CN113853949A - Method for collecting fruit tree pollen germplasm resources under field condition and sealing box - Google Patents
Method for collecting fruit tree pollen germplasm resources under field condition and sealing box Download PDFInfo
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- CN113853949A CN113853949A CN202111086819.0A CN202111086819A CN113853949A CN 113853949 A CN113853949 A CN 113853949A CN 202111086819 A CN202111086819 A CN 202111086819A CN 113853949 A CN113853949 A CN 113853949A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01D—HARVESTING; MOWING
- A01D91/00—Methods for harvesting agricultural products
- A01D91/04—Products growing above the soil
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D25/00—Details of other kinds or types of rigid or semi-rigid containers
- B65D25/02—Internal fittings
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D81/00—Containers, packaging elements, or packages, for contents presenting particular transport or storage problems, or adapted to be used for non-packaging purposes after removal of contents
- B65D81/24—Adaptations for preventing deterioration or decay of contents; Applications to the container or packaging material of food preservatives, fungicides, pesticides or animal repellants
- B65D81/26—Adaptations for preventing deterioration or decay of contents; Applications to the container or packaging material of food preservatives, fungicides, pesticides or animal repellants with provision for draining away, or absorbing, or removing by ventilation, fluids, e.g. exuded by contents; Applications of corrosion inhibitors or desiccators
- B65D81/266—Adaptations for preventing deterioration or decay of contents; Applications to the container or packaging material of food preservatives, fungicides, pesticides or animal repellants with provision for draining away, or absorbing, or removing by ventilation, fluids, e.g. exuded by contents; Applications of corrosion inhibitors or desiccators for absorbing gases, e.g. oxygen absorbers or desiccants
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Abstract
The invention provides a method for collecting fruit tree pollen germplasm resources under a field condition and a sealing box. The sealing box provided by the invention has the characteristics of portability and sealing, and the drying agent is arranged in the inner space of the sealing box and comprises anhydrous calcium chloride or silica gel. The method for collecting the fruit tree pollen germplasm resources by using the sealing box under the field condition solves the problems of collection and activity maintenance of the fruit tree pollen germplasm resources and other plant pollen germplasm resources under the field hard condition, and can simply, conveniently, quickly and efficiently collect and maintain the activity of the fruit tree pollen under the field condition.
Description
Technical Field
The invention relates to the field of collection and preservation of fruit tree pollen germplasm resources, in particular to a method for collecting fruit tree pollen germplasm resources under a field condition and a sealing box.
Background
Plant germplasm resources are strategic resources for survival and development of human society, are important guarantees for maintaining biological safety of a line, and are material bases for plant evolution, genetic research and breeding. The plant germplasm resources mainly refer to the population of the plant, and the small germplasm resources refer to plants, roots, stems, seeds, pollen and the like, and are very important for research on collection, preservation and utilization of plant germplasm resources. However, with the rapid development of urbanization and industrialization, the safety of fruit tree germplasm resources is still seriously threatened, especially the loss of wild kindred species is very striking, and the problem of collection, protection and utilization of germplasm resources is firstly solved by developing seed source technology, so that the significance of collection and storage of local varieties and wild fruit tree germplasm resources is enhanced. The pollen of the fruit tree is an important carrier carrying genetic information of the fruit tree, not only contains all the genetic information of the species, but also has rich genetic diversity, and is an important material for germplasm exchange and preservation. It can not only make early-maturing and late-maturing varieties and allopatric parents realize cross breeding, but also can store various germplasm resources in a small scale range, and is a simple, convenient and efficient way for effectively utilizing precious and rare germplasm, so that the pollen is always paid attention to in the process of collecting and protecting the germplasm resources of fruit trees.
