CN110667908B - Tissue culture seedling packaging method suitable for long-distance transportation - Google Patents
Tissue culture seedling packaging method suitable for long-distance transportation Download PDFInfo
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- CN110667908B CN110667908B CN201910997781.9A CN201910997781A CN110667908B CN 110667908 B CN110667908 B CN 110667908B CN 201910997781 A CN201910997781 A CN 201910997781A CN 110667908 B CN110667908 B CN 110667908B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B25/00—Packaging other articles presenting special problems
- B65B25/02—Packaging agricultural or horticultural products
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B51/00—Devices for, or methods of, sealing or securing package folds or closures; Devices for gathering or twisting wrappers, or necks of bags
- B65B51/04—Applying separate sealing or securing members, e.g. clips
- B65B51/06—Applying adhesive tape
- B65B51/067—Applying adhesive tape to the closure flaps of boxes
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B55/00—Preserving, protecting or purifying packages or package contents in association with packaging
- B65B55/02—Sterilising, e.g. of complete packages
- B65B55/04—Sterilising wrappers or receptacles prior to, or during, packaging
- B65B55/10—Sterilising wrappers or receptacles prior to, or during, packaging by liquids or gases
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B61/00—Auxiliary devices, not otherwise provided for, for operating on sheets, blanks, webs, binding material, containers or packages
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B61/00—Auxiliary devices, not otherwise provided for, for operating on sheets, blanks, webs, binding material, containers or packages
- B65B61/20—Auxiliary devices, not otherwise provided for, for operating on sheets, blanks, webs, binding material, containers or packages for adding cards, coupons or other inserts to package contents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B61/00—Auxiliary devices, not otherwise provided for, for operating on sheets, blanks, webs, binding material, containers or packages
- B65B61/20—Auxiliary devices, not otherwise provided for, for operating on sheets, blanks, webs, binding material, containers or packages for adding cards, coupons or other inserts to package contents
- B65B61/22—Auxiliary devices, not otherwise provided for, for operating on sheets, blanks, webs, binding material, containers or packages for adding cards, coupons or other inserts to package contents for placing protecting sheets, plugs, or wads over contents, e.g. cotton-wool in bottles of pills
Abstract
The invention discloses a tissue culture seedling packaging method suitable for long-distance transportation, which comprises the following steps: (1) preparing a culture medium; (2) grading and transferring the tissue culture seedlings; (3) packaging: laying a polyethylene or polypropylene film with good light transmittance in a foam box, placing the tissue culture seedling in the foam box, gathering the film and tightening; placing ice bags wrapped by newspapers at four corners of the foam box, and sealing after tightly covering the foam box; then the foam box is placed in a corrugated paper box for sealing and packaging; (4) and (4) storing and transporting. The invention adopts the mode of carrying the culture medium to carry out primary packaging, grope and screen the components of the culture medium and the used containers, and adopts the modes of wrapping ice bags, multilayer sealing and the like to maintain the low-temperature environment in the transportation process, thereby obviously reducing the influence of long-distance transportation on the quality of the seedlings and the later survival rate, effectively reducing the pollution rate, being convenient for inspection and quarantine, and effectively reducing the production cost.
Description
Technical Field
The invention relates to the technical field of agricultural planting, in particular to a tissue culture seedling packaging method suitable for long-distance transportation.
Background
The tissue culture seedling is a plant seedling which is obtained by utilizing a part of a plant body under the condition of manual control and has the characteristics of no influence of natural conditions, high production efficiency, good seedling consistency and the like, and is one of important seedling types in the aspects of modern horticulture, endangered plant protection, new excellent plant resource development and utilization and the like. However, the tissue culture seedlings are tender, the storage nutrients of the plant roots, stems, leaves and other nutritive organs are less, and the growth environment of the seedlings after being taken out of the bottle is changed from a high-humidity and sterile environment to a low-humidity and bacteria-bearing environment, so that the seedlings are easy to die, infect bacteria, grow badly and the like in the long-distance transportation process. Therefore, how to effectively solve the problems occurring in the long-distance transportation process of the tissue culture seedlings is a key problem for promoting the development of the flower industry including orchids.
