CN101792731A - Method for separating Chinese white poplar ovule from nucellus - Google Patents
Method for separating Chinese white poplar ovule from nucellus Download PDFInfo
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- CN101792731A CN101792731A CN201010300490A CN201010300490A CN101792731A CN 101792731 A CN101792731 A CN 101792731A CN 201010300490 A CN201010300490 A CN 201010300490A CN 201010300490 A CN201010300490 A CN 201010300490A CN 101792731 A CN101792731 A CN 101792731A
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- ovule
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- megarchidium
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Abstract
The invention discloses a method for separating Chinese white poplar ovule from nucellus, wherein female flower of the Chinese white poplar is picked and put on a gel block, bract and floral disc of the female flower are removed with the help of an anatomical lens, the ovary of the female flower is longitudinally cut open along a dorsal suture on the sidewall of the ovary, so funicle and ovule can be separated off from carpel; when the nucellus is separated, the separated funicle and the ovule can still be put on the gel block, integument of the ovule is cut open longitudinally and upwards, picked and lifted from bottom to top from the juncture of the ovule and the funicle, the step of cutting, picking and lifting the integument is repeated so that the nucellus can be exposed gradually, and in the end, the nucellus is cut off at the junction of the ovule and the integument so as to be completely separated from the integument. The method can not only avoid the influence of enzymatic method on the cultivation and growth performance of plant materials, but can also reduce the damage brought by non-enzymatic method and outstandingly maintain the original shape of materials to be separated.
Description
Technical field
The present invention relates to a kind of method of separating the Cortex Populi Tomentosae ovule, also relate to a kind of method of separating the Cortex Populi Tomentosae megarchidium simultaneously, belong to plant microscopic examination technical field.
Background technology
The ovule of Cortex Populi Tomentosae is small, and its ovule of hand and megarchidium be difficulty relatively, adopts the isolated megarchidium of non-zymolysis technique to be difficult to keep its original form.At present, mostly be zymolysis technique, finish artificial separation the (nineteen eighty-two such as the Zhou Chang paper delivered in 1998 such as the paper " the isolating research of angiosperm blastular enzyme process: the isolation technique of immobilization material and microscopic examination " delivered, Yang Hongyuan " angiosperm exsomatize fertilization and zygote culture studies progress ") of micro-structures such as plant megarchidium, blastular, ovum and zygote by means of cellulase and polygalacturonase etc.Because blastular and zygote are after enzyme is handled, may survive to it, growth and artificial culture etc. exert an adverse impact, scholars such as Yang Hongyuan propose to reduce the influence (paper " tobacco zygote isolated culture regeneration fertile plant " that the paper that Yang Hongyuan etc. delivered in 1998 " angiosperm exsomatize fertilization and zygote culture studies progress ", He Yuchi etc. delivered in 2004) of enzyme in the sepn process of ovum etc.Therefore, the novel method of research micro-structures such as separation megarchidium simple to operate still has important meaning.
Summary of the invention
The object of the present invention is to provide a kind of method of separating the Cortex Populi Tomentosae ovule, not only can keep the integrity of isolating construction, and can avoid the problems such as influence of enzyme pair cell structure in the enzyme process sepn process.
The present invention also aims to provide a kind of method of separating the Cortex Populi Tomentosae megarchidium, not only can keep the integrity of isolating construction, and can avoid the problems such as influence of enzyme pair cell structure in the enzyme process sepn process.
To achieve these goals, technical scheme of the present invention has adopted a kind of method of separating the Cortex Populi Tomentosae ovule, comprises the female flower of Cortex Populi Tomentosae is taken from female inflorescence, is placed on the blob of viscose; Under anatomical lens, bract, the floral disc of female flower are removed respectively, along the dorsal suture on the ovary sidewall ovary is vertically cut open then, can see in the locule ovule of giving birth on carpel, funicle and ovule are separated from carpel get final product.
Described blob of viscose is that chewing gum is removed the blob of viscose that forms behind the sugar, and the colloid material that also can use bubble gum, chewing gum base etc. can reach required viscosity of separation and convenient fixedly separated material all can.
