CN102144632A - Method for storing and unfreezing pinus massoniana pollen - Google Patents
Method for storing and unfreezing pinus massoniana pollen Download PDFInfo
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- CN102144632A CN102144632A CN2011100326820A CN201110032682A CN102144632A CN 102144632 A CN102144632 A CN 102144632A CN 2011100326820 A CN2011100326820 A CN 2011100326820A CN 201110032682 A CN201110032682 A CN 201110032682A CN 102144632 A CN102144632 A CN 102144632A
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Abstract
The invention discloses a method for storing and unfreezing pinus massoniana pollen. The method comprises the following steps of: storing, namely performing water culture on pinus massoniana male flowers with branches and leaves, picking off buds when the buds is to blossom, placing the buds into a kraft paper envelope, drying at constant temperature, dispersing pollen, pinking one corner of the envelope by using a needle, collecting the pollen into a brown bottle with silica gel, sealing the opening with a sealant, storing the brown bottle at normal temperature and in shade, exchanging the silica gel timely when color of the silica gel is changed, and sealing the bottle when the color of the silica gel is not changed and storing the bottle at a low temperature; and unfreezing, namely taking out the brown bottle and placing the brown bottle into a refrigerating chamber of a refrigerator for 20 to 24 hours, taking out the brown bottle and placing the brown bottle at a normal-temperature environment for 8 to 12 hours, opening the sealant and placing the brown bottle for 8 to 12 hours until the color of the silica gel is changed, or opening the opening if the color of the silica gel is not changed, adhering filter paper absorbed with water to the opening, and covering the opening by using a preservative film and sealing the opening by using a rubber band until the color of the silica gel is changed. By the method, life of the pinus massoniana pollen is prolonged; the problems of cross breeding (seed production) of pine tree seeds and flowering asynchronism of artificially auxiliary pollination in a seed orchard are solved; and utilization ratio of the pollen is increased.
Description
Technical field
The present invention relates to the storage of pollen, especially a kind of storage of masson pine pollen and defreezing method thereof.
Background technology
Masson pine (Pinus massoniana) is that the fat seeds are produced in important the holding concurrently with material in China south, and its economic worth height is of many uses, is the primary raw material seeds of industries such as China's woods slurry paper, rosin, building.In addition, its pollen, needle and bark all can be used as medicine; Pine nut oil-containing 30% can be made paint, soap and lubricating oil; Cone can refine crude oil; Pine stump can extract pine tar, can also cultivate precious traditional Chinese medicine---Poria cocos.
Guangxi is China masson pine main producing region, has the whole nation the best genetic resources and the highest productive potential.The 1980s, Guangxi has been built 200 hectares to be grown to the masson pine primary seed garden of improvement target, has brought into play important effect for promoting masson pine improved variety process.But,, cause interior male and female flowering asynchronism of seed orchard or pollen to distribute unequal phenomenon, so that seed orchard yields poorly and biennial bearing is obvious because the garden of the building clone of seed orchard reaches 464.For this reason, how efficiently to utilize pollen to become the important topic that needs to be resolved hurrily.
In southern conifer seeds, except that pinus finlaysoniana, the flowering time of masson pine the latest, and florescence difference is also bigger in planting.Therefore, in carrying out the masson pine kind and in the interspecific cross breeding process, be necessary used its pollen storage to the coming year, and the life-span of masson pine pollen has only 30 days under field conditions (factors), therefore, pollen storage has important practical significance in seed orchard artificial supplementary pollination and crossbreeding (production of hybrid seeds).
Summary of the invention
The technical problem to be solved in the present invention provides a kind of storage and defreezing method thereof of masson pine pollen, make life-span prolongation, the pollen availability of masson pine pollen improve, to solve the problems such as flowering asynchronism of male and female flowering in seed orchard artificial supplementary pollination and the crossbreeding.