With the development of open to the outside and international trade in China in recent years, the number of flower seedlings and germplasm resources for entering and exiting the border is increased, and meanwhile, the risk of pest transmission is increased. The inspection, quarantine, supervision and management work of the imported and exported flower seedlings also becomes the key point of the plant inspection and quarantine work. Quarantine of various types of imported plants including tissue culture seedlings is very strict in countries all over the world, particularly in the united states, canada, australia, korea, japan, and the like. However, because the production level, the management level and the pest control level of flowers and plants in China are different, and the rules and methods of plant quarantine in international trade are not sufficiently known by seedling export enterprises, the export is limited due to plant quarantine frequently. On the basis of knowing the plant inspection and quarantine rules, the method has great practical significance on reasonably and scientifically packaging exported nursery stocks so as to realize smooth customs clearance.
Disclosure of Invention
The invention overcomes the technical problems that in the prior art, because the tissue culture seedling is tender and has less nutrient storage nutrients of plant roots, stems, leaves and other nutritive organs, and the growth environment of the seedling is changed from a high-humidity and sterile environment to a low-humidity and bacteria-bearing environment after the seedling is taken out of a bottle, the seedling is easy to die, infect bacteria, grow badly and the like in the long-distance transportation process, and provides the packaging method of the tissue culture seedling with the culture medium, which is suitable for long-distance transportation.
In order to solve the problems, the invention adopts the following technical scheme:
a long-distance transportation and packaging method of tissue culture seedlings with culture media comprises the following steps:
(1) preparing a culture medium: preparing 1/2MS culture medium according to a set formula, subpackaging into specific containers, and sterilizing the containers with high pressure steam; after the sterilization is finished, placing the container in a clean and cool environment to solidify the culture medium for later use; the 1/2MS culture medium is prepared by the following method: reducing the dosage of macroelements in the MS culture medium to one half of the original dosage, removing sucrose components, adding 7.0-8.0 g/L of agar and 0.3-0.5 mg/L of Yipeilong bacteriostat, and adjusting the pH value to 6.0-7.0 to obtain the culture medium;
(2) grading and transferring the tissue culture seedlings: transferring the tissue culture seedlings after the rooting stage to the standby culture medium in the step (1) in an aseptic environment of a superclean workbench, inserting the roots of the tissue culture seedlings into the culture medium, and grading according to the sizes of plants; after the transfer is finished, the tissue culture seedlings can be packaged and transported, and can also be temporarily placed in a culture room;
(3) packaging: laying a polyethylene or polypropylene film with good light transmittance in a clean foam box, placing the tissue culture seedling subjected to the transfer in the step (2) in the foam box, and gathering and tightening the film; placing ice bags at four corners of the foam box, tightly covering the cover of the foam box, and sealing the joint of the cover and the box body by using transparent adhesive tapes; then the foam box is placed in a corrugated paper box for sealed packaging;
(4) storage and transportation: during storage and transportation, the carton is placed in a shady and low-temperature environment.
The container is a transparent high-temperature and high-pressure resistant polypropylene plastic box, and is in a round bowl shape, the diameter of the bottom of the container is 10-14 cm, the diameter of the top of the container is 15-22 cm, and the height of the container is 5-10 cm. The selection can be carried out according to the height of the target seedling.
Wherein, the ice bag put into the packing box needs newspaper to wrap.
Wherein the thickness of the plastic film is 5-8 threads, and the thickness of the culture medium in the container is 1 +/-0.2 cm.
Wherein, in the step (4), the environment with low shading temperature is as follows: 4-8 ℃.
Wherein in the step (1), the sterilization temperature of the high-pressure steam sterilization is 121-130 ℃, and the sterilization time is 25-30 min.
The present invention is a method of inserting tissue culture seedlings into a culture medium for packaging and transportation, rather than transporting bare-rooted seedlings. In the step (1), the culture medium is added with the bacteriostatic agent and is free of sucrose, and technical parameters such as agar dosage, pH and the like are adjusted, so that the quality of transported seedlings can be ensured, and the cost can be effectively reduced.
