CN108739800A - A kind of method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage - Google Patents
A kind of method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage Download PDFInfo
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- CN108739800A CN108739800A CN201810665453.4A CN201810665453A CN108739800A CN 108739800 A CN108739800 A CN 108739800A CN 201810665453 A CN201810665453 A CN 201810665453A CN 108739800 A CN108739800 A CN 108739800A
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- pollen
- strongly fragrant
- lonicera periclymenum
- fragrant lonicera
- periclymenum
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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
Abstract
The invention belongs to plant sexual reproduction technique fields, disclose a kind of method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage, collect the fresh pollen of strongly fragrant Lonicera periclymenum same day loose powder, detect pollen germination rate, germination rate is at 80% or more, the fresh pollen of collection is dispensed to centrifuge tube, nozzle is sealed, is placed in -20 DEG C of storages, when storing about 40d or so, pollen is taken out, its germination rate is detected, generally remains to reach 40%, at this time, other parents just enter full-bloom stage, can take out pollen, carry out artificial supplementary pollination.The present invention solves the problems, such as strongly fragrant Lonicera periclymenum crossbreeding flowering asynchronism, easy to operation, preserves at low cost and greatly prolongs the pollen tool great-hearted time；By carrying out the research of in vitro pollen low tempertaure storage, for promoting the artificial hybridization breeding work of strongly fragrant Lonicera periclymenum to be of great significance.
Description
Technical field
The invention belongs to technical field of plant propagation, are related to a kind of storage practice of plant pollen, and in particular to Yi Zhongyu
The method of Lonicera periclymenum in vitro pollen short term storage.
Background technology
Strongly fragrant Lonicera periclymenum（Lonicera fragrantissima Lindl. et Paxt.）, it is the one of Caprifoliaceae Lonicera
Kind of semi-evergreen shrubs, up to 2m, the fragrance of flowers is strong, is important aromatics flowers and trees of northern early spring, first spends rear leaf, color when blooming
Perfume (or spice) has both, and flower amount is flouring, in recent years, is often applied in vertical greening, is the extraordinary sight flower of one kind, sees fruit Evergreen garden plant
Kind.
Strongly fragrant Lonicera periclymenum has that the fragrance of flowers is strong, and early spring equal good characteristics of blooming are made in the rearing new variety of woodbine
It is available for outstanding parent material, but strongly fragrant Lonicera periclymenum is usually initially entered the full blossom stage in mid-February to March, and and its
The other plant of the closer Lonicera of affiliation（Such as Flos Lonicerae maackii（Lonicera maackii (Rupr.) Maxim.
var. maackii）Deng）, full-bloom stage is generally all in mid or late April, flowering asynchronism, limit artificial hybridization breeding work into
Row, therefore, exploring a kind of method of the strongly fragrant Lonicera periclymenum in vitro pollen of short term storage seems extremely necessary,
Ripe pollen contains whole inhereditary material information of plant, and under field conditions (factors), the service life of many pollen is very short, this
Great inconvenience is brought to research of fruit germplasm resource and crossbreeding work, while being also unfavorable for relevant plant genetic research.Into
The research of row in vitro pollen short term storage promotes the development of crossbreeding work to be all of great significance for solving flowering asynchronism.
Patent CN 201611121492.5 describes a kind of Flos Lonicerae maackii pollen cryopreservation method, by fresh Flos Lonicerae maackii pollen
From well-developed elite stand 13 on the day of blooming:It is fitted into the cryopreservation tube of 1.8ml, i.e., initially contains after adopting after 30 or so acquisitions
Water will fill when being 13%~14% or so to be preserved in polliniferous cryopreservation tube input 196 DEG C of liquid nitrogen containers of ﹣, then uses tap water
The mode of 5min of thawing is thawed.Though this method can realize the cryopreservation of pollen, more demanding to preservation condition, one
As in the case of be relatively inaccessible to, and operating process is complicated, greatly reduces the operability of pollen preservation.
