Cytological Study of Pollen-mediated Plant Transformation Method Based on GFP Observation
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- 1School of Life Sciences, Shanxi Normal University, Linfen 041004, China;
2Biotechnology Research Centre, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China;
3Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau, Ministry of Agriculture, Taiyuan 030031, China
Received date: 2013-01-30
Revised date: 2013-04-21
Online published: 2013-12-03
Abstract
Pollen-mediated gene transformation does not require tissue culture and is easy to apply. In the present study, we investigate the cytological basis for the transformation approach. Fresh pollen was taken from maize plants (Zea mays cv ‘Zhengdan 958’) and treated under ultrasonication mixed with plasmid DNA harboring green fluorescence protein (GFP) gene. Treated pollen was artificially pollinated onto silks of maize ears. The expression of GFP gene in pollen grains, pollen tubes and embryo were observed under a fluorescence microscope. Pollen grains had a strong green autoflorescent background and so were not suitable for observing the expression of exotic GFP gene. Treated pollen tubes produced strong green florescence as compared with the control. The expression of GFP gene in embryo could be used to confirm the expression of exotic genes. Our study provides cytological evidence for pollen-mediated transformation.
Cite this article
Na Li, Yi Sun, Liyan Yang . Cytological Study of Pollen-mediated Plant Transformation Method Based on GFP Observation[J]. Chinese Bulletin of Botany, 2013 , 48(6) : 616 -622 . DOI: 10.3724/SP.J.1259.2013.00616
References
崔贵梅, 孙毅, 郝曜山, 杜建中, 王亦学 (2012). 玉米花粉体外萌发方法改进及其对花粉介导转基因的作用. 植物学报 47, 155–161.
李金才, 崔彦宏, 董海荣, 王艳哲, 张丽华 (2002). 玉米花丝生长发育及受精结实能力研究进展. 河北农业大学学报25, 86-89.
孙毅, 王景雪, 崔贵梅. 超声波处理花粉介导植物基因转化方法. ZL: 99121152.9.
Sambrook J, Russell D (2002). 分子克隆实验指南 (第3版). 北京: 科学出版社.
Booy G, Krens FA, Huizing HJ (1989). Attempted pollen-mediated transformation of maize. J Plant Physiol 135, 319-324.
Chen X, Equi R, Baxter H, Berk K, Han J, Agarwal S, Zale J (2010). A high-throughput transient gene expression system for switchgrass (Panicum virgatum L.) Seedlings. Biotechnol Biofuels 3, 1-10.
Eapen S (2011). Pollen grains as a target for introduction of foreign genes into plants: an assessment. Physiol Mol Biol 17, 1-8.
Hess D, Dressler K (1989). Tumor transformation of Petunia hybrida via pollen cultured with Agrobacterium tumefaciens. Bot Acta 102, 202–207.
Luo ZX, Wu R (1989). A simple method for the transformation of rice via pollen tube pathway. Plant Mol Biol Rep 6,165–174.
Matousek J, Tupy J (1983). The release of nucleases from tobacco pollen. Plant Sci Lett 33, 83–89.
Ottenschlager I, Barinova I, Voronon V, Dahl M, Heberle-Bors E, Touraev A (1999). Green fluorescent protein (GFP) as a marker during pollen development. Transgenic Res 4, 279–294.
Roeckel P, Heizmann P, Dubois M, Dumas C (1988). Attempts to transform Zea mays via pollen grains: effect of pollen and stigma nuclease activities. Sex Plant Reprod 1, 156–163.
Sanford JC, Skubik KA, Reisch BI (1985). Attempted pollen-mediated plant transformation employing genomic donor DNA. TAG 69, 571-574.
Shou HX, Palmer RG, Wang K (2002). Irreproducibility of soybean pollen tube transformation procedure. Plant Mol Biol Rep 20, 325–332.
Suen DF, Huang AH (2007). Maize pollen coat xylanase facilitates pollen tube penetration into silk during sexual reproduction. J Biol Chem 1, 625–636.
