CN104498426A - Collection, storage, in-vitro culture and viability detection technology for acacia auriculiformis pollen - Google Patents
Collection, storage, in-vitro culture and viability detection technology for acacia auriculiformis pollen Download PDFInfo
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- CN104498426A CN104498426A CN201410778864.6A CN201410778864A CN104498426A CN 104498426 A CN104498426 A CN 104498426A CN 201410778864 A CN201410778864 A CN 201410778864A CN 104498426 A CN104498426 A CN 104498426A
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Abstract
The invention discloses a collection, storage, in-vitro culture and viability detection technology for acacia auriculiformis pollen. The technology comprises the following steps: collecting acacia auriculiformis inflorescence blooming in the morning, placing into a sulfuric acid paper bag, and drying in the shade inside at 25-30 DEG C; collecting pollen when anthers greatly crack after 10 o'clock in the next morning, and screening by virtue of a sieve; putting the collected pollen into a small glass bottle, adding dry allochroic silicagel, vacuumizing and sealing with wax; storing the sealed pollen at 25 DEG C to -25 DEG C; and germinating the pollen, namely putting the stored pollen into a liquid culture medium and germinating at 25-35 DEG C. According to the method disclosed by the invention, the technical problems of collection, storage and germination of the acacia auriculiformis pollen are solved; the germination rate of the acacia auriculiformis pollen can be significantly improved; the germination of the acacia auriculiformis pollen stored at -20 DEG C for a year is still 48%; and compared with similar artificial hybridization pollination and artificial pollination of seed orchards, the technology disclosed by the invention has great significance on improvement of the seed yield and quality.
Description
Technical field
The invention belongs to plant hybridization breeding field, be specifically related to a kind of method improving acacia auriculiformis pollen germination rate.
Background technology
Acacia auriculiformis (
acacia auriculiformis) be the plant of Mimosaceae (Mimosaceae) Acacia (Acacia Mill.).Natural distributed, in 5 ° ~ 17 °, south latitude, originates in Queensland ,Australia, Papua New Guinea south and western, her Li An island, Indonesia Ji Dao.Acacia auriculiformis biomass is large, and material is excellent, can be used as pulpwood, Building wood, furniture woods etc.; Tool nitrogen fixation, can be used for improveing soil, green barren hill etc.But Acacia seeds ubiquity the low phenomenon of setting percentage, and acacia auriculiformis is no exception, this is one of key issue of further developing of its plant husbandry of restriction.Acacia auriculiformis is solid low, and the florescence very easily affects by the bad climate such as typhoon, cold damage, causes to yield poorly and unstable, thus limits the development of acacia auriculiformis.Florescence artificial supplementary pollination promotes percentage of fertile fruit, improves seed production, is comparatively effective solution.Pollen collection, storage, vitality test technology realize the necessary sport technique segment of this measure.Therefore, the research carrying out the biology of reproduction field of acacia rachii plant is significant.
Pollen is phanerogamous microgametophyte, plays an important role in sexual propagation.The sprouting of pollen, pollen tube growth are the important steps of sexual plant reproduction, comprise the physiological process of many complexity.Pollen is that passage conveying sperm completes fertilization process with pollen tube, and the sprouting of pollen and pollen tube growth realize amphigameous key.
Acacia (Acacia) seeds ubiquity the low phenomenon of setting percentage, acacia auriculiformis (
acacia auriculiformisa. Cunn. ex Benth.) no exception, this is one of key issue of further developing of its plant husbandry of restriction.Acacia auriculiformis is solid low, and the florescence very easily affects by the bad climate such as rainwater, typhoon, causes to yield poorly and unstable, thus limits the development of acacia auriculiformis.Florescence artificial supplementary pollination promotes percentage of fertile fruit, improves seed production, is comparatively effective solution.
Summary of the invention
The object of the present invention is to provide acacia auriculiformis pollen collection, storage and isolated culture viability examination technology.
