CN108739321B

CN108739321B - Wild dendrobium nobile breeding method

Info

Publication number: CN108739321B
Application number: CN201810519586.0A
Authority: CN
Inventors: 尚迪; 刘玉珊; 孙婷; 卓明; 许震寰
Original assignee: SICHUAN ACADEMY OF BOTANICAL ENGINEERING
Current assignee: SICHUAN ACADEMY OF BOTANICAL ENGINEERING
Priority date: 2018-05-28
Filing date: 2018-05-28
Publication date: 2020-11-24
Anticipated expiration: 2038-05-28
Also published as: CN108739321A

Abstract

The invention provides a wild dendrobium propagation method, which comprises the steps of selecting a seedling bed convenient for draining, laying a sterilized fermentation degreased bark layer on the seedling bed, and arranging a sterilized first moisture-preserving layer on the fermentation degreased bark layer; cutting the wild dendrobium stem into sections, and drying in the shade until the wound is dry; flatly paving the wild dendrobium stem sections dried in the shade on the first moisturizing layer, covering a second moisturizing layer above the wild dendrobium stem sections, and starting to cover buds; after covering buds for 20-30 days, axillary buds are selected for direct transplantation. The breeding mode of the invention has low cost and simple operation. By adopting the propagation mode of the invention, the plants with necrotic roots or pathological changes of stems can be used for continuous propagation. Compared with tissue culture and rapid propagation, the method disclosed by the invention can cover buds to propagate axillary buds to germinate stout seedlings, can directly transplant the axillary buds, does not need seedling hardening, and is simple and low in cost. The survival rate after transplanting is high, the endangered wild dendrobium resources can be effectively protected, and the population quantity of the wild dendrobium resources is enlarged.

Description

Wild dendrobium nobile breeding method

Technical Field

The invention belongs to the field of plant propagation, relates to a dendrobium propagation technology, and particularly relates to a propagation method of wild dendrobium.

Background

The dendrobe resource belongs to the I-grade protective plant in China, belongs to the endangered plant resource, has less resources and low natural reproduction rate, is easy to cause virus diseases in domesticated cultivation, leads to plant death and causes resource material loss. 76 dendrobium resources exist in China, and only a few dendrobium varieties, such as dendrobium officinale, dendrobium nobile, dendrobium moniliforme and the like, are propagated in an expanded mode through an aseptic tissue culture method at present.

In the process of implementing the invention, the inventor finds that at least the following problems exist in the prior art:

1. the tissue culture propagation of the dendrobium needs corresponding facilities and aseptic system operation, although the seedling emergence amount is large, seedlings are weak, diseases are easy to occur after direct transplantation after seedling emergence, the death rate is high, and the survival rate is low;

2. strong seedlings are optimally cultured by a seedling hardening technology after the seedlings of the bottle seedlings are propagated through tissue culture and propagation and before field planting, the period is long, and the cost is high.

Disclosure of Invention

In view of the above, the invention aims to provide a wild dendrobium propagation method with high survival rate, short propagation period and low cost.

The inventor continuously reforms and innovates through long-term exploration and attempt, and multiple experiments and endeavors, and in order to solve the technical problems, the technical scheme provided by the invention is to provide a wild dendrobium propagation method, which comprises the following steps:

step a) the whole bed

Selecting a seedling bed convenient for draining, laying a sterilized fermentation degreased bark layer on the seedling bed, and arranging a sterilized first moisture-preserving layer on the fermentation degreased bark layer;

step b) cutting the stem

Cutting the wild dendrobium stem into sections, and drying in the shade until the wound is dry;

step c) covering buds

Flatly paving the wild dendrobium stem sections dried in the shade on the first moisturizing layer, covering a second moisturizing layer above the wild dendrobium stem sections, and starting to cover buds;

step d) transplanting

After covering buds for 20-30 days, axillary buds are selected for direct transplantation.

According to a preferred embodiment of the method for propagating wild dendrobium, in the step a), the diameter of the block of the fermented defatted bark is 8-18 mm.

According to a preferred embodiment of the wild dendrobium propagation method, the first moisture-retaining layer is laid by dried sphagnum or sphagnum, and the second moisture-retaining layer is laid by dried sphagnum or sphagnum.