Wang J, Li Y, Liang C (2008). Recovery of transgenic plants by pollen mediated transformation in Brassica juncea. Transgenic Res 17, 417–424.
Wang JX, Sun Y, Cui G, Hu J (2001). Transgenic maize plants obtained by pollen-mediated transformation. Acta Bot Sin 43, 275–279.
Wang W, Wang J, Yang C, Li Y, Liu L, Xu J (2007). Pollen-mediated transformation of Sorghum bicolor plants. Biotechnol Appl Biochem 48, 79–83.
Willemse MTM, Van Went JL (1984). Embryology of angiosperms, Berlin: Heidelberg, New York: Springer-Verlag. pp. 159-197.
Zhu T, Mogensen HL, Smith SE (1993). Quantiative, three dimension alanalysis of alfalfa egg cells in two genotypes: Implications for biparental plastid inheritance. Planta 100, 143-150.
李金才, 崔彦宏, 董海荣, 王艳哲, 张丽华 (2002). 玉米花丝生长发育及受精结实能力研究进展. 河北农业大学学报25, 86-89.
孙毅, 王景雪, 崔贵梅. 超声波处理花粉介导植物基因转化方法. ZL: 99121152.9.
Sambrook J, Russell D (2002). 分子克隆实验指南 (第3版). 北京: 科学出版社.
Booy G, Krens FA, Huizing HJ (1989). Attempted pollen-mediated transformation of maize. J Plant Physiol 135, 319-324.
Chen X, Equi R, Baxter H, Berk K, Han J, Agarwal S, Zale J (2010). A high-throughput transient gene expression system for switchgrass (Panicum virgatum L.) Seedlings. Biotechnol Biofuels 3, 1-10.
Eapen S (2011). Pollen grains as a target for introduction of foreign genes into plants: an assessment. Physiol Mol Biol 17, 1-8.
Hess D, Dressler K (1989). Tumor transformation of Petunia hybrida via pollen cultured with Agrobacterium tumefaciens. Bot Acta 102, 202–207.
Luo ZX, Wu R (1989). A simple method for the transformation of rice via pollen tube pathway. Plant Mol Biol Rep 6,165–174.
Matousek J, Tupy J (1983). The release of nucleases from tobacco pollen. Plant Sci Lett 33, 83–89.
Ottenschlager I, Barinova I, Voronon V, Dahl M, Heberle-Bors E, Touraev A (1999). Green fluorescent protein (GFP) as a marker during pollen development. Transgenic Res 4, 279–294.
Roeckel P, Heizmann P, Dubois M, Dumas C (1988). Attempts to transform Zea mays via pollen grains: effect of pollen and stigma nuclease activities. Sex Plant Reprod 1, 156–163.
Sanford JC, Skubik KA, Reisch BI (1985). Attempted pollen-mediated plant transformation employing genomic donor DNA. TAG 69, 571-574.
Shou HX, Palmer RG, Wang K (2002). Irreproducibility of soybean pollen tube transformation procedure. Plant Mol Biol Rep 20, 325–332.
Suen DF, Huang AH (2007). Maize pollen coat xylanase facilitates pollen tube penetration into silk during sexual reproduction. J Biol Chem 1, 625–636.
Wang J, Li Y, Liang C (2008). Recovery of transgenic plants by pollen mediated transformation in Brassica juncea. Transgenic Res 17, 417–424.
Wang JX, Sun Y, Cui G, Hu J (2001). Transgenic maize plants obtained by pollen-mediated transformation. Acta Bot Sin 43, 275–279.
Wang W, Wang J, Yang C, Li Y, Liu L, Xu J (2007). Pollen-mediated transformation of Sorghum bicolor plants. Biotechnol Appl Biochem 48, 79–83.
Willemse MTM, Van Went JL (1984). Embryology of angiosperms, Berlin: Heidelberg, New York: Springer-Verlag. pp. 159-197.
Zhu T, Mogensen HL, Smith SE (1993). Quantiative, three dimension alanalysis of alfalfa egg cells in two genotypes: Implications for biparental plastid inheritance. Planta 100, 143-150.
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