The technical solution used in the present invention is:
Acacia auriculiformis pollen collection, storage and isolated culture viability examination technology, comprise the steps:
(1) pollen collection: gather morning and opened the same day in fresh acacia auriculiformis inflorescence loading sulfuric acid paper bag, put into 25 DEG C ~ 30 DEG C indoor and dry in the shade, collect pollen when flower pesticide bursts in a large number after morning 10, with the sieved sieve of pollen;
(2) pollen storage: the pollen gathered is placed in phial, adds dry discolour silica gel, vacuumizes, seal with wax, and preserves at the pollen sealed is placed in 25 DEG C ~-25 DEG C;
(3) pollen germination: the pollen of storage is placed in liquid nutrient medium, sprouts at 25 DEG C ~ 35 DEG C.
As preferably: step (1) be in the morning dew not dry before gather the fresh inflorescence opened the same day.
As preferably: the inflorescence of collection is placed in 28 DEG C of thermostatic chambers by step (1), is laid on template and dries in the shade.
As preferably: in step (1), pollen 200 object sieve.
As preferably: pollen is placed in storage at 5 DEG C or-20 DEG C by step (2).
Liquid nutrient medium described in step (3) is configured by sucrose, boric acid and water and forms.
As preferably: consisting of of the liquid nutrient medium described in step (3): 100 ~ 200g/L sucrose, 100 ~ 300 mg/L boric acid, surplus is water.
Preferred further: consisting of of described liquid nutrient medium: 200g/L sucrose, 300 mg/L boric acid, surplus is water.
As preferably: in step (3), pollen is sprouted under 30 DEG C of constant temperature.
The invention has the beneficial effects as follows:
1, acacia auriculiformis pollen collecting method of the present invention, when thermostatic chamber dries in the shade, the period is grasped, and observes and carries out brush dust again when flower pesticide bursts completely and sieve, solve tiny this difficult problem of not easily collecting of acacia auriculiformis pollen.
2, acacia auriculiformis pollen storage method of the present invention, be placed on respectively in different environment and preserve, detected by Pollen Activity, detect along with storage time constantly increases, the Pollen Activity preserved under the environment of-20 DEG C is the strongest, solves this difficult problem of acacia auriculiformis pollen storage.
3, the boric acid containing 20% sucrose, 300ppm in the liquid nutrient medium that adopts of present method is formulated, and liquid stabilising is reliable, is particularly suitable for acacia auriculiformis pollen and leaves one's post cultivation.
4, on slide glass groove, drip 1-2 drop of liquid substratum, brushed lightly by the acacia auriculiformis pollen gathered and put on liquid nutrient medium, pollen directly can contact with oxygen, and pollen is evenly distributed, thus auriculiformis pollen can normally be sprouted.These key points for operation solve acacia rachii in-vitro pollen and cultivate germination gordian technique.
5, present method to auriculiformis pollen collection, preserve, leave one's post cultivation and viability examination, simple, the rapid and feature such as workable of method, data science is reliable, and can be completely quantitative, for raising auriculiformis breeding efficiency, there is higher actual application value, effectively can solve the problem that existing acacia rachii breeding garden nature Pod Bearing Percentage is low, improve the seed output and quality of breeding garden, enable breeding garden effectively play its due function, greatly play the economic benefit of acacia rachii seeds, social benefit and ecological benefits.
Accompanying drawing explanation
Fig. 1: for the acacia auriculiformis inflorescence of pollen collecting;
Fig. 2: the acacia auriculiformis pollen of collection;
Fig. 3: acacia auriculiformis pollen cultures 24 hours detected results of collecting the same day;
Fig. 4 :-20 DEG C storage January acacia auriculiformis pollen cultures 24 hours detected results;
Fig. 5 :-20 DEG C of storages acacia auriculiformis pollen cultures 24 hours detected results of 1 year.