The long-distance transportation and packaging method of the tissue culture seedling culture medium can be applied to most tissue culture plants, and is particularly suitable for orchids such as paphiopedilum spicatum, dendrobium officinale and the like.
The technical invention has the following characteristics:
the delivery of the tissue culture seedlings is carried out in a mode with the culture medium instead of the delivery of the bare-rooted seedlings, so that the water evaporation and the obvious reduction of the relative humidity (from nearly 100% in a culture bottle to the atmospheric relative humidity) in the transportation link can be effectively reduced, and the advantage is more obvious in the long-distance transportation process; secondly, the proper culture medium formula is designed, so that the pollution rate is reduced and the growth of seedlings is guaranteed in the transportation process; thirdly, the special containers are screened, and the adopted polypropylene containers have the characteristics of light transmission, large capacity, light weight and high temperature resistance. The light-transmitting characteristic is beneficial to the growth of the tissue culture seedlings in a period of time before packaging and after unpacking; the capacity is large, the weight is light, and the cost of long-distance transportation can be reduced; high temperature resistance, and can meet the requirement of medium sterilization (121 ℃). Fourthly, the method has the characteristics of low production cost and convenient operation. According to the invention, a large container with light weight is selected for loading, 1/2MS culture medium is selected, and the subpackaging amount (the thickness is 0.8-1.2cm) is controlled, so that the production and transportation cost can be effectively reduced, and the method is simple and convenient to operate and convenient to popularize and apply. A scientific and practical packaging mode is designed, three layers of plastic films, a foam box and a paper box are adopted for packaging, and an ice bag wrapping newspaper is used for reducing the temperature of the environment in the foam box, so that the survival time of transported tissue culture seedlings is prolonged. The whole process flow and the specific technical measures in the technical invention are suitable for long-distance transportation of most tissue culture seedlings.
Compared with the prior art, the invention has the following beneficial effects:
(1) compared with bare-rooted seedling transportation, the prior art obviously improves the quality of tissue culture seedlings and is more suitable for long-distance transportation. The transportation of the bare-rooted seedlings, namely the transportation mode without culture medium, is also the main mode of the transportation of the tissue culture seedlings at present. The bare-rooted seedling is transported, and the tissue culture seedling is directly transferred into a low-humidity sterile environment from a sterile environment with the humidity close to 100%, so that the growth vigor and the survival rate of the seedling are greatly influenced. The original growth state of the seedlings is maintained by the transportation mode with the culture medium, the influence of environmental factors in the transportation link on the tissue culture seedlings can be reduced exactly, and good growth vigor is maintained. Especially, the advantage of the transportation mode with the culture medium is more obvious for long-distance transportation.
(2) Through the improvement of technical measures, the pollution problem in the transportation process can be effectively reduced, and the good growth potential of the seedlings is maintained. The Yipelong bacteriostatic agent is added on the conventional MS basic culture medium without adding cane sugar, so that the breeding of fungi is effectively reduced, and the bacterial infection is prevented. The Yipelong is a long-acting, broad-spectrum, high-activity, novel anti-pollution bactericide special for tissue culture, has strong inhibiting and killing effects on common bacteria, fungi, algae and the like, can effectively inhibit fungi carried by original seedlings by adding the Yipelong, and can prevent the breeding of the fungi in the transportation process. In addition, by adjusting the content of major elements, the agar dosage and the pH value, the culture medium is kept to have certain hardness on the premise of ensuring the nutrition required by the normal growth of the seedlings, so as to prevent the loose or flowing of the matrix in the transportation process from influencing the state or survival of the seedlings.
(3) Effectively reducing the production cost. The invention adopts 1/2MS minimal medium, which reduces the macroelement in MS medium by half, and can reduce the production cost while ensuring the need of the tissue culture seedling for nutrient element in the transportation process; meanwhile, sucrose is not added into the culture medium, and the split charging amount of the culture medium is controlled, so that the production cost can be directly reduced; in addition, the transportation cost can be effectively reduced by adopting a large container with lighter weight to load the container.
