CN110250166A - A kind of store method of Bauhinia pollen - Google Patents
A kind of store method of Bauhinia pollen Download PDFInfo
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- CN110250166A CN110250166A CN201910489024.0A CN201910489024A CN110250166A CN 110250166 A CN110250166 A CN 110250166A CN 201910489024 A CN201910489024 A CN 201910489024A CN 110250166 A CN110250166 A CN 110250166A
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- pollen
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N3/00—Preservation of plants or parts thereof, e.g. inhibiting evaporation, improvement of the appearance of leaves or protection against physical influences such as UV radiation using chemical compositions; Grafting wax
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Agronomy & Crop Science (AREA)
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- Engineering & Computer Science (AREA)
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- Wood Science & Technology (AREA)
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention provides a kind of store methods of Bauhinia pollen, the described method comprises the following steps: (1) acquiring drying after fresh Bauhinia pollen；(2) pollen after drying is subjected to Ultra-cryofreezing preservation；(3) pollen after freezen protective is subjected to heating defrosting, the temperature that the heating is thawed successively includes 4 DEG C and 25 DEG C.The store method of Bauhinia pollen of the invention, pass through Ultra-cryofreezing preservation after drying, it thaws then in turn through 4 DEG C and 25 DEG C heatings, it extends the holding time of Bauhinia pollen and improves the germination rate of pollen after preservation, be able to solve the flower hybridization not of the same race for the Bauhinia that the florescence does not meet.
Description
Technical field
The present invention relates to Pollen preservation method fields, and in particular to a kind of store method of Bauhinia pollen.
Background technique
A kind of form of the pollen as plant germplasm, contains the various gene types of the plant, is preserving seed and friendship
The main material changed.But Pollen life-span is very short, the room temperature lower time-to-live is not grown after leaving plant, cryopreservation pollen at
Function, will make it possible pollen storing, exchange.
But after ultralow temperature preservation pollen, the germination rate of pollen is very low.
Summary of the invention
A kind of guarantor of Bauhinia pollen is provided it is an object of the invention to overcome the shortcomings of the prior art place
Deposit method.
To achieve the above object, the technical scheme adopted by the invention is as follows: a kind of store method of Bauhinia pollen, it is described
Method the following steps are included:
(1) dry after the fresh Bauhinia pollen of acquisition；
(2) pollen after drying is subjected to Ultra-cryofreezing preservation；
(3) pollen after freezen protective is subjected to heating defrosting, the temperature that the heating is thawed successively includes 4 DEG C and 25
℃。
The store method of mountain ebony pollen of the invention, by Ultra-cryofreezing preservation after drying, then in turn through 4 DEG C
It thaws with 25 DEG C of heatings, the holding time for extending mountain ebony pollen improves the germination rate of pollen after preservation.
Preferably, the dry pollen to humidity is 29%-38% in step (1).
Ultra-cryofreezing preservation is carried out after being 29%-38% by mountain ebony pollen drying to humidity, pollen after then thawing
Germination rate it is relatively high.
In step (2), the method for the Ultra-cryofreezing preservation are as follows: pollen is placed in liquid nitrogen.
Preferably, in step (3), the time that pollen thaws at 4 DEG C in the heating course of defrosting is 1-4h.
Preferably, in step (3), the time that pollen thaws at 4 DEG C in the heating course of defrosting is 1-3h.
Preferably, in step (3), the time that pollen thaws at 25 DEG C in the heating course of defrosting is 2h.
Preferably, in step (3), the method for the heating defrosting are as follows: the pollen after freezen protective is placed at 4 DEG C and is thawed
2h is subsequently placed in 25 lower defrosting 2h.
Preferably, in step (1), the dry pollen to humidity is 34%.
Preferably, in step (2), the time of pollen freezen protective in liquid nitrogen after the drying is no more than 6 months.
Preferably, in step (2), the time of pollen freezen protective in liquid nitrogen after the drying is no more than 1.5 months.
Preferably, the Bauhinia is orange Bauhinia blakeana (Bauhinia galpinii N.E.Br.), chrysanthemum sheep hoof
First (Bauhinia tomentosa L.), Chinese redbud (Bauhinia variegata L.) and mountain ebony (Bauhinia
One of purpurea L.).
The beneficial effects of the present invention are: it is of the invention the present invention provides a kind of store method of Bauhinia pollen
The store method of mountain ebony pollen is thawed by Ultra-cryofreezing preservation after drying then in turn through 4 DEG C and 25 DEG C heatings,
It is able to solve the flower hybridization not of the same race for the Bauhinia that the florescence does not meet.
