高起点温度两系高粱不育系育性转换温光特性研究
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摘要
本研究主要针对两系高粱在生产应用过程中受严格的温光条件制约，种子生产需要在特定生态点、特定时期进行，且可能出现育性的不稳定性等问题，采用人工气候室处理、大田试验、同工酶和蛋白质分析等方法，研究了两系高粱不育系的纯化稳定特性；两系高粱不育系育性转换的温光敏感时期；温度对两系高粱不育系育性转换的影响；光周期对两系不育系育性转换的影响以及育性转换的分子机理。主要结果如下：
1．两系高粱不育系“湘糯粱2S”在孕穗至抽穗期控温25℃情况下表现出稳定不育，不育株率和不育度均达100％，利用控温26℃处理后仍表现出不育的植株，砍秆再生，再生植株在28℃以上高温条件下抽穗转为可育，自交繁殖出了纯化稳定的不育系种子，对照“湘糯粱S-1”在控温25℃时即表现为可育。
2．“湘糯粱2S”育性转换的温光敏感时期为花粉母细胞形成至减数分裂期，其外形判断为旗叶叶环高出倒2叶叶环0-8厘米，即抽穗前4-10天(秋季)或3-7天(夏季)。
3．“湘糯粱2S”是一个低温不育、高温可育的温敏型不育系，其育性转换的临界温度为25℃。
4．光周期对“湘糯粱2S”育性转换有一定的促进作用，即在一定温度范围内表现出短日促进不育，长日促进可育，但不存在明显的临界光周期。
5.“湘糯粱2S”在海南三亚生态点进行冬季制种，播种期可安
排在9月卜句至11月下旬，控制抽穗期在11月下旬至1月卜句，此
时段育性表现稳定，不育株率达100%，不育度达99.8%以上。在湖南
凤凰高海拔生态点进行夏季制种，播种期适宜安排在6月_上旬，于8
月中下旬抽穗，此时育性表现稳定，不育株率达98.6%，不育度达
98.2%。在湖南长沙生态点进行繁殖，播种期适宜安排在4月下旬至
5月底，7月中旬至8月中旬抽穗，此时段日平均温度达28℃以上，
育性表现为可育或半育。
6.经同工酶和蛋白质电泳分析，“湘糯粱2S”不育材料过氧化物
酶活性比可育材料高，不育材料比可育材料多一个92KD的多肤。表
明这种不育性可能是一种受光温调节的雄性可育基因表达受到阻碍
而使花粉走向败育。
The purpose of this experiment was to deal with the problem of fertility unstability in seed production of two-line sorghum. The fertility appearance of the Dual-purpose Genie Male Sterile(GMS) line must meet the requirment for the strict thermo-photoperiod condition during the special growth period. The purification and stable characteristic and the sensitive period of GMS line of sorghum, effect of temperature and photoperiod on it's fertility transfer and molecule mechanism were studied by using phytotron, field experiment and isoenzyme and protein analysis. The results follow as:
1. The GMS line of Xiangnuoliang 2S appeared stable sterile at 25℃ at heading stage, and both of sterile plant rate and abortive pollen rate were 100%.The ratoon plants cut down from sterile plants at 26℃ displayed the fertility alternation plant at 28℃, and breed the purificatory and stable GMS line's seed. Xiangnuoliang S-1 appeared fertility at 25℃.
2. Sensitive period for fertility transfer of the Xiangnuoliang 2S ranged from pollen mother cell differentiation to meiosis, morphylogically the stage when the leaf ring of the flag leaf is 0.8cm higher then that of the second leaf from the bottom, i.e., 4-10 days(in autumn) or 3-7 days(in
summer) before heading.
3. The critical temperature of fertility transfer of Xiangnuoliang 2S was 25C, and the plant appeared sterile under lower temperature and fertility under higher temperature.
4. Photoperiod had certain effect on the fertility transfer of Xiangnuoliang 2S, The day-short was benefical to sterility but day-long to fertility.
