CN114175934B

CN114175934B - Green seedling grafting method for schisandra chinensis

Info

Publication number: CN114175934B
Application number: CN202111666703.4A
Authority: CN
Inventors: 艾军; 石广丽; 樊跃; 孙丹; 王振兴; 王英平; 刘晓颖
Original assignee: Jilin Agricultural University
Current assignee: Jilin Agricultural University
Priority date: 2021-12-31
Filing date: 2021-12-31
Publication date: 2023-05-05
Anticipated expiration: 2041-12-31
Also published as: CN114175934A; KR20230103928A

Abstract

The invention discloses a green seedling grafting method of schisandra chinensis, and relates to the technical field of asexual propagation of schisandra chinensis. It comprises the following steps: the method is characterized in that the current annual schisandra chinensis green seedling is used as a stock, the current annual schisandra chinensis new shoot, the current annual schisandra chinensis auxiliary shoot or the tissue culture green shoot is used as a scion, and the scion is grafted in a notch at the hypocotyl of the stock in a hypocotyl split grafting mode, so that the notch is fixed. The green seedlings of the current annual shizandra berry are seedlings in the two-leaf one-heart stage or the age after the two-leaf one-heart stage. The grafting method provided by the invention can improve grafting efficiency, has high survival rate, avoids the generation of the underground stems of the stock, can effectively adjust the generation position of the underground stems of the scion variety, and is suitable for breeding high-quality schisandra chinensis asexual seedlings.

Description

Green seedling grafting method for schisandra chinensis

Technical Field

The invention relates to the technical field of asexual propagation of schisandra chinensis, in particular to a green seedling grafting method of schisandra chinensis.

Background

Fructus Schisandrae chinensis (Schisandra chinensis (Turcz.) is deciduous wood vine belonging to Schisandraceae, schisandra genus, and subgenera of Schisandra japonica, and its dried berry commodity is known as Schisandra chinensis, which is a large rare land Chinese medicinal material in China. In recent years, due to the increasing market demands and the increasing serious damage of wild resources, the artificial cultivation and development of the schisandra chinensis are rapid, and the development of the efficient asexual propagation technology of the schisandra chinensis is of great significance in promoting the improvement and development of the cultivation industry of the schisandra chinensis.

Besides normal branches and tendrils on the ground, the schisandra chinensis also has specialized underground stems which mainly occur in low-node buds at the root neck part, and besides the underground transverse extension of the underground stems, the underground stems also can emit a large number of tillering branches. The sprout branches have strong growth vigor, the adventitious root system of the underground stem is undeveloped, the absorption capacity is weak, a large amount of nutrition still needs to be supplied by a mother plant, and large nutrition competition can be caused for flowering and fruiting of the schisandra chinensis, normal plant growth and the like, so that the generation of the underground stem needs to be controlled when the schisandra chinensis is cultivated. The asexual propagation technology of the schisandra chinensis is beneficial to improving the propagation efficiency and controlling the underground stems.

At present, the asexual propagation of the schisandra chinensis mainly comprises a plurality of methods such as hard branch grafting, bud grafting, tissue culture and the like, but the propagation survival rate is low, the propagation technology is complex and is not easy to operate, the underground stems of the stock and the grafted varieties compete, the generation positions of the underground stems of the variety seedlings are too low and are not easy to control, the propagation period is too short and is too concentrated, and the like, which are always in different asexual propagation technologies, so that the application of the asexual propagation technology of the schisandra chinensis and the development of fine variety are restricted.

In view of this, the present invention has been made.

Disclosure of Invention

The invention aims to provide a green seedling grafting method for schisandra chinensis, which aims to solve the technical problems. The inventor discovers that the adoption of the method for grafting the green seedlings of the hypocotyls can simultaneously overcome a plurality of bottleneck problems of low propagation survival rate, complex propagation technology, difficult operation, competition between the underground stems of the stocks and grafted varieties, difficult control of the generation positions of the underground stems of the seedlings of the varieties, too short propagation period, too concentrated and the like.

