CN111226770A - Liquid seedling hardening method for improving transplanting survival rate of tissue culture seedlings of cinnamomum kanehirae - Google Patents
Liquid seedling hardening method for improving transplanting survival rate of tissue culture seedlings of cinnamomum kanehirae Download PDFInfo
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Abstract
The invention relates to a liquid seedling hardening method for tissue culture seedlings of cinnamomum kanehirae, belonging to the technical field of plant propagation; the method comprises the following steps: (1) selecting and cleaning rooting tissue culture seedlings: selecting rooting bottle seedlings, and cleaning; (2) and (3) treatment of the rooted tissue culture seedling: grouping the rooting tissue culture seedlings, and soaking roots in seedling hardening liquid; (3) hardening seedling management: placing the hardening container filled with the rooting tissue culture seedlings in a small plastic arched shed, spraying the leaf surfaces, and hardening the seedlings; (4) transplanting: clamping the tissue culture seedling out of the seedling hardening liquid, directly planting the tissue culture seedling in a culture medium, and spraying; (5) managing five steps after transplanting; the method for exercising the tissue culture seedlings of the cinnamomum kanehirae dunn by using the seedling exercising liquid has the characteristics of simplicity and easiness in operation, does not need complex equipment, occupies a small area, can remarkably enhance the resistance of the stems and the roots of the tissue culture seedlings of the cinnamomum kanehirae dunn, reduces the phenomena of black stems and root rot of the tissue culture seedlings, improves the transplanting survival rate of the tissue culture seedlings, shortens the seedling exercising time to 5-7 days, and effectively improves the production efficiency.
Description
Technical Field
The invention relates to a liquid hardening method for tissue culture seedlings of cinnamomum kanehirae, in particular to a liquid hardening method for improving the transplanting survival rate of the tissue culture seedlings of cinnamomum kanehirae, and belongs to the technical field of plant propagation.
Background
The Cinnamomum kanehirae (Cinnamomum kanehirae) is a special evergreen broad-leaved large tree in Taiwan, is a valuable tree species with dual purposes of wood and medicine, has straight trunk, tall tree body and changeable primary leaves, and has good ornamental value; the branches and leaves are rich in essential oil, have multiple purposes of health preserving, health care, seasoning and the like, and can also be processed into various daily necessities; the trunk has special fragrance and is not easy to decay, the material is delicate, the texture is staggered, the planing is easy, the trunk is a good wood for manufacturing high-grade furniture and sculptures, and simultaneously, the trunk is also the only host of rare fungus Antrodia camphorata (Antrodia camphorata) in nature. Antrodia camphorata is a wood decay fungus which is found to grow in the hollow trunk of an Antrodia camphorata tree naturally at present, contains more than 200 triterpenes, has various drug effects of resisting tumor, virus and allergy, reducing blood sugar and blood fat, protecting liver, enhancing immunity and the like, and has extremely rare quantity and high price in the nature. The camphor tree is treasure on the whole body, related products can be developed from leaves to trunks, and the camphor tree has high economic value and great development potential.
Due to the wide application of the cinnamomum kanahirai hay, the cinnamomum kanahirai hay is excessively harvested due to the collection of the cinnamomum kanahirai hay in the past, the cinnamomum kanahirai hay is only sporadically distributed in Taiwan mountain areas, most of the cinnamomum kanahirai hay is old, wild resources are very rare, and the artificially-planted cinnamomum kanahirai hay has a wide prospect. The seeds of the cinnamomum kanehirae are difficult to obtain, have dormancy, low germination rate and long germination period, so the seedlings of the cinnamomum kanehirae are obtained by adopting methods of cuttage, layering or grafting, but the methods are very easy to be limited by external conditions such as plant raw materials, weather and the like, the propagation rate is low, and the production of the cinnamomum kanehirae seedlings is greatly restricted. The tissue culture technology has the advantages of large propagation quantity, high speed, no influence of seasons and weather conditions and the like, and can greatly improve the propagation coefficient of the cinnamomum kanehirae. However, the prior Cinnamomum kanehirae Hayata tissue culture seedling has the problem of low survival rate of ex-bottle transplantation, and even if the tissue culture seedling is acclimatized conventionally, the transplanted tissue culture seedling is very easy to have the phenomena of black stem, root rot and the like, so that the quantity of the Cinnamomum kanehirae Hayata which finally becomes seedlings is greatly reduced.
