CN115568395A - Method for improving seed setting rate of cypripedium yunnanense and obtaining full seeds - Google Patents
Method for improving seed setting rate of cypripedium yunnanense and obtaining full seeds Download PDFInfo
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- 241001176695 Cypripedium yunnanense Species 0.000 title description 4
- 239000002775 capsule Substances 0.000 claims abstract description 90
- 241001516485 Cypripedium Species 0.000 claims abstract description 48
- 230000010152 pollination Effects 0.000 claims abstract description 47
- 241000196324 Embryophyta Species 0.000 claims abstract description 35
- 238000012216 screening Methods 0.000 claims description 23
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 19
- 229910052782 aluminium Inorganic materials 0.000 claims description 19
- 238000001035 drying Methods 0.000 claims description 16
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- 241000519432 Cypripedium guttatum Species 0.000 claims description 10
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- A—HUMAN NECESSITIES
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- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/60—Flowers; Ornamental plants
- A01G22/63—Orchids
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- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a method for increasing seed setting rate and acquiring plump seeds of a national secondary protective plant, IUCN extremely-dangerous (CR) and a very small population plant-Yulong cypripedium, which is characterized in that a key phenological period of pollination of the Yulong cypripedium is optimized, pollination amount and pollination flow are controlled through artificial intervention pollination, capsules are protected, seeds are screened, and plump seeds are acquired, so that the method has important significance for the subsequent related work of seed propagation, resource conservation and the like. And has important value for the protection and expansion of the wild population of the rare or endangered species, the research of endangered mechanism, and the development of the on-site, ex-situ and near-site protection work of the species. The method has the advantages of wide application, simple and convenient operation and low cost, protects capsules to the maximum extent, collects all seeds and fills the technical blank in the field.
Description
Technical Field
The invention relates to the technical field of natural product processing, in particular to a method for improving the maturing rate of yulong cypripedium yupinicum and obtaining full seeds.
Background
Yulong cypripedium (A) and (B)Cypripedium forrestii) Is a plant of the genus Cypripedium (cymripium) of the family Orchidaceae (Orchidaceae). Collected and named in 1913 in Lijiang area of Yunnan province for botanic scientist George Forrest, is a special seed in North and West of Yunnan China, is only distributed in the North and West of Yunnan (Lijiang, shangri La), mainly grows under the forest with elevation of about 3300 m, at forest margin, on the shrub-bushy sloping field or on the open forest land, and particularly has a large population at the forest margin of the stream side.
The preservation status of cypripedium specimens worldwide was investigated by Cribb and Sandison in 1998, the total number of yulong cypripedium specimens is less than 5, the records in domestic specimen shops are more rare, and the records which can be found currently are only 1. In view of the narrow distribution area of yulong cypripedium guttatum, rare individuals and field survival conditions, yulong cypripedium guttatum is evaluated as extremely endangered (CR) (Critical endangered) in the Chinese biodiversity red famous book-higher plant volume (2013) and the Chinese higher plant threatened species famous book (2017), and the new edition of the national key protection wild plant famous book and the Yunnan province minimal population wild plant protection famous book (2021 edition) are also included in the yulong cypripedium guttatum.
The Lijiang mountain plant orchard found the colonies of Yulong cypripedium in the Yulong snow mountain natural conservation area of Lijiang in 2019, and carried out continuous monitoring work. At present, 3 colonies are found in Yulong Xueshan, the distance between every two colonies is less than hundred meters, each colony is only 20-30 plants, most of the colonies are formed by sprouting of a plurality of terminal buds of one rhizome, the individuals can be from the same or a few clones, and the genetic diversity is very low. Although the yurongyu orchid has strong asexual reproduction capability, the fructification rate is extremely low under natural conditions, and the fructification rate of a single population in 2019 and 2020 is even zero. Meanwhile, as the growing position is close to mountain stream, seasonal river channel change and water level fluctuation seriously threaten the survival of the mountain stream, and more than half of individuals in one residential colony are washed away by rivers. Aiming at the endangered current situation and the protection value, related technical means need to be explored urgently, the maturing rate of the seedlings is improved, and germplasm resources are collected for storage and breeding.
