CN114175934A

CN114175934A - Green seedling grafting method for schisandra chinensis

Info

Publication number: CN114175934A
Application number: CN202111666703.4A
Authority: CN
Inventors: 艾军; 石广丽; 樊越; 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: 2022-03-15
Anticipated expiration: 2041-12-31
Also published as: CN114175934B; 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 includes: taking the current-year Chinese magnoliavine fruit green seedling as a stock, taking the current-year Chinese magnoliavine fruit with a new shoot, an auxiliary shoot or a tissue culture green branch as a scion, and adopting a hypocotyl cleft grafting mode to graft the scion into a notch at the hypocotyl of the stock so as to fix the notch. The green seedling of the current-year schisandra chinensis is a seedling in the two-leaf one-heart period or the later age period. The grafting method provided by the invention can improve the grafting efficiency, has high survival rate, avoids the generation of the underground stem of the stock, can effectively adjust the generation position of the underground stem of the scion variety, and is suitable for breeding high-quality asexual schisandra chinensis seedlings.

Description

Green seedling grafting method for schisandra chinensis

Technical Field

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

Background

Schisandra chinensis (Turcz.) Bail is a deciduous woody vine belonging to Schisandra of Schisandraceae, Schisandra subspecies of Schisandra, and its dried berry is known as Schisandra chinensis, which is a famous and precious Chinese medicinal material in China. In recent years, due to the increasing market demand and the increasingly serious damage of wild resources, the artificial cultivation of the schisandra chinensis is developed rapidly, and the research and development of the high-efficiency asexual propagation technology of the schisandra chinensis have important significance for promoting the improved variety development of the schisandra chinensis cultivation industry.

The schisandra fruit has special underground stem, which mainly occurs in the bud of lower node in the root neck part, besides normal branches and tendrils on the ground, and the underground stem can also produce lots of sprouts besides the underground transverse extension. The tillering growth potential is strong, and the adventitious root system of the underground stem is not developed, the absorption capacity is weak, a large amount of nutrition still needs to be supplied by a mother plant, and great nutrition competition is caused to flowering and fruiting of the schisandra chinensis, normal plant growth and the like, so that the generation of the underground stem must be controlled during cultivation of the schisandra chinensis. The asexual propagation technique of schisandra chinensis is not only beneficial to improving the propagation efficiency, but also beneficial to the control of underground stems.

At present, the asexual propagation of the schisandra chinensis mainly comprises hard branch grafting, bud seedling grafting, tissue culture and other methods, and the bottleneck problems of low propagation survival rate, complex propagation technology and difficult operation, competition between the underground stem of a stock and a grafted variety, difficult control of too low generation position of the underground stem of a variety seedling, too short and too concentrated propagation period and the like exist in different asexual propagation technologies, so that the application of the asexual propagation technology of the schisandra chinensis and the development of improved varieties are restricted.

In view of this, the invention is particularly proposed.

Disclosure of Invention

The invention aims to provide a green seedling grafting method of schisandra chinensis to solve the technical problems. The inventor discovers for the first time that the adoption of the hypocotyl green seedling grafting method can simultaneously overcome a plurality of bottleneck problems of low propagation survival rate, complex propagation technology, difficulty in operation, competition between the underground stem of the stock and the grafted variety, difficulty in controlling too low generation position of the underground stem of the variety seedling, too short propagation period, too concentrated propagation period and the like.

The invention is realized by the following steps:

the invention provides a green seedling grafting method of schisandra chinensis, which comprises the following steps: taking the current-year Chinese magnoliavine fruit green seedling as a stock, taking the current-year Chinese magnoliavine fruit with a new shoot, an auxiliary shoot or a tissue culture green branch as a scion, and grafting the scion into a notch at the hypocotyl of the stock by adopting a hypocotyl cleft grafting mode to fix the notch;

the annual green Chinese magnoliavine seedling is a seedling in the period of one heart with two leaves or the later age of the period of one heart with two leaves.

The two-leaf one-heart period refers to the period when the shizandra berry seedlings grow to two complete true leaves and one new leaf (not unfolded yet). Seedlings aged after first heart of two leaves include, but are not limited to: three-leaf one-heart stage and four-leaf one-heart stage. The seedlings of the second leaf and later heart period can be grafted for a long time.