Detailed description of the invention
Fig. 1 is the mountain ebony pollen rudiment microscope enlarged drawing of the embodiment of the present invention；(a) it is sprouted for orange Bauhinia blakeana pollen
Bud microscope enlarged drawing；It (b) is chrysanthemum mountain ebony pollen rudiment microscope enlarged drawing；It (c) is mountain ebony pollen rudiment microscope
Enlarged drawing；It (d) is Chinese redbud pollen rudiment microscope enlarged drawing.
Specific embodiment
To better illustrate the object, technical solutions and advantages of the present invention, below in conjunction with specific embodiment to the present invention
It is described further.
Embodiment 1
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the described method comprises the following steps:
(1) being dried after fresh Chinese redbud (the Bauhinia variegata L.) pollen of acquisition to humidity is 34%；
(2) pollen after drying is placed in -196 DEG C of liquid nitrogen freezen protective 1.5 months；
(3) pollen after freezen protective is placed at 4 DEG C 2 hours, is subsequently placed in 2h at 25 DEG C and thaws.
Embodiment 2
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the dry pollen to humidity is 29% in step (1).
Embodiment 3
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the dry pollen to humidity is 38% in step (1).
Comparative example 1
A kind of store method of Bauhinia pollen as comparative example of the present invention, this comparative example are unique with embodiment 1
Difference are as follows: the dry pollen to humidity is 44% in step (1).
Embodiment 5
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the pollen in step (1) is orange Bauhinia blakeana (Bauhinia galpinii N.E.Br.) pollen.
Embodiment 6
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the pollen in step (1) is chrysanthemum mountain ebony (Bauhinia tomentosa L.) pollen.
Embodiment 7
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the pollen in step (1) is mountain ebony (Bauhinia purpurea L.) pollen.
Embodiment 8
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the pollen after freezen protective is placed at 4 DEG C 1 hour in step (3), 2h at 25 DEG C is subsequently placed in and thaws.
Embodiment 9
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the pollen after freezen protective is placed at 4 DEG C 3 hours in step (3), 2h at 25 DEG C is subsequently placed in and thaws.
Embodiment 10
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: the pollen after freezen protective is placed at 4 DEG C 4 hours in step (3), 2h at 25 DEG C is subsequently placed in and thaws.
Comparative example 2
A kind of store method of Bauhinia pollen as the embodiment of the present invention, this comparative example are unique with embodiment 1
Difference are as follows: the pollen after freezen protective is placed in 1-4 hours at 4 DEG C in step (3) and is thawed.
Comparative example 3
A kind of store method of Bauhinia pollen as the embodiment of the present invention, this comparative example are unique with embodiment 1
Difference are as follows: the pollen after freezen protective is placed in 2 hours at 25 DEG C in step (3) and is thawed.
Embodiment 11
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: in step (2), the time of pollen freezen protective at -180 DEG C after the drying is 3 months.
Embodiment 12
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: in step (2), the time of pollen freezen protective at -180 DEG C after the drying is 4.5 months.
Embodiment 13
A kind of store method of Bauhinia pollen as the embodiment of the present invention, the present embodiment are unique with embodiment 1
Difference are as follows: in step (2), the time of pollen freezen protective at -180 DEG C after the drying is 6 months.
Experimental example 1
Pollen viability measuring method: agar 8g/l, sucrose 220g/l, boric acid 20mg/l.As a result statistics takes 4 repetitions, often
5 visuals field are taken every repetition, totally 20 visuals field.Indoor humidity 60%, 28 DEG C of temperature.
Fresh pollen vitality test is 94.8%.
(1) pollen drying experiment
A group: fresh pollen is put in a drying box at room temperature, gradually decreases drying box humidity, from 1 hour, often
Half an hour measures drying box humidity and pollen activity.
Experimental result are as follows: in 1 to 7 hours, with the extension of time, humidity constantly declines in drying box, pollen activity
It is overall on a declining curve；After 8 hours, humidity is down to 0%, and drying at room temperature pollen completely loses vigor.It is wet after 4 hours dry
Degree is reduced to 38% hereinafter, vigor fall is obvious, and germination rate is 72.5% when 4 hours humidity 38%, 4.5 hours humidity
Germination rate is 64.9% when 34%, and germination rate is 56.9% when 5 hours humidity 29%, and germination rate is when 5.5 hours humidity 22%
11.4%.