Invention content
The other plant of the full-bloom stage of the strongly fragrant Lonicera periclymenum Lonicera closer earlier than its affiliation, causes flowering asynchronism, limit
The development artificial hybridization breeding work under natural conditions has been made, meanwhile, the Pollen life-span of strongly fragrant Lonicera periclymenum is short, is not easy to the long period
Natural storage, hinder breeding process and improve Germ-plasma resources protection.
In order to solve the above technical problems, the present invention provides a kind of method of the strongly fragrant Lonicera periclymenum in vitro pollen of short term storage, have
Effect overcomes the problem of flowering asynchronism of the strongly fragrant Lonicera periclymenum with its affiliation closer Lonicera other plant, is created for its germplasm
Newly, breeding of new variety provides reference frame.
The purpose of the present invention is be achieved through the following technical solutions：
The method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage of the present invention, acquires strongly fragrant Lonicera periclymenum pollen, is placed in pollen storage and holds
After device, pollen hold-up vessel mouth is sealed, is stored in 5 DEG C~-20 DEG C of refrigerator, storage time 40 or so.
As advanced optimizing for the method, the acquisition time of strongly fragrant Lonicera periclymenum pollen is the morning 9:00~10:00, it is adopted
Pollen should be the fresh pollen of same day loose powder.
As advanced optimizing for the method, the germination rate of the strongly fragrant Lonicera periclymenum fresh pollen of acquisition is not less than 80%.
As advanced optimizing for the method, -20 DEG C of the reserve temperature of strongly fragrant Lonicera periclymenum fresh pollen, storage time 40d
Left and right, pollen germination rate are not less than 40%.
On the basis of above-mentioned parameter optimizes, the method for the strongly fragrant Lonicera periclymenum in vitro pollen short term storage comprising with
Lower step：
1）In the morning 9:00~10:00 takes the flower of same day loose powder, and the pollen bag of loose powder is gently brushed with hairbrush, collects pollen；
2）Pollen activity is detected, the germination rate of strongly fragrant Lonicera periclymenum fresh pollen is not less than 80%；
3）The strongly fragrant Lonicera periclymenum fresh pollen of acquisition is dispensed to pollen hold-up vessel, pollen hold-up vessel mouth is sealed, be stored in-
In 20 DEG C of refrigerator, pollen activity is during which irregularly detected；
4）When storing 40d or so, pollen activity is detected, germination rate is not less than 40%.
Compared with prior art, beneficial effects of the present invention or advantage major embodiment be in the following areas：
The present invention provides a kind of method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage, by the fresh pollen of acquisition be sub-packed in from
In heart pipe, centrifuge tube nozzle is sealed, is stored in -20 DEG C of refrigerator, storage time 40d or so, by detecting the flower after storing
Powder vigor knows that preserving in vitro strongly fragrant Lonicera periclymenum pollen by the method for the invention, there is preferable vigor, pollen germination rate to be not less than
40%.The present invention solves the problems, such as strongly fragrant Lonicera periclymenum crossbreeding flowering asynchronism, and it is certain that 40d or so can guarantee that in vitro pollen has
Viability provides condition for the breeding of strongly fragrant Lonicera periclymenum, the selection and breeding work of new varieties.
The method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage of the present invention, simple and easy to do, operability is strong, using common
Just accessible -20 DEG C of chilled storages, condition simplicity easily reach, feasibility is very high typical refrigerator.
Description of the drawings
Fig. 1 is the germination rate after the in vitro fresh pollen of strongly fragrant Lonicera periclymenum is stored at different temperatures.
Fig. 2 be strongly fragrant Lonicera periclymenum dry in the shade in vitro pollen store at different temperatures after germination rate.
Specific implementation mode
With reference to embodiment, the present invention is further illustrated, and following each embodiments are merely to illustrate the present invention, right
The present invention is not restricted.
