Cuttage seedling raising method by induction of aerial roots of camellia oleifera fruits and cinnamomum camphora

Abstract

The invention relates to the technical field of forest improved variety breeding, in particular to a camellia oleifera tree aerial root induced cutting seedling raising method which comprises the following steps: S1, mashing purple soil, adding water for soaking, standing, and separating to obtain a clear solution as a basic solution; s2, a disinfectant, an aerial root induction solution and a cutting medium are prepared with the basic solution instead of water; s3, selecting a disease-free camellia oleifera tree trunk sprouting branch, pruning a cutting slip with the length of 8-10cm, and soaking the lower end of the cutting slip with an aerial root induction solution; s4, cutting the cutting slips into a cutting medium, and controlling proper temperature, humidity and illumination intensity; when the length of adventitious roots reaches 5 cm or above after cuttage is carried out for 2-3 months, bagging and seedling raising can be carried out. According to the method, water is replaced by the purple soil basic solution, and the aerial root induction solution is combined for treatment, so that the rooting rate of the cuttings is high, the root system quality is good, the transplanting survival rate is high, the transplanted seedlings grow vigorously, and the blank in the technical field of Cinnamomum tamala cutting seedling raising is filled.

Description

Technical Field

[0001] The present invention relates to the technical field of fine breeding of forest trees, and specifically relates to a method for inducing cutting seedlings with aerial roots of Cinnamomum chago, belonging to the special topic of plant protection. Background Art

[0002] Cinnamomum chago B.S.Sun & H.L.Zhao is an evergreen tall tree of the genus Cinnamomum in the Lauraceae family. This species was published as a new species by Chinese botanists Sun Bixing and Zhao Huilan in 1991. In 2021, it was listed as one of the wild plants with extremely small populations in Yunnan Province, and at the same time, it was listed as a second-class protected wild plant in the "List of National Key Protected Wild Plants 2021". Cinnamomum chago was classified in the genus Cinnamomum of the Lauraceae family in previous classifications, and the latest classification includes it in the genus Cinnamomum of the Lauraceae family. Its fruit is relatively large and the seeds are edible. Hou Min et al. published an analysis of the nutritional components and nutritional evaluation of the fruits of Cinnamomum chago in 2019. The results showed that the seeds of Cinnamomum chago contain 9 kinds of nutrients such as crude fat, protein, crude fiber, total sugar, total saponins, tannins, moisture, and vitamin C; they are composed of 9 kinds of fats, including 6 kinds of saturated fatty acids and 3 kinds of unsaturated fatty acids, and the content of unsaturated fatty acids is 5.11%; among the saturated fatty acids, the content of lauric acid is the highest, which is 68.92%; among the unsaturated fatty acids, the content of oleic acid is the highest, which is 4.43%; the protein amino acids in the kernels of Cinnamomum chago are complete in 17 kinds. It is considered that Cinnamomum chago has potential comprehensive utilization value.

[0003] Yang Huiqin studied the photosynthetic physiological changes and gene expression analysis of Cinnamomum chago seedlings in response to water stress in 2022; Mo Jinqiu studied the morphology and photosynthetic physiological characteristics of Cinnamomum chago in 2019; Li Sunling et al. published a research paper on the determination of seed viability of Cinnamomum chago in 2021, and the research believed that the germination potential of Cinnamomum chago seeds was good, but the artificial seedling raising of Cinnamomum chago should be carried out by the method of sowing seeds as soon as they are harvested; Zhang Xue et al. conducted a 120-day field survey from 2014 to 2017 and only found 5 populations in Xincun, Shunbi Township, Laguo Village in Yangbi County, Dali Prefecture, Nanmuping Village, Dangga Village, Dashi Dam in Yunlong County, and the total number of individuals was only 64. Li Shufang et al. published the investigation results of the germplasm resources of Cinnamomum chago in Yangbi County in 2020. Only 137 Cinnamomum chago plants were surveyed in Shunbi Town, Yangbi County, and only 41 of them were growing vigorously. From June 2023 to May 2024, the Dali Branch of Yunnan Forestry Investigation and Planning Institute conducted a preliminary investigation on the resource distribution of Cinnamomum chago in Dali Prefecture. In addition to the above distribution points, 3 new distribution points were added, namely Caojian Town in Yunlong County, Jinguangsi Nature Reserve in Yongping County, and Beidou Township in Yongping County. However, the number of individuals was small. So far, the number of Cinnamomum chago individuals surveyed was about 300. The population quantity was small, the population scale was small, and there were only 3 individuals in some populations. Due to the high nutritional value of Cinnamomum chago seeds and it being an excellent timber tree species, it was wantonly picked and cut by local people, the resource quantity became less and less, and it was more difficult for understory regeneration. Therefore, it was urgent to take artificial assisted breeding of Cinnamomum chago to maintain and increase the population quantity of individuals.