B group: fresh pollen is placed under room temperature, and from 1 hour, per half an hour measurement drying box humidity and pollen are living
Power.
Experimental result are as follows: in 1 to 7 hours, pollen activity is totally on a declining curve, and fall is slow, pollen at 7 hours
Germination percentage is 65.6%.Since 8 hours, downward trend was obvious, and pollen activity is lost quickly.After measured, 8,9,24,30 hours
Pollen activity is respectively 38.1%, 19.3%, 13.7%, 5.4% afterwards.
(2) pollen thawing test
A group:
It is 29%, 34%, 38%, 44% that fresh Chinese redbud pollen, which is separately dried to humidity, is placed on and is placed in -180 DEG C
Under liquid nitrogen in freezen protective 2 days, temperature programming thaw after culture detection germination rate.
The heating thaw point program of table 1
| Processing routine | Temperature gradient |
| T1 | - 180 DEG C → 25 DEG C 2h → cultures |
| T2 | - 180 DEG C → 4 DEG C 4h → 25 DEG C 2h → cultures |
| T3 | - 180 DEG C → -20 DEG C 4h → 4 DEG C 4h → 25 DEG C 2h → culture |
| T4 | - 180 DEG C → -80 DEG C 4h → -20 DEG C of 4h → 4 DEG C 4h → 25 DEG C 2h → culture |
Experimental result is as shown in table 2
2 different temperatures gradient of table and different humidity pollen germination rate (%)
| Pollen humidity % | 44 | 38 | 34 | 29 | Average value |
| T1 | 27.6 | 47.5 | 69.7 | 55.2 | 50 |
| T2 | 16.2 | 69 | 61.8 | 75.9 | 55.73 |
| T3 | 12.3 | 24.2 | 70.9 | 32.6 | 35 |
| T4 | 3.3 | 68 | 70.1 | 39.8 | 45.3 |
| Average value | 14.85 | 52.18 | 68.13 | 50.88 | -- |
In handling in 4 heating thaw point programs it can be seen from the result of table 2, T2 effect is best, and T1 times good, T3
It is worst, it is placed 4 hours from liquid nitrogen to 4 DEG C, then 2 hours thaw point gradients are placed at 25 DEG C, pollen is able to maintain highest vigor；
Humidity has higher germination rate from the pollen of 29%-38%, wherein 34% pollen is averagely sprouted in 4 gradient increased temperature processing
Rate behaves oneself best up to 68.13%.
B group:
It is 34% that fresh Chinese redbud pollen, which is separately dried to humidity, is placed on to be placed in freeze in the liquid nitrogen at -180 DEG C and protect
It deposits 6 days, culture detection germination rate after temperature programming is thawed.
Experimental result is as shown in table 3
3 different temperatures gradient of table and pollen germination rate
| Processing | Temperature gradient | Germination rate % |
| 1 | - 180 DEG C → culture | 25.7 |
| 2 | - 180 DEG C → 4 DEG C 1h → cultures | 29.2 |
| 3 | - 180 DEG C → 4 DEG C 2h → cultures | 56 |
| 4 | - 180 DEG C → 4 DEG C 3h → cultures | 34.2 |
| 5 | - 180 DEG C → 4 DEG C 4h → cultures | 48.3 |
| 6 | - 180 DEG C → 4 DEG C 1h → 25 DEG C 2h → cultures | 64.6 |
| 7 | - 180 DEG C → 4 DEG C 2h → 25 DEG C 2h → cultures | 75.3 |
| 8 | - 180 DEG C → 4 DEG C 3h → 25 DEG C 2h → cultures | 57 |
| 9 | - 180 DEG C → 4 DEG C 4h → 25 DEG C 2h → cultures | 48 |
| 10 | - 180 DEG C → 25 DEG C 2h → cultures | 41.3 |
By the experimental result of table 3 it is found that 6 days pollen of freezen protective successively passes through 4 DEG C, 25 DEG C in the liquid nitrogen at -180 DEG C
The germination rate of pollen, which is apparently higher than, after defrosting only passes through the germination rate of the pollen after 4 DEG C or 25 DEG C thaw.And from liquid nitrogen
It is placed after middle taking-up pollen at 4 DEG C 2 hours, then places 2 hours and thaw at 25 DEG C, pollen activity keeps best.