Adopt following technical scheme for solving the problems of the technologies described above the present invention: the storage practice of masson pine pollen, with masson pine male flower band branches and leaves water planting, bud is waited to open to take and is put into the ox-hide envelope, 25～28 ℃ of freeze-day with constant temperature 5～10h, pollen scatters, and envelope is used the pin pinking for one jiao, pollen is collected in the brown bottle that silica gel is housed and is sealed with fluid sealant and deposit in normal temperature lucifuge place, in time change silica gel during the silica gel variable color, be sealed in-10～-20 ℃ of low temperature when treating the silica gel nondiscolouring and preserve.
The pollen moisture content is 5～8% during the silica gel nondiscolouring.
The defreezing method of said method gained storage pollen: brown bottle is taken out be placed on cold compartment of refrigerator 20～24h earlier, take out then and be placed on normal temperature 8～12h, open fluid sealant again and place 8～12h, treat that the silica gel variable color gets final product; If bottleneck is then opened in the silica gel nondiscolouring, the filter paper of having inhaled water is attached to bottle mouth position, cover with preservative film again and seal with rubber band, treat that the silica gel variable color gets final product.
Use the storage and the defreezing method thereof of masson pine pollen of the present invention, extended to more than 1 year in 30 days that masson pine pollen is preserved by normal temperature, can effectively solve the flowering asynchronism problem of controlled pollination and the seed orchard artificial supplementary pollination of conifer seeds, also improve the pollen availability.This method equipment is simple, processing ease, can extensively be useful in scientific research, teaching and production unit, is easy to popularize in an all-round way use.
Description of drawings
Fig. 1 uses the flow sheet that the present invention handled and used masson pine pollen.
Embodiment
Embodiment 1
As shown in Figure 1, application the present invention handles and uses the production procedure of masson pine pollen mainly to comprise:
＜1〉collection of pollen and drying: at the beginning of 3 months by the end of February every year, masson pine male flower branch is waited out in collection, take back from bag with template, for preventing the pollen contamination in the seed orchard, all sprays need the water flushing, the spray of cleaning is put into different isolated areas and carries out water planting, cuts when bud is to be opened to put into and carries out drying after the ox-hide envelope seals, and drying condition is 28 ℃ of freeze-day with constant temperature 5h; Pollen scatters, and envelope is used the pin pinking for one jiao, pollen is collected in the brown bottle that silica gel is housed and with fluid sealant sealed, and the outer pollen that is scattered of bottle is clean with 75% wipes of alcohol; Brown bottle is deposited in normal temperature lucifuge place, in time change silica gel during the silica gel variable color, (the pollen moisture content is 8% after measured) reaches the storage requirement when treating the silica gel nondiscolouring.
＜2〉low temperature storage of pollen: with step＜1〉brown bottle that dry good pollen is housed seals in-20 ℃ of low temperature with fluid sealant and preserves.
＜3〉thawing of storage pollen: winter in the coming year, spring, the brown bottle that will preserve masson pine pollen earlier takes out and is placed on 8 ℃ of 24h of cold compartment of refrigerator; Take out then and be placed on normal temperature 12h, open fluid sealant again and place 12h, allow pollen in course of defrosting, recover suction, treat that the silica gel variable color gets final product; Generally speaking, south mostly is wet weather spring, and pollen can recover moisture content naturally under indoor conditions; If bottleneck is then opened in the silica gel nondiscolouring, the filter paper of having inhaled water is attached to bottle mouth position, cover with preservative film again and seal with rubber band, treat that the silica gel variable color gets final product.
