Blueberry tissue culture seedling ex-vitro rooting method

Abstract

The invention discloses an ex-vitro rooting method for blueberry tissue culture seedlings, which comprises the following steps: laying a seedbed with the thickness of 10-20cm in a flowerpot, and disinfecting a substrate; the blueberry test-tube plantlets are cut into short branch sections, and the short branch sections of the blueberries are soaked in a thiophanate-methyl solution for 8-15 min; dipping the short branch sections of the blueberries in NAA, flatly laying the short branch sections on the surface of the seedbed in the flowerpot, disinfecting after laying, and tightly sealing the opening of the flowerpot; after spreading for 7 days, spraying a WPM basic culture medium solution with the concentration of 10-25%, wherein 2g of thiophanate-methyl powder is added into each liter of the solution, to the short branch sections of the blueberries for accelerating seedlings and disinfecting, and tightly sealing the opening of the flowerpot; and spraying the blueberry branch sections with a 10-25% WPM basic culture medium solution, wherein 2g of thiophanate-methyl powder is added into each liter of the solution, every 5d to promote seedling and disinfect the blueberry branch sections. According to the method, the blueberry seedling culture cost can be reduced, the problem of low rooting rate caused by blackening of a branch base due to too high substrate humidity during cuttage is effectively solved, and the blueberry tissue culture seedling culture efficiency is greatly improved.

Description

Technical Field

[0001] The invention relates to the technical field of plant cultivation, in particular to an ex-bottle rooting method for blueberry tissue culture seedlings. Background Art

[0002] Blueberry belongs to the genus Vaccinium in the family Ericaceae and is a perennial deciduous or evergreen shrub. The tissue culture technology of blueberries is now relatively mature. However, the rooting effect of blueberry tissue culture seedlings in vitro is poor and slow, with a rooting rate of only 30%-70%, so the main method of rooting outside the test tube is used. At present, the conventional method of blueberry rooting outside the test tube is: a plastic greenhouse with a sunshade net covered on the roof is generally used, the seedling tray is placed on the seedling rack or on the ground, and a small arch shed is built on top; moss and peat are used as the cutting medium; when cutting, the test tube seedlings are cut into 5-10cm branch segments, dipped in a rooting agent and inserted into the matrix, and the insertion depth of the matrix is ​​1-2cm. Although the currently commonly used blueberry outside the test tube rooting technology is relatively mature, it still has shortcomings:

[0003] (1) High seedling raising cost: The cost of plastic greenhouses and seedling raising racks is high, and the price of moss and peat soil is also high, resulting in high seedling raising cost in the rooting stage.

[0004] (2) The utilization efficiency of test tube seedlings is low: Usually a test tube seedling is only used as a cutting branch segment, and the cutting branch segment is long, so the utilization efficiency is low.

[0005] (3) The humidity of the cutting medium is difficult to control: the branch segments are inserted deep into the cutting medium. If the humidity of the cutting medium is not well controlled, the rooting rate will be reduced. If too much water is sprayed, the cutting medium humidity will be too high, causing the part where the branch segments are inserted into the cutting medium to turn black and not take root. If too little water is sprayed, the medium humidity is low and it is also difficult for the cuttings to take root.

[0006] Therefore, how to provide a method for rooting blueberry tissue culture seedlings outside a bottle is a technical problem that those skilled in the art urgently need to solve. Summary of the invention

[0007] The purpose of the present invention is to make up for the above-mentioned shortcomings and provide a method for rooting blueberry tissue culture seedlings outside a bottle, thereby reducing the cost of blueberry seedling cultivation and greatly improving the efficiency of blueberry tissue culture seedlings.

[0008] The present invention solves the technical problem by adopting the following technical solution:

[0009] A method for rooting blueberry tissue culture seedlings outside a bottle comprises the following steps:

[0010] Step (1), seedbed preparation: the seedbed in the flowerpot is laid to a thickness of 10-20 cm and the substrate is disinfected;

[0011] Step (2), cutting the seedlings: taking the blueberry test tube seedlings out of the culture bottle, cutting them into short branches with at least one axillary bud and a length of 0.8-1.5 cm, and soaking the blueberry short branches in a 500-1000 times methyl thiophanate solution for 8-15 minutes;

[0012] Step (3), paving: dip the blueberry short branches into NAA and spread them on the surface of the seedbed in the flowerpot, spray disinfectant with 500-1000 times methyl thiophanate solution, and then seal the flowerpot tightly with white plastic film;

[0013] Step (4), accelerating seedlings: after 7 days of tiling, spray the blueberry short branches with a 10%-25% WPM basic culture medium solution containing 2g of methyl thiophanate powder per liter of solution to accelerate seedlings and disinfect, and then tightly seal the flowerpot with a white plastic film; thereafter, spray the blueberry branches with a 10%-25% WPM basic culture medium solution containing 2g of methyl thiophanate powder per liter of solution every 5 days to accelerate seedlings and disinfect.