According to a preferred embodiment of the wild dendrobium propagation method, the thickness of the first moisture-preserving layer is 3cm, and the thickness of the second moisture-preserving layer is 2 cm.

According to a preferred embodiment of the method for breeding wild dendrobium, in the step b), the upright stem of the healthy plant is cut into segments with the length of 10-15cm, and the resource variety with small plant type can be cut into segments with the length of 5cm or directly used, and dried in the shade for 1-2 days.

According to a preferred embodiment of the method for breeding wild dendrobium of the present invention, in the step b), the plants with necrotic roots or diseased stems are cut off the diseased parts, and then the stems are soaked for sterilization and dried in the shade for 2-3 days.

According to a preferred embodiment of the method for propagating wild dendrobium, in the step c), the bud-covering temperature is 28-35 ℃, and the bud-covering humidity is 80-90%. The bud-covering temperature is any one of 28 deg.C to 35 deg.C, including but not limited to 28 deg.C, 29 deg.C, 30 deg.C, 31 deg.C, 32 deg.C, 33 deg.C, 34 deg.C or 35 deg.C, preferably 30 deg.C. The humidity for covering buds is 80-90%, which refers to any humidity in the interval, including but not limited to 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, and preferably the humidity for covering buds is 85%.

According to a preferred embodiment of the method for propagating wild dendrobium, in the step c), a rooting agent is sprayed after 5 days of bud covering.

According to a preferred embodiment of the wild dendrobium propagation method, the sterilization is performed by soaking in a bactericide.

According to a preferred embodiment of the method for propagating wild dendrobium, the wild dendrobium includes dendrobium nobile lindl, dendrobium candidum, dendrobium glabratum, dendrobium candidum and dendrobium candidum.

According to a preferred embodiment of the propagation method of wild dendrobium of the present invention, the wild dendrobium is dendrobium nobile lindl, dendrobium nobile lindl or dendrobium roseum.

Compared with the prior art, one of the technical solutions has the following advantages:

a) the breeding mode of the invention has low cost and simple operation.

b) By adopting the propagation mode of the invention, the plants with necrotic roots or pathological changes of stems can be used for continuous propagation.

c) Compared with tissue culture and rapid propagation, the method disclosed by the invention can cover buds to propagate axillary buds to germinate stout seedlings, can directly transplant the axillary buds, does not need seedling hardening, and is simple and low in cost. The survival rate after transplanting is high, the endangered wild dendrobium resources can be effectively protected, and the population quantity of the wild dendrobium resources is enlarged.

d) In one embodiment of the propagation method, the stem sections of the dendrobium are dried in the shade until the wounds are dry, which is beneficial to avoiding the infection diseases of the wounds. Because the environment temperature for bud covering propagation is higher and the humidity is higher, the wound needs to be dried in the shade, so that the wound infection diseases are avoided.

e) In one embodiment of the propagation method, plants with necrotic roots or diseased stems need to be dried in the shade for 2-3 days after sterilization, so that late-stage infection diseases can be effectively reduced.

f) In one embodiment of the propagation method, the dry water moss is selected as the moisturizing material, has good moisturizing performance, long moisturizing time and good air permeability, and can maintain the stem segments of the dendrobium in a high-humidity state and promote axillary bud differentiation.

g) In one embodiment of the propagation method, the thickness of the moisturizing layer is limited, and if the thickness is too low, the substrate is easy to dry and not beneficial to axillary bud catalysis, and the moisture content of the substrate is not enough; if the thickness of the moisture retention layer is too high, the humidity is too high, and diseases are easily caused.

h) The germination rate of dendrobium nobile lindl, dendrobium glabrous, dendrobium labdanum and dendrobium roseum which are propagated by covering buds reaches more than 90 percent, and the germination rate of dendrobium candidum, dendrobium candidum and dendrobium nobile reaches more than 85 percent.

i) The survival rate of dendrobium nobile lindl, dendrobium glabrata, dendrobium labdanum and dendrobium roseum which are propagated by bud covering reaches more than 90 percent; the survival rate of the dendrobium candidum, the dendrobium officinale, the dendrobium moniliforme and the dendrobium poecilomycosum reaches more than 88 percent.

Detailed Description

The following description will be given with reference to specific examples.

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the embodiments of the present invention will be described in detail and completely with reference to the accompanying drawings. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Thus, the following detailed description of embodiments of the invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention.