Technical scheme of the present invention has also adopted a kind of method of separating the Cortex Populi Tomentosae megarchidium, may further comprise the steps
The separation of Cortex Populi Tomentosae ovule: the female flower of Cortex Populi Tomentosae is taken from female inflorescence, isolated ovule and funicle;
The separation of Cortex Populi Tomentosae megarchidium: isolated funicle and ovule are placed on the blob of viscose, and funicle is bonded on the blob of viscose; At the intersection of ovule and funicle, upwards the integument of ovule is vertically scratched lightly, at the slot place integument is chosen out gently from the bottom to top, started then, progressively expose intraovular megarchidium; Cut off in the junction of megarchidium and integument at last, megarchidium can be separated from integument fully.
Described blob of viscose is that chewing gum is removed the blob of viscose that forms behind the sugar, and the colloid material that also can use bubble gum, chewing gum base etc. can reach required viscosity of separation and convenient fixedly separated material all can.
In Cortex Populi Tomentosae female flower (as shown in Figure 1), gynoecium is made of 2 carpels, and a chase is longitudinally respectively arranged on the left and right sidewall of its ovary, is dorsal suture.When separating the Cortex Populi Tomentosae ovule, the dorsal suture of available dissecting needle (or tip of tweezers) on the ovary sidewall vertically cut ovary open, the ovary wall of cutting open raised and it is bonded at (as shown in Figure 2) on the blob of viscose; Also can be respectively two dorsal sutures on the left and right sidewall of ovary ovary is vertically cut open, and two ovary walls will cutting open raise (or further its outer wall being bonded on the blob of viscose) (as shown in Figure 3) from top to bottom, promptly can be observed ovule and carpel the position and give birth to relation.Generally having two ovules in the locule gives birth to, three ovules also can appear occasionally, three ovules etc. are big or one of them ovule volume is very little, there is not modal difference (as shown in Figure 4) etc. three big ovules with normal ovule, three ovules do not wait when big, the funicle of the ovule that volume is less is elongated, may be the sterile ovule (as shown in Figure 5) of degenerating.Can be observed after isolated ovule is dyed, transparent and exist narrow gap (as shown in Figure 6) between megarchidium and the integument, if the ovule after will dyeing is soaked overnight in transparent liquid, megarchidium is shunk strongly, gap and micropylar canal between megarchidium and the integument all enlarge markedly (as shown in Figure 7), the feature of anatropous ovule is fairly obvious, this hand for megarchidium provides foundation, it should be noted that being used for the isolating ovule of megarchidium should be the ovule of new isolated no dyeing.The isolated ovule of the inventive method can clearly be observed the adnation position of fine hair in the female flower of water planting, to the female branch water planting a couple of days after blooming, can see in locule that the surface of funicle bears elongated and intensive fine hair (as shown in Figure 8), after ovule and funicle separated from carpel, can find that elongated fine hair is to bear (as shown in Figure 9) by the surface of funicle.
Advantage of the present invention is: the inventive method both can have been avoided cultivation performance and the growth Effect on Performance of enzyme process separation to vegetable material, can reduce non-enzyme process again and separate the damage that brings, fabulous original form of having kept material to be separated; Utilize that blob of viscose is auxiliary to be separated, the part operation of alternative hand, can cushion exert oneself, the dexterous application of force, alleviate the working pressure of hand and brain; Need not to use specific apparatus such as inverted microscope, even in the relatively poor laboratory of Experimental Establishment, also can finish, and can using integral dyeing with transparent technology intranucellar blastular structure is observed, perhaps with isolated megarchidium be used for the separation of micro-structures such as blastular further.
Description of drawings
Fig. 1 is the structural representation of Cortex Populi Tomentosae female flower;
Fig. 2 be give birth to the structural representation of the ovule on carpel, along a dorsal suture ovary is vertically cut open;
Fig. 3 be give birth to the structural representation of the ovule on carpel, along two dorsal sutures ovary is vertically cut open;
Fig. 4 the synoptic diagram of the situation of giving birth to for three ovules are arranged in the locule;
Fig. 5 the synoptic diagram of the situation of giving birth to for three ovules are arranged in the locule, and ovule has dyeed;
Fig. 6 is only through the structural representation of transparent ovule of routine dyeing and short period of time;
Fig. 7 is the structural representation through the transparent ovule that spends the night;
Fig. 8 be in the locule fine hair give birth to the situation synoptic diagram;
Fig. 9 be give birth to the synoptic diagram of the fine hair on the strain handle;
Figure 10 is the structural representation of megarchidium of integument of ining succession;
Figure 11 is the structural representation of megarchidium, wherein 1. is the secondary nucleus in the blastular, 2. is the ovum in the blastular.