At present, the research on yuronyulan remains blank at home, and the same genus only aims at cypripedium flavum: (A), (B), (C) and (C)Cypripedium flavum) (iv) cypripedium guttatumCypripedium guttatum) Arytenoid macranthi (A) and (B)Cypripedium macranthum) Bar Lang Shan cypriped: (Cypripedium palangshanense) Related work such as tissue culture, sterile germination of seeds and the like is carried out.
Disclosure of Invention
In order to solve or partially solve the problems in the related technology, the invention provides a method for improving the seed setting rate of yulong cypripedium guttatum and obtaining full seeds, and the method has important significance for the subsequent related work of seed propagation, resource conservation and the like. Meanwhile, the method also has important values for the protection and the expansion of the rare or endangered species population, the research on endangered mechanisms, and the development of the on-site, ex-situ and near-site protection work of the species.
The invention provides a method for improving the maturing rate of yurongyu blue and obtaining full seeds, which comprises the following steps:
(1) Determining the plant phenological period: the Yulongyuhualan plants in the elongation period and the flowering period of the inflorescence shaft are selected as pollination objects.
(2) Artificial intervention pollination: using antistatic tweezers, the whole pollen mass on one side of a pair of stamens of the Yulongyulan plant is taken down and stuck on the stigma surface at the top end of the stigmas of the other plant.
(3) Capsule protection: the pollinated Yulong cypripedium capsules are sleeved with gauze bags, and the lower-end inflorescence axis position is fastened with an aluminum wire.
(4) Collecting capsules: collecting the Yulong cypripedium capsulatum capsule when the capsule turns into yellow brown and does not crack.
(5) Seed screening and preservation: drying capsules in a drying cabinet with the humidity of 15% at the temperature of 5 ℃ for 48h, taking out the seeds in the capsules, screening by using a 100-150-mesh screen, reserving plump seeds, and screening ineffective seeds, desquamated seed coats and impurities. Placing the screened seeds in a glass bottle with a cover, sealing and storing in a refrigerator at 4 ℃.
Further, the period of artificial intervention pollination is the flowering open period.
Further, the anti-static tweezers are ESD-15 anti-static olecranon stainless steel tweezers.
Furthermore, aiming at the colloidal adhesive part coated outside the yulongyulan pollen ball, the colloidal adhesive part is broken by the tip of a forceps, and the pollen with the exposed broken surface is adhered to the surface of the column head at the top end of the column head.
Furthermore, the capsules are required to be placed in a glass bottle when being dried, so that the capsules are convenient to take, place and measure after being dried, and the seed loss is reduced.
Furthermore, when the seeds are screened in the step (5), the edges of the screen are tapped and screened for 3-5 times, so that the seeds are prevented from flying due to violent shaking, and the screening quality is improved.
Further, the seeds are stored in the refrigerator in the step (5) for no more than 180 days at most.
Further, the gauze bag in the step (3) is a gauze bag with 5cm × 8cm and 200 meshes, and the aluminum wire is an aluminum wire with the diameter of 1 mm.
Further, in the step (5), the 100-150-mesh screen is a 120-mesh screen, and the glass bottle with the cover is a 20ml glass bottle with the cover.
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.
The invention has the beneficial technical effects that:
although the yurongyuan orchid can be propagated asexually, the fructification rate of yurongyuan orchid is extremely low under the natural condition, and the fructification rate of a single colony is even zero. According to the method, capsules are protected and seeds are screened by optimizing the optimal pollination phenological period of the cypripedium yupini, artificially intervening in pollination and controlling the pollination process, so that plump seeds are obtained, and the method has important significance for the subsequent related work of seed propagation, resource conservation and the like. Meanwhile, the method also has important values for the protection and the expansion of the rare or endangered species population, the research on endangered mechanisms, and the development of the on-site, ex-situ and near-site protection work of the species.