[0004] At present, the literature reports on the breeding technology research of Cinnamomum chago are still blank. The relevant patents are only two items, namely "Seedling raising technology for seedlings of Cinnamomum chago, a plant with extremely small populations, CN202110306143.5" and "A method for quickly breaking the dormancy of seeds of Cinnamomum chago, a characteristic resource plant, CN202110597383.5", both of which involve the seedling raising technology of Cinnamomum chago. During the investigation, it was found that the number of healthy and fruiting Cinnamomum chago individuals was extremely small, there was the phenomenon of wanton picking by humans, it was extremely difficult to collect a large number of provenances for breeding, and it was found through investigation that there were large morphological differences in the individual morphology of Cinnamomum chago in different regions. Therefore, it was necessary to carry out research on asexual propagation technology to preserve the genes of excellent individuals. Summary of the Invention

[0005] The purpose of the present invention is to solve the problem that when the provenance of Cinnamomum chago is insufficient and the amplification of the population by sowing and seedling raising is restricted, to provide a simple, effective, low-cost and highly operable cutting seedling raising method, and at the same time make up for the blank in the field of cutting seedling raising technology of Cinnamomum chago.

[0006] The purpose of the present invention is achieved through the following technical solutions: A method for inducing and cutting seedlings with aerial roots of Cinnamomum chago, comprising the following steps:

[0007] S1. Preparation of basic solution: Mash the purple soil until the soil particle diameter is less than 0.5 mm. Add 4 - 6 times the volume of water to the purple soil and soak for 1 - 2 days (stir thoroughly 1 - 2 times in the middle). The clarified liquid obtained after standing and separation is the basic solution;

[0008] S2. Preparation of disinfectant solution, aerial root induction solution and cutting substrate: Use the above-mentioned basic solution to replace water to prepare the disinfectant solution, aerial root induction solution and cutting substrate respectively;

[0009] Among them, the formula of the aerial root induction solution includes: 0.1 - 0.2% carbendazim powder, 0.5 - 1% granulated sugar, 0.2 - 0.5% gibberellic acid, 0.2 - 0.5% naphthylacetic acid rooting powder;

[0010] S3. Selection and treatment of cuttings: Select the sprouted branches of the trunk of disease-free Camphora chuga aged 0.5 - 2.0 years from February to May to trim cuttings. The length of the cuttings is 8 - 10 cm. Retain two half leaves and a pair of axillary buds at the morphological upper end. The upper cut is cut obliquely, 1 - 2 cm away from the leaf attachment. The lower cut is cut flat. Wipe off the leaves and buds that are not retained in the middle of the cuttings by hand; Soak the treated cuttings in the above-mentioned disinfectant solution for disinfection for 1.5 - 2 hours. After disinfection, soak the lower end in the aerial root induction solution, and the soaking depth is about 3 - 4 cm, and the soaking time is 1 - 1.5 hours;

[0011] S4. Cutting and management: Insert the cuttings into the cutting substrate, and the cutting depth is 3 - 5 cm. After cutting, spray the disinfectant solution, control the temperature at 25 - 30 °C, the humidity above 90%, and the light intensity at 2000 - 5000 lux; When the adventitious roots grow up to more than 5 cm after 2 - 3 months of cutting, transfer the cuttings to the seedling hardening bed for seedling raising.

[0012] Further, in step S2, the disinfectant solution is prepared with 0.2 - 0.5% carbendazim powder; The preparation method of the cutting substrate includes: Using perlite as the basic substrate, adding 1.5 - 2 times the volume of peat soil and mixing thoroughly, and adding the disinfectant solution to soak for 12 - 24 hours. Drain the water from the disinfected substrate, and control the water content of the substrate at 50 - 60%.

[0013] Further, in steps S2 and S3, the methods of disinfecting the cutting substrate and the cuttings are both: soaking and disinfecting with the disinfectant solution.