(4) The invention is suitable for long-distance transportation of tissue culture seedlings. The invention designs a scientific and practical packaging mode, adopts a plastic film, a foam box and a corrugated case to carry out three-layer packaging, and uses an ice bag wrapping newspaper to maintain the low-temperature environment in the foam box, thereby being beneficial to keeping good growth of seedlings and prolonging the survival time of the seedlings in the transportation process; meanwhile, the seedlings are prevented from being damaged by direct contact of the ice bags through newspaper wrapping. The whole process flow and the specific technical measures in the invention are suitable for long-distance transportation of most tissue culture seedlings, and are particularly suitable for orchids such as paphiopedilum albolanum and dendrobium officinale.
(5) The seedling culture packaging method is convenient for customs inspection and is beneficial to import and export trade of flower seedlings. When the seedlings are inspected and quarantined, the seedlings are often distinguished according to the appearance of the material, and then suspected plants are collected and taken into a laboratory for further inspection. According to the invention, the potential fungus pollution is effectively prevented and inhibited by measures such as adding the bacteriostatic agent Yipeilong and not adding the cane sugar; the three-layer sealed package and the low temperature environment also help to maintain a clean environment during the packaging process. In addition, the transparent plastic container and the culture medium substrate are beneficial to pollution inspection of producers and convenient for inspection work of customs personnel during customs clearance.
Detailed Description
The present invention will be further described with reference to examples and tests.
Example 1
A tissue culture seedling packing method suitable for long-distance transportation comprises the following steps:
(1) preparing a culture medium: preparing 1/2MS culture medium according to a set formula, subpackaging into specific containers, and sterilizing the containers with high pressure steam; after the sterilization is finished, placing the container in a clean and cool environment to solidify the culture medium for later use; the 1/2MS culture medium is prepared by the following method: reducing the dosage of macroelements in the MS culture medium to one half of the original dosage, removing sucrose components, adding 8.0g/L of agar and 0.3mg/L of Yipeilong bacteriostat, and adjusting the pH value to 7.0 to obtain the culture medium; the container is a transparent high-temperature and high-pressure resistant polypropylene plastic box, and is in a circular bowl shape, the diameter of the bottom of the container is 14cm, the diameter of the top of the container is 15cm, and the height of the container is 10 cm; the thickness of the culture medium in the container is 1.2 cm; the sterilization temperature of the high-pressure steam sterilization is 130 ℃, and the sterilization time is 25 min; the formulation of the 1/2MS culture medium is as follows: NH (NH) 4 NO 3 825mg/L、KNO 3 950mg/L、CaCl 2 ·2H 2 O 220mg/L、MgSO 4 ·7H 2 O 185mg/L、KH 2 PO 4 85mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 ·4H 2 O 22.3mg/L、ZnSO 4 ·7H 2 O 8.6mg/L、Na 2 MnO 4 ·2H 2 O 0.25mg/L、CuSO 4 ·5H 2 O 0.25mg/Lmg/L、CaCl 2 ·6H 2 O 0.025mg/L、FeSO 4 ·7H 2 O 27.85mg/L、Na 2 -EDTA·2H 2 37.3mg/L of O, 100mg/L of inositol, 0.5mg/L of nicotinic acid, 0.5mg/L of pyridoxine hydrochloride (vitamin B6), 0.1mg/L of thiamine hydrochloride (vitamin B1), 2.0mg/L of glycine, 8.0g/L of agar, 0.3mg/L of Yipeong bacteriostat and the pH value of 7.0;
(2) grading and transferring the tissue culture seedlings: transferring the tissue culture seedlings after the rooting stage to the standby culture medium in the step (1) in an aseptic environment of a superclean workbench, inserting the roots of the tissue culture seedlings into the culture medium, and grading according to the sizes of plants; after the transfer is finished, the tissue culture seedlings can be packaged and transported, and can also be temporarily placed in a culture room;
(3) packaging: laying a polypropylene film with good light transmittance in a clean foam box, placing the tissue culture seedlings subjected to the transfer in the step (2) in the foam box, and gathering and tightening the film; placing ice bags packaged by newspaper at four corners of the foam box, tightly covering the cover of the foam box, and sealing the joint of the cover and the box body by using transparent adhesive tapes; then placing the foam box in a corrugated paper box for sealing and packaging; the thickness of the plastic film is 5 filaments;
(4) storage and transportation; during storage and transportation, the carton is placed in a shady and low-temperature environment at 8 ℃.
