CN102523785B - Application of safranine T in seed dye mark tracing aspect - Google Patents
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- CN102523785B CN102523785B CN 201210016548 CN201210016548A CN102523785B CN 102523785 B CN102523785 B CN 102523785B CN 201210016548 CN201210016548 CN 201210016548 CN 201210016548 A CN201210016548 A CN 201210016548A CN 102523785 B CN102523785 B CN 102523785B
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Abstract
The invention discloses application of safranine T in a seed dye mark tracing aspect. According to the application disclosed by the invention, a seed dye marking operation is carried out by using the safranine T, so that the seed is free from damage, the activity of the seed is not influenced, and the dyeing effect is lasting. The application disclosed by the invention can be applied to researching plant seed dispersal and seed bank in various types of ecosystems, such as farmland ecosystem, rivers and lakes ecosystem, grassland ecosystem, forest ecosystem and the like.
Description
Technical field
The present invention relates to a kind of new purposes of safranine T, be specifically related to safranine T and be applied to the spike of seed dye marker.
Background technology
Seed dispersal, diffusion etc. have linked the end of maternal plant cyclostage and the foundation of their offspring populations, think extensively that now it has deep effect to vegetation structure.Seed dispersal has a lot of meanings to plant population, group and ecosystem biology, it not only influences dynamically growing with continuing of population, and the heredity districtization influences interbiotic interaction, can also change richness and the distribution of species, and then the structure of group is exerted an influence.And seed dispersal also has very important meaning to the weeds fauna in the agricultural ecosystem.Study that various media generally adopt botanizing seed bank quantity and spatial distribution dynamically to propagation, the diffusion influence of plant seed or with the method for bead simulation.These methods can disclose various routes of transmission qualitatively to be influenced dynamically to the dissemination of plant seed with to plant species word bank space, can not determine the propagation ratio of ruderal plant seed, also be difficult to the group in plant seed propagation and each ecosystem is linked up.Primary reason is that the seed dispersal circulation is complicated: become between the strain formation a lot of intermediate steps and process are arranged in seed generation and a new generation; Another reason is, seed dispersal is difficult to follow the tracks of, and the researcher is difficult to follow the tracks of seed dispersal that maternal plant produces to new place, thereby also is difficult to determine their destiny.In recent years, the application of some new technologies and method is carried out the mark spike with the seed of propagating, and seed and seedling and maternal plant corresponding aspects are being shown good prospect.These mark tracer techniques comprise: labelled with radioisotope method, fluorochrome label method, stable isotope analysis and molecular genetic marker etc.Use these methods and successful mark often need big grain, contain the seed that enriches endosperm, and these detections that are labeled seed need special instrument and complicated operations comparatively, use inconvenient, time-consuming taking a lot of work, wherein fluorochrome label method and radioisotope exist safety and problem of environmental pollution especially, in addition, the cost of these methods is all expensive, thereby their range of application is restricted.
Summary of the invention
To the objective of the invention is the defective that exists in the prior art in order solving, a kind of method that adopts safranine T to carry out the spike of seed dye marker to be provided.
In order to achieve the above object, the invention provides the application of a kind of safranine T aspect the spike of seed dye marker.Safranine T may further comprise the steps when the seed dyeing that comes off for prematurity:
(1) preparation safranine T dyeing liquor: get safranine T or safranine T and ethanol, add water, mix, safranine T is fully dissolved, preparation safranine T dyeing liquor; The concentration expressed in percentage by weight of safranine T is 0.5～5% in the described safranine T dyeing liquor, and the concentration expressed in percentage by volume of ethanol is 0～70%;
(2) seed dye marker: before the plant seed maturation comes off, get the safranine T dyeing liquor of preparation in the step (1), the original position of plant spray or repeatedly brushing safranine T dyeing liquor on plant seed, dyed redness until seed.
Safranine T may further comprise the steps when being used for ripe seed dyeing:
(1) preparation safranine T dyeing liquor: get safranine T or safranine T and ethanol, add water, mix, safranine T is fully dissolved, preparation safranine T dyeing liquor; The concentration expressed in percentage by weight of safranine T is 0.5～5% in the described safranine T dyeing liquor, and the concentration expressed in percentage by volume of ethanol is 0～70%;
(2) seed dye marker: gather ripe plant seed, after indoor natural wind is done, place the safranine T dyeing liquor of step (1) preparation to soak 0.5～5 hour, pull out then and drain dyeing liquor, plant seed is namely dyed and is redness behind the room-dry.