Fruit tree pollen is itself a living granule rich in starch and fat as well as certain hormones. In a natural state, the pollen life is not long, the inventor collects ten varieties of flowers by taking sweet cherries as an example, the average vitality of the flowers is lost by about 40 percent after the flowers are placed for 1 day in a room temperature environment, and 70 percent of vitality is lost after the flowers are placed for two days. Of course, the vitality of the fruit tree pollen is determined by the genetic characteristics of the fruit tree pollen, and environmental factors, particularly temperature and humidity conditions, are also main influence factors. Also taking sweet cherry as an example, pollen is stripped immediately after the flowers are collected, and the pollen is placed in a dry environment at room temperature for 1 day, so that the vitality of the pollen is only lost by about 10 percent, and 50 percent of the vitality is still maintained after the pollen is placed for 1 week. The dried pollen can maintain the original activity more than 50% after being placed for 1 week at 5 ℃, can be stored for 1 year at-80 ℃ and can be stored for a long time at-196 ℃. However, in the process of collecting the wild fruit tree germplasm resources, the work of collecting and preserving the fruit tree pollen is difficult and serious due to the severe wild environment and the rough conditions. The study of the invention finds that most flowers collected in the field can only be placed in the collecting bag firstly due to the rough conditions, and the activity loss of the flowers collected in the field under the sunny weather conditions is measured in hours in the collecting bag. The activity is lost by about 20 percent after the mixture is placed for 2 hours, and the activity is lost by more than 50 percent after the mixture is placed for 8 hours. If the flowers are rotted and deteriorated after raining or under humid conditions, the collected flowers begin to rot and deteriorate after 4-6 hours, as shown in fig. 1-2. The value and the meaning of the field pollen resource collection in the fruit tree germplasm resource collection process are greatly reduced and even become invalid. Because the collection work of the wild fruit tree germplasm resources is limited by the environment and conditions, and the time, the manpower and the material resources are limited, the pollen cannot be timely and properly treated, so that the collection and the maintenance of the activity of the fruit tree pollen in the wild environment become a difficult problem of the collection work of the germplasm resources.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for collecting fruit tree pollen germplasm resources under the field condition and a sealing box.
Specifically, the present invention provides, first, a sealing case including: and a drying agent is arranged in the inner space of the sealing box, and the drying agent comprises anhydrous calcium chloride or silica gel. The invention discovers that the invention selects light materials and drying agents with high water absorption capacity, such as anhydrous calcium chloride and the like, which are more suitable for hard conditions in the field; the sealing box is convenient to carry, the subsequent treatment is very simple and convenient, the sealing box can be placed at room temperature and can be used for direct express mailing, and the requirement for field flower treatment is reduced to the maximum extent.
According to the sealing box provided by the invention, the volume of the sealing box is 1200-1800 cm3The bottom area is 550-850 cm2(ii) a The sealed box is preferably a sealed plastic box. The flower drying device adopts the light and thin sealed plastic box, so that the flowers can be fully contacted with the drying agent, meanwhile, the flowers can be better fixed, and the friction damage to the flowers in the field collecting process can be reduced. Preferably, the sealed plastic box is an A4 sealed plastic box, and the A4 sealed plastic boxThe length X width X height of the resin composition is 31cm X22.8 cm X2.2 cm. The fruit tree pollen germplasm resources collected by one or two A4 sealed plastic boxes can meet the pollination and research dosage, and the collection efficiency of the wild fruit tree germplasm resources is greatly improved.
According to the sealing box provided by the invention, the drying agent is anhydrous calcium chloride.
According to the sealing box provided by the invention, the weight of the drying agent is 180-220 g; and/or the ratio of the drying agent to the volume is 180-220 g: 1200-1800 cm3。
According to the sealing box provided by the invention, the drying agent is divided into at least two parts, wherein at least 80-95 wt% of the drying agent can be taken out of the sealing box.
According to the sealing box provided by the invention, the desiccant is placed in the sealing box in a flat way: preferably, the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in a sealing box, and filter paper is placed on the contact surface of the nylon gauze bag and the flowers; preferably, the nylon gauze bag is 500 meshes. The invention adopts the nylon mesh bag and the filter paper to fully isolate the contact of the flowers and the drying agent, so that the friction damage and the like caused to the flowers in the field collecting and carrying process are reduced to the maximum extent, in addition, the filter paper can ensure the stable and uniform dehydration process of the drying agent to the flowers, and the influence of unbalanced drying of the flowers on the pollen activity is avoided.
The invention also provides a method for collecting fruit tree pollen germplasm resources by using the sealed box under the field condition, which comprises the step of treating the collected flowers by using the sealed box.
According to the method for collecting the fruit tree pollen germplasm resources by using the sealed box under the field condition, the flowers are tiled on the sealed box in a single-layer mode; preferably, when the water content of the flower is reduced to 30-45%, 80-95 wt% of the drying agent is taken out.