The prior researches on the yulong cypripedium yupini, particularly the related technologies aiming at improving the fructification rate and obtaining seeds, are not reported. The method has the advantages of wide application, simple and convenient operation and low cost, protects capsules to the maximum extent, collects all seeds and fills the technical blank in the field.
Drawings
FIG. 1 shows flower forms of Yulongyulan in different phenological stages (wherein: a shows bud formation in bud stage, b shows long-term rachis of rachis, c shows flower opening, d shows sepals in late flowering stage withering);
FIG. 2 shows the form of the capsule for natural fruit setting and artificial pollination intervention of cypripedium yulongatum in the application of the invention (wherein: a. Shows the capsule which is aborted under the natural fruit setting state; b. Shows the capsule which is plump after artificial pollination intervention);
fig. 3 shows the appearance of the mature capsule of cypripedium yuronatum in the present application (wherein: a. Shows the uncracked capsule; b. Shows the microcracked capsule and fracture).
Detailed Description
Alternative embodiments of the present application will be described in more detail below with reference to the accompanying drawings. While the drawings show alternative embodiments of the present application, it should be understood that the present application may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the present disclosure to those skilled in the art.
The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used in this application and the appended claims, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It should also be understood that the term "and/or" as used herein refers to and encompasses any and all possible combinations of one or more of the associated listed items.
The method for improving the maturing rate of yurongyuan orchid and acquiring full seeds of the yurongyuan orchid of the invention is described in detail by combining the accompanying drawings as follows:
the method for improving the seed setting rate of the cypripedium yupini and acquiring the plump seeds in the application of the invention comprises the following steps:
(1) Determining the plant phenological period: yulongyulan plants in the elongation period (5 days) and the blooming period (13 days) of the inflorescence shaft are selected as pollinating objects.
(2) Artificial intervention pollination: using anti-static tweezers to take off the whole pollen mass at one side of a pair of stamens of the Yulongyulan plant and stick the pollen mass on the stigma surface at the top end of the other plant.
(3) Capsule protection: the pollinated Yulong cypripedium capsules are sleeved with gauze bags, and the lower-end inflorescence axis position is fastened with an aluminum wire.
(4) Collecting capsules: when the capsule turns into yellow brown and does not crack, the Yulong cypripedium capsule is collected. Because the seeds of the cypripedium yuronatum are extremely small, the seeds are difficult to be completely collected after the capsules are cracked, and the tiny seeds are greatly influenced by the external environment and are easy to enter deep dormancy or inactivation, so that the subsequent breeding work is influenced.
(5) Seed screening and preservation: putting the capsule in a drying cabinet with the humidity of 15% and the temperature of 5 ℃ for drying for 48h, putting and metering the capsule in a glass bottle when the capsule is dried, taking and placing the capsule after drying, reducing seed loss, taking the seeds in the capsule out, screening by using a screen with 100 to 150 meshes, retaining full seeds, and screening out ineffective seeds, fallen seed coats and impurities. Placing the screened seeds in a glass bottle with a cover, sealing and storing in a refrigerator at 4 ℃.
In one embodiment of the present application, the period of said artificial intervention pollination is the flowering stage.
In one embodiment of the present application, the anti-static tweezers are ESD-15 anti-static olecranon stainless steel tweezers, which prevent electrostatic interference, and meanwhile, the bent tweezers are convenient to operate, and damage to flower structures is reduced.
In one embodiment of the present application, the outside of the yulongyulan pollen ball is wrapped with a gummy part, which is broken by the tip of a forceps, and the pollen with the exposed broken surface is stuck on the surface of the head of the stigma.
In one embodiment of the present application, the capsule should be dried in a glass bottle, which facilitates the taking and placing of the capsule after drying and reduces the loss of seeds.
In one embodiment of the present invention, when the seeds are screened in step (5), the edges of the screen are tapped and the screening is performed for 3-5 times, so that the scattering of the seeds caused by severe shaking is prevented, and the screening quality is improved.
In one embodiment of the present application, the seeds are stored in the refrigerator in step (5) for a maximum of not more than 180 days.