＜4〉pollen germination rate detects: adopt the culture fluid method, and specific as follows
1. prepare culture fluid: take by weighing boric acid 0.5g, glucose 5g, anhydrous calcium chloride 0.03g, be settled to 100mL after adding the dissolving of a small amount of aquae destillata respectively, promptly get best culture fluid " glucose (or sucrose) 5%+0.5% boric acid+0.03%CaCl
2(pH5.6～6.0) ";
2. pollen is cultivated: splash into 1～2 culture fluid in the slide of band groove, be placed in the culture fluid with the sticking a small amount of pollen of dissecting needle and stir 28 ℃ of dark 24h of cultivation that preserve moisture;
3. detect: with 100 times of biology microscope sem observation pollen, the pollen tube growth of the blodynamic pollen of tool, extended length is got 3 about 100 pollen in the above visual field greater than the diameter of pollen, calculates germination rate.
＜5〉land for growing field crops artificial pollination: through step＜3〉thaw and absorb water finish after, pollen is sucked rubber pipette bulb, seal stand-by with fluid sealant; Carry out the 1st artificial pollination in the sunny morning breezy, concrete steps are to open the pollination sack, aim at female flower spray 1 to 2 time, seal immediately; Finish the 2nd pollination the 2nd to 3 day the morning.
Embodiment 2
＜1〉collection of pollen and drying: 25 ℃ of freeze-day with constant temperature 10h, pollen moisture content are 6%.
＜2〉low temperature storage of pollen :～10 ℃ of low temperature are preserved.
＜3〉thawing of storage pollen: 4 ℃ of 21h of cold compartment of refrigerator, normal temperature 8h opens fluid sealant again and places 10h.
Other are substantially with embodiment 1.
Embodiment 3
＜1〉collection of pollen and drying: 26 ℃ of freeze-day with constant temperature 7h, pollen moisture content are 5%.
＜2〉low temperature storage of pollen :-15 ℃ of low temperature are preserved.
＜3〉thawing of storage pollen: 6 ℃ of 23h of cold compartment of refrigerator, normal temperature 10h opens fluid sealant again and places 9h.
Other are substantially with embodiment 1.
Embodiment 4
＜1〉collection of pollen and drying: 28 ℃ of freeze-day with constant temperature 8h, pollen moisture content are 7%.
＜2〉low temperature storage of pollen :-17 ℃ of low temperature are preserved.
＜3〉thawing of storage pollen: 4 ℃ of 20h of cold compartment of refrigerator, normal temperature 9h opens fluid sealant again and places 8h.
Other are substantially with embodiment 1.
Embodiment 1 to 4 masson pine pollen germination rate testing result such as table 1:
Table 1 masson pine pollen germination rate testing result
Annotate: control group system preserves be placed on indoor black out place behind the fresh pollen collecting chamber inner drying in brown bottle.
As shown in Table 1, use the germination rate of pollen of the present invention after 1 year and reach more than 60%, and the germination rate of control group is 0, illustrates that storage of the present invention and defreezing method thereof are feasible, has effectively prolonged the life-span of masson pine pollen.
In addition, the inventor to drying after different reserve temperature conditions study result such as table 2:
The masson pine pollen germination rate of the different reserve temperature conditions of table 2
Annotate: 0 day result is meant the germination rate before the refrigeration of dry back.
By table 2 as seen, the pollen germination rate that normal temperature is preserved descended nearly 30% in 72 days, germination rate reduces to about 10% in the time of 144 days, be defective pollen, promptly the holding time under normal temperature condition is 72～144 days (holding time of the fresh pollen of undried is 20～30 days) to masson pine pollen after drying.By contrast, the following 358 days average germination rate of pollen of-20 ℃ deepfreeze condition also has 62.2%, has only descended 15.1%.-20 ℃ therefore, be impose a condition the down preferable reserve temperature of horse hair pollen pini of the present invention.
In sum, adopt storage of the present invention and defreezing method, can make masson pine pollen reach desirable germination rate,, opened up a new way for the crossbreeding (production of hybrid seeds) of conifer seeds simultaneously for research and production provide a cover complete pollen handling technique.