At present, most of traditional seedling hardening methods for tissue culture seedlings adopt a two-step method of putting bottle seedlings in natural light of a greenhouse for 5-20 days and gradually opening a bottle opening for transition (generally 3-7 days) to harden seedlings, the time consumption is long, the seedling hardening method cannot enhance the resistance of stems and roots of the tissue culture seedlings of cinnamomum kanehirai dunn, the transplanting survival rate cannot be guaranteed, and the death rate is high. The transplanting survival rate of the tissue culture seedlings of the cinnamomum kanehirae dunn reported at present is about 60%, so that the search for a seedling hardening method capable of improving the resistance and the transplanting survival rate of the tissue culture seedlings of the cinnamomum kanehirae dunn is urgent.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides a liquid seedling hardening method for improving the resistance of tissue culture seedlings of camphor trees, solves the problem that the tissue culture seedlings of camphor trees are easy to die during transplanting, can obviously improve the survival rate of the transplanted tissue culture seedlings of camphor trees, increases the seedling rate, improves the production efficiency of seedlings and meets the market demand.
In order to achieve the purpose of the invention, the technical scheme adopted by the invention is as follows:
a liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata comprises the following steps:
(1) selecting and cleaning rooting tissue culture seedlings: selecting the rooting tissue culture seedlings of the cinnamomum kanehirai dunn which have the rooting culture time of 25-40 days, 2-6 developing leaves, 1-6 white roots and the plant height of 3-7 cm, clamping the seedlings by using forceps, washing off a root culture medium by using clear water, and cleaning by using the clear water;
(2) and (3) treatment of the rooted tissue culture seedling: taking 20-30 cleaned rooting tissue culture seedlings as 1 group, aligning roots, soaking the roots in a seedling exercising container filled with 25-45 mL of seedling exercising liquid, and finishing the rooting seedlings in the container by using tweezers to enable all the plants to be upright and ensure that all the roots are immersed in the seedling exercising liquid;
(3) hardening seedling management: placing a seedling hardening container filled with the rooting tissue culture seedlings of the cinnamomum kanehirae dunn in a small plastic arched shed, and spraying small water on the leaf surfaces of the tissue culture seedlings in the morning, at noon and in the afternoon by using a spraying pot, wherein the spraying time is 1-2 s each time, so that the surfaces of the leaves are uniformly covered with water drops, a proper seedling hardening environment is provided, and the seedling hardening time is 5-7 days;
(4) transplanting: clamping the tissue culture seedlings from the seedling hardening liquid by using tweezers, directly planting the tissue culture seedlings in a culture medium with the water content of 70-80% without cleaning, wherein the culture density is 4cm multiplied by 4cm, and spraying for 1-2 s by using a spray can after planting to uniformly cover the surfaces of the leaves with water drops;
(5) managing after transplanting: placing the transplanted tissue culture seedlings in proper humidity, controlling the temperature to be 18-35 ℃, lighting intensity to be 3000-7000 lx, preventing the matrix from being too wet, not watering when the matrix still contains water, watering when the matrix is dry and not dry, and using the standard that the watering amount is 70% of the matrix is wet;
the seedling exercising liquid in the step (2) is a water solution of commercial universal quick-acting rooting powder or strong rooting powder with the concentration of 1-3 g/L;
the seedling exercising container in the step (2) is a cylindrical full-transparent glass container with the bottom diameter of 6-7 cm, the inner diameter of a bottle opening of 5-6 cm, the height of 9-10.5 cm, the thickness of 0.2-0.3 cm and the volume of 220-400 mL;
the bracket of the small plastic arched shed in the step (3) is a semicircular iron bracket, the length of the bottom surface is 150-170 cm, the width of the bottom surface is 120-130 cm, the radius (height) of a circle is 80-90 cm, and a plastic film with the thickness of 0.06-0.1 mm is covered on the bracket;
the suitable seedling exercising environment in the step (3) is as follows: the temperature is 15-32 ℃, the illumination is 1500-5000 lx, and 3-10 containers with the volume of 250-400 mL and filled with clear water are uniformly placed in the plastic arched shed according to the number of the rooted seedlings so as to keep the humidity balance in the arched shed;
the culture medium in the step (4) is peat coal: perlite ═ 1: 1-1.5;