[0014] Furthermore, in the step (1), the seedling medium is soil, and the flower pot is filled with soil as a seedbed. The thickness of the soil in the flower pot is 10-20 cm, and the soil is sprayed with 200-500 times mancozeb solution to disinfect the medium.

[0015] Furthermore, in step (2), each test tube seedling with a proliferation period of 45 days can be cut into 6-12 short branch segments of 0.8-1.5 cm.

[0016] Furthermore, the method further comprises step (5), temperature and humidity control: placing the flower pots in a small arch shed, covering the top of the small arch shed with a sunshade net with a shading rate of 60%, and by adjusting the opening and closing of the small arch shed and spraying water into the small arch shed, spraying water on the matrix and blueberry branches in the flower pots every day after cutting, so as to control the air humidity in the flower pots at 80%-100% and the temperature at 16-30°C.

[0017] Furthermore, the method further comprises step (6), hardening the seedlings and placing them in pots for care: when the leaves at the top of the seedlings begin to unfold after taking root, gradually open the plastic film that closes the mouth of the flowerpot, and simultaneously open the two ends of the small arch shed. After hardening the seedlings for 25-30 days, place the rooted blueberry seedlings directly in the nutrient pots and provide shade for care.

[0018] The invention discloses a method for rooting blueberry tissue culture seedlings outside a bottle, which has the following beneficial effects:

[0019] (1) Reducing the cost of raising seedlings: The substrate used in the present invention is soil, which is much cheaper than moss and peat. In addition, no greenhouse or seedling rack is required, which is equivalent to deducting the cost of the greenhouse and seedling rack. The price of small arch sheds and flower pots is relatively low, which can greatly reduce the cost of raising blueberry seedlings.

[0020] (2) Improving the utilization rate of test tube seedlings: The length of the branch segments of tissue culture test tube seedlings commonly used in the conventional method of blueberry seedling cultivation is 5-10 cm, while that of the present invention is 0.8-1.5 cm. Therefore, the present invention increases the utilization rate of the branch segments of blueberry test tube seedlings by 6-12 times, greatly improving the efficiency of blueberry tissue culture seedlings.

[0021] (3) The substrate humidity and air humidity are easier to control: On the one hand, the blueberry test tube seedlings are spread flat on the substrate surface, and water can be sprayed on the substrate frequently to maintain a high humidity. The high humidity of the substrate will not affect the rooting, thus avoiding the problem of blackening of the part where the blueberry branches are inserted into the substrate due to excessive substrate humidity, resulting in a low rooting rate. On the other hand, flower pots are used as seedling containers, and the mouth of the flower pots is tightly sealed with white plastic film to ensure a high air humidity in the flower pots.

[0022] (4) Temperature is easier to control: Use a small arch shed combined with a closed flower pot. If the temperature is too high, the cutting temperature can be adjusted by ventilating the small arch shed, sprinkling water inside and outside the small arch shed, covering the roof or removing the shade net. Temperature control is quick, simple and easy. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The present invention is a flow chart of the method for rooting blueberry tissue culture seedlings outside a bottle. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] The purpose of the present invention is to provide a method for rooting blueberry tissue culture seedlings outside a bottle, thereby reducing the cost of blueberry seedling cultivation and improving the efficiency of blueberry tissue culture seedling cultivation. Of course, the method can also be applied to rooting tissue culture seedlings of other plants outside a test tube, thereby reducing the cost of tissue culture seedling cultivation and improving the efficiency of tissue culture seedling cultivation.

[0026] Example 1

[0027] like Figure 1 As shown, the present embodiment discloses a method for rooting blueberry tissue culture seedlings outside a bottle, comprising the following steps:

[0028] Step (1), soil is used as a seedling medium, a plastic flower pot with a diameter of 45 cm is filled with soil as a seedling bed, the thickness of the soil in the flower pot is 10-20 cm, and the soil is sprayed with 200-500 times mancozeb solution to disinfect the medium; the flower pot is used as a blueberry tissue culture seedling seedling container, which is very convenient to manage, and the soil is used as the seedling medium, which reduces the cost of the seedling medium. The present invention does not impose any specific restrictions on the material and size of the flower pot.