In the bud seedling grafting method at the present stage, a ventral grafting mode is adopted, so that the rootstock is thin (the diameter is less than 1mm), is difficult to control, is easy to cleave the rootstock, causes unnecessary loss of the rootstock, and has the problem of low grafting efficiency. And the stock grafted by the bud seedling is the bud seedling in the cotyledon flattening period, the grafting proper period is short, and a large number of seedlings are difficult to propagate in the short grafting proper period. In addition, the method for grafting the bud seedlings also has the problem that the removal of the sprouts germinated from cotyledon buds is time-consuming and labor-consuming.

The inventors have found that cleft grafting is easier by selecting a suitable rootstock thickness. The selected rootstock is thicker, so that cleft grafting operation is easier, and grafting efficiency is high. The problem of survival rate can be solved by adopting the mode of taking the annual green branch with leaves as the scion. The current-year leafy green branch serving as the scion has the advantages of quick healing of the stock and the scion and easy survival, and the grafting survival rate reaches 96 percent.

And the hypocotyl grafting is adopted, so that rootstock sprouting and underground stems are not generated, and the problem of competition between the rootstock and a grafted variety is solved. In addition, the grafting method provided by the invention can control the generation 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 the preferred embodiment of the invention, before grafting, the scion is kept with an upper bud, a young shoot on the bud is cut with 0.5-1.0 cm, and the leaves are cut with 1/3-1/2. It should be noted that the length of the scion can be determined according to the specific requirements of the generation position of the underground stem of the grafted seedling, when a plurality of buds exist, other buds except the upper buds and leaves are all removed, only the upper buds germinate to form a plant, and the underground stem is generated at the corresponding position, so that the subsequent management of the underground stem is facilitated.

For example, 0.5cm, 0.6cm, 0.7cm, 0.8cm, 0.9cm or 1.0cm of cut is left on the sprouts. The upper part of the bud is cut to leave a certain length of new shoots which have the protection effect on the bud.

In an alternative embodiment, the robust young shoots of the elite variety or strain are selected as the scions, which are preferably transported at any time, if needed, and should be kept at a low temperature, moist, and fresh to increase survival rate.

In the preferred embodiment of the invention, if the scion has multiple buds, one upper bud is kept, the rest lower buds are removed from the base part 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 shape. The length of the double-inclined-plane wedge is 0.5-0.8 cm, and the double-inclined-plane wedge is smooth in inclined plane and small and uniform in angle. The cutting is beneficial to better fit the wedge-shaped inclined plane of the scion and the cut of the stock, so that the cambium of the scion and the cut of the stock can be better aligned. If the thicknesses of the scion and the rootstock are not consistent, the scion and the rootstock are aligned to one side.

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

In a preferred embodiment of the invention, before grafting, the current-year green schisandra chinensis seedlings are watered thoroughly, the lower parts of two cotyledons of hypocotyls of the current-year green schisandra chinensis seedlings are transversely cut, the upper parts of stocks are removed, a notch is cut in the middle of the hypocotyls, and the length of the notch is equal to or slightly shorter than that of a scion cutting surface.

Definition of the middle of hypocotyl: including but not limited to the approximate middle of the hypocotyl visible to the naked eye.

In an alternative embodiment, cleft grafting is performed when the thickness of the hypocotyl reaches 1.5-3.0 mm. For example 2.0mm thick.

In a preferred embodiment of the invention, the scion is inserted into the incision, the cambium layers of both the scion and the rootstock are aligned, and the incision is then fixed. The interface can be bound and fixed by a grafting clip or a thread rope.

In a preferred embodiment of the invention, the grafted plant further comprises light management, temperature management and humidity management after grafting. The grafted schisandra chinensis grafted seedlings can heal wounds more quickly through management after grafting, and adaptation to external environments is accelerated.

In a preferred embodiment of the present invention, the lighting management comprises: after grafting, the seedling tray bearing the grafted seedlings is protected from light within 3-4d, then cultivation is carried out in a moderate light supplement mode, and normal illumination is recovered from the day of light protection to 10-12 d.

In an optional embodiment, the moderate light supplement means: shading with shading net with light transmittance of 50-70% at noon, and performing illumination compensation in the morning and afternoon.

In a preferred embodiment of the present invention, the temperature management comprises: controlling the cultivation temperature of 25-30 ℃ in the daytime and 15-20 ℃ at night during the wound healing period of the grafted seedlings; ventilating after 20 days to harden the seedlings.