Example 2
A tissue culture seedling packing method suitable for long-distance transportation comprises the following steps:
(1) preparing a culture medium: preparing 1/2MS culture medium according to a set formula, subpackaging into specific containers, and sterilizing the containers with high pressure steam; after the sterilization is finished, placing the container in a clean and cool environment to solidify the culture medium for later use; the 1/2MS culture medium is prepared by the following method: reducing the dosage of macroelements in the MS culture medium to one half of the original dosage, removing sucrose components, adding 7.0g/L of agar and 0.5mg/L of Yipeilong bacteriostat, and adjusting the pH value to 6.0 to obtain the culture medium; the container is a transparent high-temperature and high-pressure resistant polypropylene plastic box, and is in a circular bowl shape, the diameter of the bottom of the container is 14cm, the diameter of the top of the container is 15cm, and the height of the container is 10 cm; the thickness of the culture medium in the container is 0.8 cm; the sterilization temperature of the high-pressure steam sterilization is 121 ℃, and the sterilization time is 30 min; the formulation of the 1/2MS culture medium is as follows: NH (NH) 4 NO 3 825mg/L、KNO 3 950mg/L、CaCl 2 ·2H 2 O 220mg/L、MgSO 4 ·7H 2 O 185mg/L、KH 2 PO 4 85mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 ·4H 2 O 22.3mg/L、ZnSO 4 ·7H 2 O 8.6mg/L、Na 2 MnO 4 ·2H 2 O 0.25mg/L、CuSO 4 ·5H 2 O 0.25mg/Lmg/L、CaCl 2 ·6H 2 O 0.025mg/L、FeSO 4 ·7H 2 O 27.85mg/L、Na 2 -EDTA·2H 2 37.3mg/L of O, 100mg/L of inositol, 0.5mg/L of nicotinic acid, 0.5mg/L of pyridoxine hydrochloride (vitamin B6), 0.1mg/L of thiamine hydrochloride (vitamin B1), 2.0mg/L of glycine, 7.0g/L of agar, 0.5mg/L of Yipeong bacteriostat and the pH value of 6.0;
(2) grading and transferring the tissue culture seedlings: transferring the tissue culture seedlings after the rooting stage to the standby culture medium in the step (1) in an aseptic environment of a superclean workbench, inserting the roots of the tissue culture seedlings into the culture medium, and grading according to the sizes of plants; after the transfer is finished, the tissue culture seedlings can be packaged and transported, and can also be temporarily placed in a culture room;
(3) packaging: laying a polyethylene film with good light transmittance in a clean foam box, placing the tissue culture seedlings subjected to the transfer in the step (2) in the foam box, and gathering and tightening the film; placing ice bags packaged by newspaper at four corners of the foam box, tightly covering the cover of the foam box, and sealing the joint of the cover and the box body by using transparent adhesive tapes; then placing the foam box in a corrugated paper box for sealing and packaging; the thickness of the plastic film is 8 filaments;
(4) storage and transportation; during storage and transportation, the carton is placed in a shady and low-temperature environment at 4 ℃.