In the invention, when the flowers are collected, the flower stalks are preferably left for 1-2 cm.
According to the invention, the preferable drying agent is a nylon gauze bag filled with anhydrous calcium chloride, the nylon gauze bag is 400-600 meshes, the nylon gauze bag is preferably provided with a bag opening, and 200g of anhydrous calcium chloride is filled in the nylon gauze bag; and (3) putting the collected flowers into a sealed box, and keeping 20g of anhydrous calcium chloride when the water content of the flowers is reduced to 30-45%.
The method for collecting fruit tree pollen germplasm resources by using the sealed box under the field condition further comprises the following steps of collecting and preserving pollen: and taking out the flowers in the sealed box, manually stripping the anthers or carrying out crushing treatment, wherein the rotation speed of the crushing treatment is 8000-12000 r/min, the time is 1-5 min, and sieving the crushed flowers through a 30-70-mesh sieve to obtain the anthers.
In the invention, the anther is preferably dried at 18-30 ℃ for 4-6 hours and then stored at low temperature; preferably, the low-temperature preservation is refrigeration at-60 to-90 ℃ or preservation in liquid nitrogen.
In the invention, the flowers placed in the sealed box can be carried with the collector in the field collection process. After the resources are collected and returned to the premises on the same day, the sealed box is placed at room temperature for 24-36h, then the anhydrous calcium chloride in the nylon mesh bag in the sealed box is taken out, only 20g of anhydrous calcium chloride is left, and the box is covered again. The sealed box can be mailed to a special unit for pollen collection by express and then stored for a long time, and can also be stored for 5-6 days at room temperature at the station, and the sealed box can be returned to the unit for pollen collection and then stored for a long time after the resource collection task is finished.
The method can be used for rapidly collecting wild fruit tree pollen germplasm resources, and the influence of the external environment on the flower and the pollen vitality is reduced to the maximum extent by simply processing the flower. Meanwhile, the method has the advantages of simple treatment method, small volume and convenient carrying, and can collect fruit tree germplasm resources of as many varieties as possible in a small space. In addition, the fruit tree pollen germplasm resources collected by the method are simple and convenient to post-process, and various problems encountered in the field germplasm resource collection and preservation process due to crude conditions are further solved.
The method for collecting fruit tree pollen germplasm resources by using the sealing box under the field condition comprises the following steps:
1) preparing materials in advance:
a4 sealed plastic box, length × width × height: 31cm × 22.8cm × 2.2 cm;
a nylon gauze bag made of a 500-mesh nylon gauze, wherein one wide edge is 1/2 reserved as a bag opening, 200g of anhydrous calcium chloride is filled in the bag opening, and the bag opening is sealed;
flatly laying the nylon gauze bag filled with anhydrous calcium chloride into an A4 sealing box, placing filter paper with the size of A4 paper on the nylon gauze bag, and sealing for later use;
2) collecting flowers:
collecting flowers of fruit trees in the field, and reserving 1-2cm of flower stalks; opening the A4 sealing box, spreading the flowers on the filter paper in a single layer manner, and sealing for storage;
3) carrying and post-processing of flowers:
the flowers put into the sealed box are carried with one during the field collection process; and after the resources are collected and returned to the place on the same day, the sealing box is placed at room temperature for 24-36h, then the anhydrous calcium chloride in the nylon mesh bag in the sealing box is taken out, only 20g of anhydrous calcium chloride is left, and the box is covered again. The sealed box is mailed to a special unit by express delivery to collect pollen and then is stored for a long time, or is stored for 5-6 days at room temperature at the place, and the pollen is collected and stored after the collection task is finished;
4) collecting and preserving pollen:
taking out the flowers in the sealed box, manually stripping the anthers or crushing the anthers for 2min at 10000r/min, sieving the crushed material with a 50-mesh sieve to remove impurities, and collecting the anthers; drying the anther at 25 ℃ for 4-6 h, collecting pollen, wrapping with parchment paper, and storing in an ultra-low temperature refrigerator at-80 ℃ or liquid nitrogen for later use.
The invention also provides the sealing box or the method for collecting the fruit tree pollen germplasm resources under the field condition, and the application of the sealing box or the method for collecting the fruit tree pollen germplasm resources under the field condition in collection and preservation of the fruit tree pollen germplasm resources; preferably, the fruit trees comprise one or more of sweet cherries, apples, pears and strawberries.