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

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

The invention has the following beneficial effects:

the method adopts the current-year-old schisandra green seedling as the stock, the current-year-old schisandra with new shoots, minor shoots or tissue culture green branches as the scion, and adopts the lower billet shaft cleft grafting mode to graft the scion in the notch at the lower billet shaft of the stock. The seedling in the first heart period and later of two leaves has larger thickness, is easier to split and graft, and improves the grafting efficiency.

The current-year leafy green branch serving as the scion has the advantages of quick healing of the stock and the scion, easy survival and higher grafting survival rate. And the adoption of hypocotyl grafting does not produce rootstock sprout tillers and underground stems, and does not have the competition between the rootstock and the grafted variety. In addition, the grafting method provided by the invention can control the generation 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 needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1a is a drawing of a hardwood grafting object of Schisandra chinensis; FIG. 1b is a diagram of a green branch grafted entity; FIG. 1c is a diagram of a grafted seedling; FIG. 1d is a diagram of a green seedling grafted entity;

FIG. 2a is a diagram of the generation of subterranean stems from a green-branch grafted plant rootstock; FIG. 2b is a diagram showing the condition that a tissue culture plant produces a large number of subterranean stems; FIG. 2c is a graph showing the condition of a green grafted plant with little or no subterranean stem (similar to the hard shoot grafting and bud grafting results);

FIG. 3 is a schematic diagram of grafting green seedlings of Schisandra chinensis.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The features and properties of the present invention are described in further detail below with reference to examples.

Example 1

The embodiment provides a method for grafting green seedlings of schisandra chinensis, which is shown in fig. 1d and fig. 3 and comprises the following steps:

1. stock cultivation

The current-year green seedling which is sown and propagated is adopted as a stock, a 50-hole seedling tray is utilized for protected field seedling culture 60 days before grafting, the substrate is required to be loose, fertile and rich in organic matters, and the soil-borne root diseases are sterilized. The fertilizer and water are enhanced and the weeding management is enhanced in the whole process. When the seedling grows to the period of two leaves and one heart and the thickness of the hypocotyl reaches about 2.0mm, grafting can be carried out.

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 the grafted seedlings grow into seedlings in the same year.

2. Hypocotyl cleft grafting

Before grafting, the matrix of the stock seedling is watered thoroughly and grafted under the condition of shading. The sharp double-sided blade is used during grafting, the hand and the blade are kept clean, and the blade needs to be replaced in time after becoming dull.

The method selects the new shoots growing well of good varieties or strains as the scions, and the scions are preferably adopted at any time, and if long-distance transportation is needed, the low-temperature, moisture-keeping and fresh-keeping work is needed to improve the survival rate. During grafting, only one superior bud is reserved in the scion, the length of the scion is determined according to the specific requirements of the generation position of the underground stem of the grafted seedling, 0.5-1.0 cm is reserved on the young shoot of the bud in a shearing mode, and 1/3-1/2 of the leaf is reserved 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 0.5-0.8 cm, the inclined plane is smooth, and the angle is small and uniform.

And (4) transversely cutting the lower parts of two cotyledons of the hypocotyls of the stock seedlings, and removing the upper parts of the stocks. Splitting a notch in the middle of the hypocotyl, wherein the length of the notch is equivalent to the length of the cut surface of the scion, carefully inserting the scion, aligning cambiums of the scion and the stock, aligning the scion and the stock to one side when the thicknesses of the scion and the stock are inconsistent, leaving a space of about 1.0mm on the cut surface of the scion, and fixing the notch by using a grafting clip. Grafting one plant in each hole, and keeping the scion leaves of the grafted plant moist before grafting all the stocks in each seedling-raising plate.

Except for the young shoots, the annual secondary shoots or tissue culture green shoots can be used for grafting, and when the young shoots are used as scions, the leaf buds of the harvested young shoots are ensured not to be differentiated by flower buds.

3. Post-grafting management

3.1 after the illumination management grafting, the seedling culture plate is placed in a small well-sealed arched shed, the light shielding management is carried out 3d in the front, then the shading net with 70% of light transmittance is adopted to shade in the noon, the illumination is properly carried out in the morning and afternoon, and the shading net is removed after 10d, so that the illumination is normal.

3.2 after the grafting of the green seedlings of the Chinese magnoliavine fruit is carried out under the humidity management, the relative humidity of the environment is kept to be more than 95% in the first 10 days. Ventilating 1-2 times every morning or evening after 10d while keeping the air humidity in the small shed not lower than 90%.