Wherein, be used for the seed source of dyeing in farmland weed, forest-tree, give birth to or water plants in the natural pond, specifically derive from following plant: grass family such as Japanese amur foxtail, amur foxtail Wang grass, the weeds rice, wild oat, Paspalum distichum, barnyard grass, long awns barnyard grass, lady's-grass, faber bristlegrass herb, green foxtail, pearl millet, Bermuda grass, siberian wildrye, perennial ryegrass, Korea lawn grass, the caput grass, long awns caput grass, gunn is grass firmly, bromegrass, annual bluegrass, roegneria kamoji, junglerice, reed, reed, hispid arthraxon, bent grass, the weeds rice, Caryophyllaceae such as ox chickweed, the adhesive hair grasswort, polygonaceae such as sheep hoof, rumex dentatus, the willow leaf knotweed, Ranunculaceae such as Ranunculus sceleratus, fennel fennel garlic, Chenopodiaceae such as little lamb's-quarters, Scrophulariaceae such as Mazus japonicus, resupinate woodbetony leaf or root, the Persian Veronica, wall speedwell, Veronica, the north water speedwell, Oenotheraceae such as false loosestrife, composite family such as Herb of Common Nipplewort, common sow thistle, the mugwort wormwood artemisia, lyrate hemistepta herb, Siberian cocklebur, Solidago Canadensis, little fluffy grass, Radix Conyzae sumatrensis, the spoon dish, annual fleabane herb, Amaranthaceae such as Amaranthus retroflexus, the root of bidentate achyranthes, Malvaceae such as piemarker, Labiatae such as benbie, Rubiaceae such as clearvers, Cruciferae such as penny cress, shepherd's purse, sedge family such as Fischer grass, needle spikesedge herb, dark green sedge, the flat Sha of ball fringe, beach louse grass, kyllinga brevifolia, difformed galingale herb, Pittosporaceae such as pittosporum tobira, Magnoliaceae is as having a smile on one's face, Buxaceae such as little leaf boxwood, Ulmaceae such as elm, Tiliaceae such as lime tree, Aceraceae such as trident maple, acer monoes, the shank tool, Juglandaceae such as blue or green money willow, Chinese ash, the rose family such as potentilla chinensis; Towards the sky potentilla chinensis, Euphorbiaceae such as humid euphorbia, big humid euphorbia, spot humid euphorbia, Lythraceae such as mexicana, Umbelliferae such as cnidium monnieri etc.
The present invention has the following advantages compared to existing technology: safranine T is safranin O again, is a kind of natural dyeing agent of extracting from the safflower column cap.The employing safranine T carries out the seed dye marker, and is harmless to seed, do not influence the activity of seed, and Color is lasting.It is simple to operate to utilize the present invention that seed is carried out dye marker, and cost is low, and is pollution-free.Can be applicable to study that plant seed in the multiple ecosystems such as field ecosystem, the rivers and lakes ecosystem, forest ecosystem is propagated and the research of seed bank.
Embodiment
Below in conjunction with example the present invention is described in further details.
Embodiment 1
(by the end of May) used the safranine T dyeing liquor that the seed of wheatland Zhong Wang grass plant is carried out the original position dye marker before the real maturation of Zai Wang grass seed came off, use banister brush to dip in the safranine T dyeing liquor during dyeing Wang grass seed is brushed repeatedly, until the faint yellow Wang grass seed of script is dyed equal level dyeing to red.
Embodiment 2
1. ripening in summer the weed seed maturing stage, collection wheat field weed Wang grass (
Beckmannia syzigachne), Japanese amur foxtail (
Alopecurus japonicus) ripe son real, after indoor natural wind is done, pull out after 2 hours with the immersion of safranine T dyeing liquor, be placed on room-dry.Originally faint yellow De Wang grass, real being dyed of Japanese amur foxtail are redness.
2. the comparison of the weight of weed seed, germination rate comparison and floating ability before and after being labeled
The mensuration of seed weight: the thousand kernel weight before and after the weighing grass seed is labeled, repeat 5 times.