Embodiment
Embodiment 1
The method that present embodiment separates the Cortex Populi Tomentosae ovule may further comprise the steps:
The preparation of blob of viscose: get 2 chewing gums and put into warm water and rub, embathe repeatedly, with the materials such as sugar in the abundant flush away chewing gum, washed blob of viscose is kneaded, the sticking visual field central authorities that are placed on anatomical lens are standby;
The separation of Cortex Populi Tomentosae ovule: the female flower of Cortex Populi Tomentosae is taken from female inflorescence, sticking being placed on the blob of viscose, under anatomical lens, the bract of female flower and the floral disc of oblique cup-shaped are removed, along a dorsal suture ovary wall is vertically cut open, raise then and its outer wall is bonded on the blob of viscose, as seen have two ovules in the locule and give birth to (as shown in Figure 2), the funicle under the ovule is long and sturdy, funicle and ovule is separated from carpel get final product.
Embodiment 2
The method that present embodiment separates the Cortex Populi Tomentosae ovule may further comprise the steps:
The preparation of blob of viscose: get 3 chewing gums and put into warm water and rub, embathe repeatedly, with the materials such as sugar in the abundant flush away chewing gum, washed blob of viscose is kneaded, the sticking visual field central authorities that are placed on anatomical lens are standby;
The separation of Cortex Populi Tomentosae ovule: the female flower of Cortex Populi Tomentosae is taken from female inflorescence, sticking being placed on the blob of viscose, under anatomical lens, the bract of female flower and the floral disc of oblique cup-shaped are removed, along two dorsal sutures ovary is vertically cut open, and two ovary walls are raised be bonded on the blob of viscose, the funicle branch of just observing under each ovule wears the base portion (as shown in Figure 3) of being born in two carpels, and a side of each funicle is free, one side and carpel adhesion separate funicle and ovule and to get final product from carpel.
Embodiment 3
The method that present embodiment separates the Cortex Populi Tomentosae megarchidium may further comprise the steps:
Adopt conventional separation method to separate the Cortex Populi Tomentosae ovule;
Adopt the Dai Shi phenodin of dilution to dye to isolated ovule, and in volume ratio is the mixed solution of 1: 1 dehydrated alcohol and dimethylbenzene, carry out transparently, after film-making on the depression slide, observe, to understand the constitutional features (as shown in Figure 6) of ovule;
Then with new isolating ovule and the sticking blob of viscose that is placed on of funicle, after fixing, intersection at ovule and funicle, upwards the integument of ovule is vertically scratched lightly with dissecting needle, select one from the bottom to top the integument of ovule to be scratched gently more in its vicinity after the same method to number place, with dissecting needle the integument between two slots is chosen out from the bottom to top gently, start, the integument outer wall that starts is bonded on the blob of viscose, just can progressively expose intraovular megarchidium, the junction (being the chalaza place) of megarchidium and integument is cut off with dissecting needle in the megarchidium bottom at last, just megarchidium can be separated from integument fully.
Embodiment 4
The method that present embodiment separates the Cortex Populi Tomentosae megarchidium may further comprise the steps:
The method of step (1) and step (2) is with the method for embodiment 1;
Adopting the Dai Shi phenodin of dilution to dye to isolated ovule, after the dyeing, it is transferred on the depression slide, is transparent the spending the night of mixed solution of 10: 1 dehydrated alcohol and dimethylbenzene through volume ratio, observes the constitutional features (as shown in Figure 7) of ovule after the film-making;
Then with new isolating ovule and the sticking blob of viscose that is placed on of funicle, after fixing, intersection at ovule and funicle, upwards the integument of ovule is vertically scratched lightly with dissecting needle, select one from the bottom to top the integument of ovule to be scratched gently more in its vicinity after the same method to number place, with dissecting needle the integument between two slots is chosen out from the bottom to top gently, start, the integument outer wall that starts is bonded on the blob of viscose, just can progressively expose intraovular megarchidium (as shown in figure 10, can see that megarchidium links to each other at the chalaza place with integument), the junction (being the chalaza place) of megarchidium and integument is cut off with dissecting needle in the megarchidium bottom at last, just megarchidium can be separated from integument fully.
