obtained from the primary culture, 3 ~ 6 multiparous stems , And the multipotent stem is grown as a single dominant trunk when subcultured with a basic medium in which no growth hormone is added.

④ root induction

Growth of more than 2 cm in the in vitro proliferated stem was induced by root induction by treatment with 0.2 ~ 0.5 mg / L IBA in 1 / 2WPM primary medium or 1 / 2WPM primary medium. More than 98% of the rooting occurs.

⑤ Soil transplantation (acclimatization)

The young plants roosted in the cabin are removed from the roots by tap water and transferred to a mixture of peatmoss, perlite and vermiculite, and are grown in a purifying greenhouse maintained at a temperature of 20 to 25 ° C with a relative humidity of 90% When incubated, it activates more than 94%.

⑥ Root cutting

Roots of paulownia seedlings were sown at the end of October and stored in a refrigerator maintained at 2-4 ℃ for about 4 months. Then, in the following spring and March, When cultivated in a purifying greenhouse maintained at 20 ~ 25 ℃, more than 98% of stems were produced and seedling production was possible.

Hereinafter, the present invention will be described with reference to specific examples. However, the scope of rights of cultivation and rooting of a paulownia species similar to that of the present invention is not limited to the embodiments described below.

Example 1 Surface sterilization

Seedlings of paulownia thunbergii were collected from April to May and stored in a refrigerator maintained at 4 ℃ for surface sterilization.

The sections were prepared so that the leaves were removed and 2 ~ 4 pieces of each animal were adhered. A few drops of tween 20 were added as a surfactant, and the pieces were washed with tap water several times by shaking and bubbling.

After that, the cells were sterilized in 70% ethanol for 30 seconds to 1 minute and 2% sodium hypochlorite (NaClO) for 5 to 8 minutes on a clean bench, and then washed and washed 5 times or more with sterilized distilled water.

≪ Example 2 > Stem induction and proliferation

The medium was cured by 2% sucrose treatment and 0.3% phytagel in MS (Murashige and Skoog, 1962) and WPM (Lloyd and McCown, 1981) basal media and placed in a 15 × 2.5 cm glass test tube with 8 ml of medium Lt; / RTI >

The stems were grown by adding 0.1-0.1 mg / L BA, 3% sucrose, and 0.3% phytagel medium to 13x7.5 cm glass culture bottles into WPM medium using the in vitro stem derived from the primary culture.

The incubation was carried out in a culture room maintained at a temperature of 24 ± 2 ° C and a light intensity of 40 μmol m ^-2 s ^-1 for 16 hours per day with illumination of a cool white fluorescent lamp. The cells were grown in fresh media every 3 to 4 weeks.

The following culture conditions were carried out in the same manner as the conditions of this Example.

<Example 3> Root induction

(2% sucrose, 0.7% agar) in 0.5% to 1.0 mg / L IBA, 2% sucrose, 0.7% agar or 1 / 2WPM medium supplemented with 1 / Respectively.

Rooting was observed after 1 week, and the rooted individuals were transferred to the artificial soil after 3 to 4 weeks of cultivation to purify them.

&Lt; Example 4 > Purification of soil transplantation)

The rooted individuals were carefully taken out, the baby was removed with tap water, the peatmoss perlite and vermiculite were transplanted into the culture soil mixed with the equivalent volume ratio (1: 1: 1 v / v / v) Or more.

Ventilated once or twice a day, irrigated periodically to avoid drying, and the purified seedlings were transferred to larger vessels and cultivated.

&Lt; Example 5 >

At the end of October, the root of the paulownia seedlings was partially shredded and mixed with wet sand so as not to dry, and then stored in a refrigerator maintained at 2 to 4 ° C.

The root of 0.5cm in diameter stored in the end of March the following year was cut into 10cm and the horizontal axis was cut horizontally from the greenhouse into the upper soil. After cutting, watering was done and watering was periodically performed so that the cuttings did not dry out.

After 3 weeks, shoots rose and 4-6 weeks later seedlings were grown in the form of stem breaks.

The propagation method according to the present invention can be applied to the propagation of various kinds of hardwood species in a variety of species, and in particular, the young tissue of the aged tree is propagated through a rejuvenation process There are advantages to be able to.

In addition, seedlings such as paulownia trees in the seongnam-myeon can be planted by cutting the roots in a short-cut manner by using the root-cutting method, which is advantageous for the preservation and restoration of the native species with historical characteristics.

Claims (5)

Paulownia coreana ) in the middle of October and stored in a refrigerator maintained at 2-4 ° C; And
And the step of cutting the stored roots in the sand in the spring of the following year to produce seedlings ( Paulownia &lt; RTI ID = 0.0 &gt; Coreana ) breeding methods. [2] The method according to claim 1, wherein the seedlings are produced by cultivating roots sculpted in sandy soil in a purifying greenhouse maintained at 20 ~ 25 [deg.] C. 3. The method according to claim 1 or 2, wherein the paulownia tree is Sung Sam Mun Paulownia coreana . A paulownia plant propagated by the method of claim 1 or 2 coreana ) . The paulownia as set forth in claim 4, wherein the paulownia tree is Sung Sam Mun Paulownia coreana .

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