5. Seeding stage of hybrid seed production in Sanya, Hainan of the Xiangnuoliang 2S in winter was from the third ten days of September to November, heading stage was from the third ten days of November to next January. The fertility of plant appeared stable during this period, the sterile plant rate reached to 100%, the abortive pollen rate reached to 99.8%. Seeding stage of hybrid seed production for high altitude in summer was from the first ten days of June to the second ten days of August in Fenhuang, Hunan. The fertility of plant appeared stable during this period, the sterile plant rate reached over 98.6%, the abortive pollen rate reached over 98.2%. Seeding stage of multipication in Changsha, hunan was from the third ten days of April to May, heading stage was from the second ten days of July to August. Xiangnuoliang 2S was fertility when temperature was over 28C during this period.
6. Activity of peroxide enzyme of sterile material of the Xiangnuoliang 2S was higher than that of other fertility material, and the sterile material
increased 92KD polypeptide through analysising to isoenzyme and protein's electrophoresis. The results showed that this sterile might be male fertility gene's expression in thermo-photosensitivity modulatory was set back leading to pollen abortivity.
1．两系高粱不育系“湘糯粱2S”在孕穗至抽穗期控温25℃情况下表现出稳定不育，不育株率和不育度均达100％，利用控温26℃处理后仍表现出不育的植株，砍秆再生，再生植株在28℃以上高温条件下抽穗转为可育，自交繁殖出了纯化稳定的不育系种子，对照“湘糯粱S-1”在控温25℃时即表现为可育。
2．“湘糯粱2S”育性转换的温光敏感时期为花粉母细胞形成至减数分裂期，其外形判断为旗叶叶环高出倒2叶叶环0-8厘米，即抽穗前4-10天(秋季)或3-7天(夏季)。
3．“湘糯粱2S”是一个低温不育、高温可育的温敏型不育系，其育性转换的临界温度为25℃。
4．光周期对“湘糯粱2S”育性转换有一定的促进作用，即在一定温度范围内表现出短日促进不育，长日促进可育，但不存在明显的临界光周期。
5.“湘糯粱2S”在海南三亚生态点进行冬季制种，播种期可安
排在9月卜句至11月下旬，控制抽穗期在11月下旬至1月卜句，此
时段育性表现稳定，不育株率达100%，不育度达99.8%以上。在湖南
凤凰高海拔生态点进行夏季制种，播种期适宜安排在6月_上旬，于8
月中下旬抽穗，此时育性表现稳定，不育株率达98.6%，不育度达
98.2%。在湖南长沙生态点进行繁殖，播种期适宜安排在4月下旬至
5月底，7月中旬至8月中旬抽穗，此时段日平均温度达28℃以上，
育性表现为可育或半育。
6.经同工酶和蛋白质电泳分析，“湘糯粱2S”不育材料过氧化物
酶活性比可育材料高，不育材料比可育材料多一个92KD的多肤。表
明这种不育性可能是一种受光温调节的雄性可育基因表达受到阻碍
而使花粉走向败育。
The purpose of this experiment was to deal with the problem of fertility unstability in seed production of two-line sorghum. The fertility appearance of the Dual-purpose Genie Male Sterile(GMS) line must meet the requirment for the strict thermo-photoperiod condition during the special growth period. The purification and stable characteristic and the sensitive period of GMS line of sorghum, effect of temperature and photoperiod on it's fertility transfer and molecule mechanism were studied by using phytotron, field experiment and isoenzyme and protein analysis. The results follow as:
1. The GMS line of Xiangnuoliang 2S appeared stable sterile at 25℃ at heading stage, and both of sterile plant rate and abortive pollen rate were 100%.The ratoon plants cut down from sterile plants at 26℃ displayed the fertility alternation plant at 28℃, and breed the purificatory and stable GMS line's seed. Xiangnuoliang S-1 appeared fertility at 25℃.
2. Sensitive period for fertility transfer of the Xiangnuoliang 2S ranged from pollen mother cell differentiation to meiosis, morphylogically the stage when the leaf ring of the flag leaf is 0.8cm higher then that of the second leaf from the bottom, i.e., 4-10 days(in autumn) or 3-7 days(in
summer) before heading.
3. The critical temperature of fertility transfer of Xiangnuoliang 2S was 25C, and the plant appeared sterile under lower temperature and fertility under higher temperature.
4. Photoperiod had certain effect on the fertility transfer of Xiangnuoliang 2S, The day-short was benefical to sterility but day-long to fertility.