the suitable humidity in the step (5) means: when the environmental humidity is higher than 65%, the transplanted tissue culture seedlings do not need special moisture preservation treatment; when the environmental humidity is lower than 65%, the transplanted tissue culture seedlings need to be placed in a small plastic arched shed for moisturizing, and 3-10 containers with the volume of 250-400 mL and filled with clear water are uniformly placed in the arched shed so as to maintain the humidity balance in the arched shed.
Compared with the prior art, the invention has the following beneficial effects:
the liquid rooting and seedling hardening method has the characteristics of simplicity and easiness in operation, does not need complex equipment, occupies a small area, can obviously enhance the resistance of the stems and roots of the tissue culture seedlings of the cinnamomum kanehirai dunn, obviously reduces the phenomena of black stems, rotten roots and the like of the transplanted tissue culture seedlings, ensures that the transplanting survival rate of the tissue culture seedlings reaches more than 90 percent, shortens the seedling hardening time to 5-7 days, and greatly improves the production efficiency.
Detailed Description
The present invention is further illustrated in detail by the following examples, which are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.
Example 1
A liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata comprises the following steps:
(1) selection and cleaning of rooted tissue culture seedlings
Selecting the rooting tissue culture seedlings of the cinnamomum kanehirai dunn which have the rooting culture time of 25-30 days, 2-3 developing leaves, 1-4 white roots and the plant height of 3-5 cm, clamping the seedlings by using forceps, washing off a root culture medium by using clear water, and washing for 1 time by using the clear water;
(2) preparing a seedling exercising liquid: purchasing commercially available quick-acting rooting powder (produced by Zhengzhou Perster agriculture science and technology Co., Ltd.), weighing 1g, dissolving in 1000mL of clear water, and preparing into 1g/L seedling hardening liquid;
(3) selecting a seedling exercising container: selecting a cylindrical full-light-transmitting glass container with the bottom diameter of 6cm, the inner diameter of a bottle opening of 5.5cm, the height of 9.5cm, the thickness of 0.2-0.3 cm and the volume of 230mL as a seedling exercising container;
(4) and (3) subpackaging the seedling hardening liquid: 30mL of seedling exercising liquid is subpackaged in each seedling exercising container;
(5) and (3) treatment of the rooted tissue culture seedling: taking 25 cleaned rooting tissue culture seedlings as 1 group, aligning roots, putting the groups into 1 seedling hardening container, finishing the rooting seedlings in a glass bottle by using tweezers to enable all the plants to be upright, and simultaneously ensuring that all root systems are immersed in seedling hardening liquid;
(6) hardening seedling management: placing a seedling exercising container filled with cinnamomum kanehirae nakai rooting tissue culture seedlings in a small plastic arched shed with the bottom surface length of 150cm, the bottom surface width of 120cm and the circular radius (height) of 85cm, and uniformly placing 5 containers with the volume of 300mL and filled with clear water in the plastic arched shed; and (3) carrying out small spraying treatment on the leaf surfaces of the tissue culture seedlings in the morning and afternoon by using a spraying pot, wherein the spraying time is 1-2 s each time, so that the surfaces of the leaves are uniformly covered with water drops. Controlling the temperature to be 23-30 ℃, illuminating 2000-4000 lx, and hardening the seedlings for 7 days;
(7) preparing a transplanting substrate: mixing peat and perlite at a ratio of 1: 1 with clear water to obtain a matrix with water content of 70%, and packaging in seedling hardening trays with planting density of 4cm × 4 cm;
(8) and (3) planting tissue culture seedlings: clamping the tissue culture seedlings out of the seedling hardening liquid by using tweezers, directly planting the tissue culture seedlings in the culture medium, and spraying water for 1-2 s by using a watering can after planting to uniformly cover the surfaces of the leaves with water drops;
(9) managing after transplanting: the transplanting month is 4 months, the air humidity is more than 80%, the transplanted tissue culture seedlings are placed in an environment with the temperature of 22-33 ℃ and the illumination intensity of 3000-5000 lx, special moisture preservation treatment is not needed, water is sprayed for 1 time every 1-2 days, the water is watered when the matrix is dry, and the watering amount is based on that the matrix is completely wetted by 70%.