A frame with the height of 50cm away from the ground is arranged in the greenhouse, so that water draining and daily management are facilitated, the seedling flat plate is horizontally placed on the frame, and the seedling bed is built. The seedling flat plate is provided with water leakage holes, so that redundant water can be conveniently drained.

Soaking the fermented defatted bark in 500 times of 50% carbendazim wettable powder for 3 hr for sterilization, and spreading on seedling plate. The diameter of the fermented defatted bark is 8-18 mm. The untreated bark contains grease, harmful bacteria and the like, the fermentation can remove the harmful bacteria and eggs, the degreasing mainly adopts high-temperature high-pressure cooking to remove lipid substances which are not removed in the fermentation and are not beneficial to plant growth, and simultaneously, the water retention and the fertility of the bark are enhanced. The bark after degreasing and fermentation is not easy to rot and acidify, the air permeability is good, and the bark substrate which is not subjected to degreasing and fermentation treatment is easy to cause diseases such as plant soft rot.

Soaking dried sphagna or dried moss in 500 times of 50% carbendazim wettable powder for 1 hr, sterilizing, spreading on fermented defatted bark, and making into first moisture-retaining layer. The dry moss or moss refers to the material used for the moisture-retaining layer, and is not the real state in the sprouting process. In the bud covering process, water is sprayed to the dry sphagnum or the dry sphagnum to keep the humidity of the dry sphagnum or the dry sphagnum. In this example, dry moss was used as the first moisture-retaining layer material. The dry water moss has good moisture retention, long moisture retention time and good air permeability, and can maintain the high-moisture state of the upright stem and promote the differentiation of axillary buds. The thickness of the dried sphagnum or sphagnum laid on the fermented defatted bark is preferably 3 cm. If the thickness of the dried sphagnum or the dried sphagnum is less than 3cm, the moisture of the moisturizing material is insufficient, the dried sphagnum or the dried sphagnum is easy to dry, and the catalysis of axillary buds is not facilitated; if the thickness is higher than 3cm, the humidity in the moisturizing material is very high, and diseases are easily caused. The dried moss refers to a dried moss which has lost physiological functions and is used only as a moisturizing material in the present example. The dry moss refers to moss which is dried and loses physiological functions, and is used as a moisturizing material in the embodiment of the invention.

Cutting the wild dendrobium stem into segments of 10-15cm, wherein the cut stem needs no diseases. The resource variety with small plant type can be cut into about 5cm or directly used as the upright stem. And drying the cut upright stem segments of the wild dendrobium in the shade for 1-2 days to dry the cut wounds. Because the environment temperature for bud covering propagation is higher and the humidity is higher, the wound needs to be dried in the shade, so that the wound infection diseases are avoided.

If the material comprises plants with necrotic roots or diseased stems, the stem sections with the diseased parts cut off are soaked in 500 times of 70 percent thiophanate methyl powder for sterilization, and the stem sections are dried in the shade for 2 to 3 days after sterilization so as to reduce later-stage infection diseases.

Spreading the wild herba Dendrobii stem section after drying in the shade on the first moisture-preserving layer, and covering a second moisture-preserving layer with thickness of 1-2cm on the wild herba Dendrobii stem section. The second moisture-preserving layer and the first moisture-preserving layer are made of the same materials and have the same sterilization mode. Spraying water to the moisture retention layer to keep the moisture retention layer moist, and starting to cover the buds. The temperature for covering buds is 28-35 ℃, and the humidity for covering buds is 80-90%. The bud-covering temperature is any one of 28 deg.C to 35 deg.C, including but not limited to 28 deg.C, 29 deg.C, 30 deg.C, 31 deg.C, 32 deg.C, 33 deg.C, 34 deg.C or 35 deg.C, in this embodiment, the bud-covering temperature is 30 deg.C. The humidity for covering buds is 80-90%, which refers to any humidity in the interval, including but not limited to 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, and the humidity for covering buds is 85% in this embodiment. The embodiment is to cover buds in a greenhouse shed, and the temperature and humidity control of the matrix has good facility guarantee.

And 5 days after the bud covering starts, spraying a rooting agent, wherein the main components of the rooting agent comprise naphthylacetic acid, sodium indolebutyrate, trace elements and the like. The method for spraying the rooting agent is to add 400ml of water to 1g of the rooting agent. The rooting agent belongs to a commercial product, and related commodities can be inquired by using main components on a Chinese pesticide information network and related websites.