Claims (3)
1. the storage practice of a masson pine pollen, it is characterized in that: with masson pine male flower band branches and leaves water planting, bud is waited to open to take and is put into the ox-hide envelope, 25～28 ℃ of freeze-day with constant temperature 5～10h, pollen scatters, and envelope is used the pin pinking for one jiao, pollen is collected in the brown bottle that silica gel is housed and is sealed with fluid sealant and deposit in normal temperature lucifuge place, in time change silica gel during the silica gel variable color, be sealed in-10～-20 ℃ of low temperature when treating the silica gel nondiscolouring and preserve.
2. the storage practice of masson pine pollen according to claim 1 is characterized in that: the pollen moisture content is 5～8% during described silica gel nondiscolouring.
3. according to the defreezing method of the method gained storage pollen of claim 1, it is characterized in that: brown bottle is taken out be placed on cold compartment of refrigerator 20～24h earlier, take out then and be placed on normal temperature 8～12h, open fluid sealant again and place 8～12h, treat that the silica gel variable color gets final product; If bottleneck is then opened in the silica gel nondiscolouring, the filter paper of having inhaled water is attached to bottle mouth position, cover with preservative film again and seal with rubber band, treat that the silica gel variable color gets final product.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103385238A (en) * | 2013-07-01 | 2013-11-13 | 镇江瑞繁农艺有限公司 | Method for keeping water lily pollen viability |
| CN104351170A (en) * | 2014-09-23 | 2015-02-18 | 江苏省林业科学研究院 | Shrub willow pollen preservation method |
| CN105104362A (en) * | 2015-08-13 | 2015-12-02 | 广西壮族自治区中国科学院广西植物研究所 | Simple long-term storage preservation method for pitaya pollen |
| CN106472487A (en) * | 2016-09-30 | 2017-03-08 | 湖南省茶叶研究所（湖南省茶叶检测中心） | A kind of collection storage practice of tea plant pollen |
| CN108719281A (en) * | 2018-07-24 | 2018-11-02 | 广西壮族自治区林业科学研究院 | A kind of cryopreservation method of Pollen of Masson Pine |
| CN113575413A (en) * | 2021-09-06 | 2021-11-02 | 棕榈生态城镇发展股份有限公司 | Breeding method of new variety of michelia figo |
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| CN101129174A (en) * | 2007-09-06 | 2008-02-27 | 安徽省华鼎生物科技有限公司 | Process for preparing organic selenium pine pollen |
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- 2011-01-30 CN CN2011100326820A patent/CN102144632B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101129174A (en) * | 2007-09-06 | 2008-02-27 | 安徽省华鼎生物科技有限公司 | Process for preparing organic selenium pine pollen |
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| 徐进等: "不同贮藏方法及光照对马尾松花粉活力的影响", 《南京林业大学学报》, vol. 22, no. 3, 31 December 1998 (1998-12-31), pages 71 - 74 * |
| 顾培英: "精制马尾松花粉的营养和加工储藏技术", 《经济林研究》, vol. 16, no. 1, 31 December 1998 (1998-12-31), pages 15 - 18 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103385238A (en) * | 2013-07-01 | 2013-11-13 | 镇江瑞繁农艺有限公司 | Method for keeping water lily pollen viability |
| CN104351170A (en) * | 2014-09-23 | 2015-02-18 | 江苏省林业科学研究院 | Shrub willow pollen preservation method |
| CN105104362A (en) * | 2015-08-13 | 2015-12-02 | 广西壮族自治区中国科学院广西植物研究所 | Simple long-term storage preservation method for pitaya pollen |
| CN106472487A (en) * | 2016-09-30 | 2017-03-08 | 湖南省茶叶研究所（湖南省茶叶检测中心） | A kind of collection storage practice of tea plant pollen |
| CN108719281A (en) * | 2018-07-24 | 2018-11-02 | 广西壮族自治区林业科学研究院 | A kind of cryopreservation method of Pollen of Masson Pine |
| CN113575413A (en) * | 2021-09-06 | 2021-11-02 | 棕榈生态城镇发展股份有限公司 | Breeding method of new variety of michelia figo |
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