Embodiment 1
The present invention is described in further detail by taking the strongly fragrant Lonicera periclymenum in the botanical garden of Xian District, Shanxi Province as an example.Described in the present embodiment
The method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage, in vitro pollen used are the fresh pollen of strongly fragrant Lonicera periclymenum.
1）Harvest pollen
Pollen-March 10 is harvested, growth selection is healthy and strong in the garden of Xi'an botanical garden, the strongly fragrant Lonicera periclymenum plant of no disease and pests harm,
In the morning 9:00~10:00 takes the flower of same day loose powder, is placed on template, takes back laboratory rapidly, is gently brushed and dissipated with hairbrush
The pollen bag of powder collects pollen.
2）Measure pollen water content
The water content of the fresh pollen of acquisition is measured by the prior art, measurement result shows that the water content of fresh pollen is
90%。
3）Detect pollen activity
The vigor of pollen is measured by the prior art（TTC decoration methods）, measurement result shows that the germination rate of fresh pollen is
96.28%。
4）Storage
Fresh pollen is dispensed to centrifuge tube, nozzle is sealed, is respectively placed in typical refrigerator freezing chamber（-20℃）, refrigerating chamber（5
℃）And room temperature storage.
5）Detect pollen germination rate
The pollen germination rate under different reserve temperatures is detected respectively when storing 5d, 10d, 21d, 31d and 40d, as a result such as Fig. 1 institutes
Show.From figure 1 it appears that under different reserve temperatures, the germination rate and storage time of pollen are negatively correlated, in room temperature, 5
DEG C and -20 DEG C under the conditions of, with the extension of storage time, the fresh pollen germination rate of strongly fragrant Lonicera periclymenum reduces, in room temperature storage
When 11d, when pollen germination rate is reduced to 0,5 DEG C of storage 31d, pollen germination rate is reduced to 0, after storing 40d at -20 DEG C, germination rate
Remain at 40.63%, illustrate for fresh pollen, -20 DEG C be its short term storage best mode.
6）Human assistance crossbreeding
May 19 pollen that stored in -20 DEG C, germination rate is 40.63% was taken out, other woodbines are manually awarded
Powder.
Embodiment 2
The present invention is described in further detail by taking the strongly fragrant Lonicera periclymenum in the botanical garden of Xian District, Shanxi Province as an example.Described in the present embodiment
The method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage, in vitro pollen used are the pollen that dries in the shade of strongly fragrant Lonicera periclymenum.
1）Harvest pollen
Pollen-March 10 is harvested, growth selection is healthy and strong in the garden of Xi'an botanical garden, the strongly fragrant Lonicera periclymenum plant of no disease and pests harm,
In the morning 9:00~10:00 takes the flower of same day loose powder, is placed on template, takes back laboratory rapidly, is gently brushed and dissipated with hairbrush
The pollen bag of powder collects pollen.
2）Measure pollen water content
After the pollen of the acquisition 48h disposed within that dries in the shade, the water content of pollen is measured by the prior art, measurement result is shown,
Dry in the shade pollen water content be 55%.
3）Detect pollen activity
Pass through the prior art（TTC decoration methods）The vigor for the pollen that dries in the shade is measured, measurement result shows that the germination rate for the pollen that dries in the shade is
86.32%。
4）Storage
The pollen that will dry in the shade is dispensed to centrifuge tube, seals nozzle, is respectively placed in typical refrigerator freezing chamber（-20℃）, refrigerating chamber（5
℃）And room temperature storage.
5）Detect pollen germination rate
The pollen germination rate under different reserve temperatures is detected respectively when storing 5d, 10d, 21d, 31d and 40d, as a result such as Fig. 2 institutes
Show.From figure 2 it can be seen that under different reserve temperatures, the germination rate and storage time of pollen are negatively correlated, in room temperature, 5
DEG C and -20 DEG C under the conditions of, with the extension of storage time, the strongly fragrant Lonicera periclymenum pollen germination rate after drying in the shade reduces, room temperature store
When hiding 11d, when pollen germination rate is reduced to 0,5 DEG C of storage 31d, pollen germination rate is reduced to 0, after storing 40d at -20 DEG C, sprouts
Rate is 36.17%, is illustrated for the strongly fragrant Lonicera periclymenum in vitro pollen after drying in the shade, -20 DEG C be its short term storage best mode.