Seed germination rate is measured: the filter paper of diameter 10 cm is layered in the culture dish of same diameter, uses water-soaked filter paper, and weed seed dye marker unmarked with 100 evenly is placed on the filter paper respectively then, add a cover, repeat, be placed on 20 ± 1 ℃, light for 5 times: in the plant incubator of dark=12:12.Whole experimental session keeps filter paper moistening.Counted the weed seed quantity of once sprouting in per 2 days, till weed seed is no longer sprouted.
Dyeing Hou De Wang grass and Japanese amur foxtail seed all increase a little than undyed weight, but difference not significantly (table 1) showing that the dyeing processing has only increased the weight of these two kinds of weed seeds a little, can not change their floatation characteristic.The germination rate of dyeing Hou De Wang grass, Japanese amur foxtail seed reduces a little than undyed, and difference is remarkable (table 1) not, shows that dyeing has no significant effect the germination rate of these two kinds of weed seeds, i.e. dyeing does not damage their vigor, dyeing liquor safety.
Weight and germination rate are relatively before and after table 1 Wang grass, the dyeing of Japanese amur foxtail seed
Annotate: it is not remarkable that the alphabetical identical table behind the mean value is shown on 0.05 level difference.
Embodiment 3
The safranine T dyeing liquor of preparation variable concentrations: get safranine T 0.5g, 1g, 2g, 3g respectively, 4g, 5g join in the 100ml ethanolic solution of 50% concentration expressed in percentage by volume, mix, and safranine T is fully dissolved, and are prepared into the safranine T dyeing liquor.The Wang grass seed is soaked in the different safranine T dyeing liquors respectively, pulls out after 1 hour, be placed on indoor natural seasoning after, originally light color De Wang grass seed is dyed and is redness, and the germination rate before and after the dyeing of Bi Jiao Wang grass seed changes.
1% safranine T dyeing liquor of preparation different ethanol concentration: get safranine T 1g, add concentration expressed in percentage by volume respectively and be 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70% 100ml ethanolic solution, mix, safranine T is fully dissolved, be prepared into the safranine T dyeing liquor.The Wang grass seed is soaked in the different safranine T dyeing liquors respectively, pulls out after 1 hour, be placed on indoor natural seasoning after, originally light color De Wang grass seed is dyed and is redness, and the germination rate before and after the dyeing of Bi Jiao Wang grass seed changes.
When germination rate is measured, Wang grass seed branch interspersed among be equipped with in the plate of moistening filter paper, each dull and stereotyped 50 seed repeats, plate is placed on 20 ± 1 ℃, light for 5 times: in the plant incubator of dark=12:12.Whole experimental session keeps filter paper moistening.Counting was once sprouted De Wang grass seed quantity in per 2 days, till seed is no longer sprouted.
Table 2 uses the germination rate of variable concentrations safranine T dyeing De Wang grass seed to compare
Annotate: (it is not remarkable that ± alphabetical identical table after SE) is shown on 0.05 level difference for mean value.
Table 3 uses the germination rate of the safranine T dyeing De Wang grass seed of different percentage concentration ethanol preparations to compare
Annotate: (it is not remarkable that ± alphabetical identical table after SE) is shown on 0.05 level difference for mean value.
Use the germination rate of safranine T dyeing liquor dyeing Hou Wang grass of variable concentrations for there not being significant difference, the safranine T Ke of 0.5%-5% be described with the dyeing of Yong Yu Wang grass, and the germination (table 2) of Bu Ying Xiang Wang grass after dyeing.
Use the germination rate of 1% safranine T dyeing liquor dyeing Hou Wang grass of different concentration ethanol preparation for there not being significant difference, water is described, or concentration expressed in percentage by volume is not higher than the preparation (table 3) that 70% ethanol may be used to dyeing liquor.
Embodiment 4
Gather ripe weeds rice autumn in the farmland, after drying, seed soaked with the safranine T dyeing liquor pull out after 1 hour, be placed on indoor natural seasoning after, script light color weeds rice is dyed is redness.The relatively germination rate variation of weeds rice dyeing front and back behind the seed drying.In addition the weeds rice of dyeing is imbedded respectively in rice field 5cm, 10 cm, the 15cm soil, observed the change color of dyeing weeds rice after 1 year.