The invention has the beneficial effects that:
1. the used materials are the least, the weight is the lightest, and the physical consumption of field collecting activities is reduced to the maximum extent.
2. The processing method is simple, the complex work is carried out before and after the field collection activity, the time requirement in the collection activity is reduced, and the field collection efficiency is improved.
3. The flower germplasm is dehydrated and dried by adopting anhydrous calcium chloride with strong water absorption capacity, and the flower is tiled in a single layer, so that the contact area with a drying agent is large, and the drying effect is best.
4. The protection effect on the flower germplasm is the best, and besides the good sealing effect, the A4 sealing box can better fix the single-layer tiled flowers, so that the friction damage to the flowers in the field carrying process is reduced.
5. The fruit tree germplasm resources are convenient to collect as many as possible, and from the pollen collection amount, the fruit tree pollen resources collected by one to two A4 sealed boxes can meet most of scientific research and pollination requirements. The fruit tree germplasm resources which are most collected in one field investigation are realized.
6. The collected germplasm of the flowers of the fruit trees is convenient for post-processing, if the field germplasm resources are collected for a short time (<6 days), the collected flowers can be placed in an A4 sealed box at a station room temperature, and can return to a unit for collecting and storing pollen after the collection activity is finished; if the field germplasm resources are collected for a long time (>6 days), the A4 sealed box can be directly sent to a unit by express mail to collect and store the pollen.
7. The method for collecting and maintaining the vigor of the fruit tree pollen germplasm can improve the collection efficiency of field fruit tree germplasm resources, can ensure the vigor of the pollen of the fruit tree germplasm, further ensures the collection effect, can realize the purpose of benefiting the research work of multiple fruit tree germplasm for many years by one-time field general survey, and has important significance for the collection, storage and utilization work of the fruit tree germplasm resources.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a graph showing the effect of the time of placing in the sealed bag on the pollen viability of the flowers of fruit trees.
FIG. 2 is a diagram of the fruit tree flowers collected under humid conditions in the sealed bag of the present invention, which is placed at room temperature for 6 hours.
FIG. 3 is a sealing box diagram of fruit tree pollen germplasm resources collected under field conditions in an embodiment of the invention, wherein A is a sealing plastic box (x y x y x y x y x y x y x y x y x y x y x y x y x y; b is a nylon mesh bag with the thickness of 500m and a bag opening; and C, a spare sealing box (containing filter paper, a nylon mesh bag and 200g of anhydrous calcium chloride) is used before collection of fruit tree pollen germplasm resources.
FIG. 4 is a diagram of fruit tree pollen germplasm resources collected in the field laid in a sealed box in the embodiment of the invention.
FIG. 5 is a diagram of strawberry flower germplasm resources collected in the field in a sealed box for 24 hours in the embodiment 1 of the present invention.
FIG. 6 is a diagram of sweet cherry flower germplasm resources collected in the field in a sealed box for 6 days in the embodiment 2 of the invention.
FIG. 7 shows that the germplasm resources of the apple flowers collected in the field are placed in a sealed box for 7 days in the embodiment 3 of the invention.
FIG. 8 is a graph of a sweet cherry flower of comparative example 2 dried in a larger (12L) sealed box for 24 hours in accordance with the practice of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and the following embodiments are used for illustrating the present invention and are not intended to limit the scope of the present invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products available from regular distributors, not indicated by the manufacturer.
As a preferable scheme, the method for efficiently collecting and maintaining the pollen viability of the fruit trees in the field, provided by the invention, comprises the following steps:
1. preparing in advance:
a4 sealed plastic box (length × width × height: 31cm × 22.8cm × 2.2 cm);
one wide edge of the bag made of 500-mesh nylon gauze is left half as a bag opening, 200g of anhydrous calcium chloride is filled before field germplasm resources are collected, and the bag opening is sealed by a staple;
the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in an A4 sealing box, a piece of filter paper with the size of A4 paper is placed on the nylon gauze bag, and the sealing box is closed for standby. As shown in FIG. 3, the filter paper, the nylon mesh bag and the anhydrous calcium chloride in the mesh bag are arranged in the sealed box from top to bottom.
