Kanamycin was used to screen To seeds of the variety Dongnong 46 transformed by means of pollen-tube method. The results showed that 400 mg·L^-1 kanamycin could inhibit growth of non-transgenic plants, and 2 posi...Kanamycin was used to screen To seeds of the variety Dongnong 46 transformed by means of pollen-tube method. The results showed that 400 mg·L^-1 kanamycin could inhibit growth of non-transgenic plants, and 2 positive plants were gotten combined with Gus dyeing and PCR detection. It is proved that this method is economic and effective in preliminary screening the transgenic plants.展开更多
[Objective] This research aimed to explore the method to increase conversion rate of pollen-tub pathway in Chinese cabbage.[Method] Chinese cabbage varieties Yuqing and No.03 were used as materials for the selection o...[Objective] This research aimed to explore the method to increase conversion rate of pollen-tub pathway in Chinese cabbage.[Method] Chinese cabbage varieties Yuqing and No.03 were used as materials for the selection of germination buffer and parameters for ultrasonication.[Result] The optimal buffer for pollen germination of Chinese cabbage was 200 g/L sucrose ＋ 100 mg/L boric acid ＋ 200 mg/L calcium nitrate,the preferred ultrasonic processing power was 150 W,processing time was 5 s,interval time was 5 s and processing frequency was 8.Three T1-generation plants were obtained through selection with 200 mg/L kanamycin.[Conclusion] This research laid foundation for the further genetic transformation of Chinese cabbage.展开更多
The green fluorescent protein (GFP) gene from the jellyfish Aequorea victoria as a vital reporter for gene expression in plants is considered to have several advantages over other reporter genes. The pBIN35S-mGFP4 pla...The green fluorescent protein (GFP) gene from the jellyfish Aequorea victoria as a vital reporter for gene expression in plants is considered to have several advantages over other reporter genes. The pBIN35S-mGFP4 plasmid DNA has been introduced into cotton embryos by the pollen-tube pathway method. A transformed seedling has been verified according to its GFP-related fluorescence and Southern blotting analysis. The results provided direct and convincing facts in cytology and molecular biology for the pollen-tube pathway method, an efficient transformation technique used in plants.展开更多
β-glucuronidase (GUS) gene (carried by pBI121 plasmid) has been introduced into hexaploid wheat c. v. Xiaoshan No. 3 with the pollen-tube pathway method. Five transgenic plants are selected from 106 T0 plants. All of...β-glucuronidase (GUS) gene (carried by pBI121 plasmid) has been introduced into hexaploid wheat c. v. Xiaoshan No. 3 with the pollen-tube pathway method. Five transgenic plants are selected from 106 T0 plants. All of these plants are identified by the Southern hybridization, and the GUS activity can be detected by using fluorogenic assay and histochemical stain.展开更多
Elongation of the pollen tube in pistil is essential for delivering sperms into the female gametophyte in sexual plant reproduction. Recently, a group of cell wall enzymes, pectin methylesterases （PMEs）, have been i...Elongation of the pollen tube in pistil is essential for delivering sperms into the female gametophyte in sexual plant reproduction. Recently, a group of cell wall enzymes, pectin methylesterases （PMEs）, have been identified as playing an important role in this process. This article reviews the new understanding of the roles of PMEs in regulating pollen tube growth.展开更多
The origin and the recent progress of pollen_tube pathway method (PTPM) is described. The results of transforming wheat during 1990 to 1994 are reported briefly. They are: (ⅰ) Transformation frequencies were 3%-6%, e...The origin and the recent progress of pollen_tube pathway method (PTPM) is described. The results of transforming wheat during 1990 to 1994 are reported briefly. They are: (ⅰ) Transformation frequencies were 3%-6%, even reached 13.5%; (ⅱ) the genetic expression of four continuous generations obtained from single individuals of transgenic plants by self_fertilization was investigated. Plants with higher expression of foreign gene were found in every generation (from T1 to T4), but a phenomenon of gene silence, in which the genotype contained foreign gene but did not express, was also observed. In addition, from the angle of fertilization biology the transformation mechanism of PTPM (or PTDM) was analyzed and discussed. It was considered that the essential principle of this method was just like that of protoplast method.展开更多
文摘Kanamycin was used to screen To seeds of the variety Dongnong 46 transformed by means of pollen-tube method. The results showed that 400 mg·L^-1 kanamycin could inhibit growth of non-transgenic plants, and 2 positive plants were gotten combined with Gus dyeing and PCR detection. It is proved that this method is economic and effective in preliminary screening the transgenic plants.
基金Supported by Scientific and Technological Project in Shanxi Province(021034)Scientific Research Project of Shanxi Academy of Agri-cultural Sciences (YGX-01)~~
文摘[Objective] This research aimed to explore the method to increase conversion rate of pollen-tub pathway in Chinese cabbage.[Method] Chinese cabbage varieties Yuqing and No.03 were used as materials for the selection of germination buffer and parameters for ultrasonication.[Result] The optimal buffer for pollen germination of Chinese cabbage was 200 g/L sucrose ＋ 100 mg/L boric acid ＋ 200 mg/L calcium nitrate,the preferred ultrasonic processing power was 150 W,processing time was 5 s,interval time was 5 s and processing frequency was 8.Three T1-generation plants were obtained through selection with 200 mg/L kanamycin.[Conclusion] This research laid foundation for the further genetic transformation of Chinese cabbage.
文摘The green fluorescent protein (GFP) gene from the jellyfish Aequorea victoria as a vital reporter for gene expression in plants is considered to have several advantages over other reporter genes. The pBIN35S-mGFP4 plasmid DNA has been introduced into cotton embryos by the pollen-tube pathway method. A transformed seedling has been verified according to its GFP-related fluorescence and Southern blotting analysis. The results provided direct and convincing facts in cytology and molecular biology for the pollen-tube pathway method, an efficient transformation technique used in plants.
文摘β-glucuronidase (GUS) gene (carried by pBI121 plasmid) has been introduced into hexaploid wheat c. v. Xiaoshan No. 3 with the pollen-tube pathway method. Five transgenic plants are selected from 106 T0 plants. All of these plants are identified by the Southern hybridization, and the GUS activity can be detected by using fluorogenic assay and histochemical stain.
基金Supported by the Talent Introduction Program of China Agricultural University. Publication of this paper is supported by the National Natural Science Foundation of China （30624808） and Science Publication Foundation of the Chinese Academy of Sciences.
文摘Elongation of the pollen tube in pistil is essential for delivering sperms into the female gametophyte in sexual plant reproduction. Recently, a group of cell wall enzymes, pectin methylesterases （PMEs）, have been identified as playing an important role in this process. This article reviews the new understanding of the roles of PMEs in regulating pollen tube growth.
文摘The origin and the recent progress of pollen_tube pathway method (PTPM) is described. The results of transforming wheat during 1990 to 1994 are reported briefly. They are: (ⅰ) Transformation frequencies were 3%-6%, even reached 13.5%; (ⅱ) the genetic expression of four continuous generations obtained from single individuals of transgenic plants by self_fertilization was investigated. Plants with higher expression of foreign gene were found in every generation (from T1 to T4), but a phenomenon of gene silence, in which the genotype contained foreign gene but did not express, was also observed. In addition, from the angle of fertilization biology the transformation mechanism of PTPM (or PTDM) was analyzed and discussed. It was considered that the essential principle of this method was just like that of protoplast method.
