甘蔗花粉离体萌发研究
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摘要
为筛选适宜栽培种甘蔗花粉离体萌发的培养基配方,建立有效的离体萌发体系,以糖用甘蔗主栽品种ROC22花粉为材料,通过单因素试验探索满足甘蔗花粉离体萌发的基本条件,然后利用L_(16)(4~5)正交试验,寻求甘蔗花粉离体萌发的最适宜培养基配方,在此基础上进一步探讨不同温度对甘蔗花粉萌发的影响。在单因素试验中,培养基以刚好凝固时为最佳。蔗糖、硼酸和硝酸钙在一定范围内对甘蔗花粉萌发起促进作用,超过一定浓度则起抑制作用。硫酸镁作用不稳定,硝酸钾影响较小。正交试验中,蔗糖对甘蔗花粉离体萌发率和花粉管生长速率影响最大,呈显著水平,硼酸次之,硫酸镁影响最小。花粉萌发的适宜温度为30℃,萌发率和花粉管长度分别为89.83%和143.03μm。200 g/L蔗糖+400 mg/L硼酸+100 mg/L硝酸钙+400 mg/L硫酸镁+1 g/L琼脂,在30℃下培养花粉萌发率最高,花粉管生长状态良好,这种方法可快速有效鉴定不同甘蔗花粉活力差异。
The pollens of sugarcane cultivar ROC22 were used as the main experimental material to seek for the basic conditions of the pollen germination in vitro by a single factor experiment, and the most appropriate culture medium formula was explored by an orthogonal experiment L_(16)(4~5) in order to investigate the effects on sugarcane pollen germination under different temperatures. In the single factor experiment, the culture medium solidified slightly had the best effect. In certain range of concentration, sucrose, boric acid and calcium nitrate might promote pollen germination and inhibit pollen germination over the critical concentration. In addition, magnesium sulfate worked unsteadily, and the effect of potassium nitrate was little. In the orthogonal experiment, sucrose influenced significantly on the pollen germination rate in vitro and the pollen tube growth rate, boric acid took the second place, magnesium sulfate had little effect. The optimum temperature for pollen germination was 30 ℃, the pollen germination rate was 89.93% and the pollen tube length was 143.03 μm. It is concluded that, under 30 ℃, the pollen had the highest germination rate and the pollen tube grew well using the best culture medium formulas(200 g/L sucrose, 400 mg/L boric acid, 100 mg/L calcium nitrate, 400 mg/L magnesium sulfate and 1 g/L agar), which might help to identify the difference of various sugarcane pollen efficiently.
The pollens of sugarcane cultivar ROC22 were used as the main experimental material to seek for the basic conditions of the pollen germination in vitro by a single factor experiment, and the most appropriate culture medium formula was explored by an orthogonal experiment L_(16)(4~5) in order to investigate the effects on sugarcane pollen germination under different temperatures. In the single factor experiment, the culture medium solidified slightly had the best effect. In certain range of concentration, sucrose, boric acid and calcium nitrate might promote pollen germination and inhibit pollen germination over the critical concentration. In addition, magnesium sulfate worked unsteadily, and the effect of potassium nitrate was little. In the orthogonal experiment, sucrose influenced significantly on the pollen germination rate in vitro and the pollen tube growth rate, boric acid took the second place, magnesium sulfate had little effect. The optimum temperature for pollen germination was 30 ℃, the pollen germination rate was 89.93% and the pollen tube length was 143.03 μm. It is concluded that, under 30 ℃, the pollen had the highest germination rate and the pollen tube grew well using the best culture medium formulas(200 g/L sucrose, 400 mg/L boric acid, 100 mg/L calcium nitrate, 400 mg/L magnesium sulfate and 1 g/L agar), which might help to identify the difference of various sugarcane pollen efficiently.
引文
[1]骆君骕.甘蔗学[M].广州:中国轻工业出版社, 1984:156-220.
[2]李奇伟,陈子云,梁洪.现代甘蔗改良技术[M].广州:华南理工大学出版社, 2000:19-42.
[3]左丹丹,明军,刘春.植物花粉生活力检测技术进展[J].安徽农业科学, 2007, 35(16):4742-4745.
[4]李富生,林位夫,何顺长,等.不同无性系蔗茅抽穗开花特性的观察和花粉贮藏条件的研究[J].植物生理学通讯,2005, 41(2):171-174.
