CN117859598A - Method for safely preserving, purifying and rejuvenating local rice germplasm resources - Google Patents
Method for safely preserving, purifying and rejuvenating local rice germplasm resources Download PDFInfo
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- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 63
- 235000009566 rice Nutrition 0.000 title claims abstract description 63
- 230000003716 rejuvenation Effects 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 36
- 240000007594 Oryza sativa Species 0.000 title 1
- 238000012360 testing method Methods 0.000 claims abstract description 82
- 241000209094 Oryza Species 0.000 claims abstract description 62
- 238000004321 preservation Methods 0.000 claims abstract description 42
- 238000009395 breeding Methods 0.000 claims abstract description 32
- 230000001488 breeding effect Effects 0.000 claims abstract description 32
- 238000000746 purification Methods 0.000 claims abstract description 28
- 238000003306 harvesting Methods 0.000 claims abstract description 25
- 201000010099 disease Diseases 0.000 claims abstract description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 18
- 239000000463 material Substances 0.000 claims abstract description 12
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- 238000000926 separation method Methods 0.000 claims abstract 2
- 238000009331 sowing Methods 0.000 claims description 45
- 230000035784 germination Effects 0.000 claims description 23
- 235000013339 cereals Nutrition 0.000 claims description 21
- 241000196324 Embryophyta Species 0.000 claims description 18
- 238000002791 soaking Methods 0.000 claims description 17
- 238000003860 storage Methods 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 238000002955 isolation Methods 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 12
- 241000878007 Miscanthus Species 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 9
- 238000004140 cleaning Methods 0.000 claims description 6
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- 241000238631 Hexapoda Species 0.000 claims description 5
- 230000009418 agronomic effect Effects 0.000 claims description 5
- 239000003337 fertilizer Substances 0.000 claims description 5
- 238000010899 nucleation Methods 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 4
- 229910052801 chlorine Inorganic materials 0.000 claims description 4
- 239000000460 chlorine Substances 0.000 claims description 4
- 238000009826 distribution Methods 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 4
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 3
- 229910052782 aluminium Inorganic materials 0.000 claims description 3
- 238000007791 dehumidification Methods 0.000 claims description 3
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- 238000000605 extraction Methods 0.000 claims description 3
- 239000011888 foil Substances 0.000 claims description 3
- 239000012535 impurity Substances 0.000 claims description 3
- 238000011068 loading method Methods 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 3
- 235000016709 nutrition Nutrition 0.000 claims description 3
- 230000035764 nutrition Effects 0.000 claims description 3
- 230000000644 propagated effect Effects 0.000 claims description 3
- 238000005057 refrigeration Methods 0.000 claims description 3
- 239000002689 soil Substances 0.000 claims description 3
- 230000001133 acceleration Effects 0.000 claims 1
- 238000009355 double cropping Methods 0.000 claims 1
- 238000011161 development Methods 0.000 abstract description 3
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Abstract
The invention relates to the technical field of germplasm preservation, in particular to a method for safely preserving, purifying and rejuvenating local rice germplasm resources, which comprises the steps of constructing a germplasm resource low-temperature preservation bank, isolating a test nursery, identifying a propagation nursery and purifying and rejuvenating nursery; randomly extracting a preset number of seeds from the acquired local germplasm resources to serve as test seeds, and acquiring image information of the test seeds for checking during harvesting; according to the characteristic features of the original records, a single spike representing a group is harvested as seeds for breeding and identifying germplasm in a mature period, the shape features of local germplasm resources are undefined or the field expressive separation of the resources is changed seriously, the local germplasm resources are divided into a plurality of different resources according to the characteristic features according to the field phenotypes, the single spike is respectively collected for purification, rejuvenation and observation and comparison, so that the seeds which can be stored and utilized at low temperature are obtained, the genetic diversity of the germplasm resources can be protected as comprehensively as possible, original research materials are provided for improving the stress tolerance and the disease and pest resistance of the bred rice varieties, and the sustainable development of crops is promoted.
Description
Technical Field
The invention relates to the technical field of germplasm resource preservation, in particular to a method for safely preserving, purifying and rejuvenating local rice germplasm resources.
Background
Seeds are carriers of genetic materials and are bands for species continuation and variety maintenance characteristic; the quality of seeds is a primary factor in crop production. The rice is an important crop for guaranteeing the grain safety of China, the rice has long history, and rich local rice varieties form local variety distribution with very special local rice due to regional characteristics, history leather, eating habits, customs and other reasons.