The invention is realized in the following way:

the invention provides a green seedling grafting method of schisandra chinensis, which comprises the following steps: taking a green seedling of the current annual schisandra chinensis as a stock, taking a new shoot with leaves, a side shoot or a tissue culture green shoot of the current annual schisandra chinensis as a scion, and adopting a hypocotyl split grafting mode to enable the scion to be grafted in a notch at the hypocotyl of the stock, and fixing the notch;

the green seedlings of the current annual shizandra berry are seedlings in two-leaf one-heart stage or in the age after the two-leaf one-heart stage.

The two-leaf one-heart period refers to the period when the schisandra chinensis seedlings grow to two complete true leaves and one new leaf (not yet developed). Seedlings of the age after the two-leaf one-heart period include, but are not limited to: three leaves and one heart stage, four leaves and one heart stage. Seedlings of the age after the two-leaf and one-heart period can be grafted for a long time.

In the bud seedling grafting method at the present stage, a belly grafting mode is adopted, so that the stock is thinner (the diameter is smaller than 1 mm), the control is difficult, the unnecessary loss of the stock caused by the stock being easily split, and the problem of low grafting efficiency exists. And the stock grafted by the bud seedling is the bud seedling in the cotyledon flattening period, the grafting adaptation period is short, and a large number of seedlings are difficult to propagate in the short grafting adaptation period. In addition, the bud grafting method also has the problem that the removal of the tillers of the cotyledon bud germination is time-consuming and labor-consuming.

The inventors found that by selecting the appropriate thickness of the stock, cleft grafting is easier. The selected stock is thicker, easier to split grafting operation and high in grafting efficiency. The survival rate can be solved by adopting the mode that the green branches with leaves in the current year are used as scions. The annual green branch with leaves as scion has the advantages of quick healing of the scion and easy survival, and the grafting survival rate reaches 96 percent.

The hypocotyl grafting is adopted, so that the rootstock tillers and underground stems are not generated, and the problem of competition between the rootstock and the grafted variety is solved. In addition, the grafting method provided by the invention can control the occurrence position of the underground stem of the grafted seedling by adjusting the height of the scion bud, thereby being more convenient for the management of the underground stem.

In a preferred embodiment of the invention, before grafting, the scion is reserved with an upper bud, and the new tip on the bud is sheared by 0.5-1.0 cm, and 1/3-1/2 of the blade is sheared. It should be noted that, the length of the scion can be determined according to the specific requirement of the occurrence position of the underground stem of the grafted seedling, when a plurality of buds exist, all buds except the upper buds are removed together with the leaves, only the upper buds germinate to form plants, and the underground stem is generated at the corresponding position, so that the management of the subsequent underground stem is facilitated.

For example, 0.5cm, 0.6cm, 0.7cm, 0.8cm, 0.9cm or 1.0cm is sheared from the shoots. The upper part of the bud is sheared with a certain length of new shoots, which has a protective effect on the bud.

In an alternative embodiment, the healthy and strong young shoots of the fine variety or strain are selected as scions, and the scions are preferably used as the scions are selected, and the scions are required to be transported along with the picking process, and are required to be kept wet and fresh at low temperature so as to improve the survival rate.

In a preferred embodiment of the invention, if the scion has a plurality of buds, one upper bud is reserved, the rest of the lower buds are cut off from the base together with the leaves, and the lower end of the scion is cut into a double-bevel wedge shape.

In an alternative embodiment, the lower end of the scion is cut into a double bevel wedge. The length of the double-inclined-plane wedge is 0.5-0.8 cm, and the double-inclined-plane wedge is smooth and has small and uniform angle. The cutting is beneficial to better fit between the wedge-shaped inclined plane of the scion and the notch of the stock, so that cambium of the scion and the notch of the stock can be aligned better. And if the thicknesses of the scions and the stocks are inconsistent, aligning one side.

In an alternative embodiment, the upper end of the dual bevel wedge of the scion is left with a 0.8-1.2mm cut as the white. The arrangement can increase the contact area of the scion and the cambium of the stock, and is beneficial to the formation and healing of meristematic tissues.