After the buds are covered for 20-30 days, axillary buds can be selected for direct transplanting without hardening seedlings.

The breeding mode of the invention has low cost and simple operation, and can also utilize plants with necrotic roots or pathological changes of stems to continue breeding. Compared with tissue culture and rapid propagation, the method has the advantages of lower cost of the bud-sealing propagation mode, simplicity, no need of hardening seedlings, stout axillary bud seedlings, direct transplantation, high survival rate after transplantation, effective protection of endangered wild dendrobium resources and expansion of the population quantity of the wild dendrobium resources.

To further illustrate the technical effects of the present invention, the following description is made with reference to comparative test data.

Test one: and (4) transplanting survival rate tests of different seedling exercising methods.

Four seedling exercising treatments are designed as controls by taking dendrobium nobile tissue culture seedlings as test materials. In the following treatments, the first to fourth treatments were control groups, and the fifth treatment was an experimental group.

Processing one: and (5) taking out to a greenhouse 17 days before transplanting, sealing bottles, hardening seedlings for 15 days, and then opening bottles and hardening seedlings for 2 days.

And (5) processing: directly moving the seedlings to a greenhouse from a tissue culture room before transplanting, and directly transplanting the seedlings without greenhouse seedling hardening.

And (3) treatment III: after inoculation, the seedlings grow in a tissue culture room for 40 days, are transferred to a greenhouse for 30 days, and then are taken out of bottles for transplantation.

And (4) treatment: after inoculation, the seedlings grow in a tissue culture room for 30 days, are transferred to a greenhouse for 40 days, and then are taken out of bottles for transplantation.

And (5) processing: the dendrobium nobile lindl is used as a test material, and axillary buds which are propagated by bud covering are directly transplanted by adopting the embodiment of the invention.

Test survey items and survey recording standards:

a, survival rate:

the number of surviving seedlings in the whole cell is recorded every 5 days and 8 times of continuous observation, and all the plants which are not completely dead are regarded as the surviving seedlings, but the seedlings whose stem base part is withered or died but the upper part is not dead are regarded as dead seedlings.

b, character investigation:

and 5, surveying after transplanting, randomly measuring the stem thickness of 10 plants in each cell, and taking the average value as a cell observation value. The experimental data are shown in table 1.

As can be seen from Table 1, the survival rate of the seedling hardening test after tissue culture is low, and the best treatment for the survival rate is only 46%. And the second step is to treat the second step, the second step is to directly take out from a tissue culture room for transplantation, and test-tube seedlings are difficult to adapt to environmental changes, are easy to generate soft rot and other diseases, and are extremely easy to dehydrate, wither and die. The third and fourth treatments are serious in pollution in the environment of a large greenhouse, and the temperature in the bottle is easy to rise under the condition that the bottle cover is sealed, so that the bottle seedlings are dehydrated and withered or the leaf tip parts are withered, and the resistance and the survival rate are greatly reduced. And fifthly, the survival rate of directly transplanted axillary buds propagated by the bud-covering method is highest.

Table 1: different seedling hardening methods and transplanting survival rate of bud-covering propagation

Statistics of stem thickness data of different seedling exercising methods and sprouting propagation are shown in table 2.

As can be seen from Table 2, the stem diameter of the plant treated in one of the different seedling exercising manners is 2.13 cm. The stems of the axillary bud sprouting seedlings propagated by covering buds are 3.43cm thick, and the axillary bud sprouting seedlings obtained by covering buds are obviously stronger than tissue culture seedlings.

Table 2: different seedling hardening methods and stem thickness of bud-covering propagation

And (2) test II: germination rate of different varieties in germination-covering propagation.

The germination rate was counted by using the upright stems of 8 different dendrobium as the test material and the bud-covering propagation method of this example 1, and the statistical results are shown in table 3.

The statistical results in table 3 show that the vertical stems are adopted to cover the buds for propagation, the propagation amount is large, 2-5 axillary buds can be generated on each section of the vertical stems, and the axillary bud germination rate is high.

Wherein the germination rate of Dendrobium nobile, herba Pileae Scriptae, herba Botrychii and herba Dendrobii Rugosae bred by bud covering is more than 90%. The germination rate of the dendrobium candidum, the dendrobium officinale, the dendrobium moniliforme and the dendrobium poecilomikovii can reach more than 85 percent.