5. Seeding stage of hybrid seed production in Sanya, Hainan of the Xiangnuoliang 2S in winter was from the third ten days of September to November, heading stage was from the third ten days of November to next January. The fertility of plant appeared stable during this period, the sterile plant rate reached to 100%, the abortive pollen rate reached to 99.8%. Seeding stage of hybrid seed production for high altitude in summer was from the first ten days of June to the second ten days of August in Fenhuang, Hunan. The fertility of plant appeared stable during this period, the sterile plant rate reached over 98.6%, the abortive pollen rate reached over 98.2%. Seeding stage of multipication in Changsha, hunan was from the third ten days of April to May, heading stage was from the second ten days of July to August. Xiangnuoliang 2S was fertility when temperature was over 28C during this period.
6. Activity of peroxide enzyme of sterile material of the Xiangnuoliang 2S was higher than that of other fertility material, and the sterile material
increased 92KD polypeptide through analysising to isoenzyme and protein's electrophoresis. The results showed that this sterile might be male fertility gene's expression in thermo-photosensitivity modulatory was set back leading to pollen abortivity.
引文
1．卢庆善主编．高梁学．北京：中国农业出版社，1999，11：179～184
2．洪亚辉主编．植物外源DNA直接导入技术．长沙：湖南科学技术出版社，2000．4：107～153
3．袁隆平．农作物两系法杂种优势利用的现状与前景．科技导报，1997(12)：8～10
4．黄群策，孙敬三．我国农作物两系法杂种优势育种和应用的进展．科技导报，1997(12)：11～14
5．刘自华，贺际春．我国高粱育种方向探讨．作物杂志，1996(2)：19～21
6．卢庆善，孙毅，华泽田主编．农作物杂种优势．北京：中国农业科技出版社，2001，10：290～322
7. Murty-U-R. Breeding two line hybrids in sorghum bicolou(L). Moench Cereal Research Communication,1995, 23 (4): 397～402
8．汤文光，杨光立，李光林．两系法高粱两用不育系选育及应用研究．农作物两系法杂种优势利用国际学术讨论会论文集．1997，9：206～214
9．汤文光，陈叔君，李光林等．两系法杂交高粱新品种湘糯粱(S)1号的选育与应用．湖南农业科学，1994(5)：4～6
10．汤文光，刘海军，尹颛斌等．两系高粱杂种优势分析．湖南农业科学，2000(5)：16～17
11．牛天堂，张福耀，吴树彪等．高粱无融合生殖系SSA—1和296B的选育．作物杂志，1994(1)：5～6
12．张福耀，孟春刚，阎喜梅等．高粱SSA—1无融合生殖特性及其遗传分析．作物学报，1997，23(1)：89～94
13．张福耀，平俊爱，程庆军等．无融合生殖研究及其在高粱育种中的应用前景．杂粮作物，2000，20(3)：3～6
14．王成社，杨建设，刘平社．无融合在作物改良中的应用．作物杂志，1996(3)：17～19
15．罗耀武，阎学忠，陈士林等．高粱同源四倍体及四倍体杂交种．遗传学报，1985，12(5)：339～343
16．蒋观敏，罗耀武，李喆浩．同源四倍体高粱不育系和保持系的生物学及生理特性研究．作物学报，2000，26(4)：444～448
17．张继益，蒋观敏，罗耀武．高粱同源四倍体及其杂交种的细胞遗传学研究．华北农学报，1997，12(3)：1～6