In the preferred embodiment of the invention, the green seedling of the annual schisandra chinensis is watered thoroughly before grafting, the lower parts of two cotyledons of the hypocotyl of the green seedling of the annual schisandra chinensis are transversely cut, the upper part of the stock is removed, a notch is split in the middle of the hypocotyl, and the length of the notch is equal to or slightly shorter than the scion cutting surface.

Definition of hypocotyl intermediate above: including but not limited to the approximate intermediate location of the macroscopic hypocotyl.

In an alternative embodiment, the cleaving is performed until the hypocotyl thickness reaches 1.5 to 3.0 mm. For example 2.0mm thick.

In a preferred embodiment of the invention, the scion is inserted into the incision, aligned with the cambium of both the scion and the stock, and the interface is then secured. The interface can be bound and fixed by grafting clips or ropes.

In a preferred embodiment of the invention, the method further comprises illumination management, temperature management and humidity management after grafting. The grafted seedlings of the shizandra berry can heal wounds more quickly by managing the grafted seedlings, and the grafted seedlings of the shizandra berry are suitable for external environments.

In a preferred embodiment of the application of the present invention, the illumination management comprises: after grafting, the seedling raising tray carrying grafted seedlings is processed in a light-shading way within 3-4d, then is cultivated in a moderate light-supplementing way, and normal illumination is recovered from the light-shading day to 10-12 d.

In an alternative embodiment, moderate light filling refers to: the noon adopts a shading net with the light transmittance of about 50-70% to shade, and the illumination compensation is carried out in the morning and afternoon.

In a preferred embodiment of the present invention, the temperature management includes: controlling the cultivation temperature in the daytime to be 25-30 ℃ and the cultivation temperature in the evening to be 15-20 ℃ during the wound healing period of the grafted seedlings; and (5) ventilation and seedling hardening after 20 d.

In a preferred embodiment of the application of the present invention, humidity management comprises: after grafting, the environmental humidity is kept above 95% for the first 10 days, ventilation is carried out for 1-2 times in the early morning or evening every day after 10 days, and the air humidity is kept above 90%.

In the preferred embodiment of the invention, the method also comprises the cultivation of the stock before grafting, and the seedling is cultivated by using a seedling cultivation tray 50-65 days before grafting.

The invention has the following beneficial effects:

the invention adopts the green seedling of the current annual schisandra chinensis as a stock and adopts the new shoot with leaves, the auxiliary shoot or the tissue culture green shoot of the current annual schisandra chinensis as a scion, and adopts a lower billet shaft split grafting mode to enable the scion to be grafted in a notch at the hypocotyl of the stock. The seedlings with two leaves and one core period and later have larger thickness, are easier to split grafting operation, and improve grafting efficiency.

The annual green branch with leaves is used as a scion, has the advantages of quick healing of the scion and easy survival, and has higher grafting survival rate. And the hypocotyl grafting is adopted, so that the rootstock tillers and underground stems are not generated, and the competition between the rootstock and the grafted variety is not generated. In addition, the grafting method provided by the invention can control the occurrence position of the underground stem of the grafted seedling by adjusting the height of the scion bud, thereby being more convenient for the management of the underground stem.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1a is a diagram of a hard branch grafting entity of Schisandra chinensis; FIG. 1b is a view of a green branch grafting material; FIG. 1c is a view of a sprout grafting entity; FIG. 1d is a green seedling grafting physical diagram;

FIG. 2a is a diagram showing the generation of an underground stem by a stock of a green branch grafted plant; FIG. 2b is a diagram of conditions in which tissue culture plants produce a large number of subterranean stems; FIG. 2c is a plot of the condition of little or no underground stem production of green seedling grafted plants (similar to hard branch grafting, bud grafting results);

fig. 3 is a schematic view of green seedling grafting of schisandra chinensis.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.

The features and capabilities of the present invention are described in further detail below in connection with the examples.