Example 2
A liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata comprises the following steps:
(1) selection and cleaning of rooted tissue culture seedlings
Selecting the cinnamomum kanehirae dungs which have the rooting culture time of 30-35 days, 3-5 developing leaves, 2-5 white roots and the plant height of more than 3.5cm, rooting and tissue culturing the seedlings, clamping the seedlings out by using tweezers, washing a root culture medium by using clear water, and washing for 1 time by using the clear water;
(2) preparing a seedling exercising liquid: purchasing commercially available strong rooting powder (produced by Shandong Sheng blue Biotech Co., Ltd.), weighing 1.2g, dissolving in 1000mL of clear water, and preparing into 1.2g/L seedling hardening liquid;
(3) selecting a seedling exercising container: selecting a cylindrical full-light-transmitting glass container with the bottom diameter of 7cm, the bottleneck inner diameter of 6cm, the height of 10.5cm, the thickness of 0.2-0.3 cm and the volume of 375mL as a seedling exercising container;
(4) and (3) subpackaging the seedling hardening liquid: each seedling exercising container is filled with 40mL of seedling exercising liquid;
(5) and (3) treatment of the rooted tissue culture seedling: taking 30 cleaned rooting tissue culture seedlings as 1 group, aligning roots, putting the groups into 1 seedling hardening container, finishing the rooting seedlings in a glass bottle by using tweezers to enable all the plants to be upright, and simultaneously ensuring that all root systems are immersed in seedling hardening liquid;
(6) hardening seedling management: placing a seedling exercising container filled with cinnamomum kanehirae nakai rooting tissue culture seedlings in a small plastic arched shed with the bottom surface length of 150cm, the bottom surface width of 120cm and the circular radius (height) of 85cm, and uniformly placing 8 containers with the volume of 300mL and filled with clear water in the plastic arched shed; carrying out small spraying treatment on the surfaces of the green seedlings in the morning, at noon and in the afternoon by using a spraying pot, wherein the spraying time is 1-2 s each time, so that water drops are uniformly covered on the surfaces of the leaves; controlling the temperature to be 20-30 ℃, illuminating 1500-4000 lx, and hardening the seedlings for 6 days;
(7) preparing a transplanting substrate: mixing peat and perlite at a ratio of 1: 1.3 with clear water to water content of 80%, and packaging in seedling tray with planting density of 4cm × 4 cm;
(8) and (3) planting tissue culture seedlings: clamping the tissue culture seedlings out of the seedling hardening liquid by using tweezers, directly planting the tissue culture seedlings in the culture medium, and spraying water for 1-2 s by using a watering can after planting to uniformly cover the surfaces of the leaves with water drops;
(9) managing after transplanting: transplanting the seedlings in 11 months with air humidity lower than 65%, placing the transplanted tissue culture seedlings in a small plastic arched shed for moisturizing, and uniformly placing 8 containers with the volume of 300mL and filled with clear water in the arched shed; controlling the temperature at 18-30 ℃, illuminating at 2000-5000 lx, spraying water for 1-2 times every day, and watering when the matrix is dry, wherein the watering amount is based on that the matrix is completely wetted by 70%.