With the rapid development of commercial breeding, local rice seed resources are gradually replaced by new bred varieties due to the disadvantages of yield, taste and the like, local varieties are rapidly reduced, and variety characteristics of the local resources are also lost. In recent years, local germplasm resources with regional characteristics and strong stress tolerance are less and less due to the change of climate conditions and breeding requirements, and the protection of the resources is urgently needed to be enhanced manually.
Disclosure of Invention
The invention overcomes the defects of the prior art and provides a method for safely preserving, purifying and rejuvenating local rice germplasm resources.
The technical scheme adopted by the invention for achieving the purpose is as follows:
the invention discloses a method for safely preserving, purifying and rejuvenating local rice germplasm resources, which comprises the following steps:
obtaining local germplasm resources through a preset channel, constructing a germplasm resource low-temperature conservation library in an ecological area with the same or similar growth, isolating a test nursery, identifying a breeding nursery and purifying a rejuvenating nursery;
randomly extracting a preset number of seeds from the acquired local germplasm resources to serve as test seeds, and acquiring image information of the test seeds for checking during harvesting; sowing and transplanting all the test seeds uniformly according to the planting time of the rice in one season;
after a preset time period, harvesting a single spike at a maturity stage according to the originally recorded characteristic characteristics of the single spike as seeds for breeding and identifying germplasm, dividing the local germplasm resources into a plurality of different resources according to the characteristic characteristics of the local germplasm resources or separating the local resource agronomic characteristics seriously, respectively harvesting the single spike according to the field phenotype, purifying, rejuvenating and observing and comparing the single spike to obtain the test-seed harvested seeds;
during propagation identification, the sowing period, the heading period, the maturation period, the light temperature, the plant height, the ear length, the effective ear number, the ear extraction degree, the ear color, the leaf sheath color, the leaf fluff, the sword leaf length, the sword leaf width, the sword leaf angle, the stem thickness, the stem angle, the lodging property, the miscanthus length, the miscanthus color and the miscanthus distribution are examined in the field growth process, and the grain number, the seed setting rate, the thousand grain weight, the grain length, the grain width, the grain thickness, the seed coat color and the grain shape are examined after the maturation and the harvest;
for resources which need to be purified and rejuvenated, after the first year of trial planting, resources which are not grown in the final field due to low vitality are selected, and original seeds are transplanted after indoor seedling raising; for resources with serious population mixing, single plants are reserved for planting and observing after trial planting, and the resources with partial diseases are reserved for breeding again after single-pot planting.
Specifically, the low-temperature preservation library is mainly used for low-temperature safe preservation of germplasm resources;
the isolation test nursery is used for the first year test of newly collecting rice germplasm resources, observing the growth condition and light temperature characteristics of the germplasm resources and eliminating quarantine diseases and insect pests;
the identification breeding nursery is divided into three relatively separated fields, and the rice is divided into the early, middle and late according to the basic observation of the first year, and the basic agronomic characters are observed and recorded after the rice is sowed in seasons;
the purification rejuvenation nursery is used for the appearance after the first year trial: the materials with low seed activity, serious group mixing and part of diseases are singly bred after single spike seed collection, and the purification rejuvenation nursery consists of two parts of indoor seedling culture and outdoor breeding.
Specifically, the low-temperature storage warehouse is divided into two independent environments with the environmental temperature capable of being set to-4 ℃ and 4 ℃, the relative humidity is controlled below 40%, and the environment is a closed environment and the automatic refrigeration and dehumidification equipment is provided.
Specifically, each nursery of the isolation test nursery, the identification breeding nursery and the purification rejuvenation nursery is independently isolated by a net-shaped enclosing wall with the height of 3 meters; the isolation test nursery, the identification breeding nursery and the purification rejuvenation nursery are also provided with a framework paved with an anti-bird net.