Example 3
A tissue culture seedling packing method suitable for long-distance transportation comprises the following steps:
(1) preparing a culture medium: preparing 1/2MS culture medium according to a set formula, subpackaging into specific containers, and sterilizing the containers with high pressure steam; after the sterilization is finished, placing the container in a clean and cool environment to solidify the culture medium for later use; the 1/2MS culture medium is prepared by the following method: reducing the dosage of macroelements in the MS culture medium to one half of the original dosage, removing sucrose components, adding 7.5g/L of agar and 0.4mg/L of Yipeilong bacteriostat, and adjusting the pH value to 6.5 to obtain the culture medium; the container is transparentThe high-temperature and high-pressure resistant polypropylene plastic box is in a round bowl shape, the diameter of the bottom of the box is 12cm, the diameter of the top of the box is 20cm, and the height of the box is 8 cm; the thickness of the culture medium in the container is 1 cm; the sterilization temperature of the high-pressure steam sterilization is 125 ℃, and the sterilization time is 28 min; the formulation of the 1/2MS culture medium is as follows: NH (NH) 4 NO 3 825mg/L、KNO 3 950mg/L、CaCl 2 ·2H 2 O 220mg/L、MgSO 4 ·7H 2 O 185mg/L、KH 2 PO 4 85mg/L、KI 0.83mg/L、H 3 BO 3 6.2mg/L、MnSO 4 ·4H 2 O 22.3mg/L、ZnSO 4 ·7H 2 O 8.6mg/L、Na 2 MnO 4 ·2H 2 O 0.25mg/L、CuSO 4 ·5H 2 O 0.25mg/Lmg/L、CaCl 2 ·6H 2 O 0.025mg/L、FeSO 4 ·7H 2 O 27.85mg/L、Na 2 -EDTA·2H 2 37.3mg/L of O, 100mg/L of inositol, 0.5mg/L of nicotinic acid, 0.5mg/L of pyridoxine hydrochloride (vitamin B6), 0.1mg/L of thiamine hydrochloride (vitamin B1), 2.0mg/L of glycine, 7.5g/L of agar, 0.4mg/L of Yipeilong bacteriostat and 6.5 of pH value;
(2) grading and transferring the tissue culture seedlings: transferring the tissue culture seedlings after the rooting stage to the standby culture medium in the step (1) in an aseptic environment of a superclean workbench, inserting the roots of the tissue culture seedlings into the culture medium, and grading according to the sizes of plants; after the transfer is finished, the tissue culture seedlings can be packaged and transported, and can also be temporarily placed in a culture room;
(3) packaging: laying a polyethylene film with good light transmittance in a clean foam box, placing the tissue culture seedlings subjected to the transfer in the step (2) in the foam box, and gathering and tightening the film; placing ice bags packaged by newspaper at four corners of the foam box, tightly covering the cover of the foam box, and sealing the joint of the cover and the box body by using transparent adhesive tapes; then placing the foam box in a corrugated paper box for sealing and packaging; the thickness of the plastic film is 7 filaments;
(4) storage and transportation; during storage and transportation, the carton is placed in a shady and low-temperature environment at 6 ℃.
Control group 1
The way of transporting the tissue culture seedlings of control 1 was different from that of example 1 in that the tissue culture seedlings of control 1 were placed in a plastic container without a culture medium, and a wet paper towel was laid under the container. Namely: the improved bare-rooted seedlings are transported.
The mode of transporting the tissue culture seedlings of the control group 2 is basically the same as that of the example 1, except that the culture medium adopted by the control group 2 is not added with the Yipelong bacteriostat and contains sucrose, the dosage of the sucrose is 30g/L, and the rest components and the content thereof are the same as those of the 1/2MS culture medium of the example 1.
Control group 3
The mode of transporting the tissue culture seedlings of the control group 3 was substantially the same as that of example 1, except that in the step (3) packaging of the control group 3, 4 wrapped ice bags were not placed at the four corners of the foam box, and two wrapped ice bags were placed only at the opposite two corners.
Control group 4
The way of transporting the tissue culture seedlings of the control group 4 was substantially the same as that of example 1, except that the agar content in the medium formulation used in the control group 4 was 5.5g/L, and the remaining components and their contents were the same as those in the 1/2MS medium of example 1.
Control group 5
The transportation mode of the tissue culture seedlings of the control group 5 is basically the same as that of the example 1, except that the culture medium formula adopted by the control group 5 is MS culture medium, and the split charging thickness of the culture medium is 1.5 cm.