Example 3
The effect of the conventional seedling exercising method is compared with that of the liquid seedling exercising method.
1. Selecting 30 bottles (20 plants in each bottle) of the cinnamomum kanehirae root tissue culture bottle seedlings with rooting culture time of 30-35 days, 3-6 developing leaves, 2-5 white roots and plant height of more than 3.5cm, and hardening the seedlings by respectively adopting a conventional seedling hardening method and a liquid seedling hardening method (15 bottles in each method).
(1) A conventional seedling exercising method: placing the bottle seedlings in natural light of a greenhouse for 7 days; opening 1/4 bottle mouth, and standing for 2 days; opening the bottle mouth to 1/2, and standing for 3 days; finally, the bottle mouth is completely opened and the bottle is placed for 2 days. The hardening time is 14 days.
(2) Liquid seedling hardening method: the bottle seedling treatment method is the same as the steps 1-6 in the example 2, and the seedling hardening time is 7 days.
2. The transplanting month is 11 months, and the steps of transplanting substrate preparation, tissue culture seedling planting method, post-transplanting management and the like are the same as the steps 7-9 in the example 2. The tissue culture seedlings of each seedling hardening method are planted with 100 plants, and the process is repeated for 3 times. The results of the transplantation are shown in Table 1.
TABLE 1 Effect of different seedling exercising methods on the growth of Cinnamomum kanehirae Hayata tissue culture seedlings
As can be seen from table 1, in the seedling hardening-out period, the culture medium contamination rate by the conventional seedling hardening-out method is high, close to 50%, and contaminated mixed bacteria in the culture medium may erode the tissue culture seedlings of cinnamomum kanehirai dunn and generate toxins, which may have adverse effects on the tissue culture seedlings; and by adopting the liquid seedling hardening method, the tissue culture seedlings are not polluted by mixed bacteria because the culture medium is washed away, and the pollution rate is 0 percent. Secondly, because the hardening-seedling period is 11 months, the air humidity is low and is mostly below 65%, and the leaves of the tissue culture seedlings of the cinnamomum kanehirai dunn are tender, the leaves are very easy to dehydrate when the bottle mouth is opened by 1/4 and 1/2 by the conventional hardening-seedling method, and then the leaf wilting rate is 36.6%; and the liquid seedling hardening method adopts a method of spraying and replenishing water on the leaf surface, so that the wilting number of the leaves is very small and is only 3.3 percent, and the quality of the tissue culture seedlings before transplantation is better ensured. Thirdly, compared with the liquid seedling hardening method, the black stem phenomenon is more likely to occur after the tissue culture seedling of the cinnamomum kanahirai dunn tree adopting the conventional seedling hardening method is transplanted, the black stem rate reaches 41%, which is caused by the fact that the tissue culture seedling wilts leaves too much in the seedling hardening period, and the phenomenon that the whole tissue culture seedling wilts and loses water and has black stems is more likely to occur once the wilting number of the leaves of the plant exceeds 2/3. Once the whole plant loses water or the black stem part reaches above 1/2, the plant is difficult to survive, so that the transplanting survival rate of the conventional seedling hardening method is lower: 62 percent; although the liquid seedling hardening method can also generate partial black stem plants, the black stem proportion of the plants is small, the influence on the whole plants is small, and the plants basically have no wilting phenomenon of the whole plants, so that higher survival rate can be obtained after transplanting: 91 percent. In conclusion, the liquid seedling hardening method can shorten the seedling hardening time by half for the tissue culture seedlings of the cinnamomum kanehirae, and meanwhile, the transplanting survival rate is improved by about 30 percent, which is superior to the conventional seedling hardening method.