[5]杜玉虎,张绍铃,姜雪婷,等.果梅花粉离体萌发及花粉管生长特性研究[J].西北植物学报, 2006, 26(9):1846-1852.
[6]朱展望,张改生,牛娜.小麦花粉的离体萌发研究[J].麦类作物学报, 2007, 27(1):12-15.
[7]王岚岚,游捷,俞红强.月季花粉离体萌发液体培养基组分的优化[J].河北农业大学学报, 2008, 31(3):42-45.
[8]周耀辉,黄启尧,杨业后.甘蔗花粉低温贮存与生活力[J].甘蔗糖业, 1994(2):7-10.
[9]王开良,姚小华,任华东,等.余甘子花粉萌发与贮藏特性研究[J].江西农业大学学报, 2006, 28(1):69-73.
[10]杜纪红,叶正文,苏明申,等.桃花粉离体萌发和花粉管生长特性研究[J].西北植物学报, 2011, 31(1):64-71.
[11] Fei S, Nelson E. Estimation of pollen viability, shedding pattern, and longevity of creeping bentgrass on artificial media[J]. Crop Science, 2003, 43(6):2177-2181.
[12] Feruzan D, Goksel O, Ozlem D. In vitro pollen germination of some plant species in basic in basic culture medium[J]. Journal of Cell and Molecular Biology, 2004, 3:71-76.
[13]金爱红,褚立民,徐东青,等.钙对葱兰花粉萌发和花粉管生长的影响[J].湖北农业科学, 2005(3):91-93.
[14]罗凤霞,周威,杨春起,等.水仙花粉离体萌发温度和培养液研究[J].辽宁林业科技, 2007(5):13-14, 25, 57.
[15]赵丽娟,李淑娟,于金海,等.小黑杨花粉离体萌发和细胞学分析[J].林业科学, 2011, 47(6):36-41.
[16]潘瑞炽.植物生理学[M]. 4版.北京:高等教育出版社,2001:31, 260.
[17]刘雪莲,李庆玲,秦佳梅,等.不同培养基对紫丁香花粉离体萌发的影响[J].东北林业大学学报, 2010, 38(7):61-63.
[18]蒲光兰,周兰英,李瑾宵,等.温度、蔗糖和硼酸对麻疯树花粉离体萌发的研究[J].西北林学院学报, 2011, 26(3):55-58.
[19] Zhao J, Yu F, Liang S, et al. Changes of calcim distribution in egg cells zygotes and two-celled pro-embryos of rice(Oryza sativa L.)[J]. Sexual Plant Reproduction, 2002, 14(6):331-337.
[20] Tian H Q, Zhang Z, Russell S D. Isolation of the male germ unit organization and function in tobacco(Nicotana tabacum L.)[J]. Plant Cell Reports, 1998, 18:143-147.
[21] Zhang Z, Tian H Q, Russell S D. Localization of myosin on sperm-cell-associated membranes of tobacco(Nicotana tabacun L.)[J]. Protoplasma, 1999, 208:123-128.
[22]垄明,曹宗巽.钙和钙调素对花粉萌发和花粉管生长的调控[J].植物生理学通讯, 1995, 31(5):321-328.
[23]孙颖,孙大业.花粉萌发和花粉管生长发育的信号转导[J].植物学报, 2001, 43(12):1211-1217.
[24] Tian H Q, Kuang A, Musgrave M E, et al. Calcium distribution in fertile and sterile anthers of a photoperiod-sensitive genic male-sterile rice[J]. Plana, 1998, 204(2):183-192.
[25] Heslop-Harrison J. Pollen germination and pollen-tube growth[J]. International Review of Cytology, 1987, 107:1-78.
[26]周峰,胡后祥,金玉峰,等.夜间温度对光周期诱导甘蔗开花的影响[J].西南农业学报, 2013, 26(1):99-102.
[27]王丽萍,马丽.提高甘蔗远缘杂交后代花粉育性的研究[J].亚热带农业研究, 2005, 1(1):1-3.
[2]李奇伟,陈子云,梁洪.现代甘蔗改良技术[M].广州:华南理工大学出版社, 2000:19-42.