[0014] Further, in step S3, the cuttings need to be cleaned of stains before disinfection, and the method of cleaning stains includes: Soak the cuttings in the washing powder water for 20 - 30 minutes, stir to remove the stains, and rinse thoroughly.

[0015] Further, in step S4, the cutting medium is filled into the seedling-raising bag for use. The caliber of the seedling-raising bag is 6 cm, the height is 10 cm, the filling depth of the cutting medium is 4 - 6 cm, and 3 - 4 cuttings are inserted into each seedling-raising bag.

[0016] Further, in step S4, the specific method of spraying the disinfectant after cutting is: spraying a disinfectant with a concentration of 0.2% on the leaf surface of the cuttings.

[0017] Further, in step S4, the specific method of raising seedlings in the hardening bed is: transferring the cuttings and the cutting medium around them as a whole into the hardening bed for hardening, and continuously changing the humidity, temperature, and light intensity until they are the same as the outdoor conditions.

[0018] The beneficial effects of the present invention are as follows:

[0019] 1. The present invention fills the blank in the field of cutting seedling raising technology for Cinnamomum pedunculatum var. szemois.

[0020] 2. The present invention uses purple soil base liquid to replace water and combines it with the treatment of aerial root induction liquid. The cuttings have a high rooting rate, good root quality, high transplanting survival rate, and the seedlings grow vigorously after transplantation.

[0021] 3. The materials used in the present invention are low in price, easy to obtain, reusable, and have low input costs.

[0022] 4. The cutting management process of the present invention requires little manual intervention, is simple to manage, and has strong operability. Description of the Drawings

[0023] Figure 1 It is a picture of adventitious aerial roots; when cutting for 20 days, a large number of lenticels have been produced in the phloem area below the leaf attachment of the cuttings in the left picture, and adventitious aerial roots occur here. In the right picture, some cuttings have produced adventitious aerial roots. Detailed Embodiments

[0024] The technical solution of the present invention will be further described in detail below with reference to the drawings, but the protection scope of the present invention is not limited to the following.

[0025] Example 1

[0026] The method for inducing adventitious aerial roots and cutting seedlings of Cinnamomum pedunculatum var. szemois includes the following steps:

[0027] Step 1: Preparation of the adventitious aerial root induction box for cuttings

[0028] Prepare a 120L transparent storage box with dimensions of 70 cm in length, 40 cm in width, and 40 cm in height. Make 2 holes on each of the four side faces of the box, 15 cm and 25 cm vertically upward from the bottom respectively, with each hole having a diameter of 0.5 cm, and the interval between the two holes being 10 - 20 cm, which are used as air exchange channels. Make 3 holes on the long side face of the box, 20 cm vertically upward from the bottom, with a hole diameter of 1.1 cm. Use a PVC pipe with a diameter of 1 cm to cross through as a support beam, and lay a seedling tray (the size can be appropriately cut) on the beam. Lay a layer of 5 cm thick sponge on the bottom of the box and on the upper seedling tray respectively to maintain the humidity in the box. Cover the storage box lid during seedling cultivation. Before cutting, all the above-mentioned equipment is sprayed and disinfected with a 0.5% carbendazim solution. The sponge is soaked in carbendazim disinfectant water and then wrung out appropriately until it doesn't drip, and then it can be used.

[0029] Step 2: Preparation of treatment solution

[0030] The treatment solution includes a disinfectant solution and an adventitious root induction solution. The preparation method is as follows: Collect weathered and dried purple soil, and use a hammer to crush it into fine soil with a particle diameter of less than 0.5 mm. Soak it in tap water, add 4 - 6 times the volume of water according to the volume of the purple soil, and soak for 1 - 2 days. Stir it thoroughly 1 - 2 times in the middle to make the elements in the soil fully dissolve in the water. After soaking, let it stand for the sediment to settle, and keep the clarified water for later use. The water obtained above will be used to prepare the disinfectant solution for the cutting substrate, cuttings, etc. and the basic solution for the adventitious root induction solution of the cuttings. Preparation of the disinfectant solution: Prepare a solution with a concentration of 0.2 - 0.5% using the basic solution and carbendazim powder; Preparation of the adventitious root induction solution: Based on the basic solution, add 0.1 - 0.2% carbendazim powder, 0.5 - 1% granulated sugar, 0.2 - 0.5% gibberellic acid solution, and 0.2 - 0.5% naphthaleneacetic acid rooting powder, and stir thoroughly. The dosage depends on the actual situation.