2. Collecting flowers:
collecting flowers of the fruit trees in the big balloon period in the field, and reserving 1-2cm of flower stalks. The A4 box was opened, the flowers were spread in a single layer on filter paper, approximately 250 flowers per box, the sealed box was closed, and the bag was placed in a backpack. As shown in FIG. 4, the field collected pollen germplasm resources of different fruit trees of different tree species are laid in different sealed boxes to be easily stacked and carried.
3. Carrying and post-processing of flowers:
the flowers put in the sealed box can be carried about in the field collecting process. After the resources are collected and returned to the premises on the same day, the sealed box is placed at room temperature for 24-36 hours, then the anhydrous calcium chloride in the nylon mesh bag in the sealed box is taken out, only 20g of anhydrous calcium chloride is left, and the box is covered again. The sealed box can be mailed to a special unit for pollen collection by express and then stored for a long time, and can also be stored for 5-6 days at room temperature at the station, and the sealed box can be returned to the unit for pollen collection and then stored for a long time after the resource collection task is finished.
4. Pollen collection and preservation
Taking out the flowers in the sealed box, and manually stripping the anthers; or pulverizing with a small pulverizer at 10000 rpm for 2min, sieving with 50 mesh sieve to remove impurities, and collecting anther. Drying anther at 25 deg.C for 4-6 hr, collecting pollen, wrapping with parchment paper, and storing in ultralow temperature refrigerator (-80 deg.C) or liquid nitrogen (-196 deg.C).
As a preferable scheme of the invention, a drying agent is arranged in the inner space of the sealing box, and the drying agent comprises anhydrous calcium chloride; the volume of the sealing box is 1200-1800 cm3The bottom area is 550-850 cm2(ii) a The weight of the drying agent is 180-220 g. The drying agent is divided into two parts, wherein 80-95 wt% of the drying agent can be taken out of the sealing box. The desiccant is placed in a sealing box in a flat way: preferably, the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in a sealing box, and filter paper is placed on the contact surface of the nylon gauze bag and the flowers; preferably, the nylon gauze bag is 500 meshes.
Example 1 method suitable for field Collection and preservation of strawberry pollen viability
The embodiment provides a method suitable for field collection and strawberry pollen viability maintenance, which comprises the following specific steps:
1. preparing in advance:
a4 sealed plastic box (length × width × height: 31cm × 22.8cm × 2.2 cm);
filling 200g of anhydrous calcium chloride into a bag made of a 500-mesh nylon gauze, and sealing the bag opening by using a staple;
the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in an A4 sealing box, a piece of filter paper with the size of A4 paper is placed on the nylon gauze bag, and the sealing box is closed for standby.
2. Collecting flowers:
collecting big flower buds of strawberries in the Taihang mountain area of the southern wing, and reserving flower stalks for 1-2 cm. The A4 box was opened, the flowers were spread in a single layer on filter paper, approximately 300 flowers per box, the sealed box was closed, and the bag was placed in a backpack.
3. Carrying and post-processing of flowers:
the flowers placed in the sealed box are carried about for about 6 hours in the field collecting process. And after the resources are collected and returned to the place on the same day, the sealed box is placed at room temperature for 24 hours, the water content of the flowers is reduced to 32%, the anhydrous calcium chloride in the nylon mesh bag in the sealed box is taken out, only 20g of anhydrous calcium chloride is left, and the box is covered again. The sealing box adopts the express delivery mode to send back the unit. As shown in fig. 5, the strawberry flower germplasm resources were placed in the sealed box for 24 hours.
4. Pollen collection and preservation
Taking out the flowers in the sealed box, manually stripping off the anthers, drying the anthers at the constant temperature of 25 ℃ for 4 hours, collecting pollen, measuring the pollen activity (65.67 percent after collecting for 5 days), wrapping the pollen with parchment paper, and storing the wrapped pollen in liquid nitrogen (-196 ℃) for later use.
Example 2 method suitable for the field Collection and preservation of sweet cherry pollen viability
The embodiment provides a method suitable for collecting and maintaining the activity of sweet cherry pollen in the field, which comprises the following specific steps:
1. preparing in advance:
a4 sealed plastic box (length × width × height: 31cm × 22.8cm × 2.2 cm);
filling 200g of anhydrous calcium chloride into a bag made of a 500-mesh nylon gauze, and sealing the bag opening by using a staple;
the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in an A4 sealing box, a piece of filter paper with the size of A4 paper is placed on the nylon gauze bag, and the sealing box is closed for standby.