[3]左丹丹,明军,刘春.植物花粉生活力检测技术进展[J].安徽农业科学, 2007, 35(16):4742-4745.
[4]李富生,林位夫,何顺长,等.不同无性系蔗茅抽穗开花特性的观察和花粉贮藏条件的研究[J].植物生理学通讯,2005, 41(2):171-174.
[5]杜玉虎,张绍铃,姜雪婷,等.果梅花粉离体萌发及花粉管生长特性研究[J].西北植物学报, 2006, 26(9):1846-1852.
[6]朱展望,张改生,牛娜.小麦花粉的离体萌发研究[J].麦类作物学报, 2007, 27(1):12-15.
[7]王岚岚,游捷,俞红强.月季花粉离体萌发液体培养基组分的优化[J].河北农业大学学报, 2008, 31(3):42-45.
[8]周耀辉,黄启尧,杨业后.甘蔗花粉低温贮存与生活力[J].甘蔗糖业, 1994(2):7-10.
[9]王开良,姚小华,任华东,等.余甘子花粉萌发与贮藏特性研究[J].江西农业大学学报, 2006, 28(1):69-73.
[10]杜纪红,叶正文,苏明申,等.桃花粉离体萌发和花粉管生长特性研究[J].西北植物学报, 2011, 31(1):64-71.
[11] Fei S, Nelson E. Estimation of pollen viability, shedding pattern, and longevity of creeping bentgrass on artificial media[J]. Crop Science, 2003, 43(6):2177-2181.
[12] Feruzan D, Goksel O, Ozlem D. In vitro pollen germination of some plant species in basic in basic culture medium[J]. Journal of Cell and Molecular Biology, 2004, 3:71-76.
[13]金爱红,褚立民,徐东青,等.钙对葱兰花粉萌发和花粉管生长的影响[J].湖北农业科学, 2005(3):91-93.
[14]罗凤霞,周威,杨春起,等.水仙花粉离体萌发温度和培养液研究[J].辽宁林业科技, 2007(5):13-14, 25, 57.
[15]赵丽娟,李淑娟,于金海,等.小黑杨花粉离体萌发和细胞学分析[J].林业科学, 2011, 47(6):36-41.
[16]潘瑞炽.植物生理学[M]. 4版.北京:高等教育出版社,2001:31, 260.
[17]刘雪莲,李庆玲,秦佳梅,等.不同培养基对紫丁香花粉离体萌发的影响[J].东北林业大学学报, 2010, 38(7):61-63.
[18]蒲光兰,周兰英,李瑾宵,等.温度、蔗糖和硼酸对麻疯树花粉离体萌发的研究[J].西北林学院学报, 2011, 26(3):55-58.
[19] Zhao J, Yu F, Liang S, et al. Changes of calcim distribution in egg cells zygotes and two-celled pro-embryos of rice(Oryza sativa L.)[J]. Sexual Plant Reproduction, 2002, 14(6):331-337.
[20] Tian H Q, Zhang Z, Russell S D. Isolation of the male germ unit organization and function in tobacco(Nicotana tabacum L.)[J]. Plant Cell Reports, 1998, 18:143-147.
[21] Zhang Z, Tian H Q, Russell S D. Localization of myosin on sperm-cell-associated membranes of tobacco(Nicotana tabacun L.)[J]. Protoplasma, 1999, 208:123-128.
[22]垄明,曹宗巽.钙和钙调素对花粉萌发和花粉管生长的调控[J].植物生理学通讯, 1995, 31(5):321-328.
[23]孙颖,孙大业.花粉萌发和花粉管生长发育的信号转导[J].植物学报, 2001, 43(12):1211-1217.
[24] Tian H Q, Kuang A, Musgrave M E, et al. Calcium distribution in fertile and sterile anthers of a photoperiod-sensitive genic male-sterile rice[J]. Plana, 1998, 204(2):183-192.
[25] Heslop-Harrison J. Pollen germination and pollen-tube growth[J]. International Review of Cytology, 1987, 107:1-78.
[26]周峰,胡后祥,金玉峰,等.夜间温度对光周期诱导甘蔗开花的影响[J].西南农业学报, 2013, 26(1):99-102.
[27]王丽萍,马丽.提高甘蔗远缘杂交后代花粉育性的研究[J].亚热带农业研究, 2005, 1(1):1-3.