(3) pollen, which thaws, saves experiment
It is 34% that Chinese redbud fresh pollen, which is dried to humidity, pollen activity after detection fresh pollen and drying, be stored in-
In 180 DEG C of liquid nitrogen, every one and a half months according to thaw routine: -180 DEG C → 4 DEG C 2h → 25 DEG C 2h → cultures detect pollen germination
Rate, as a result such as table 4.
Influence of the time of 4 cryopreservation of table to pollen germination rate
Table 4 shows that the method according to the invention stores pollen, after the pollen activity of storage 1 after half a month is relative to drying
The vigor of pollen is hardly suffered a loss.6 months pollen activities of storage are still able to maintain 40% or more, after pollen activity is dry
64.4% before storage, still it is able to maintain preferable vigor.Illustrate that method of the invention is able to extend the holding time of pollen, moreover it is possible to protect
The germination rate of pollen after card saves.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of range is protected, although the invention is described in detail with reference to the preferred embodiments, those skilled in the art should
Understand, it can be with modification or equivalent replacement of the technical solution of the present invention are made, without departing from the essence of technical solution of the present invention
And range.
Claims (10)
1. a kind of store method of Bauhinia pollen, which is characterized in that the described method comprises the following steps:
(1) dry after the fresh Bauhinia pollen of acquisition；
(2) pollen after drying is subjected to Ultra-cryofreezing preservation；
(3) pollen after freezen protective is subjected to heating defrosting, the temperature that the heating is thawed successively includes 4 DEG C and 25 DEG C.
2. the method according to claim 1, wherein the dry pollen to humidity is 29%- in step (1)
38%.
3. method according to claim 1 or 2, which is characterized in that in step (3), pollen in the heating course of defrosting
The time thawed at 4 DEG C is 1-4h.
4. according to the method described in claim 3, it is characterized in that, pollen is 4 in the heating course of defrosting in step (3)
The time thawed at DEG C is 1-3h.
5. the method according to claim 1, wherein in step (2), the method for the Ultra-cryofreezing preservation
Are as follows: pollen is placed in liquid nitrogen.
6. the method according to claim 1, wherein in step (3), method that the heating is thawed are as follows: will be cold
The pollen frozen after saving is placed at 4 DEG C the 2h that thaws, and is subsequently placed at 25 DEG C the 2h that thaws.
7. the method according to claim 1, wherein the dry pollen to humidity is 34% in step (1).
8. the method according to claim 1, wherein the pollen after the drying is cold in liquid nitrogen in step (2)
Freeze the time saved no more than 6 months.
9. the method according to claim 1, wherein the pollen after the drying is cold in liquid nitrogen in step (2)
Freeze the time saved no more than 1.5 months.
10. the method according to claim 1, wherein the Bauhinia is orange Bauhinia blakeana, chrysanthemum sheep hoof
One of first, Chinese redbud and mountain ebony.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104206378A (en) * | 2014-09-23 | 2014-12-17 | 江苏省林业科学研究院 | Willow pollen ultralow-temperature storage method |
| CN105104361A (en) * | 2015-06-30 | 2015-12-02 | 中国林业科学研究院亚热带林业研究所 | Camellia oleifera pollen ultra low temperature storage method |
| US20170238535A1 (en) * | 2015-06-25 | 2017-08-24 | Accelerated Ag Technologies, Llc | Pollen field conditioning and preservation method |
-
2019
- 2019-06-05 CN CN201910489024.0A patent/CN110250166A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104206378A (en) * | 2014-09-23 | 2014-12-17 | 江苏省林业科学研究院 | Willow pollen ultralow-temperature storage method |
| US20170238535A1 (en) * | 2015-06-25 | 2017-08-24 | Accelerated Ag Technologies, Llc | Pollen field conditioning and preservation method |
| CN105104361A (en) * | 2015-06-30 | 2015-12-02 | 中国林业科学研究院亚热带林业研究所 | Camellia oleifera pollen ultra low temperature storage method |
Non-Patent Citations (4)
| Title |
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| 傅鸿妃: "植物花粉超低温保存研究进展", 《长江蔬菜》 * |
| 胡柔璇 等: "洋紫荆的栽培技术研究", 《绿色科技》 * |
| 陈世昌 等: "《植物组织培养》", 30 June 2016, 重庆大学出版社 * |
| 陈伟峰: "羊蹄甲种间杂交不亲和初探", 《西北植物学报》 * |
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Application publication date: 20190920 |