2. Collecting flowers:
collecting flowers of cherries in large balloon period in Yanshan Qinhuang island mountain area, and reserving flower stalks for 1-2 cm. The A4 box was opened, the flowers were spread in a single layer on filter paper, approximately 250 flowers per box, the sealed box was closed, and the bag was placed in a backpack.
3. Carrying and treating flowers:
the flowers placed in the sealed box are carried about for about 6 hours in the field collecting process. And after the resources are collected and returned to the place on the same day, the sealed box is placed at room temperature for 24 hours, the water content of the flowers is reduced to 42%, the anhydrous calcium chloride in the nylon mesh bag in the sealed box is taken out, only 20g of anhydrous calcium chloride is left, and the box is covered again. The capsules were stored at the premises for 5 days at room temperature.
4. Pollen collection and preservation
After 5 days, after the field resources are collected and returned to the unit, the flowers in the sealed box are taken out, the flowers are crushed for 2 minutes at 10000r/min by a small crusher, the crushed materials are screened by 50m to remove impurities, and the anthers are collected. Drying anther at 25 deg.C for 4 hr, collecting pollen, measuring pollen activity (72.15% after 7 days), wrapping with parchment paper, and storing in liquid nitrogen (-196 deg.C) for use. As shown in figure 6, the sweet cherry flower germplasm resource is placed in the sealed box for 6 days.
Example 3 method suitable for field Collection and preservation of apple pollen viability
The embodiment provides a method suitable for field collection and apple pollen viability maintenance, which comprises the following specific steps:
1. preparing in advance:
a4 sealed plastic box (length × width × height: 31cm × 22.8cm × 2.2 cm);
filling 200g of anhydrous calcium chloride into a bag made of a 500-mesh nylon gauze, and sealing the bag opening by using a staple;
the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in an A4 sealing box, a piece of filter paper with the size of A4 paper is placed on the nylon gauze bag, and the sealing box is closed for standby.
2. Collecting flowers:
collecting flowers of apple in big balloon period in Yanshan Qinhuang island mountain area, and reserving 1-2cm of flower stalk. The A4 box was opened, the flowers were spread in a single layer on filter paper, approximately 200 flowers per box, the sealed box was closed, and the bag was placed in a backpack.
3. Carrying and treating flowers:
the flowers placed in the sealed box are carried about for about 6 hours in the field collecting process. And after the resources are collected and returned to the place on the same day, the sealed box is placed at room temperature for 24 hours, the anhydrous calcium chloride in the nylon mesh bag in the sealed box is taken out, only 20g of anhydrous calcium chloride is left, and the box is covered again. The capsules were stored at the premises for 5 days at room temperature.
4. Pollen collection and preservation
After 5 days, the field resources are collected and returned to the unit, the flowers in the sealed box are taken out, and the anthers are manually stripped. Drying anther at 25 deg.C for 4 hr, collecting pollen, measuring pollen activity (81.4% after 7 days), wrapping with parchment paper, and storing in liquid nitrogen (-196 deg.C). As shown in figure 7, the germplasm resources of the apple flowers are placed in the sealed box for 7 days.
Example 4 method for wild collection and preservation of pollen viability of pear trees
The embodiment provides a method suitable for field collection and pear pollen viability maintenance, which comprises the following specific steps:
1. preparing in advance:
a4 sealed plastic box (length × width × height: 31cm × 22.8cm × 2.2 cm);
filling 200g of anhydrous calcium chloride into a bag made of a 500-mesh nylon gauze, and sealing the bag opening by using a staple;
the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in an A4 sealing box, a piece of filter paper with the size of A4 paper is placed on the nylon gauze bag, and the sealing box is closed for standby.
2. Collecting flowers:
collecting the pear flowers in the big ballonflower stage in the southern Taihang mountain area of the wing, and keeping the pedicel of the pear 1-2 cm. The A4 box was opened, the flowers were spread in a single layer on filter paper, approximately 200 flowers per box, the sealed box was closed, and the bag was placed in a backpack.