[0029] Step (2), cutting the seedlings: taking out 30 blueberry test tube seedlings from the culture bottle, cutting them into short branch segments with at least one axillary bud and a length of 0.8-1.5 cm, in this embodiment, cutting them into 244 short branch segments in total, and soaking the blueberry short branch segments in 500-1000 times methyl thiophanate solution for 8-15 minutes; cutting the blueberries into very short branch segments, usually each test tube seedling with a proliferation period of 45 days can be cut into 6-12 short branch segments, which can increase the utilization rate of the blueberry test tube seedlings by 6-12 times, greatly improving the efficiency of blueberry tissue culture seedlings.

[0030] Step (3), spreading: dip the blueberry short branch segments in NAA and spread them on the surface of the seedbed in the flowerpot, spray disinfection with 500-1000 times methyl thiophanate solution, and then tightly seal the flowerpot with a white plastic film; the cutting method is changed from conventional insertion into the matrix to spreading, which can prevent the cutting branch segments from rotting after being inserted into the matrix, thereby reducing the rooting rate, and is conducive to the cutting operation of the short branch segments.

[0031] Step (4), seedling promotion: after 7 days of flattening, spray the blueberry short branches with a 10%-25% WPM basic culture medium solution with 2g of methyl thiophanate powder added per liter of solution to promote seedling promotion and disinfection, and then seal the flower pot mouth tightly with a white plastic film; thereafter, spray the blueberry branches with a 10%-25% WPM basic culture medium solution with 2g of methyl thiophanate powder added per liter of solution every 5 days to promote seedling promotion and disinfection. Spraying the 10%-25% WPM basic culture medium solution can provide nutrients for the blueberry short branches like tissue culture propagation, and promote the growth of the blueberry short branches; at the same time, adding 2g of methyl thiophanate powder per liter of spray solution can effectively prevent the blueberry short branches from being contaminated by fungi.

[0032] Step (5), temperature and humidity control: place the flower pots in a small arch shed, cover the top of the small arch shed with a sunshade net with a shading rate of 60%, adjust the opening and closing of the small arch shed and spray water into the small arch shed, spray water on the matrix and blueberry branches in the flower pots every day after cutting, control the air humidity in the flower pots at 80%-100%, and control the temperature at 16-30°C.

[0033] Step (6), hardening and potting the seedlings: On the 13th to 20th day, the base and slightly upper part of the blueberry branch segments began to take root, and on the 30th day, a total of 223 branch segments took root, with a rooting rate of 91.39% and a seedling success rate of 743%. When the leaves at the top of the seedlings began to unfold after taking root, the plastic film that closed the mouth of the flower pot was gradually opened, and the two ends of the small arch shed were opened simultaneously. After hardening the seedlings for 25 to 30 days, the rooted blueberry seedlings were directly placed in the nutrient pots and shaded for care.

[0034] The present invention has the following advantages: (1) Reducing the cost of raising seedlings: The substrate used in the present invention is soil, and the price of soil is much lower than that of moss and peat. At the same time, no greenhouse and seedling rack are needed, which is equivalent to deducting the cost of the greenhouse and seedling rack. The price of small arch sheds and flower pots is relatively low, which can greatly reduce the cost of raising blueberry seedlings. (2) Improving the utilization rate of test tube seedlings: The length of the branch segment of tissue culture test tube seedlings commonly used in the conventional method of raising blueberry seedlings is 5-10 cm, while the length of the branch segment of the present invention is 0.8-1.5 cm. Therefore, the present invention increases the utilization rate of the branch segment of blueberry test tube seedlings by 6-12 times, greatly improving the efficiency of blueberry tissue culture seedlings. (3) The humidity of the substrate and the air are easier to control: On the one hand, the blueberry test tube seedlings are spread flat on the surface of the substrate, and water can be sprayed on the substrate frequently to maintain a high humidity of the substrate. The high humidity of the substrate will not affect rooting, avoiding the problem of low rooting rate caused by the blackening of the part where the blueberry branches are inserted into the substrate due to excessive humidity of the substrate; on the other hand, flower pots are used as seedling containers, and the mouth of the flower pots is tightly sealed with white plastic film to ensure a high air humidity in the flower pots. (4) The temperature is easier to control: A small arch shed is used in combination with a closed flower pot. If the temperature is too high, the cutting temperature can be adjusted by ventilating the small arch shed, sprinkling water inside and outside the small arch shed, covering the roof or removing the shade net. Temperature control is quick and easy.