Embodiment
Below in conjunction with embodiment, the present invention is further illustrated, but be not limited thereto.
embodiment 1
In auriculiformis the flowers are in blossom time early August, before 8 o'clock, gather fresh inflorescence be placed in sulfuric acid paper bag, tile in 28 DEG C of thermostatic chambers and dry in the shade, to be seen burst completely to flower pesticide after, brush dust sieving for standby after the next morning 10, the groove of slide glass drips suitable liquid nutrient medium (200g/L sucrose+300 mg/L boric acid) 1-2 drip, the pollen gathered is brushed lightly on liquid medium within respectively, slide glass is put into culture dish cultivate, put into two moistening filter paper in culture dish, keep culture temperature 30 DEG C.Cultivate 3,6,9,12, after 24, by the sprouting number of observation by light microscope pollen, pollen tube length and pollen tube number.The acacia auriculiformis pollen that the same day collects, can be observed pollen germination rate is after 24 hours of incubation 94%, the longest 6 times of reaching pollen diameter of pollen tube length, and a pollen can sprout at most 6 pollen tubes.The pollen purity of the acacia auriculiformis collected is very high, does not almost have other impurity.Fig. 1 is the acacia auriculiformis inflorescence for pollen collecting.Fig. 2 is the pollen of the acacia auriculiformis of collection under opticmicroscope.Fig. 3 is that the acacia auriculiformis in-vitro pollen collecting the same day cultivates viability examination result.
embodiment 2
In acacia auriculiformis in-vitro pollen germination process, the groove of slide glass drips suitable liquid nutrient medium (200g/L sucrose+300 mg/L boric acid) 1-2 drip, by gather and at room temperature, preserved the pollen in January in 5 DEG C ,-20 DEG C environment and instead do not brushed lightly on liquid medium within, slide glass is put into culture dish cultivate, put into two moistening filter paper in culture dish, keep culture temperature 30 DEG C.Cultivate 3,6,9,12, after 24, by the sprouting number of observation by light microscope pollen, pollen tube length and pollen tube number.Preserve the acacia auriculiformis pollen in January; after at room temperature storage, 5 DEG C of storages ,-20 DEG C of storages, the pollen germination rate cultivated after 24 hours is followed successively by 28%, 42%, 63%; the longest 3 times of reaching pollen diameter of pollen tube length, a pollen can sprout at most 3 pollen tubes.Fig. 4 is that-20 DEG C of acacia auriculiformis in-vitro pollens preserving January cultivate viability examination result.
embodiment 3
In acacia auriculiformis in-vitro pollen germination process, the groove of slide glass drips suitable liquid nutrient medium (200g/L sucrose+300 mg/L boric acid) 1-2 drip, by gather and at room temperature, preserved in 5 DEG C ,-20 DEG C environment 1 week, the pollen in January, 1 year instead do not brush lightly on liquid medium within, slide glass is put into culture dish cultivate, put into two moistening filter paper in culture dish, keep culture temperature 30 DEG C.Cultivate 3,6,9,12, after 24, by the sprouting number of observation by light microscope pollen, pollen tube length and pollen tube number.Preserve the acacia auriculiformis pollen of 1 year; after at room temperature storage, 5 DEG C of storages ,-20 DEG C of storages, the pollen germination rate cultivated after 24 hours is followed successively by 13%, 28%, 48%; the longest 2 times of reaching pollen diameter of pollen tube length, a pollen can sprout at most 2 pollen tubes.Fig. 5 is that-20 DEG C of storages acacia auriculiformis in-vitro pollen of 1 year cultivates viability examination result.
Claims (9)
1. acacia auriculiformis pollen collection, storage and isolated culture viability examination technology, comprise the steps:
(1) pollen collection: gather morning and opened the same day in fresh acacia auriculiformis inflorescence loading sulfuric acid paper bag, put into 25 DEG C ~ 30 DEG C indoor and dry in the shade, collect pollen when flower pesticide bursts in a large number after morning 10, by sieve;
(2) pollen storage: the pollen gathered is placed in phial, adds dry discolour silica gel, vacuumizes, seal with wax, and preserves at the pollen sealed is placed in 25 DEG C ~-25 DEG C;
(3) pollen germination: the pollen of storage is placed in liquid nutrient medium, sprouts at 25 DEG C ~ 35 DEG C.