3. Carrying and post-processing of flowers:
the flowers placed in the sealed box are carried about for about 6 hours in the field collecting process. And after the resources are collected and returned to the place on the same day, the sealed box is placed at room temperature for 24 hours, the water content of the flowers is reduced to 45%, the anhydrous calcium chloride in the nylon mesh bag in the sealed box is taken out, only 20g of anhydrous calcium chloride is left, and the box is covered again. The sealing box adopts the express delivery mode to send back the unit.
4. Pollen collection and preservation
Taking out the flowers in the sealed box, manually stripping off the anthers, drying the anthers at the constant temperature of 25 ℃ for 4 hours, collecting pollen, measuring the pollen activity (75.4 percent after collecting for 5 days), wrapping the pollen with parchment paper, and storing the wrapped pollen in liquid nitrogen (-196 ℃) for later use.
Comparative example 1
Continuous observation on the pollen viability of the flowers shows that the sweet cherry flowers collected in field in sunny weather lose about 20 percent of the pollen viability after being placed in a sealed bag for 2 hours, and lose more than 50 percent of the pollen viability after being placed in a sealed bag for 8 hours (as shown in figure 1).
If the flowers collected in the field are in rainy days or under humid conditions, the flowers will rot and deteriorate after being placed in a sealed bag for 4-6 hours (as shown in figure 2).
Comparative example 2
Sweet cherry flowers collected in the field are placed in a large sealed box (12L, 400g of allochroic silica gel and 40g of flowers) for 24 hours, and due to the fact that the space is large, the drying speed of the flowers is low, the drying is not uniform, petals of the flowers are opened, powder is scattered, and the pollen collecting work and the pollen vitality maintaining work are not facilitated. (as shown in fig. 8).
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. The sealing box is characterized in that a drying agent is arranged in the inner space of the sealing box, and the drying agent comprises anhydrous calcium chloride or silica gel.
2. The capsule according to claim 1, wherein the capsule has a volume of 1200-1800 cm3The bottom area is 550-850 cm2(ii) a The sealing box is preferably a sealing plastic box; more preferably, the sealed plastic box is an A4 sealed plastic box, and the A4 sealed plastic box has a length, a width and a height of 31cm, 22.8cm and 2.2 cm.
3. The capsule according to claim 1 or 2, characterized in that said desiccant is anhydrous calcium chloride.
4. The capsule according to claim 3, wherein the desiccant has a weight of 180 to 220 g; and/or the presence of a gas in the gas,the ratio of the drying agent to the volume is 180-220 g: 1200-1800 cm3。
5. The capsule according to claim 4, wherein the desiccant is divided into at least two portions, wherein at least 80-95 wt% of the desiccant is removable from the capsule.
6. The capsule according to claim 4, characterized in that the desiccant is placed in a flat manner inside the capsule: preferably, the nylon gauze bag filled with anhydrous calcium chloride is flatly laid in a sealing box, and filter paper is placed on the contact surface of the nylon gauze bag and the flowers; preferably, the nylon gauze bag is 500 meshes.
7. A method for collecting fruit tree pollen germplasm resources by using a sealed box under field conditions is characterized by comprising the following steps: treating the harvested flowers with a sealed box according to any one of claims 1 to 6.
8. The method of claim 7, wherein said flowers are tiled onto the capsule in a single layer; preferably, when the water content of the flower is reduced to 30% -45%, 80-95 wt% of the drying agent is taken out.
9. The method for collecting fruit tree pollen germplasm resources by using a sealed box under the field condition as claimed in claim 8, further comprising the steps of collecting and preserving pollen: taking out the flowers in the sealed box, manually stripping the anthers or carrying out crushing treatment, wherein the rotation speed of the crushing treatment is 8000-12000 r/min, the time is 1-5 min, and sieving the crushed flowers through a 30-70-mesh sieve to obtain the anthers; preferably, the anther is dried at 18-30 ℃ for 4-6 h and then stored at low temperature; preferably, the low-temperature preservation is refrigeration at-60 to-90 ℃ or preservation in liquid nitrogen.
10. Use of a sealed box according to any one of claims 1 to 6 or a sealed box for collection of fruit tree pollen germplasm resources under field conditions according to any one of claims 7 to 9 in the collection and preservation of fruit tree pollen germplasm resources under field conditions; preferably, the fruit trees comprise one or more of sweet cherries, apples, pears and strawberries.
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