[0035] Comparative Example 1

[0036] Comparative Example 1 is a method for rooting blueberries in a bottle using a traditional method, comprising the following steps:

[0037] Step (1), building a seedling greenhouse: building a plastic greenhouse, covering the roof with a layer of sunshade net, and pulling another layer of sunshade net inside the greenhouse.

[0038] Step (2), build a seedling rack: the seedling rack is 12m long, 1.8m wide and 0.8m high, and a plastic film is built on the seedling rack to form an arch shed.

[0039] Step (3), select the seedling container and substrate: select a 70-hole plug tray as the seedling container and moss as the cutting substrate.

[0040] Step (4), test tube seedling cuttings: 30 test tube seedling branches are provided, each branch is 5-10 cm long. When cutting, the base of the seedling is clamped with tweezers, dipped in 1000 mg / L NAA and inserted into the moss, with an insertion depth of 1.0-1.5 cm; after the plug tray is full, the plug tray is placed on the seedling rack, sprayed with 500-1000 times carbendazim solution, covered with arch film, and disinfected with 500-1000 times carbendazim and thiophanate-methyl solution alternately every 7 days after cutting.

[0041] Step (5), temperature and humidity control: the temperature in the arch shed is controlled below 35°C, and the air humidity is controlled at 80%-100%.

[0042] Step (6), rooting and transplanting: roots began to emerge at the base and slightly above the blueberry branch segment on the 13th day. On the 30th day, a total of 26 branches took root, with a rooting rate of 86.67%. The blueberry seedlings can be directly placed in the nutrient pot for care for two months.

[0043] Comparative Example 2

[0044] Comparative Example 2 is a method for rooting blueberries in a bottle using a traditional method, comprising the following steps:

[0045] Step (1), building a seedling greenhouse: building a plastic greenhouse, covering the roof with a layer of sunshade net, and pulling another layer of sunshade net inside the greenhouse.

[0046] Step (2), build a seedling rack: the seedling rack is 12m long, 1.8m wide and 0.8m high, and a plastic film is built on the seedling rack to form an arch shed.

[0047] Step (3), select the seedling container and substrate: select a 42×42×5 cm square plate as the seedling container, and the cutting substrate is peat + perlite (mixed in a ratio of 10:1).

[0048] Step (4), test tube seedling cuttings: 30 test tube seedling branches are provided, each branch is 5-10 cm long. When cutting, the base of the seedling is clamped with tweezers, dipped in 1000 mg / L NAA and inserted into the matrix, with an insertion depth of 1.0-1.5 cm; after the cuttings are full, the square plate is placed on the seedling rack, sprayed with 500-1000 times carbendazim solution, covered with arch film, and disinfected with 500-1000 times carbendazim and thiophanate-methyl solution alternately every 7 days or so after cutting.

[0049] Step (5), temperature and humidity control: the temperature in the arch shed is controlled below 35°C, and the air humidity is controlled at 80%-100%.

[0050] Step (6), rooting and transplanting: roots began to emerge at the base and slightly above the blueberry branch segment on the 15th day. On the 30th day, 25 branches had taken root, with a rooting rate of 83.33%. The blueberry seedlings were directly placed in the nutrient pots for care for two months.

[0051] The utilization rates and seedling rates of the blueberry test tube seedlings of Example 1, Comparative Example 1 and Comparative Example 2 are shown in Table 1.

[0052]

[0053] Table 1

[0054] By comparing the blueberry test tube seedling branch segment utilization rate, rooting rate and seedling rate of Example 1, Comparative Example 1 and Comparative Example 2, Example 1 cuts 1 test tube seedling into 6-12 branch segments for cutting, and 30 test tube seedling branches are cut into 244 segments, with a branch segment utilization rate of 813%, which is equivalent to a large number of propagation. On the 30th day, a total of 223 seedlings took root, and the seedling rate was 743.33%. Comparative Examples 1 and Comparative Examples 2 only used 1 test tube seedling as 1 branch segment for cutting, without propagation, and the seedling rates were 86.67% and 83.33% respectively. Therefore, the test tube seedling branch segment utilization rate of Example 1 of the present invention is increased by 713% compared with the traditional seedling raising method Comparative Examples 1 and Comparative Examples 2, and the seedling rates are increased by 656.66% and 660% respectively.