Embodiment 5
The method that present embodiment separates the Cortex Populi Tomentosae megarchidium may further comprise the steps:
The method of step (1) and step (2) is with the method for embodiment 1;
(3) new isolating ovule and funicle are adhered with blob of viscose, after fixing, intersection at ovule and funicle, upwards the integument of ovule is vertically scratched lightly with dissecting needle, select one from the bottom to top the integument of ovule to be scratched gently more in its vicinity after the same method to number place, with dissecting needle the integument between two slots is chosen out from the bottom to top gently, start, the integument outer wall that starts is bonded on the blob of viscose, just can progressively expose intraovular megarchidium, the junction (being the chalaza place) of megarchidium and integument is cut off with dissecting needle in the megarchidium bottom at last, just megarchidium can be separated from integument fully.
Can adopt the Dai Shi phenodin of dilution to dye to isolated megarchidium, after the dyeing, it is transferred on the depression slide, through volume ratio is that the mixed solution of 5: 1 dehydrated alcohol and dimethylbenzene is transparent, and with after the cedar oil mounting, can carry out film-making and observe, as shown in figure 11, can observe in the blastular secondary nucleus 1. with ovum 2..
Claims (4)
1. method of separating the Cortex Populi Tomentosae ovule, it is characterized in that: described method comprises to be taken the female flower of Cortex Populi Tomentosae from female inflorescence, be placed on the blob of viscose; Under anatomical lens, bract, the floral disc of female flower are removed respectively, along the dorsal suture on the ovary sidewall ovary is vertically cut open then, can see in the locule ovule of giving birth on carpel, funicle and ovule are separated from carpel get final product.
2. a kind of method of separating the Cortex Populi Tomentosae ovule according to claim 1 is characterized in that: described blob of viscose is that chewing gum is removed the blob of viscose that forms behind the sugar.
3. a method of separating the Cortex Populi Tomentosae megarchidium is characterized in that: said method comprising the steps of
(1) separation of Cortex Populi Tomentosae ovule: the female flower of Cortex Populi Tomentosae is taken from female inflorescence, isolated ovule and funicle;
(2) separation of Cortex Populi Tomentosae megarchidium: isolated funicle and ovule are placed on the blob of viscose, and funicle is bonded on the blob of viscose; At the intersection of ovule and funicle, upwards the integument of ovule is vertically scratched lightly, at the slot place integument is chosen out gently from the bottom to top, started then, progressively expose intraovular megarchidium; Cut off in the junction of megarchidium and integument at last, megarchidium can be separated from integument fully.
4. a kind of method of separating the Cortex Populi Tomentosae megarchidium according to claim 3 is characterized in that: described blob of viscose is that chewing gum is removed the blob of viscose that forms behind the sugar.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2010103004909A CN101792731B (en) | 2010-01-20 | 2010-01-20 | Method for separating Chinese white poplar ovule from nucellus |
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| CN2010103004909A CN101792731B (en) | 2010-01-20 | 2010-01-20 | Method for separating Chinese white poplar ovule from nucellus |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108486036A (en) * | 2018-02-06 | 2018-09-04 | 保定学院 | A kind of separation method of Chinese white poplar blastular |
| CN109310061A (en) * | 2016-03-31 | 2019-02-05 | 日本烟草产业株式会社 | The method that substance is imported into plant |
| CN109725411A (en) * | 2017-10-27 | 2019-05-07 | 河南科技大学 | A kind of anatomical lens sample fixing device and anatomical lens |
| CN112881112A (en) * | 2021-01-07 | 2021-06-01 | 河南科技大学 | In-situ microdissection method for sweet potato fruits |
-
2010
- 2010-01-20 CN CN2010103004909A patent/CN101792731B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109310061A (en) * | 2016-03-31 | 2019-02-05 | 日本烟草产业株式会社 | The method that substance is imported into plant |
| CN109725411A (en) * | 2017-10-27 | 2019-05-07 | 河南科技大学 | A kind of anatomical lens sample fixing device and anatomical lens |
| CN109725411B (en) * | 2017-10-27 | 2021-01-01 | 河南科技大学 | Sample fixing device for dissecting mirror and dissecting mirror |
| CN108486036A (en) * | 2018-02-06 | 2018-09-04 | 保定学院 | A kind of separation method of Chinese white poplar blastular |
| CN108486036B (en) * | 2018-02-06 | 2020-07-17 | 保定学院 | Method for separating Chinese white poplar embryo sac |
| CN112881112A (en) * | 2021-01-07 | 2021-06-01 | 河南科技大学 | In-situ microdissection method for sweet potato fruits |
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| CN101792731B (en) | 2012-01-25 |
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