3.3 the proper temperature for managing the wound healing of the grafted seedlings is about 25 ℃ in the daytime, and the highest temperature is not higher than 30 ℃; the temperature is about 20 ℃ at night, the lowest temperature is not lower than 15 ℃, and the healing can be good after 10 days generally. Ventilating after 20 days to acclimatize the seedlings so as to gradually adapt to the open field environment conditions.

3.4 seedling transplanting and management

The schisandra grafted seedlings after full hardening can be transplanted into a nutrition pot with a proper size, the matrix is required to be loose and fertile and is rich in organic matters, the grafted seedlings are intensively placed in a seedling raising area in the open field after being transplanted, a shading net with 70% of light transmittance is adopted to shade, and the matrix is kept moist. And (5) removing the shading net after 10d, and regularly supplementing the nitrogen, phosphorus and potassium composite water-soluble fertilizer according to the nutritional requirements. In the management process, attention needs to be paid to the prevention and control of powdery mildew, for example, the wettable powder of the triadimefon is adopted for prevention and control.

Example 2

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

1. stock cultivation

The current-year green seedling which is sowed and propagated is adopted as a stock, a seedling tray with 72 holes is utilized for protective field seedling culture 65 days before grafting, the substrate is required to be loose, fertile and rich in organic matters, and sterilization treatment is carried out on soil-borne root diseases. The fertilizer and water are enhanced and the weeding management is enhanced in the whole process. When the seedling grows to the four-leaf one-heart stage, the thickness of the hypocotyl reaches about 2.5mm, and then grafting can be carried out.

2. Hypocotyl cleft grafting

Selecting the strong-growing current-year secondary shoots of the excellent variety or strain as scions. During grafting, only one superior bud is reserved in the scion, the length of the scion is determined according to the specific requirements of the generation position of the underground stem of the grafted seedling, 0.5-1.0 cm is reserved on the young shoot of the bud in a shearing mode, and 1/3-1/2 of the leaf is reserved 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 0.5-0.8 cm, the inclined plane is smooth, and the angle is small and uniform.

3. Post-grafting management

3.1 light management

After grafting, the seedling culture plate is placed in a small well-sealed arched shed, light-shading management is carried out for 3d in the front, shading is carried out for 70% of light transmittance in the afternoon, illumination is properly carried out in the morning and afternoon, and the shading net is removed after 10d, so that illumination is normal.

3.2 humidity management

After the green Chinese magnoliavine seedling is grafted, the relative humidity of the environment is kept above 95% for the first 10 days. Ventilating 1-2 times every morning or evening after 10d while keeping the air humidity in the small shed not lower than 90%.

3.3 temperature management

The proper temperature for wound healing of the grafted seedlings is about 25 ℃ in the daytime, and the maximum temperature is not higher than 30 ℃; the temperature is about 20 ℃ at night, the lowest temperature is not lower than 15 ℃, and the healing can be good after 10 days generally. Ventilating after 20 days to acclimatize the seedlings so as to gradually adapt to the open field environment conditions.

3.4 seedling transplanting and management

The schisandra grafted seedlings after full hardening can be transplanted into a nutrition pot with a proper size, the matrix is required to be loose and fertile and is rich in organic matters, the grafted seedlings are intensively placed in a seedling raising area in the open field after being transplanted, a shading net with 70% of light transmittance is adopted to shade, and the matrix is kept moist. And (5) removing the shading net after 10d, and regularly supplementing the nitrogen, phosphorus and potassium composite water-soluble fertilizer according to the nutritional requirements. In the management process, attention needs to be paid to the control of powdery mildew, for example, wettable powder is adopted for control.

Comparative example 1

This comparative example was propagated asexually by hard-shoot grafting (FIG. 1 a).

Comparative example 2

This comparative example refers to the bud grafting method of CN101194570B for vegetative propagation (fig. 1 c).

Comparative example 3

This comparative example was carried out using the method of green branch cleft grafting for vegetative propagation (FIG. 1 b).

Comparative example 4

This comparative example refers to the layering propagation method of CN105210638B for vegetative propagation.

Comparative example 5

This comparative example was carried out by vegetative propagation using a tissue culture method (FIG. 2 b).