Specifically, before sowing test seeds, soaking all test seeds in clear water for 18-24 hours, soaking the test seeds in 0.3% strong chlorine essence for 8 hours, cleaning the test seeds in clear water, soaking the test seeds in clear water for 12 hours, draining off water, keeping the test seeds moist, placing the test seeds in an environment at 30 ℃ for accelerating germination for 24-36 hours, observing that white short roots emerge from the test seeds, and sowing the test seeds in a seedling bed;
before transplanting the test seeds, the test field is leveled and the disease, pest and weed control is carried out, but the original growth environment of local resources is poorer, so that the growth characteristics of the test seeds are truly reflected, and chemical fertilizers are not needed in the planting process;
the method comprises the steps of transplanting test seed field blocks into grids according to 6 inch x 8 inch lines, inserting each test seed at grid crossing points during transplanting, planting 2 rows of each variety, 10 plants in each row, 20 plants in total, spacing one row between varieties, reserving 3 grids between every 10 rows as walkways, and inserting a number plate at a first right position of each variety.
Specifically, the seeds harvested by trial seeding are dried in the sun until the safe moisture content is below 13% in the current year of harvesting, stored in a low-temperature storage warehouse at 4 ℃, and then propagated and identified in the next year;
the propagation identification is carried out according to the light-sensitive characteristics of the varieties observed and collected in the first year trial planting, the varieties can be planted in the middle and lower reaches of Yangtze river according to seasons in the middle and lower reaches of Yangtze river, the sowing time of early rice is 3 months and 25-31 days, the transplanting time is 30 days after sowing, the sowing and transplanting time of middle rice is the same as that of isolated trial planting rice, the sowing time of late rice is 6 months and 20-25 days, and the transplanting time is 25-30 days after sowing;
wherein, the seed soaking and germination accelerating of the seeds before sowing are the same as the test rice seeds, the seed soaking time is reduced by 12 hours as a whole before sowing late rice due to higher air temperature, namely, the seed soaking time is reduced by 6 hours before and after the strong chlorine refining treatment;
the preparation before isolation test and propagation identification transplanting is basically the same, chemical fertilizers are not needed, 2-3 times of ploughing is needed before leveling, and the purity of the planting in the season resource community is ensured not to be influenced by grain falling;
transplanting grids according to 6 inch-6 inch mark lines, transplanting each resource variety at grid crossing points, planting 6 lines of each variety, 8 lines of each variety, 48 lines of each variety, leaving 3 lines of each 8 lines as walkways, and inserting a number plate at a first right position of each variety;
all propagation identification resources should be harvested immediately after the propagation identification resources reach maturity in the field, and the propagation identification resources only collect 4*6 =24 strains of which the outermost circle is removed from each cell for storage; and the harvested fresh seeds are immediately sun-dried or dried at a temperature of not higher than 40 ℃ until the safe moisture of the seeds is lower than 13%.
Specifically, for the resources with low vigor and failure in a large Tian Chengmiao area, the original resources are taken to be soaked, germinated and then sowed in a nutrition soil small pot with the diameter of 10cm and the height of 8cm, the small pot is placed in an artificial climate box or an artificial climate chamber, the temperature is 30 ℃ and the humidity is 70 percent, the culture is carried out until the seedlings are 10cm long, then the seedlings are moved to natural conditions to grow to two leaves and one core, and then transplanted to a field for normal growth;
immersing low-vigor seeds to accelerate germination, observing germination conditions, and prolonging germination time according to whether seedlings emerge;
harvesting single spike seeds in the current year after the rejuvenation and propagation of the seeds with low vigor succeed, and carrying out propagation identification on the field in the next year;
for resources with serious population mixing, single spikes for trial planting and seed reserving need to be numbered continuously, the sowing is the same as other propagation identification materials, and the single spikes are planted in adjacent cells of the same field to observe the characters; if the phenotype of each cell is neat during rejuvenation and purification and is consistent with that of the original sowing sample, respectively harvesting and storing the seeds; if the community still has the mixing phenomenon during the test, the material is purified and is not a germplasm resource meeting the preservation requirement, and the material is eliminated.
Specifically, the resources with diseases are subjected to single-pot isolated planting in the whole growth period, then single plants are harvested for field propagation identification in the second year, and the seeds are subjected to disinfection and sterilization treatment before sowing;
the seed quantity of the resource preservation is not less than 200g after the seed is derived from propagation identification;
harvesting seeds, carrying out impurity cleaning, detecting germination rate, drying, and storing in a low-temperature warehouse after meeting the storage requirement; the resource requirement of the low-temperature warehouse preservation is that the net seeds have a germination rate higher than 90 percent and a water content of the seeds lower than 7 percent;
the seeds are stored in two parts after reaching the warehouse-in condition, an aluminum foil bag or a low-temperature-resistant plastic bottle is used, a resource information bar code label is attached to the bag or the bottle, and the two parts of resources are stored in two low-temperature storage libraries at 4 ℃ and-4 ℃ respectively.