18．梁凤山，罗耀武．多倍体及其在农业生产中的应用．国外农学—杂粮作物，1999，19(2)：20～23
19．裴新澍．多倍体诱导与育种．上海：上海科学技术出版社，1963．1：95～126
20. Blakeslee, A.F. & A.G. Avery. Methods of inducing doubling of chromosomesin plants. Jour. Hered. 1937,28:393～411
21．卢庆善，宋仁本，卢峰．高粱组织培养研究进展．国外农学—杂
粮作物，1998，18(2)：30～34
22．蔡忠杰，宋十二本，邹剑秋．辽宁省高梁育种工作的现状及展望．杂粮作物，2002，22(1)：11～13
23．中国农科院品资所主编．全国高粱品种资源目录．北京：中国农业出版社，1998．11．
24．徐瑞洋编著．高粱品质育种．北京：农业出版社．1987．5：209～214
25．韦耀明，李田银，李三棉等．高赖氨酸高粱265—1Y选育研究初报．山西农业科学，1995，23(1)：3～6
26．黎大爵，廖馥荪编著．甜高粱及其利用．北京：科学出版社．1992．8：56～77
27．汤文光，刘海军．用袁隆平学术思想指导高粱育种．袁隆平学术思想研究．长沙：湖南科学技术出版社，2000．4：161～167
28．袁隆平．杂交水稻育种的战略设想[J]．杂交水稻，1987(1)：1～3
29．袁隆平．杂交水稻超高产育种．科学中国人，1996，(1)：14～16
2．洪亚辉主编．植物外源DNA直接导入技术．长沙：湖南科学技术出版社，2000．4：107～153
3．袁隆平．农作物两系法杂种优势利用的现状与前景．科技导报，1997(12)：8～10
4．黄群策，孙敬三．我国农作物两系法杂种优势育种和应用的进展．科技导报，1997(12)：11～14
5．刘自华，贺际春．我国高粱育种方向探讨．作物杂志，1996(2)：19～21
6．卢庆善，孙毅，华泽田主编．农作物杂种优势．北京：中国农业科技出版社，2001，10：290～322
7. Murty-U-R. Breeding two line hybrids in sorghum bicolou(L). Moench Cereal Research Communication,1995, 23 (4): 397～402
8．汤文光，杨光立，李光林．两系法高粱两用不育系选育及应用研究．农作物两系法杂种优势利用国际学术讨论会论文集．1997，9：206～214
9．汤文光，陈叔君，李光林等．两系法杂交高粱新品种湘糯粱(S)1号的选育与应用．湖南农业科学，1994(5)：4～6
10．汤文光，刘海军，尹颛斌等．两系高粱杂种优势分析．湖南农业科学，2000(5)：16～17
11．牛天堂，张福耀，吴树彪等．高粱无融合生殖系SSA—1和296B的选育．作物杂志，1994(1)：5～6
12．张福耀，孟春刚，阎喜梅等．高粱SSA—1无融合生殖特性及其遗传分析．作物学报，1997，23(1)：89～94
13．张福耀，平俊爱，程庆军等．无融合生殖研究及其在高粱育种中的应用前景．杂粮作物，2000，20(3)：3～6
14．王成社，杨建设，刘平社．无融合在作物改良中的应用．作物杂志，1996(3)：17～19
15．罗耀武，阎学忠，陈士林等．高粱同源四倍体及四倍体杂交种．遗传学报，1985，12(5)：339～343
16．蒋观敏，罗耀武，李喆浩．同源四倍体高粱不育系和保持系的生物学及生理特性研究．作物学报，2000，26(4)：444～448
17．张继益，蒋观敏，罗耀武．高粱同源四倍体及其杂交种的细胞遗传学研究．华北农学报，1997，12(3)：1～6
18．梁凤山，罗耀武．多倍体及其在农业生产中的应用．国外农学—杂粮作物，1999，19(2)：20～23
19．裴新澍．多倍体诱导与育种．上海：上海科学技术出版社，1963．1：95～126
20. Blakeslee, A.F. & A.G. Avery. Methods of inducing doubling of chromosomesin plants. Jour. Hered. 1937,28:393～411
21．卢庆善，宋仁本，卢峰．高粱组织培养研究进展．国外农学—杂
粮作物，1998，18(2)：30～34
22．蔡忠杰，宋十二本，邹剑秋．辽宁省高梁育种工作的现状及展望．杂粮作物，2002，22(1)：11～13
23．中国农科院品资所主编．全国高粱品种资源目录．北京：中国农业出版社，1998．11．
24．徐瑞洋编著．高粱品质育种．北京：农业出版社．1987．5：209～214
25．韦耀明，李田银，李三棉等．高赖氨酸高粱265—1Y选育研究初报．山西农业科学，1995，23(1)：3～6
26．黎大爵，廖馥荪编著．甜高粱及其利用．北京：科学出版社．1992．8：56～77
27．汤文光，刘海军．用袁隆平学术思想指导高粱育种．袁隆平学术思想研究．长沙：湖南科学技术出版社，2000．4：161～167
28．袁隆平．杂交水稻育种的战略设想[J]．杂交水稻，1987(1)：1～3
29．袁隆平．杂交水稻超高产育种．科学中国人，1996，(1)：14～16
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