In one embodiment of the present application, the gauze bag in step (3) is a 5cm × 8cm,200 mesh gauze bag, and the aluminum wire is an aluminum wire with a diameter of 1 mm.
In an embodiment of the invention, in the step (5), the 100 to 150-mesh screen is a 120-mesh screen, and the glass bottle with the cover is a 20ml glass bottle with the cover.
For the sake of clarity, the following examples are given in detail.
Example 1
A method of increasing the fruiting rate of yurongyuan orchid and obtaining full seeds, the method comprises the following steps:
(1) Determining a phenological period: the habitation period of yulongyunacanthus was determined in 2019 and 2020 and divided into four stages, wherein the flower and leaf synchronization period indicates 10 days for bud formation, 5 days for inflorescence axis elongation, 13 days for flower opening, and 5 days for the end flower period from the withering of the sepals to the withering of the lipflaps (fig. 1).
(2) Artificial intervention pollination: in 5 months of 2021, carrying out artificial intervention pollination on 3 habitats of Yulongyuhuan, 20 individuals under each habitats, taking down the whole pollen mass on one side of a pair of stamens of the Yulongyuhuan plant by using an ESD-15 model forceps, pricking the colloidal viscous part outside the pollen mass by using the tip of the forceps, adhering the pollen exposed on the fracture surface to the head surface of the head of the other plant, and removing the labial flaps by using the forceps. The pollination time of individuals under each colony is divided into 4 groups according to different phenological periods, and each group comprises 5 individuals.
(3) Protection of capsules: a5 cm multiplied by 8cm gauze bag with 200 meshes is adopted, a pollinated Yulong cypripedium capsule is sleeved, and the lower end inflorescence axis position is fastened by an aluminum wire with the diameter of 1 mm.
(4) Collecting capsules: before the cracking of the Yulong cypripedium capsule, the appearance of the Yulong cypripedium capsule is changed into yellow brown for harvesting.
(5) Seed screening and preservation: drying the capsule in a drying cabinet with the humidity of 15% and the temperature of 5 ℃ for 48h, and then taking out the seeds in the capsule for sampling and counting. And (4) screening by using a 120-mesh screen, tapping the edge of the screen, keeping full seeds, and screening off peeled seed coats and impurities. This step was repeated 5 times, and the screened seeds were placed in 20ml glass bottles with caps, sealed and stored in a refrigerator at 4 ℃.
Comparative example 1
According to the method for improving the seed setting rate of yulong cypripedium yupinicum and obtaining plump seeds provided by the embodiment 1, the packing mode of capsules is changed, aluminum foil is adopted for packing, and the specific process is as follows: the open period of the cypripedium yunnanense is selected for pollination. Taking down the whole pollen mass on one side of a pair of stamens of the Yulongyulan plant, pricking the outside of the pollen mass by using the tip of a tweezers to wrap a colloid viscous part, and adhering the pollen exposed on the fractured surface to the head surfaces of the tops of stigmas of different plants. Aluminum foil with the thickness of 0.05mm is cut into a square with the thickness of 15cm multiplied by 15cm, the pollinated Yulong cypripedium capsule is wrapped, the lower end is tightly twisted and is tightly tied by an aluminum wire with the thickness of 1 mm. The volume of the packaging cavity is kept to be 1.5cm multiplied by 2.5cm, and enough space is reserved for the growth of capsules. Before the cracking of the Yulong cypripedium capsule, the appearance of the Yulong cypripedium capsule is changed into yellow brown for harvesting. Drying, sampling and counting, screening seeds and storing the screened seeds.
Comparative example 2
According to the method for improving the seed setting rate of cypripedium yupinicum and acquiring full seeds provided by the embodiment 1, manual pollination intervention is not performed, and the specific process is as follows: observing the wild Yulongyulan cypripedium colonies, carrying out no artificial pollination intervention on 10 individuals below each colony, bagging capsules by adopting 5cm multiplied by 8cm and 200-mesh gauze bags after lipflaps wither, and fastening the positions of lower inflorescence axes by aluminum wires with the diameter of 1 mm. Before the cracking of the Yulong cypripedium capsule, the appearance of the Yulong cypripedium capsule is changed into yellow brown for harvesting. Drying, sampling, counting, screening seeds and storing the screened seeds.