Specifically, the method comprises the steps of loading seeds into a breathable package, placing the breathable package in equipment with the ambient temperature of 15 ℃ and the relative humidity of less than 15%, and drying the seeds at a slow speed so as not to damage the seeds in the drying process.
Specifically, the 4 ℃ low-temperature preservation resource is firstly used for monitoring and using the vitality, the-4 ℃ low-temperature preservation warehouse is used for backup, and the-4 ℃ low-temperature preservation warehouse resource is started after the 4 ℃ low-temperature preservation resource is used up or the vitality is lost.
The invention solves the technical defects existing in the background technology, and has the following beneficial effects: the method selects varieties with stable and consistent genetic characteristics and adaptability for propagation identification and safe preservation through isolation test and purification rejuvenation, can ensure that the genetic diversity of local rice is not lost, is beneficial to providing original research materials for researching the excellent characteristics of local rice germplasm resources, improving the disease and pest resistance and adaptability of the existing rice varieties and promoting the sustainable development of crops.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other embodiments of the drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a flow chart of a method for safely preserving, purifying and rejuvenating local rice germplasm resources.
Detailed Description
In order that the above-recited objects, features and advantages of the present invention will be more clearly understood, a more particular description of the invention will be rendered by reference to the appended drawings and appended detailed description. It should be noted that, in the case of no conflict, the embodiments of the present application and the features in the embodiments may be combined with each other.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, however, the present invention may be practiced in other ways than those described herein, and therefore the scope of the present invention is not limited to the specific embodiments disclosed below.
As shown in FIG. 1, the invention discloses a method for safely preserving, purifying and rejuvenating local rice germplasm resources, which comprises the following steps:
s102: obtaining local germplasm resources through a preset channel, constructing a germplasm resource low-temperature conservation library in an ecological area with the same or similar growth, isolating a test nursery, identifying a breeding nursery and purifying a rejuvenating nursery;
s104: randomly extracting a preset number of seeds from the acquired local germplasm resources to serve as test seeds, and acquiring image information of the test seeds for checking during harvesting; sowing and transplanting all the test seeds uniformly according to the planting time of the rice in one season;
s106: after a preset time period, harvesting a single spike which can represent the whole of the single spike in a mature period according to the characteristic characteristics of the original record of the single spike as seeds for breeding and identifying germplasm, dividing the local germplasm resources with undefined characteristics or severely separated resource field performance characteristics into a plurality of different resources according to the characteristic characteristics of the field phenotype, respectively harvesting the single spike for purification, rejuvenation and observation comparison to obtain test-seed harvest seeds;
s108: during propagation identification, the distribution of seeding time, heading time, mature time, growth time, light temperature property, plant height, spike length, effective spike number, spike extraction degree, spike shape, leaf color, leaf sheath color, leaf fluff, sword leaf length, sword leaf width, sword leaf angle, stem thickness, stem angle, lodging property, miscanthus length, miscanthus color and miscanthus is examined in the growth process, and the grain number, fruiting rate, thousand grain weight, grain length, grain width, grain thickness, seed coat color and grain shape character of each spike are observed after the mature harvest;
s110: for resources which need to be purified and rejuvenated, after trial planting in the first year, the resources which are not grown in the field finally due to low vitality are transplanted to the field after indoor seedling raising is carried out; for resources with serious population mixing, after trial planting, single plants are left for planting respectively, and then field characters are observed; and (3) carrying out single-pot planting maturing and seed reserving propagation on part of resources with diseases in the whole growth period.
It should be noted that, the sources of local germplasm resources mainly include the following modes: seeds that they have remained in or before planting are collected from farmers or breeders, or are taken from a storage or the like. Sowing is generally carried out on 15-20 days of 5 months, and transplanting is carried out on 6-10 days of 6 months.