Example 1

The embodiment provides a green seedling grafting method of schisandra chinensis, which is shown with reference to fig. 1d and fig. 3, and comprises the following steps:

1. stock cultivation

The current annual green seedlings which are sowed and propagated are used as the stocks, the 50-point seedling tray is used for carrying out protected field seedling culture before grafting for 60 days, the matrix is required to be loose and fertile, organic matters are rich, and soil-borne root diseases are sterilized. And the fertilizer and water and weeding management are enhanced in the whole process. When the seedlings grow to the period of two leaves and one heart, grafting can be carried out when the thickness of hypocotyls reaches about 2.0 mm.

In addition, in other embodiments, green seedlings of different ages after the two-leaf one-heart period can be used for long-term grafting on the premise of ensuring that grafted seedlings grow into seedlings in the current year.

2. Hypocotyl split joint

The matrix of the stock seedling is watered thoroughly before grafting, and grafting is carried out under the condition of shading. When grafting, a sharp double-sided blade is used, so that the hand and the blade are kept clean, and the blade needs to be replaced in time after the blade becomes blunt.

The healthy and strong young shoots of good varieties or strains are selected as scions, and the scions are preferably used along with the picking, and if long-distance transportation is needed, the low-temperature, moisture-preserving and fresh-keeping work is needed to improve the survival rate. When grafting, the scion only retains one upper bud, the length of the scion is determined according to the specific requirement of the underground stem occurrence position of the grafted seedling, the new tip on the bud is sheared by 0.5-1.0 cm, and 1/3-1/2 of the leaf is sheared according to the size of the leaf. If the scion is provided with a plurality of buds, the rest lower buds and the leaves are cut off from the base part, the lower end of the scion is cut into a double-inclined-plane wedge shape of 0.5cm to 0.8cm, and the inclined plane is smooth, small and uniform in angle.

And (3) transversely cutting the lower parts of the two cotyledons of the hypocotyl of the stock seedling, and removing the upper part of the stock. And splitting a notch in the middle of the hypocotyl, wherein the length of the notch is equivalent to that of a scion cutting surface, carefully inserting the scion, aligning the cambium of the scion and the cambium of the stock, aligning one side when the thicknesses of the scion and the stock are inconsistent, leaving about 1.0mm on the scion cutting surface, and fixing the interface by using a grafting clip. One plant is grafted at each hole, and the grafted plant scion leaves are kept moist before all the stocks of each seedling tray are grafted.

Besides the young shoots, the annual side shoots or the tissue culture green shoots can be used for grafting, and when the young shoots are used as scions, the leaf buds of the picked young shoots are ensured to be not differentiated.

3. Post-grafting management

3.1, after illumination management grafting, placing the seedling raising tray in a small closed arch shed, carrying out light-shielding management in the first 3 days, then shading by a shading net with 70% of light transmittance in the noon, carrying out appropriate illumination in the morning and afternoon, and removing the shading net after 10 days to ensure that the illumination is normal.

3.2 humidity management after grafting of green seedlings of shizandra berry, the relative humidity of the environment is maintained to be more than 95% for the first 10 days. Ventilation is carried out 1-2 times a day in the early morning or evening after 10d, taking care to keep the air humidity in the small arch shed to be no less than 90%.

3.3, managing the proper temperature for wound healing of the grafted seedlings to be about 25 ℃ in daytime, and not higher than 30 ℃ at maximum; the night is about 20 ℃, the lowest temperature is not lower than 15 ℃, and the healing is good after 10 days. And (5) ventilation hardening off after 20d, so that the seedlings are gradually adapted to the exposed environment conditions.

3.4 seedling transplanting and management

The grafted schisandra chinensis seedling after full seedling hardening can be transplanted in a nutrition pot with a proper size by soil lump, the matrix is required to be loose and rich, the grafted schisandra chinensis seedling is rich in organic matters, the grafted schisandra chinensis seedling is concentrated in a seedling raising area on the open field after being transplanted, and the grafted schisandra chinensis seedling is covered by a shading net with 70% of light transmittance, so that the matrix is kept moist. And after 10d, removing the shading net, and periodically supplementing the nitrogen, phosphorus and potassium compound water-soluble fertilizer according to the nutrition requirement. In the management process, the powdery mildew is prevented and treated, such as by adopting the triadimefon wettable powder.