[0031] Step 3: Preparation and treatment of cutting substrate

[0032] The cutting substrate is based on perlite, add 1.5 - 2 times the volume of peat soil and mix thoroughly, and soak it in the disinfectant solution for 12 - 24 hours. After disinfection, drain the water from the substrate, control the water content of the substrate at 50 - 60%, and fill the substrate into black seedling plastic bags with a diameter of 6 cm and a height of 10 cm, with a filling depth of 5 cm. The filled seedling bags are neatly arranged at the bottom of the induction box for later use. The upper layer will be carried out after the cuttings in the bottom layer are cut.

[0033] Step 4: Selection and treatment of cuttings

[0034] Select disease-free tea fruit camphor tree trunk sprouted branches for pruning cuttings from February to May, with the cuttings being 0.5 - 2.0 years old. The length of the cuttings is 8 - 10 cm. Retain two half-leaves and a pair of axillary buds at the morphological upper end. The upper cut is made obliquely, about 1.0 - 2.0 cm away from the leaf attachment point. The lower cut is made flat. Wipe off the leaves and buds that are not retained in the middle of the cuttings by hand. Bundle the trimmed cuttings in a certain quantity with a rubber band, soak them in washing powder water for 20 - 30 min, stir appropriately to remove stains, and then rinse the washing powder water off with tap water after taking them out. The preliminarily treated cuttings are soaked in the above-mentioned disinfectant solution again for 1.5 - 2 h. After disinfection, gently shake off the water from the cuttings and let them dry in a cool place. Immerse the lower end of the cuttings in the air root induction solution, with a depth of about 3 - 4 cm, and the soaking time is 1.0 - 1.5 h.

[0035] Step 5: Cuttage and Management

[0036] Insert the cuttings treated in Step 3 into the nutrient bags filled with substrate, using the direct insertion method. Insert 3 - 4 cuttings into each bag, and the cutting depth is one-third of the cutting. The upper layer is cut in the same way. After cutting, use a sprayer to spray the leaves with a disinfectant solution with a concentration of 0.2%. Cover the box lid, and lay two layers of 70% sunshade nets on the box body. Place the induction box in the greenhouse, and control the indoor temperature at 25 - 30 °C. At this time, the temperature in the induction box can reach 25 - 30 °C, and the humidity is above 90%. About 20 days after cutting, callus will be produced on the cuticle stomata of the exposed part of the cuttings, and some cuttings will start to produce aerial adventitious roots (such as Figure 1 ), about 40 days after cutting, a large number of aerial adventitious roots will start to be produced in the middle of the cuttings. About 2 - 3 months after cutting, the adventitious roots will be more than 5 cm long. After the root tips insert into the substrate, the whole nursery bag can be transferred to the hardening bed.

[0037] Step 6: Hardening and Management of Rooted Seedlings

[0038] After the aerial adventitious roots fall and insert into the substrate, transfer the whole nursery bag to the hardening bed in the greenhouse for hardening. The bottom of the hardening bed is covered with a plastic film, and the whole is a trough bed. Lay 5 cm thick river sand on the bed surface. Pour the river sand thoroughly with a 0.2% carbendazim solution. Place the transferred nursery bags on the river sand, and cover a layer of plastic film about 10 cm above the cuttings to maintain humidity. Lay a layer of 70% sunshade net on the hardening bed. After about 20 days of hardening, gradually uncover the plastic films on both sides of the hardening bed appropriately to gradually reduce the humidity and temperature in the bed. After the growth of the rooted seedlings is basically stable, transplant the rooted seedlings into larger nutrient bags for routine management.

[0039] Comparative Example 1

[0040] Comparative Example 1 and Example 1 adopted the same induced cutting seedling raising method, the only difference being that Comparative Example 1 did not prepare the basic solution of purple soil and directly used water; and also did not use the aerial root induction solution, but dipped the lower cut end of the disinfected cutting in naphthalene acetic acid rooting powder and then carried out cutting. The cutting result was that aerial adventitious roots could not be produced and the rooting rate was also low, with only 3.3 adventitious roots per cutting on average.

[0041] Comparative Example 2

[0042] Comparative Example 2 and Example 1 adopted the same induced cutting seedling raising method, the only difference being that Comparative Example 2 did not prepare the basic solution of purple soil and directly used water. The cutting result was that aerial adventitious roots could not be produced and the rooting rate was also low, with only 1.9 adventitious roots per cutting on average.