Specifically, the low-temperature preservation library is mainly used for low-temperature safe preservation of germplasm resources;
the isolation test nursery is used for the first year test of rice germplasm resources newly collected, observing growth conditions, light-temperature characteristics and eliminating quarantine diseases and insect pests of the germplasm resources;
the identification breeding nursery is divided into three relatively separated fields, and the rice is divided into the early, middle and late according to the basic observation of the first year, and the basic agronomic characters are observed and recorded after the rice is sowed in seasons;
the purification rejuvenation nursery is used for the appearance after the first year trial: the materials with low seed activity, serious group mixing and part of diseases are singly bred after single spike seed collection, the purification and rejuvenation nursery consists of two parts of indoor seedling culture and outdoor breeding, and the outdoor breeding is divided into two modes of field breeding and single pot breeding.
The isolation test nursery is used for the first year test of new collection of rice germplasm resources, basic growth conditions and light temperature characteristics of the resources are observed, and some quarantine diseases and insect pests are eliminated; the breeding nursery is identified and mainly divided into three relatively separated fields, and the rice is divided into the early, middle and late according to the basic observation of the first year, and the basic agronomic characters are recorded by observation after the rice is sowed in seasons.
Specifically, the low-temperature storage warehouse is divided into two independent environments with the environmental temperature capable of being set to-4 ℃ and 4 ℃, the relative humidity is controlled below 40%, and the environment is a closed environment and the automatic refrigeration and dehumidification equipment is provided.
Specifically, each nursery of the isolation test nursery, the identification breeding nursery and the purification rejuvenation nursery is independently isolated by a net-shaped enclosing wall with the height of 3 meters; the isolation test nursery, the identification breeding nursery and the purification rejuvenation nursery are paved with the bird preventing net with the 2.5-meter high frame, so that the observation and the record of the characters in the heading and flowering period are facilitated, and the feeding of birds in the grouting maturity period can be prevented. .
All the fields are required to be leveled, the irrigation and drainage are convenient, the fertility is uniform, and drought and inundation risks are avoided.
Specifically, before sowing test seeds, soaking all test seeds in clear water for 18-24 hours, soaking the test seeds in 0.3% strong chlorine essence for 8 hours, cleaning the test seeds in clear water, soaking the test seeds in clear water for 12 hours, draining off water, keeping the test seeds moist, placing the test seeds in an environment at 30 ℃ for accelerating germination for 24-36 hours, observing that white short roots emerge from the test seeds, and sowing the test seeds in a seedling bed;
before transplanting the test seeds, the test field is required to be leveled and the disease, pest and control is required to be carried out, but because the original growth environment of local resources is poor and the number of the seeds is high, the growth characteristics of the seeds are truly reflected, lodging is avoided, and chemical fertilizers are not required to be used in the planting process;
the method comprises the steps of transplanting test seed field blocks into grids according to 6 inch x 8 inch lines, inserting each test seed at grid crossing points during transplanting, planting 2 rows of each variety, 10 plants in each row, 20 plants in total, spacing one row between varieties, reserving 3 grids between every 10 rows as walkways, and inserting a number plate at a first right position of each variety.
Specifically, the seeds harvested by trial seeding are dried in the sun until the safe moisture content is below 13% in the current year of harvesting, stored in a low-temperature storage warehouse at 4 ℃, and then propagated and identified in the next year;
the breeding identification is carried out according to the light-sensitive characteristics of the varieties observed and collected in the first year trial planting, for example, the varieties can be planted in the middle and lower reaches of Yangtze river according to seasons in the morning, the middle and late regions, the sowing time of early rice is 3 months and 25-31 days, the transplanting time is 30 days after sowing, the sowing and transplanting time of middle rice is the same as that of isolated trial planting rice, the sowing time of late rice is 6 months and 20-25 days, and the transplanting time is 25-30 days after sowing;
wherein, the seed soaking and germination accelerating of seeds before sowing are identical to those of the test rice, and the seed soaking time is reduced by 12 hours due to higher air temperature before sowing late rice, and the seed sowing quantity is 80-100 seeds by breeding and identification;
the preparation before isolation test and propagation identification transplanting is basically the same, chemical fertilizers are not needed, 2-3 times of ploughing is needed before leveling, and the purity of the planting in the season resource community is ensured not to be influenced by grain falling;
transplanting grids according to 6 inch-6 inch mark lines, transplanting each resource variety at grid crossing points, planting 6 lines of each variety, 8 lines of each variety, 48 lines of each variety, leaving 3 lines of each 8 lines as walkways, and inserting a number plate at a first right position of each variety;
all propagation identification resources should be harvested immediately after the propagation identification resources reach maturity in the field, and the propagation identification resources only collect 4*6 =24 strains of which the outermost circle is removed from each cell for storage; and the harvested fresh seeds are immediately sun-dried or dried at a temperature of not higher than 40 ℃ until the safe moisture of the seeds is lower than 13%.