Comparative example 3
According to the method for improving the seed setting rate of cypripedium yupinicum and acquiring full seeds provided by the embodiment 1, the self-pollination is carried out on the cypripedium yupinicum seeds, and the specific process is as follows: selecting the blooming period of the Yulongyuhualan flowers for pollination, taking down the whole pollen mass on one side of a pair of stamens of the Yulongyuhualan plants, breaking the colloidal viscous part outside the pollen mass by using the tip of a forceps, and adhering the pollen with the exposed broken surface to the column head surface at the top end of the column head of the same plant. The capsule is bagged by a 5cm multiplied by 8cm and 200 mesh gauze bag, and the position of the lower inflorescence axis is fastened by an aluminum wire with the diameter of 1 mm. Before the cracking of the Yulongyulan capsule, the appearance of the Yulongyulan capsule is changed into yellow brown for harvesting. Drying, sampling and counting, screening seeds and storing the screened seeds.
Comparative example 4
According to the method for improving the seed setting rate and obtaining the plump seeds of cypripedium yuronatum provided by the embodiment 1, the capsule harvesting time is delayed, harvesting is carried out when the top end of the capsule is microcracked, and capsule protection is not carried out, and the specific process is as follows: selecting the blooming period of the Yulongyuhualan flowers for pollination, taking down the whole pollen mass on one side of a pair of stamens of the Yulongyuhualan plants, breaking the colloidal viscous part outside the pollen mass by using the tips of tweezers, and adhering the pollen with the exposed broken surfaces to the column head surfaces at the top ends of the column heads of different plants. Harvesting after the top of the Yulongyulan capsule is slightly cracked. Drying and sampling counting the seeds, screening the seeds and storing the screened seeds.
Comparative example 5
According to the method for improving the maturing rate of the yuronyulan magnolia and obtaining the full seeds provided by the embodiment 1, the manual intervention pollination amount is increased, and the specific process is as follows: selecting the blooming period of the Yulongyuhualan flowers for pollination, taking down two pollen masses of a pair of stamens of the Yulongyuhualan plants, breaking the outside of the pollen masses by using the tips of tweezers to wrap colloidal viscous parts, and adhering the pollen with exposed broken surfaces to the column head surfaces at the top ends of different plant stigmas. The capsule is bagged by a 5cm multiplied by 8cm and 200 mesh gauze bag, and the position of the lower inflorescence axis is fastened by an aluminum wire with the diameter of 1 mm. Before the cracking of the Yulongyulan capsule, the appearance of the Yulongyulan capsule is changed into yellow brown for harvesting. Drying, sampling and counting, screening seeds and storing the screened seeds.
Comparative example 6
According to the method for improving the seed setting rate of yulong cypripedium guttatum and acquiring full seeds provided by the embodiment 1, the manual intervention pollination amount is reduced in equal proportion, and the specific process is as follows: selecting the blooming period of Yulongyulan flowers for pollination, taking down the whole pollen mass on one side of a pair of stamens of the flowers, placing the pollen mass in a 6mm glass culture dish, pricking the colloidal viscous part outside the pollen mass by using the tip of a forceps, peeling and respectively keeping 1/2 and 1/4 of the pollen mass, and adhering the pollen mass on the column head surfaces of the top ends of different plant stigmas. The capsule is bagged by a 5cm multiplied by 8cm and 200 mesh gauze bag, and the position of the lower inflorescence axis is fastened by an aluminum wire with the diameter of 1 mm. Before the cracking of the Yulong cypripedium capsule, the appearance of the Yulong cypripedium capsule is changed into yellow brown for harvesting. Drying, sampling and counting, screening seeds and storing the screened seeds. For the above examples 1 and 2 and comparative examples 1 to 5, the expansion rate and volume of the capsule of Yulong cypripedium grandiflorum are counted, and the fructification rate, the total amount of single-fruit seeds and the number of filled seeds are counted by a sampling method.