When germination rate is measured, weeds rice branch interspersed among be equipped with in the plate of moistening filter paper, each dull and stereotyped 50 seed repeats, plate is placed on 20 ± 1 ℃, light for 5 times: in the plant incubator of dark=12:12.Whole experimental session keeps filter paper moistening.Counted the weed seed quantity of once sprouting in per 2 days, till the weeds rice is no longer sprouted.
Finally, germination rate before the dyeing of weeds rice is 76.2 ± 9.5%, and using the germination rate after the safranine T dyeing liquor dyes is 74.8 ± 10.4%, the weeds rice germination rate there was no significant difference before and after the dyeing, illustrate that the safranine T dyeing liquor is good to the safety of weeds rice, does not influence its germination and emergence.Experiment finds, the weeds rice of safranine T dyeing in imbedding rice field 5cm, 10 cm, 15cm soil after 12 months institute's red colouration all clear and legible, illustrate that this dyeing can keep lastingly.
Embodiment 5
Be collected in the lime tree samara in the string bag of falling into prior setting after autumn, maturation came off, samara is pulled out after 3 hours with the immersion of safranine T dyeing liquor, after being placed on indoor natural seasoning, elm seed and fruit wing are all dyed is redness, and relatively the germination rate of elm seed dyeing front and back changes.
When germination rate is measured, the seed before and after the elm dyeing divided respectively to intersperse among be equipped with in the plate of moistening filter paper, 25 in every ware repeats, plate is placed on 25 ± 1 ℃, light for 4 times: in the plant incubator of dark=12:12.Whole experimental session keeps filter paper moistening.Break in the seed coat 1mm as the seed sprouting standard so that radicle is prominent, in the germination process every day observed and recorded, note to keep filter paper moistening, when continuous 2 d do not have new seed to germinate, be considered as germinateing and finish, chitting piece number before and after the statistics dyeing calculates germination rate, germination index.Computing formula is as follows:
Subnumber * 100 % are planted experimentally in the total germinative number of germination rate (G)=seed/confession
Germination index (G
i)=∑ (G
t/ D
t)
Wherein: G
t-at the germination rate of time t; D
t-fate accordingly germinates
All in the time of 2-4 days, whole germination process has continued about 15 days to elm seed sprouting peak, dyeing front and back, and the germination rate before the dyeing is 89 ± 9.5%, germination index 6.2 ± 1.4%; Using the germination rate after the dyeing of safranine T dyeing liquor is 84 ± 3.3%, germination index 5.7 ± 0.6%, and there are no significant the difference of elm percentage of seedgermination, the germination index before and after the dyeing illustrates that the safranine T dyeing liquor is good to the elm seed safety, does not influence its germination and emergence.
Embodiment 6
Gather ripe Pedicularis palustris L seed in the fall, behind the natural air drying seed soaked with the safranine T dyeing liquor and pull out after 1 hour, be placed on after the indoor natural seasoning seed namely quilt dyed and be redness.Relatively the germination rate before and after the dyeing of Pedicularis palustris L seed changes.
When germination rate is measured, Pedicularis palustris L seed branch interspersed among be equipped with in the plate of moistening filter paper, each dull and stereotyped 50 seed repeats, plate is placed on 20 ± 1 ℃, light for 5 times: in the plant incubator of dark=12:12.Whole experimental session keeps filter paper moistening.Counted the weed seed quantity of once sprouting in per 2 days, the chitting piece number before and after statistics dyes respectively till the Pedicularis palustris L seed is no longer sprouted, calculating germination rate.
The Pedicularis palustris L seed germination rate is 69 ± 7.8% before the dyeing, using the germination rate after the safranine T dyeing liquor dyes is 65 ± 8.2%, elm Pedicularis palustris L percentage of seedgermination there was no significant difference before and after the dyeing, illustrate that the safranine T dyeing liquor is good to the Pedicularis palustris L seed safety, does not influence its germination and emergence.
Embodiment 6
At the end of spring and the beginning of summer or gather ripe following table real (fruit and seed) autumn, behind the natural air drying the practical safranine T dyeing liquor of son soaked after 1 hour and pull out, be placed on after the indoor natural seasoning seed and namely dyed and be redness.