Example 2

The embodiment provides a green seedling grafting method of schisandra chinensis, which comprises the following steps:

1. stock cultivation

The current annual green seedlings which are sowed and propagated are used as the stocks, the seedlings are cultivated in a protected field by using a 72-point seedling cultivation tray 65d before grafting, the matrix is required to be loose and fertile, organic matters are rich, and soil-borne root diseases are sterilized. And the fertilizer and water and weeding management are enhanced in the whole process. When the seedlings grow to a four-leaf one-heart period, grafting can be carried out when the hypocotyl thickness reaches about 2.5 mm.

2. Hypocotyl split joint

The current annual side shoots with strong growth of good varieties or strains are selected as scions. When grafting, the scion only retains one upper bud, the length of the scion is determined according to the specific requirement of the underground stem occurrence position of the grafted seedling, the new tip on the bud is sheared by 0.5-1.0 cm, and 1/3-1/2 of the leaf is sheared according to the size of the leaf. If the scion is provided with a plurality of buds, the rest lower buds and the leaves are cut off from the base part, the lower end of the scion is cut into a double-inclined-plane wedge shape of 0.5cm to 0.8cm, and the inclined plane is smooth, small and uniform in angle.

3. Post-grafting management

3.1 light management

After grafting, placing the seedling raising tray in a small well-closed arch shed, performing light-shielding management in the first 3 days, then shading by a shading net with 70% of light transmittance in the noon, and properly illuminating in the morning and afternoon, and removing the shading net after 10 days to enable illumination to be normal.

3.2 humidity management

After the green seedlings of the schisandra chinensis are grafted, the relative humidity of the environment is maintained to be more than 95% for the first 10 days. Ventilation is carried out 1-2 times a day in the early morning or evening after 10d, taking care to keep the air humidity in the small arch shed to be no less than 90%.

3.3 temperature management

The proper temperature for wound healing of grafted seedlings is about 25 ℃ in daytime, and the highest temperature is not higher than 30 ℃; the night is about 20 ℃, the lowest temperature is not lower than 15 ℃, and the healing is good after 10 days. And (5) ventilation hardening off after 20d, so that the seedlings are gradually adapted to the exposed environment conditions.

3.4 seedling transplanting and management

The grafted schisandra chinensis seedling after full seedling hardening can be transplanted in a nutrition pot with a proper size by soil lump, the matrix is required to be loose and rich, the grafted schisandra chinensis seedling is rich in organic matters, the grafted schisandra chinensis seedling is concentrated in a seedling raising area on the open field after being transplanted, and the grafted schisandra chinensis seedling is covered by a shading net with 70% of light transmittance, so that the matrix is kept moist. And after 10d, removing the shading net, and periodically supplementing the nitrogen, phosphorus and potassium compound water-soluble fertilizer according to the nutrition requirement. In the management process, the powdery mildew is prevented and treated, such as wettable powder.

Comparative example 1

The comparative example uses the method of hard branch grafting for asexual propagation (FIG. 1 a).

Comparative example 2

This comparative example was propagated asexually with reference to the sprout grafting method in CN101194570B (fig. 1 c).

Comparative example 3

This comparative example was propagated asexually using the method of green branch cleavage (FIG. 1 b).

Comparative example 4

This comparative example was subjected to asexual propagation by referring to the layering propagation method in CN 105210638B.

Comparative example 5

This comparative example was propagated asexually using tissue culture (FIG. 2 b).

Experimental example 1

The schisandra chinensis plants obtained by propagation in the above example 1 and comparative examples 1 to 5 are shown in fig. 1, and the survival rate statistics are shown in table 1.