[0043] The experimental results of the above Example 1, Comparative Example 1 and Comparative Example 2 are summarized in the following table:

[0044]

[0045] The above are only the preferred embodiments of the present invention. It should be understood that the present invention is not limited to the form disclosed herein, should not be regarded as excluding other embodiments, but can be used in various other combinations, modifications and environments, and can be changed within the scope of the concept described herein through the above teachings or the techniques or knowledge in related fields. And any changes and modifications made by those skilled in the art without departing from the spirit and scope of the present invention shall fall within the protection scope of the appended claims of the present invention.

Claims

1. A method for inducing aerial root cuttings of Cinnamomum camphora, characterized in that: The following steps are involved: S1. Preparation of base liquid: crush the purple soil to make the diameter of soil particles less than 0.5mm, add 4-6 times the volume of purple soil into water and soak for 1-2 days, let it stand and separate the clear liquid obtained as the base liquid; S2, preparation of disinfectant, aerial root inducing solution and cutting medium: using the base solution instead of water to prepare disinfectant, aerial root inducing solution and cutting medium respectively; The formula of the aerial root inducing solution includes: 0.1-0.2% carbendazim powder, 0.5-1% sugar, 0.2-0.5% gibberellic acid, and 0.2-0.5% naphthylacetic acid rooting powder; S3, cutting selection and treatment: select 0.5-2.0 year old disease-free tea fruit camphor tree trunk sprout branches to prune cuttings from February to May, the cutting length is 8-10cm, the upper end of the morphology retains two and a half leaves and a pair of axillary buds, the upper end of the cut is cut obliquely, 1-2cm away from the leaf attachment, the lower end of the cut is cut flat, and the leaves and buds that are not retained in the middle of the cutting are wiped off by hand; after the treated cuttings are disinfected, the lower end is soaked in the aerial root induction solution, the soaking depth is about 3-4cm, and the soaking time is 1-1.5h; S4, cutting and management: the cuttings are inserted into the cutting medium at a depth of 3-5 cm, disinfectant is sprayed after cutting, the temperature is controlled at 25-30°C, the humidity is above 90%, and the light intensity is 2000-5000 lux; when the adventitious roots are more than 5 cm long after 2-3 months of cutting, the cuttings are transferred to the seedling hardening bed for seedling cultivation.

2. A method for raising seedlings of Cinnamomum camphora by aerial root induction cuttings according to claim 1, characterized in that: In step S2, the disinfectant is prepared from 0.2-0.5% carbendazim powder; the preparation method of the cutting matrix includes: taking perlite as the base matrix, adding 1.5-2 times the volume of peat soil to mix thoroughly, adding disinfectant to soak for 12-24 hours, draining the water from the disinfected matrix, and controlling the moisture content of the matrix to be 50-60%.

3. A method for raising seedlings of Cinnamomum camphora by aerial root induction cuttings according to claim 2, characterized in that: In step S2 and step S3, the method for disinfecting the cutting substrate and the cuttings is: immersing and disinfecting them in the disinfectant.

4. A method for raising seedlings of Cinnamomum camphora by aerial root induction cuttings according to claim 1, characterized in that: In step S3, the cuttings need to be cleaned before disinfection, and the method for cleaning the stains includes: soaking the cuttings in detergent water for 20-30 minutes, stirring to remove stains, and rinsing.

5. The method for raising seedlings of Cinnamomum camphora by aerial root induction cutting according to claim 1, characterized in that: In step S4, the cutting matrix is ​​put into a seedling bag for use. The diameter of the seedling bag is 6 cm and the height is 10 cm. The filling depth of the cutting matrix is ​​4-6 cm. Each seedling bag is used for cutting 3-4 cuttings.

6. The method for raising seedlings of Cinnamomum camphora by aerial root induction cuttings according to claim 1, characterized in that: In step S4, spraying disinfectant after cuttings specifically includes: spraying disinfectant with a concentration of 0.2% on the leaf surfaces of the cuttings.

7. The method for raising seedlings of Cinnamomum camphora by aerial root induction cutting according to claim 1, characterized in that: In step S4, the specific method of raising seedlings in the seedling hardening bed is: transferring the cuttings and the cutting matrix around the cuttings as a whole to the seedling hardening bed for hardening, and constantly changing the humidity, temperature, and light intensity until they are the same as outdoor conditions.

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