Specifically, for the resources with low vigor and failure in a large Tian Chengmiao area, the original resources are taken to be soaked, germinated and then sowed in a nutrition soil small pot with the diameter of 10cm and the height of 8cm, the small pot is placed in an artificial climate box or an artificial climate chamber, the temperature is 30 ℃ and the humidity is 70 percent, the culture is carried out until the seedlings are 10cm long, then the seedlings are moved to natural conditions to grow to two leaves and one core, and then transplanted to a field for normal growth;
judging whether the germination accelerating time is prolonged according to the germination accelerating condition;
it should be noted that the germination may be prolonged more than normal for 24 to 48 hours.
Harvesting a single spike of seeds in the current year after the rejuvenation and propagation of the seeds with low vigor succeed, and carrying out propagation identification on the field in the next year;
for resources with serious population mixing, single spikes for trial planting and seed reserving need to be numbered continuously, the sowing is the same as other propagation identification materials, and the single spikes are planted in adjacent cells of the same field to observe the characters; if the phenotype of each cell is neat during rejuvenation and purification and is consistent with that of the original sowing sample, respectively harvesting and storing the seeds; if the community still has the mixing phenomenon during the test, the material is not purified, is not a germplasm resource meeting the preservation requirement, and is eliminated.
Specifically, after the whole growth period of the resources of the plant diseases and insect pests is found in the test, single plants are harvested for field propagation identification in the second year, and the seeds are subjected to disinfection and sterilization treatment before sowing;
the seed quantity of the resource preservation is not less than 200g after the seed is derived from propagation identification;
harvesting seeds, carrying out impurity cleaning, detecting germination rate, drying, and storing in a low-temperature warehouse after meeting the storage requirement; the resource requirement of the low-temperature warehouse preservation is that the net seeds have a germination rate higher than 90 percent and a water content of the seeds lower than 10 percent;
the method for detecting the germination rate of the rice seeds is carried out by adopting a method in GB/T3543.4-1995, and the water content is measured by adopting a method in GB/T3543.6-1995.
The seeds are stored in two parts after reaching the warehouse-in condition, an aluminum foil bag or a low-temperature-resistant plastic bottle is used, a resource information bar code label is attached to the bag or the bottle, and the two parts of resources are respectively stored in two low-temperature storage banks at 4 ℃ and-4 ℃ for medium and long term storage.
Specifically, the method comprises the steps of loading clean seeds into a breathable package, placing the breathable package in equipment with the ambient temperature of 15 ℃ and the relative humidity of less than 15%, and drying the seeds at a slow speed so as not to damage the activity of the seeds in the drying process.
Specifically, the 4 ℃ low-temperature preservation resource is firstly used for monitoring and using the vitality, the-4 ℃ low-temperature preservation warehouse is used for backup, and the-4 ℃ low-temperature preservation warehouse resource is started after the 4 ℃ low-temperature preservation resource is used up or the vitality is lost.
In the several embodiments provided in this application, it should be understood that the disclosed apparatus and method may be implemented in other ways. The above described device embodiments are only illustrative, e.g. the division of the units is only one logical function division, and there may be other divisions in practice, such as: multiple units or components may be combined or may be integrated into another system, or some features may be omitted, or not performed.
The units described above as separate components may or may not be physically separate, and steps shown as units may or may not be single units; can be singly executed or combined to finish; some or all of the units may be selected according to actual needs to achieve the purpose of the solution of this embodiment.
In addition, each functional unit in each embodiment of the present invention may be integrated in one processing unit, or each unit may be separately used as one unit, or two or more units may be integrated in one unit; the integrated units may be implemented in hardware or in hardware plus software functional units.
The foregoing is merely illustrative embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily think about variations or substitutions within the technical scope of the present invention, and the invention should be covered. Therefore, the protection scope of the invention is subject to the protection scope of the claims.