Claims (7)
1. A liquid seedling hardening method for improving transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata is characterized in that: the method comprises the following steps:
(1) selecting and cleaning rooting tissue culture seedlings: selecting the rooting tissue culture seedlings of the cinnamomum kanehirai dunn which have the rooting culture time of 25-40 days, 2-6 developing leaves, 1-6 white roots and the plant height of 3-7 cm, clamping the seedlings by using forceps, washing off a root culture medium by using clear water, and cleaning by using the clear water;
(2) and (3) treatment of the rooted tissue culture seedling: taking 20-30 cleaned rooting tissue culture seedlings as 1 group, aligning roots, soaking the roots in a seedling exercising container filled with 25-45 mL of seedling exercising liquid, and finishing the rooting seedlings in the container by using tweezers to enable all the plants to be upright and ensure that all the roots are immersed in the seedling exercising liquid;
(3) hardening seedling management: placing a seedling hardening container filled with the rooting tissue culture seedlings of the cinnamomum kanehirae dunn in a small plastic arched shed, and spraying small water on the leaf surfaces of the tissue culture seedlings in the morning, at noon and in the afternoon by using a spraying pot, wherein the spraying time is 1-2 s each time, so that the surfaces of the leaves are uniformly covered with water drops, a proper seedling hardening environment is provided, and the seedling hardening time is 5-7 days;
(4) transplanting: clamping the tissue culture seedlings from the seedling hardening liquid by using tweezers, directly planting the tissue culture seedlings in a culture medium with the water content of 70-80% without cleaning, wherein the culture density is 4cm multiplied by 4cm, and spraying for 1-2 s by using a spray can after planting to uniformly cover the surfaces of the leaves with water drops;
(5) managing after transplanting: placing the transplanted tissue culture seedlings in proper humidity at the temperature of 18-35 ℃, the illumination intensity of 3000-7000 lx, the matrix is not too wet, watering is not needed when the matrix still contains water, watering is carried out when the matrix is dry or not dry, and the watering amount is based on that the matrix is 70% wet.
2. The liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata as claimed in claim 1, wherein: the seedling exercising liquid in the step (2) is a water solution of commercial universal quick-acting rooting powder or strong rooting powder with the concentration of 1-3 g/L.
3. The liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata as claimed in claim 1, wherein: the seedling exercising container in the step (2) is a cylindrical full-light-transmitting glass container with the bottom diameter of 6-7 cm, the inner diameter of a bottle opening of 5-6 cm, the height of 9-10.5 cm, the thickness of 0.2-0.3 cm and the volume of 220-400 mL.
4. The liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata as claimed in claim 1, wherein: the support of the small plastic arched shed in the step (3) is a semicircular iron support, the length of the bottom surface is 150-170 cm, the width of the bottom surface is 120-130 cm, the radius of the circle is 80-90 cm, and a plastic film with the thickness of 0.06-0.1 mm is covered on the support.
5. The liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata as claimed in claim 1, wherein: the suitable seedling exercising environment in the step (3) is as follows: the temperature is 15-32 ℃, the illumination is 1500-5000 lx, and 3-10 containers with the volume of 250-400 mL and filled with clear water are uniformly placed in the plastic arched shed according to the number of the rooted seedlings so as to keep the humidity balance in the arched shed.
6. The liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata as claimed in claim 1, wherein: the culture medium in the step (4) is peat and perlite which are mixed at a ratio of 1: 1-1.5.
7. The liquid seedling hardening method for improving the transplanting survival rate of tissue culture seedlings of Cinnamomum kanehirae Hayata as claimed in claim 1, wherein: the proper humidity in the step (5) refers to: when the environmental humidity is higher than 65%, the transplanted tissue culture seedlings do not need special moisture preservation treatment; when the environmental humidity is lower than 65%, the transplanted tissue culture seedlings need to be placed in a small plastic arched shed for moisturizing, and 3-10 containers with the volume of 250-400 mL and filled with clear water are uniformly placed in the arched shed so as to maintain the humidity balance in the arched shed.
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