Table 1 shows the effect comparison of the asexual propagation modes of Schisandra chinensis

As can be seen from fig. 1a to 1d, fig. 2a to 2c and table 1, the hard branch grafting method in comparative example 1 has a problem of low grafting survival rate. In contrast, the bud grafting method in comparative example 2 has thinner stock, is not easy to operate, and has low grafting efficiency; the stock is bud seedling in cotyledon flattening period, and the grafting adaptation period is short; removing the tillers of the germination of cotyledon buds is time-consuming and labor-consuming. Referring to fig. 2a, the method of split grafting of green branches in comparative example 3 has a large workload of removing tillers after grafting; the underground stems of the stock are produced, the control is difficult, and the method is not suitable for large-area popularization and utilization. The propagation method of the pressing strips in the comparative example 4 needs 2 years to grow seedlings, and the underground stems have low production positions and are not easy to control. Referring to FIG. 2b, the tissue culture method of comparative example 5 has too short internode, resulting in too low a position of underground stem production, which is extremely difficult to control. And referring to fig. 2c, the hypocotyl grafting performed in embodiment 1 can improve grafting efficiency, increase survival rate, avoid generation of stock underground stems, effectively adjust the occurrence position of scion variety underground stems, and is suitable for breeding high-quality schisandra chinensis asexual seedlings.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The green seedling grafting method of the schisandra chinensis is characterized by comprising the following steps of: taking a green seedling of the current annual schisandra chinensis as a stock, taking a new shoot with leaves, a side shoot or a tissue culture green shoot of the current annual schisandra chinensis as a scion, and adopting a hypocotyl split grafting mode to enable the scion to be grafted in a notch at the hypocotyl of the stock, and fixing the notch;

the green seedlings of the current annual schisandra chinensis are seedlings in two-leaf one-heart period or in the age after the two-leaf one-heart period;

before grafting, reserving an upper bud, shearing tender tips on the bud by 0.5-1.0 cm, and shearing leaves by 1/3-1/2;

if the scion has a plurality of buds, reserving an upper bud, cutting off the rest lower buds and the leaves from the base part, and cutting off the lower end of the scion into a double-inclined-plane wedge shape;

the length of the double-inclined-plane wedge is 0.5-0.8 cm, and the double-inclined-plane wedge inclined plane is smooth;

a cutting opening with the diameter of 0.8-1.2mm is reserved at the upper end of the double-inclined-plane wedge shape of the scion to serve as a reserved white;

before grafting, the green seedling of the current annual Chinese magnoliavine fruit is watered thoroughly, the lower parts of two cotyledons of the hypocotyl of the green seedling of the current annual Chinese magnoliavine fruit are transversely cut, the upper part of the stock is removed, a notch is split in the middle of the hypocotyl, and the length of the notch is equal to or slightly shorter than that of a scion cutting surface;

and splitting when the hypocotyl thickness reaches 1.5-3 mm.

2. The method of grafting green seedlings of schisandra chinensis according to claim 1, wherein the scion is inserted into the incision, the cambium of both scion and stock is aligned, and then the interface is fixed.

3. The method for grafting green seedlings of schisandra chinensis according to claim 2, wherein the grafted schisandra chinensis further comprises illumination management, temperature management and humidity management after grafting.

4. A method of grafting green seedlings of schisandra chinensis according to claim 3, wherein the illumination management comprises: after grafting, the seedling raising tray carrying grafted seedlings is processed in a light-shading way within 3-4d, then is cultivated in a moderate light-supplementing way, and normal illumination is recovered from the light-shading day to 10-12 d.

5. The method for grafting green seedlings of schisandra chinensis according to claim 4, wherein the moderate light supplement means: the noon is covered by a shading net with 50-70% of light transmittance, and the illumination compensation is carried out in the morning and afternoon.

6. A green seedling grafting method of schisandra chinensis according to claim 3, wherein the temperature management comprises: controlling the cultivation temperature in the daytime to be 25-30 ℃ and the cultivation temperature in the evening to be 15-20 ℃ during the wound healing period of the grafted seedlings; and (5) ventilation and seedling hardening after 20 d.

7. A method of grafting green seedlings of schisandra chinensis according to claim 3, wherein the humidity management comprises: after grafting, the environmental humidity is kept above 95% for the first 10 days, ventilation is carried out for 1-2 times in the early morning or evening every day after 10 days, and the air humidity is kept above 90%.

8. The method for grafting green seedlings of schisandra chinensis according to claim 1, wherein the method further comprises cultivation of stock before grafting, and seedling cultivation is carried out by using a seedling cultivation tray 50-65 days before grafting.