[0055] The invention is a set of brand-new blueberry seedling raising technology explored on the basis of traditional seedling raising technology, and the invention has the following obvious effects: first, the branch segment of tissue culture test tube seedlings commonly used in blueberry seedling raising is 5-10 cm, while that of the invention is 0.8-1.5 cm, so the invention improves the utilization rate of the branch segment of the blueberry test tube seedling by 6-12 times, and greatly improves the efficiency of blueberry tissue culture seedling raising; second, the substrate used in the invention is soil, and the price of soil is much lower than that of moss and peat, and no greenhouse and seedling raising frame are needed, which is equivalent to deducting the cost of the greenhouse and the seedling raising frame, and the price of the small arch shed and the flower pot is relatively low, so the cost of blueberry seedling raising can be greatly reduced; third, the substrate humidity and air humidity of the invention are easier to control. On the one hand, the blueberry test tube seedling branches are laid flat on the surface of the substrate, and water can be sprayed on the substrate to maintain a high humidity of the substrate, thereby avoiding the problem of low rooting rate caused by blackening of the part where the blueberry branches are inserted into the substrate due to excessive humidity of the substrate. On the other hand, a flower pot is used as a seedling container, and the mouth of the flower pot is tightly sealed with a white plastic film to ensure a high air humidity in the flower pot. Fourth, the temperature of the present invention is easier to control. By using a small arch shed combined with a closed flower pot, if the temperature is too high, the cutting temperature can be adjusted by ventilating the small arch shed, sprinkling water inside and outside the small arch shed, covering the roof or removing the sunshade net, and the temperature control is quick, simple and easy.

[0056] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for rooting blueberry tissue culture seedlings outside a bottle, characterized in that: The steps include: Step (1), seedbed preparation: the seedbed in the flowerpot is laid to a thickness of 10-20 cm and the substrate is disinfected; Step (2), cutting the seedlings: taking the blueberry test tube seedlings out of the culture bottle, cutting them into short branches with at least one axillary bud and a length of 0.8-1.5 cm, and soaking the blueberry short branches in a 500-1000 times methyl thiophanate solution for 8-15 minutes; Step (3), paving: dip the blueberry short branches into NAA and spread them on the surface of the seedbed in the flowerpot, spray disinfectant with 500-1000 times methyl thiophanate solution, and then seal the flowerpot tightly with white plastic film; Step (4), accelerating seedlings: after 7 days of tiling, spray the blueberry short branches with a 10%-25% WPM basic culture medium solution containing 2g of methyl thiophanate powder per liter of solution to accelerate seedlings and disinfect, and then tightly seal the flowerpot with a white plastic film; thereafter, spray the blueberry branches with a 10%-25% WPM basic culture medium solution containing 2g of methyl thiophanate powder per liter of solution every 5 days to accelerate seedlings and disinfect.

2. A method for rooting blueberry tissue culture seedlings outside a bottle according to claim 1, characterized in that: In the step (1), the seedling medium is soil, and the flower pot is filled with soil as a seedbed. The thickness of the soil in the flower pot is 10-20 cm, and the soil is sprayed with 200-500 times mancozeb solution to disinfect the medium.

3. A method for rooting blueberry tissue culture seedlings outside a bottle according to claim 1, characterized in that: In the step (2), each test tube seedling with a proliferation period of 45 days can be cut into 6-12 short branch segments of 0.8-1.5 cm.

4. A method for rooting blueberry tissue culture seedlings outside a bottle according to claim 1, characterized in that: The method also includes step (5), temperature and humidity control: placing the flower pots in a small arch shed, covering the top of the small arch shed with a sunshade net with a shading rate of 60%, and spraying water on the substrate and blueberry branches in the flower pots every day after cutting to control the air humidity in the flower pots at 80%-100% and the temperature at 16-30°C by adjusting the opening and closing of the small arch shed and spraying water into the small arch shed.

5. A method for rooting blueberry tissue culture seedlings outside a bottle according to claim 4, characterized in that: The method also includes step (6), hardening the seedlings and placing them in pots for care: when the leaves at the top of the seedlings begin to unfold after taking root, gradually open the plastic film that seals the mouth of the flowerpot, and simultaneously open the two ends of the small arch shed. After hardening the seedlings for 25-30 days, place the rooted blueberry seedlings directly in the nutrient pots and provide shade for care.

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