Claims (10)
1. The method for safely preserving, purifying and rejuvenating local rice germplasm resources is characterized by comprising the following steps of:
obtaining local germplasm resources through a preset channel, constructing a germplasm resource low-temperature conservation library in an ecological area with the same or similar growth, isolating a test nursery, identifying a propagation nursery and purifying a rejuvenation nursery;
randomly extracting a preset number of seeds from the obtained local rice germplasm resources to serve as test seeds, and obtaining image information of the test seeds for checking during harvesting; and the sowing time of sowing and transplanting all the test seeds in the double-cropping rice area is 5 months 15-20 days according to the planting time of the rice in one season, and the transplanting time is 20-25 days after sowing;
after a preset time period, harvesting a single spike at the maturity according to the originally recorded characteristic characteristics of the single spike as seeds for breeding and identifying germplasm, dividing local germplasm resources into a plurality of different resources according to the characteristic characteristics of the field phenotype for purification, rejuvenation and observation comparison according to the characteristic characteristics of the local germplasm resources, wherein the shape characteristics of the local germplasm resources are undefined or the field expressive separation of the resources is serious, so as to obtain test-seed harvested seeds;
during propagation identification, the distribution of seeding time, heading time, mature time, growth period, light temperature property, plant height, spike length, effective spike number, spike extraction degree, spike shape, leaf color, leaf sheath color, leaf fluff, sword leaf length, sword leaf width, sword leaf angle, stem thickness, stem angle, lodging property, miscanthus length, miscanthus color and miscanthus is examined in the growth process, and the grain number per spike, fruiting rate, thousand grain weight, grain length, grain width, grain thickness, seed coat color and grain shape are examined after the mature harvest;
part of resources need to be purified and rejuvenated after trial planting in the first year, and the method comprises the following conditions: after trial planting in the first year, carrying out indoor seedling raising on resources with low vitality and which cannot be planted in the final field, and transplanting the resources to the field for breeding; for resources with serious population mixing, after the test, respectively reserving single plants for continuous number planting observation according to the current annual character of the test; and (4) re-reserving seeds for reproduction after single-pot full-growth period planting is carried out on the resources with diseases.
2. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein:
the low-temperature preservation warehouse is mainly used for low-temperature safe preservation of germplasm resources;
the isolation test nursery is used for the first year test of newly collecting rice germplasm resources, observing the growth condition and light temperature characteristics of the germplasm resources and eliminating quarantine diseases and insect pests;
the identification breeding nursery is divided into three relatively separated fields, and the rice is divided into the early, middle and late according to the basic observation of the first year, and the basic agronomic characters are observed and recorded after the rice is sowed in seasons;
the purification rejuvenation nursery is used for the appearance after the first year trial: the materials with low seed activity, serious group mixing and diseases are singly bred after single spike seed collection, the purification and rejuvenation nursery consists of two parts of indoor seedling culture and outdoor breeding, and the outdoor breeding is divided into two modes of field breeding and single pot breeding.
3. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein: the low-temperature storage warehouse is divided into two independent environments with the environmental temperature capable of being set to-4 ℃ and 4 ℃, the relative humidity is controlled below 40%, and the environment is a closed environment and provided with automatic refrigeration and dehumidification equipment.
4. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 2, wherein: each nursery of the isolation test nursery, the identification breeding nursery and the purification rejuvenation nursery is independently isolated by a net-shaped enclosing wall with the height of 3 meters; the isolation test nursery, the identification breeding nursery and the purification rejuvenation nursery are paved with frames of bird-preventing nets.
5. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein:
before sowing test seeds, soaking all test seeds in clear water for 18-24 hours, soaking the test seeds in 0.3% strong chlorine essence for 8 hours, cleaning the test seeds with clear water, soaking the test seeds in clear water for 12 hours, draining off water, keeping the test seeds moist, placing the test seeds in an environment of 30 ℃ for accelerating germination for 24-36 hours, observing that the test seeds are exposed to white, and sowing the test seeds in a seedling bed;
before transplanting test seeds, the test field needs to be ploughed for 2-3 times and then leveled and subjected to disease and pest control, and chemical fertilizers are not recommended to be used in the planting process because the original growth environment of local resources is poor to truly reflect the growth characteristics of the local resources;
the method comprises the steps of transplanting test seed field blocks into grids according to 6 inch x 8 inch lines, inserting each test seed at grid crossing points during transplanting, planting 2 rows of each variety, 10 plants in each row, 20 plants in total, spacing one row between varieties, reserving 3 grids between every 10 rows as walkways, and inserting a number plate at a first right position of each variety.
6. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein:
the seeds harvested by trial seeding are dried in the sun until the safe moisture is below 13% in the current year of harvesting, stored in a low-temperature storage warehouse at 4 ℃, and then propagated and identified in the next year;
the propagation identification is carried out according to the light-sensitive characteristics of the varieties observed and collected in the first year trial planting, the regions at the middle and lower reaches of the Yangtze river can be planted in seasons in the morning, the middle and the late regions, the sowing time of early rice is 3 months and 25-31 days, the transplanting time is 30 days after sowing, the sowing and transplanting time of middle rice is the same as that of isolated trial planting rice, the sowing time of late rice is 6 months and 20-25 days, and the transplanting time is 25-30 days after sowing;
wherein, the seed soaking and germination accelerating of seeds before sowing are identical to those of the test rice, and the seed soaking time is reduced by 12 hours due to higher air temperature before sowing late rice, and the seed sowing quantity is 80-100 seeds by breeding and identification;
flattening the field before propagation, identification and transplanting, and preparing the same-isolation test seeds;
transplanting grids according to 6 inch-6 inch mark lines, transplanting each resource variety at grid crossing points, planting 6 lines of each variety, 8 lines of each variety, 48 lines of each variety, leaving 3 lines of each 8 lines as walkways, and inserting a number plate at a first right position of each variety;
all propagation identification resources should be harvested immediately after the propagation identification resources reach maturity in the field, the propagation identification resources only collect 4*6 =24 plants of which the outermost circle is removed from each cell, and seeds are harvested for low-temperature preservation; and the harvested fresh seeds are immediately sun-dried or dried at a temperature of not higher than 40 ℃ until the safe moisture of the seeds is lower than 13%.
7. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein:
for the resources with low vigor and failure to grow in a large Tian Chengmiao area, the original resources are taken for seed soaking and germination acceleration, then are sowed in a nutrition soil small pot with the diameter of 10cm and the height of 8cm, are placed in an artificial climatic chamber or an artificial climatic chamber, are cultured in an environment with the temperature of 30 ℃ and the humidity of 70% until the seedlings grow to 10cm and then are moved to natural conditions for growing to two leaves and one core, and are transplanted to a field for normal growth;
immersing low-vigor seeds to accelerate germination, observing germination conditions, and prolonging germination time according to whether seedlings emerge;
harvesting single spike seeds in the current year after the rejuvenation and propagation of the seeds with low vigor succeed, and carrying out propagation identification on the field in the next year;
for resources with serious population mixing, all single spikes of trial planting and seed reserving are required to be numbered continuously, sowing in the second year is the same as other propagation identification materials, and planting in adjacent cells of the same field to observe the characters; if the phenotype of each cell is neat during rejuvenation and purification and is consistent with that of the original sowing sample, respectively harvesting and storing the seeds; if the community still has the mixing phenomenon during the test, the material is not purified, is not a germplasm resource meeting the preservation requirement, and is eliminated.
8. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein:
the resources with diseases are planted in a single pot in the whole growth period, then the single plant is harvested for field propagation identification in the second year, and the seeds are sterilized before sowing;
the seed quantity of the resource preservation is not less than 200g after the seed is derived from propagation identification;
harvesting seeds, carrying out impurity cleaning, detecting germination rate, drying, and storing in a low-temperature warehouse after meeting the storage requirement; the resource requirement of the low-temperature warehouse preservation is that the net seeds have a germination rate higher than 90 percent and a water content of the seeds lower than 10 percent;
the seeds are stored in two parts after reaching the warehouse-in condition, and are sealed and packaged by using an aluminum foil bag or a low-temperature-resistant plastic bottle and the like, a resource information bar code label is attached to the bag or the bottle, and the two parts of resources are respectively stored in two low-temperature storage libraries at 4 ℃ and-4 ℃.
9. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein:
the method comprises the steps of loading seeds into a breathable package, placing the breathable package in equipment with the ambient temperature of 15 ℃ and the relative humidity of less than 15%, and carrying out slow drying so as not to damage the vitality of the seeds in the drying process.
10. A method for safe preservation and purification and rejuvenation of local rice germplasm resources according to claim 1, wherein: the 4 ℃ low-temperature preservation resource is firstly used for monitoring and using the vitality, the-4 ℃ low-temperature preservation library is used for backup preservation, and the-4 ℃ low-temperature preservation library resource is started after the 4 ℃ low-temperature preservation resource is used up or the vitality is lost.
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