Experimental example 1

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

TABLE 1 comparison of asexual reproduction mode effect of Schisandra chinensis

As can be seen from fig. 1a to 1d, fig. 2a to 2c and table 1, the method for grafting a hard branch in comparative example 1 has a problem of low survival rate of grafting. The bud seedling grafting method in the comparative example 2 is thin in stock, difficult to operate and low in grafting efficiency; the stock is a cotyledon flattening stage bud seedling, and the grafting optimum period is short; removing the sprouts germinated from the cotyledon buds is time-consuming and labor-consuming. Referring to fig. 2a, in the method for green branch cleft grafting in comparative example 3, the workload of removing sprout tillers after grafting is large; the generation of the rootstock underground stem is difficult to control and is not suitable for large-area popularization and utilization, and the like. Comparative example 4 the shoot-pressing propagation method required seedling formation for 2 years, and the generation position of underground stems was low and could not be easily controlled. As shown in FIG. 2b, the tissue culture method of comparative example 5 has too short internodes, resulting in too low generation site of subterranean stems, which are very difficult to control. As shown in fig. 2c, the hypocotyl grafting in example 1 can improve the grafting efficiency, has high survival rate, avoids the generation of the underground stem of the stock, can effectively adjust the generation position of the underground stem of the scion variety, and is suitable for breeding high-quality asexual seedlings of schisandra chinensis.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A green seedling grafting method of schisandra chinensis is characterized by comprising the following steps: taking the current-year Chinese magnoliavine fruit green seedling as a stock, taking the current-year Chinese magnoliavine fruit with a new shoot, an auxiliary shoot or a tissue culture green branch as a scion, and grafting the scion into a notch at the hypocotyl of the stock by adopting a hypocotyl cleft grafting mode to fix the notch;

the annual schisandra chinensis green seedling is a seedling in the period of two leaves and one heart or the age after the period of two leaves and one heart.

2. The schisandra chinensis green seedling grafting method according to claim 1, wherein before grafting, the scion is provided with an upper bud, a tender shoot on the bud is cut for 0.5-1.0 cm, and a leaf is cut for 1/3-1/2.

3. The method for grafting schisandra chinensis green seedlings according to claim 2, wherein if the scion has a plurality of buds, one upper bud is retained, the remaining 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;

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

preferably, a cut of 0.8-1.2mm is reserved at the upper end of the double-bevel wedge of the scion as a white residue.

4. The schisandra chinensis green seedling grafting method according to claim 1, wherein before grafting, the current-year schisandra chinensis green seedling is watered thoroughly, the lower parts of two cotyledons of the hypocotyl of the current-year schisandra chinensis green seedling are transversely cut, the upper part of the stock is removed, a cut is split in the middle of the hypocotyl, and the length of the cut is equal to or slightly shorter than the cutting surface of the scion;

preferably, cleft grafting is performed when the hypocotyl thickness reaches 1.5-3 mm.

5. The method for grafting green seedlings of schisandra chinensis according to claim 1 or 4, wherein the scion is inserted into the incision, cambiums of both the scion and the rootstock are aligned, and then the incision is fixed.

6. The green seedling grafting method of schisandra chinensis according to claim 5, further comprising light management, temperature management and humidity management after grafting.

7. The green seedling grafting method of schisandra chinensis according to claim 6, wherein the light management comprises: after grafting, the seedling tray bearing the grafted seedlings is protected from light within 3-4d, then cultivation is carried out in a moderate light supplementing mode, and normal illumination is recovered from the day of light protection to 10-12 d;

preferably, the moderate supplementary lighting means: shading with shading net with light transmittance of 50-70% at noon, and performing illumination compensation in the morning and afternoon.

8. The green seedling grafting method of schisandra chinensis according to claim 6, wherein the temperature management comprises: controlling the cultivation temperature of 25-30 ℃ in the daytime and 15-20 ℃ at night during the wound healing period of the grafted seedlings; ventilating after 20 days to harden the seedlings.

9. The green seedling grafting method of schisandra chinensis according to claim 6, wherein the humidity management comprises: after grafting, keeping the environmental humidity above 95% for the first 10 days, and ventilating for 1-2 times in the early morning or evening after 10 days, and keeping the air humidity above 90%.

10. The green seedling grafting method of schisandra chinensis according to claim 1, further comprising cultivation of a rootstock before grafting, wherein seedling is raised by using a seedling raising tray 50-65 days before grafting.