TABLE 1 influence of manual intervention pollination of Yulongyulan in different phenological stages on setting percentage and number of single fruit seeds (example 1)
Table 2 influence of different pollination modes on the maturing rate and the number of single fruit seeds of yurongyuan orchid
TABLE 3 influence of different capsule protection modes on the yield and quantity of seeds of Yulong cypripedium
TABLE 4 Effect of different pollination on the setting percentage and the number of seeds per fruit of Yulong-cypripedium
As can be seen from the table 1, the artificial pollination intervention on the Yulongyulan has different capsule expansion rate, capsule volume, plump seed number and maturing rate. The flowers and leaves of the yulongyulan are in the same period, the stigma pollen in the bud period is not completely developed, and effective capsules cannot be produced after pollination; the pollen of the flower at the elongation stage of the inflorescence axis is mature, the stigma receptivity is improved, and partial effective capsules can be generated; the activity of stigma and pollen at the end of flowering period is reduced, and even if effective capsules can be produced, the quantity is small. The selection of flower blooming period as an intervention in the manual pollination of cypripedium yuronatum is most advantageous for capsule production. In addition, because the wild habitats of the yulongyulan are few, the number of individuals under each habitats is small, and the individuals are mostly distributed near stream valleys in deep mountain forest lands, the exploration is difficult and the accessibility is poor; and because the whole florescence is not long and the phenological condition is advanced or delayed along with the climate change, the artificial intervention pollination can be carried out in the elongation stage of the inflorescence axis under the condition of non-ideal conditions for replacement.
As can be seen from Table 2, the Yulongyulan is extremely difficult to form capsule and fruit under natural conditions, and if the seeds need to be obtained, artificial pollination intervention is necessary (figure 2). Compared with artificial self-pollination and cross-pollination, although the total amount of seeds is not greatly different, the capsules for self-pollination are smaller, the number of plump seeds is greatly reduced, and the maturing rate is obviously reduced, so that the cross-pollination should be selected when the artificial intervention pollination is carried out on the Yulongyulan cypripedium, and the maturing rate and the seed yield are ensured.
As can be seen from table 3, when the yulong cypripedium capsulatum capsule is protected, the breathable gauze bag is preferably selected, and although the aluminum foil can preserve plant seeds to the maximum extent, the inside of the aluminum foil is relatively closed, so that temperature aggregation in a wrapping cavity can be caused along with the change of environmental temperature, and plant seeds can be inactivated due to high temperature; in addition, because the capsule and the seeds generate water vapor by respiration, the sealed environment is easy to cause the fruit pods and the seeds to be mildewed and rotted, and the total amount of the single-fruit seeds, the full seed amount and the seed setting rate are greatly reduced. It can also be seen from table 3 that although the capsule expansion rate and the capsule volume are slightly different from the embodiment of selecting a gauze bag to protect the capsule, the seed loss of the yurongyuan is reduced because the seed of the yurongyuan cypress is very small and the capsule is easy to spread once the capsule is cracked (fig. 3).
As can be seen from Table 4, the pollination amount of the cypripedium yuronatum is a single pollen ball on one side of a pair of stamens, and after the pollination amount is increased, the differences of the capsule volume, the total amount of single-fruit seeds, the number of plump seeds and the seed set rate are small. With the decrease of the pollination amount, the capsule volume, the total amount of the seeds of the single fruit and the number of the filled seeds are all reduced, which is unfavorable for the seed harvest of the yuronyulan. The research on the correlation between artificial pollination amount and capsule production and seed production in orchids is still few, and the research in cypripedium is still blank. Therefore, the invention has higher innovation for the explanation of the technical steps, and can be referred to and implemented by related technicians in the process of artificial intervention pollination of the congeneric species.