2. acacia auriculiformis pollen collection according to claim 1, storage and isolated culture viability examination technology, is characterized in that: step (1) be in the morning dew not dry before gather the fresh inflorescence opened on the same day.
3. acacia auriculiformis pollen collection according to claim 1, storage and isolated culture viability examination technology, is characterized in that: the inflorescence of collection is placed in 28 DEG C of thermostatic chambers by step (1), is laid on template and dries in the shade.
4. acacia auriculiformis pollen collection according to claim 1, storage and isolated culture viability examination technology, is characterized in that: in step (1), pollen 200 object sieve.
5. acacia auriculiformis pollen collection according to claim 1, storage and isolated culture viability examination technology, is characterized in that: step (2) is preserved at pollen being placed in 5 DEG C or-20 DEG C.
6. acacia auriculiformis pollen collection according to claim 1, storage and isolated culture viability examination technology, is characterized in that: the liquid nutrient medium described in step (3) is configured by sucrose, boric acid and water and forms.
7. acacia auriculiformis pollen collection according to claim 6, storage and isolated culture viability examination technology, it is characterized in that: consisting of of the liquid nutrient medium described in step (3): 100 ~ 200g/L sucrose, 100 ~ 300 mg/L boric acid, surplus is water.
8. acacia auriculiformis pollen collection according to claim 7, storage and isolated culture viability examination technology, is characterized in that: consisting of of described liquid nutrient medium: 200g/L sucrose, 300 mg/L boric acid, and surplus is water.
9. acacia auriculiformis pollen collection according to claim 1, storage and isolated culture viability examination technology, is characterized in that: in step (3), pollen is sprouted under 30 DEG C of constant temperature.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104872111A (en) * | 2015-05-22 | 2015-09-02 | 桂林嘉佰农农业科技有限公司 | Storing method of pollens subjected to soft x ray irradiation |
| CN105176910A (en) * | 2015-10-28 | 2015-12-23 | 山东省林业科学研究院 | Superior dry-land willow tree pollen in-vitro germination culture medium and method for determining viability of dry-land willow pollen |
| CN106212267A (en) * | 2016-08-04 | 2016-12-14 | 四川省林业科学研究院 | A kind of Alnus cremastogyne staminate inflorescence rapid draing and pollen efficient collecting method |
| CN107494522A (en) * | 2017-08-29 | 2017-12-22 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Spontaneum pollen low-temperature storage method |
| WO2019010360A1 (en) * | 2017-07-06 | 2019-01-10 | Accelerated Ag Technologies, Llc | Pollen preservation method |
| RU2799580C2 (en) * | 2017-07-06 | 2023-07-06 | Акселерейтед Эй Джи Текнолоджиз, Ллк | Pollen preservation method |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104872111A (en) * | 2015-05-22 | 2015-09-02 | 桂林嘉佰农农业科技有限公司 | Storing method of pollens subjected to soft x ray irradiation |
| CN105176910A (en) * | 2015-10-28 | 2015-12-23 | 山东省林业科学研究院 | Superior dry-land willow tree pollen in-vitro germination culture medium and method for determining viability of dry-land willow pollen |
| CN106212267A (en) * | 2016-08-04 | 2016-12-14 | 四川省林业科学研究院 | A kind of Alnus cremastogyne staminate inflorescence rapid draing and pollen efficient collecting method |
| WO2019010360A1 (en) * | 2017-07-06 | 2019-01-10 | Accelerated Ag Technologies, Llc | Pollen preservation method |
| RU2799580C2 (en) * | 2017-07-06 | 2023-07-06 | Акселерейтед Эй Джи Текнолоджиз, Ллк | Pollen preservation method |
| CN107494522A (en) * | 2017-08-29 | 2017-12-22 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Spontaneum pollen low-temperature storage method |
| CN107494522B (en) * | 2017-08-29 | 2020-05-19 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Low-temperature storage method for sugarcane pollen |
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