It is found through experiments that, when carrying out short term storage to strongly fragrant Lonicera periclymenum pollen with the method, pollen water content stores it
Hide influential effect it is little, therefore, may be selected fresh pollen is directly stored, the method had both avoided dry in the shade during to spending in vitro
The influence of powder vigor, simple and easy to do, operability is strong, and solves the problems, such as flowering asynchronism in Eclectics's breeding.
Further narration is done to the present invention above in conjunction with embodiment, but present invention is not limited to the embodiments described above,
One skilled in the relevant art within the scope of knowledge, can also make without departing from the purpose of the present invention
Various change.
Claims (5)
1. a kind of method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage, which is characterized in that acquire strongly fragrant Lonicera periclymenum pollen, be placed in pollen
After hold-up vessel, pollen hold-up vessel mouth is sealed, is stored in 5 DEG C~-20 DEG C of refrigerator, 40 d of storage time or so.
2. the method for strongly fragrant Lonicera periclymenum in vitro pollen short term storage according to claim 1, which is characterized in that strongly fragrant Lonicera periclymenum flower
The acquisition time of powder is the morning 9:00~10:00, picked flowers powder should be the fresh pollen of same day loose powder.
3. the method for strongly fragrant Lonicera periclymenum in vitro pollen short term storage according to claim 1, which is characterized in that the Yu Xiang of acquisition
The germination rate of honeysuckle fresh pollen is not less than 80%.
4. the method for strongly fragrant Lonicera periclymenum in vitro pollen short term storage according to claim 2, which is characterized in that strongly fragrant Lonicera periclymenum is new
- 20 DEG C of the reserve temperature of fresh flower powder, storage time 40d or so, pollen germination rate is not less than 40% after storage.
5. the method for strongly fragrant Lonicera periclymenum in vitro pollen short term storage according to claim 1, which is characterized in that including following step
Suddenly：
1）In the morning 9:00~10:00 takes the flower of same day loose powder, and the pollen bag of loose powder is gently brushed with hairbrush, collects pollen；
2）Pollen activity is detected, the germination rate of strongly fragrant Lonicera periclymenum fresh pollen is not less than 80%；
3）The strongly fragrant Lonicera periclymenum fresh pollen of acquisition is dispensed to pollen hold-up vessel, pollen hold-up vessel mouth is sealed, be stored in-
In 20 DEG C of refrigerator, pollen activity is during which irregularly detected；
4）When storing 40d or so, pollen is taken out, detects pollen activity, pollen germination rate is not less than 40%.
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| CN201810665453.4A CN108739800A (en) | 2018-06-26 | 2018-06-26 | A kind of method of strongly fragrant Lonicera periclymenum in vitro pollen short term storage |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110024777A (en) * | 2018-12-22 | 2019-07-19 | 吉林省农业科学院 | A kind of short-term preservation method of sunflower powder |
| CN110558224A (en) * | 2019-08-30 | 2019-12-13 | 台州市农业科学研究院 | Method for collecting, identifying and storing tulip pollen and tulip crossbreeding method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110024777A (en) * | 2018-12-22 | 2019-07-19 | 吉林省农业科学院 | A kind of short-term preservation method of sunflower powder |
| CN110024777B (en) * | 2018-12-22 | 2021-03-26 | 吉林省农业科学院 | Short-term preservation method of sunflower pollen |
| CN110558224A (en) * | 2019-08-30 | 2019-12-13 | 台州市农业科学研究院 | Method for collecting, identifying and storing tulip pollen and tulip crossbreeding method |
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