In summary, the method for improving the maturing rate of yulong cypripedium guttatum and obtaining full seeds provided by the invention has important functions in promoting the maturing rate of yulong cypripedium guttatum and obtaining full seeds in all the steps, wherein the steps (1) to (4) are particularly critical for improving the maturing rate of yulong cypripedium guttatum and obtaining full seeds. Therefore, the invention carries out artificial intervention pollination, controls pollination amount and pollination flow, protects capsules and screens seeds to obtain plump seeds by researching and judging the pollination key phenological period of the cypripedium yunnanense, and well solves the key technical problems of increasing the seed setting rate and obtaining plump seeds. Has important significance for the subsequent development of related work such as seed propagation, resource conservation and the like. The method has important values for the protection and the expansion of rare or endangered species communities, the research on endangered mechanisms, the development of on-site, ex-situ and near-site protection work of species, and fills the blank of research work on secondary protection plants, IUCN (critical recovery) and minimum population plants in the country and the technical field.
Having described embodiments of the present application, the foregoing description is intended to be exemplary, not exhaustive, and not limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen in order to best explain the principles of the embodiments, the practical application, or improvements made to the technology in the marketplace, or to enable others of ordinary skill in the art to understand the embodiments disclosed herein.
Claims (9)
1. A method for improving the seed set rate of cypripedium yunatum and obtaining full seeds is characterized by comprising the following steps:
(1) Determining the plant phenological period: selecting Yulongyuhualan plants in the elongation period and the flowering period of the inflorescence shaft as pollination objects;
(2) Artificial intervention pollination: using anti-static tweezers to take down the whole pollen mass on one side of a pair of stamens of the Yulongyulan plant and stick the pollen mass on the stigma surface at the top end of the stigmas of the other plant;
(3) Protection of capsules: the pollinated Yulong cypripedium capsules are sleeved with gauze bags, and the lower-end inflorescence axis position is fastened by aluminum wires;
(4) Collecting capsules: collecting the Yulong cypripedium capsulatum capsule when the capsule turns into yellow brown and does not crack;
(5) Seed screening and preservation: putting capsules in a drying cabinet with the humidity of 15% and the temperature of 5 ℃ for drying for 48h, taking out seeds in the capsules, screening by using a 100-to 150-mesh screen, retaining plump seeds, screening invalid seeds, desquamated seed coats and impurities, putting the screened seeds in glass bottles with covers, sealing and storing in a refrigerator with the temperature of 4 ℃.
2. The method of improving the maturing rate of yurongyu blue and obtaining full seeds of claim 1 wherein the period of manual intervention pollination is the blooming period.
3. The method of claim 1 wherein the anti-static tweezers are ESD-15 anti-static eagle beak stainless steel tweezers.
4. The method for improving the maturing rate of yulong cypripedium guttatum and obtaining the full seeds of claim 1, wherein the colloidal adhesive part is wrapped outside the yulong cypripedium guttatum pollen ball, the colloidal adhesive part is broken through the tip of a tweezers, and pollen exposed at the broken surface is adhered to the column head surface at the top end of the column head.
5. The method for improving the seed set rate and obtaining full seeds of cypripedium yuronatum according to claim 1, wherein the capsules are dried and placed in glass bottles, so that the capsules can be conveniently taken, put and metered after being dried, and the seed loss is reduced.
6. The method for improving the maturing rate of yurongyuan cypripedium and obtaining full seeds of claim 1, wherein when the seeds are screened in the step (5), the edges of the screen are tapped and the screening is carried out for 3 to 5 times, so that the seeds are prevented from flying off due to violent shaking, and the screening quality is improved.
7. The method of claim 1 wherein the seeds are stored in a freezer for a maximum of 180 days in step (5).
8. The method for improving the maturing rate of yuronyuzuyulan and obtaining full seeds of claim 1, wherein the gauze bag in the step (3) is a 5cm x 8cm 200-mesh gauze bag, and the aluminum wire is an aluminum wire with the diameter of 1 mm.
9. The method for improving the maturing rate of yurongyuan cypripedium and obtaining full seeds according to claim 1, wherein in the step (5), the 100-150-mesh screen is a 120-mesh screen, and the glass bottle with the cover is a 20ml glass bottle with the cover.
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