With dyeing and the seed that not have to dye bury respectively in the dark soil of paddy field 10cm, keep the 5cm depth of water in the rice field, after burying 30 days, 90 days, 150 days, the situation of change that the seed of every kind dyeing and non-dyeing digs out 20 above observed and recorded seed colors respectively.Seed digs out and cleans the back 30 ± 2
ZeroC drying box inner drying is after 10-15 days, if naked eyes can go out the difference of the seed of dyeing and non-dyeing respectively, then get 5 seeds and under identical conditions, these seeds are taken pictures for every kind, use Adobe Photoshop 6.0 softwares that the color of seed is carried out analysis and distinguishing then.
As shown in Table 4, the weed seed of the dyeing when burying 30 days in all tables is all clear and legible, has only the weed seed of only a few to fade when burying 90 days, and burying still has can clearly differentiating of seed dyeing over half and non-dyeing after 150 days.
Table 4.The color of the plant seed of dyeing is with the variation of buried time
Annotate: weed seed buries in paddy field, buried depth 10cm, and keep 5 centimetres the depth of water in the rice field.(± lowercase different table after 1SD) is shown in significant difference on 0.05 level to mean value.
Claims (4)
1. the application of safranine T aspect the spike of seed dye marker; Described seed source is given birth to or water plants in farmland weed, forest-tree, natural pond.
2. the application of safranine T according to claim 1 aspect the spike of seed dye marker is characterized in that: may further comprise the steps:
(1) preparation safranine T dyeing liquor: get safranine T or safranine T and ethanol, add water, mix, safranine T is fully dissolved, preparation safranine T dyeing liquor; The concentration expressed in percentage by weight of safranine T is 0.5～5% in the described safranine T dyeing liquor, and the concentration expressed in percentage by volume of ethanol is 0～70%;
(2) seed dye marker: before the plant seed maturation comes off, get the safranine T dyeing liquor of preparation in the step (1), the original position of plant spray or repeatedly brushing safranine T dyeing liquor on plant seed, dyed redness until seed.
3. the application of safranine T according to claim 1 aspect the spike of seed dye marker is characterized in that: may further comprise the steps:
(1) preparation safranine T dyeing liquor: get safranine T or safranine T and ethanol, add water, mix, safranine T is fully dissolved, preparation safranine T dyeing liquor; The concentration expressed in percentage by weight of safranine T is 0.5～5% in the described safranine T dyeing liquor, and the concentration expressed in percentage by volume of ethanol is 0～70%;
(2) seed dye marker: gather ripe plant seed, after indoor natural wind is done, place the safranine T dyeing liquor of step (1) preparation to soak 0.5～5 hour, pull out then and drain dyeing liquor, plant seed is namely dyed and is redness behind the room-dry.
4. the application of safranine T according to claim 1 aspect the spike of seed dye marker, it is characterized in that: described seed source is in grass family, Caryophyllaceae, Ranunculaceae, Chenopodiaceae, Scrophulariaceae, Oenotheraceae, composite family, Amaranthaceae, Malvaceae, Labiatae, Rubiaceae, Cruciferae, sedge family, Pittosporaceae, Magnoliaceae, Buxaceae, Ulmaceae, Tiliaceae, Aceraceae, Juglandaceae, the rose family, Euphorbiaceae, Lythraceae or samphire.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3267010A (en) * | 1962-04-16 | 1966-08-16 | Udylite Corp | Electrodeposition of copper from acidic baths |
| CN101002679A (en) * | 2007-01-26 | 2007-07-25 | 中国人民解放军第三军医大学第一附属医院 | Tracing method for medulla desmohemoblast stem cell |
| CN101536629A (en) * | 2009-03-17 | 2009-09-23 | 北京市植物园 | Method for causing seeds of cypripedium macranthum to sprout |
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2012
- 2012-01-19 CN CN 201210016548 patent/CN102523785B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3267010A (en) * | 1962-04-16 | 1966-08-16 | Udylite Corp | Electrodeposition of copper from acidic baths |
| CN101002679A (en) * | 2007-01-26 | 2007-07-25 | 中国人民解放军第三军医大学第一附属医院 | Tracing method for medulla desmohemoblast stem cell |
| CN101536629A (en) * | 2009-03-17 | 2009-09-23 | 北京市植物园 | Method for causing seeds of cypripedium macranthum to sprout |
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