Table 3: germination rate

Variety of (IV) C	Number of erect stems	Number of upright stems of germinated axillary buds	Germination percentage (%)
				Dendrobium stem	72	70	97%
Herba Dendrobii	60	55	92%
				Herba Dendrobii	62	58	94%
Rose dendrobium stem	42	40	95%
				Dendrobium officinale Kimura et Migo	56	48	86%
Dendrobium officinale Kimura et Migo	38	33	87%
				Caulis Dendrobii	42	36	86%
Herba Dendrobii	68	60	88%

And (3) test III: survival rate of different varieties of bud-covering propagation.

The number of survived seedlings was recorded 30 days after transplanting by using the bud-covering propagation method of this example with the upright stems of 8 different dendrobium as test materials, and all plants that did not die completely were considered as survived seedlings, but seedlings whose stem bases died or died but whose upper parts did not die were considered as dead seedlings, and the statistical data is shown in table 4.

As can be seen from Table 4, the survival rate of dendrobium nobile lindl, dendrobium glabrata, dendrobium labdanum and dendrobium nobile lindl which are propagated by bud covering reaches over 90 percent; the survival rate of the dendrobium candidum, the dendrobium officinale, the dendrobium moniliforme and the dendrobium poecilomycosum reaches more than 88 percent.

Table 4: survival rate

Variety of (IV) C	Total amount of axillary buds sprouting	Number of surviving seedlings after transplantation	Survival rate
				Dendrobium stem	186	180	97%
Herba Dendrobii	111	104	94%
				Herba Dendrobii	120	112	93%
Rose dendrobium stem	88	80	91%
				Dendrobium officinale Kimura et Migo	138	122	88%
Dendrobium officinale Kimura et Migo	68	60	88%
				Caulis Dendrobii	78	70	89%
Herba Dendrobii	196	175	89%

The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.

Claims

1. A wild dendrobium propagation method, the wild dendrobium comprises any one of dendrobium nobile, dendrobium bibract, dendrobium roseum, dendrobium glabrous, dendrobium candidum and dendrobium candidum, and is characterized by comprising the following steps: step a) the whole bed

a) Selecting a seedling bed convenient for draining, laying a sterilized fermentation degreased bark layer on the seedling bed, and arranging a sterilized first moisture-preserving layer on the fermentation degreased bark layer; the first moisture-preserving layer is formed by paving dry sphagna or dry sphagna;

step b) cutting the stem

b) Cutting the wild dendrobium stem into sections, and drying in the shade until the wound is dry;

step c) covering buds

c) Flatly paving the wild dendrobium stem sections dried in the shade on the first moisturizing layer, covering a second moisturizing layer above the wild dendrobium stem sections, and starting to cover buds; the second moisture-keeping layer is formed by paving dry sphagna or dry sphagna;

step d) transplanting

2. The wild dendrobium propagation method of claim 1, wherein in the step a), the diameter of the block of the fermented defatted bark is 8-18 mm.

3. The wild dendrobium propagation method of claim 1, wherein the thickness of the first moisture retaining layer is 3cm, and the thickness of the second moisture retaining layer is 2 cm.

4. The wild dendrobium propagation method of claim 1, wherein in the step b), the erect stem of the healthy dendrobium is cut into segments with the length of 10-15cm, the resource variety with small plant type can be cut into segments with the length of 5cm or the erect stem can be directly used, and the wild dendrobium is dried in the shade for 1-2 days; the plant with necrotic root or diseased stem is prepared through cutting the diseased part, soaking stem, sterilizing and drying in the shade for 2-3 days.

5. The propagation method of wild dendrobium nobile lindl according to claim 1, wherein in the step c), the temperature for covering buds is 28-35 ℃, and the humidity for covering buds is 80-90%.

6. The propagation method of wild dendrobium nobile lindl as claimed in claim 1, wherein in the step c), a rooting agent is sprayed 5 days after covering buds.

7. The wild dendrobium propagation method of claim 1, wherein the sterilization is performed by soaking in a bactericide.

8. The wild dendrobium propagation method of claim 1, wherein the wild dendrobium is dendrobium nobile lindl, dendrobium nobile or dendrobium roseum.