1. Transport tissue culture seedling test
And simulating the long-distance transportation environment for paphiopedilum albolanum and dendrobium officinale according to the tissue culture seedling transportation modes provided by the embodiments 1 to 3 and the comparison groups 1 to 5, shading indoors, storing at normal temperature for 10 days, and transplanting. The growth conditions of the tissue culture seedlings after unpacking and the survival rates after transplanting are shown in the following table 1.
TABLE 1 growth vigor and survival rate of transplanted seedlings after normal temperature preservation for 10 days
As can be seen from Table 1, the group 1 cultured seedlings of the control group suffered from severe wilting, poor growth potential and low survival rate after transplantation, which is related to water loss during storage; the control group 2 has obvious pollution to seedling culture bacteria and fungi, and seriously influences the quality of seedlings and the survival rate in the later period; the phenomenon of yellow leaves and excessive growth of the tissue culture seedlings in the control group 3 indicates that the two ice bags are not enough to maintain the low-temperature environment within 10 days, and further the quality and the survival rate of the seedlings are influenced. The culture medium of the control group 4 is easy to loose and overturn, and is not beneficial to transportation, thereby influencing the consistency and survival rate of tissue culture seedlings. The growth of the tissue culture seedlings in the control group 5 was good, but the production cost and the transportation cost were significantly increased. The leaves of the tissue culture seedlings of the embodiments 1 to 3 are light green, do not wither, have good growth vigor and high survival rate (more than 98%), and the production cost of the embodiments 1 to 3 is low. The tissue culture seedling packing method can ensure that the tissue culture seedlings keep good growth potential in the long-distance transportation process (more than 10 days), and has outstanding technical effects.
The above description is intended to describe in detail the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the claims of the present invention, and all equivalent changes and modifications made within the technical spirit of the present invention should fall within the scope of the claims of the present invention.
Claims (2)
1. A tissue culture seedling packing method suitable for long-distance transportation is characterized by comprising the following steps:
(1) preparing a culture medium: preparing 1/2MS culture medium according to a set formula, subpackaging into specific containers, and sterilizing the containers with high pressure steam; after the sterilization is finished, placing the container in a clean and cool environment to solidify the culture medium for later use; the 1/2MS culture medium is prepared by the following method: reducing the dosage of macroelements in the MS culture medium to one half of the original dosage, removing sucrose components, adding 7.0-8.0 g/L of agar and 0.3-0.5 mg/L of Yipeilong bacteriostat, and adjusting the pH value to 6.0-7.0 to obtain the culture medium;
(2) grading and transferring the tissue culture seedlings: transferring the tissue culture seedlings after the rooting stage to the standby culture medium in the step (1) in an aseptic environment of a superclean workbench, inserting the roots of the tissue culture seedlings into the culture medium, and grading according to the sizes of plants; after the transfer is finished, the tissue culture seedlings can be packaged and transported, and can also be temporarily placed in a culture room;
(3) packaging: laying a polyethylene or polypropylene film with good light transmittance in a clean foam box, placing the tissue culture seedling subjected to the transfer in the step (2) in the foam box, and gathering and tightening the film; placing ice bags at four corners of the foam box, tightly covering the cover of the foam box, and sealing the joint of the cover and the box body by using transparent adhesive tapes; then the foam box is placed in a corrugated paper box for sealing and packaging;
(4) storage and transportation; during storage and transportation, the carton is placed in a shady and low-temperature environment;
the container is a transparent, high-temperature and high-pressure resistant polypropylene plastic box and is in a round bowl shape; the diameter of the bottom of the container is 10-14 cm, the diameter of the top of the container is 15-22 cm, and the height of the container is 5-10 cm;
the ice bag put into the foam box needs to be wrapped by newspaper;
the thickness of the film is 5-8 filaments, and the thickness of the culture medium in the container is 1 +/-0.2 cm;
in the step (4), the shady low-temperature environment is as follows: 4-8 ℃.
2. The method for packaging tissue culture seedlings suitable for long-distance transportation according to claim 1, wherein in the step (1), the sterilization temperature of the high-pressure steam sterilization is 121-130 ℃, and the sterilization time is 25-30 min.
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