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Populus tomentosa embryo sac isolation method

A separation method and technology of Populus tomentosa, applied in the field of separation and extraction of plant embryo sac, can solve problems such as poor dyeing effect of film production, and achieve the effects of overcoming poor dyeing effect, huge application value and improving work efficiency.

Active Publication Date: 2018-09-04
BAODING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, it overcomes the problem of poor staining effect in the microtubule immunofluorescence localization experiment of Populus tomentosa embryo sac, and improves the work efficiency of observation of microtubule skeleton changes and organelle nucleoid behavior during the induction of female gametes in Populus tomentosa

Method used

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  • Populus tomentosa embryo sac isolation method
  • Populus tomentosa embryo sac isolation method
  • Populus tomentosa embryo sac isolation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The female flowering branches of Populus tomentosa were collected in November 2015 from the Nursery of Guanxian County, Shandong Province, a national key forest tree breeding base, and flowering was carried out hydroponically in the greenhouse. The flowering non-pollinated and 1-2d florets in the middle of the female inflorescence of Populus tomentosa at different developmental stages were selected as experimental materials.

[0031] Ovule collection and enzymatic hydrolysis

[0032] Select a floret with mature female gametes but not pollinated, which corresponds to the eight-nucleated embryo sac at the developmental stage of the flower. Usually the inflorescence is 3-4cm long, yellow-green, and the stigma is four-forked, slightly red; pollination 1-2d after flowering little flowers. After removing the bracts and discs of the female flowers with pointed tweezers, take some of the female flowers and put them in a petri dish. Then, under the dissecting mirror, fix the o...

Embodiment 2

[0077] The experimental materials were purchased from Changshouhua, which grew well in the greenhouse of Huaniaoyu Market on the East Second Ring Road of Baoding City. Changshouhua is a perennial succulent herb whose flowering period is from February to May. Flowers were collected from 2 days before flowering to 1 week after flowering. After removing the petals, pedicels and calyxes, the ovary is exposed, and the ovules are taken out with a dissecting needle and set aside.

[0078] Enzymatic hydrolysis of ovules and observation

[0079] According to the characteristics of Changshouhua ovules and embryo sacs and the characteristics of different enzymes, 9 kinds of enzyme solutions were prepared, and the prepared enzyme solutions were stored in a refrigerator at 4°C overnight for precipitation. For each experiment, just take 1-2mL and put it into a 4mL small centrifuge tube, and place the centrifuge tube in an ice box with ice cubes, pick the ovules with a dissecting needle an...

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Abstract

The invention relates to a populus tomentosa embryo sac isolation method. An enzymatic hydrolysis-dissection method and a suck-and-blow method are combined, the complete embryo sac can be obtained byisolating the fresh populus tomentosa ovule, and the yield is as high as 8.7%; the mixed enzyme liquid for screening the populus tomentosa ovule comprises the components of hemicellulase, cellulose, pectase, mannitol, and BSA, after enzymatic hydrolysis processing, through acetic red dyeing and transparency processing, the product is observed under an inverted microscope, a complete and clear formand an internal structure of the populus tomentosa embryo sac can be observed, the populus tomentosa embryo sac is in a long elliptical shape, a size is about 2.52 [mu]m, the populus tomentosa embryosac is unitegmic, egg apparatus cells exist at a micropyle end, a chalazal end has a structure which is similar with that of antipodal cells. The technical scheme can be used for isolating the embryosac of Kalanchoe blossfeldiana.

Description

technical field [0001] The invention relates to a step of separating and extracting plant embryo sacs, in particular to separating and extracting Populus tomentosa and Longevity Flower. Background technique [0002] For animals and lower plants, it is relatively easy to conduct experiments on sperm and egg cells in vitro, because in the process of fertilization, the fusion of sperm cells and egg cells is completed in the state of separation or half separation from the mother, so the obtained research The results are also more in-depth. However, higher plants are different. The egg cells are hidden deep in the ovules in the ovary, which brings great difficulties to the research on the development mechanism and in vitro operation of egg cells. In vitro fertilization technology, which utilizes isolated sperm cells and egg cells to induce fusion in vitro and reproduce the fertilization process, eliminates the interference of somatic cells and tissues, and makes it possible to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
CPCC12N5/04C12N2509/00C12N2509/10
Inventor 崔彬彬魏俊杰张妍曹柳青朱维红王晨汤飞飞孙宇涵沈一岚
Owner BAODING UNIV
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