甘蔗茎径增粗的细胞学基础研究
摘要
甘蔗是广西重要的经济作物。原生分生组织是甘蔗地上部分一切组织形态发生中心，因此，研究原生分生组织的结构和生长分化对于阐明甘蔗的增粗机理具有重要意义。
本实验通过对6个不同甘蔗品种(粤糖91／976、月。粤糖86／368、桂糖11号、新台糖16号、美国CP80和农林8号)茎尖生长点采用压片，石蜡连续切片，半薄切片，超薄切片和酶组织化学定位等方法进行研究，取得主要结果如下：
1)甘蔗一天内(白天)不同时间都存在细胞分裂，大部分品种均在10:00出现细胞分裂高峰期，12:00～14:00是细胞分裂的低谷。早熟品种细胞分裂高峰期出现的时期早一些，晚熟品种则晚一些。甘蔗茎径和各生长时期细胞分裂指数呈正相关。
2)甘蔗生长锥原生分生组织具有明显的细胞—组织学区域
甘蔗茎尖生长锥原生分生组织可以划分为四个区域：原套原始细胞区、原体原始细胞区、髓分生区以及周缘分生区。不同区域之间细胞核及细胞的长、宽、面积、灰度值、液泡化程度指标差异显著，这些指标是甘蔗茎尖生长锥可以分区的重要依据。
从ATP酶的分布看，生长锥不同区域细胞ATP酶活性分布不一样，周缘分生区细胞ATP酶分布密度最大，其次到原体原始细胞区再到原套原始细胞区，髓分生区ATP酶分布相对较少。这也是甘蔗茎尖分区的重要依据。
3)周缘分生区细胞在甘蔗整个生育期都维持比较高的分裂频率，
是由原始细胞产生的苗端最活跃的、直接参加到苗的形态形成的分生
组织。
的甘蔗茎尖不同区域细胞ATP酶活性分布特征不同
原生分生组织ATP酶的活性主要分布在细胞核和线粒体上，而细
胞膜和小的液泡膜内则呈负反应。原形成层细胞的细胞核和线粒体上
ATP酶活性都比较强，细胞膜也显出了很高的活性。
甘蔗茎尖具有初生们粗分生组织，这些组织细胞的细胞核、细胞
膜、线粒体、胞间连丝上ATP酶活性都明显比周围的细胞高。
甘蔗节上的细胞ATP酶的活性比节间的高，这与甘蔗茎的增粗有
密切的关系。
Sugarcane is one of the most important economic crops in Guangxi. It is very meaningful to study the thickening mechanism by studying on the structure of protomerism, growth and differentiation, which formed at the time histomorpho I ogy.
In this experiment six sugarcane varieties: Yuetang91/976, Yangtang86/368, GT11, TT16, CP80, Nonglin 8 were used. The pressed disc, paraffin, semi slice, ultramicrotomy and Enzyme histology chemical locat i on methods were applied in the experiment. The results were followings:
1. Cell division took place in anytime and at 10 AM cell it reached the climax for most sugarcane varieties.
2. The breadth of growing tip was positively relative to the sugarcane stem.
3. In the view of cell morphology and the location of ATP Enzyme, the growing prick of sugarcane had significant cell-histology areas, which were tunica -primary cell zone (the growing tip had only one layer of tunica), corpus-primary cell zone, peripheral meristem zone and pith zone.
4. The indexes such as, cell nuclear in the growing tips, the length, breadth, areas and intensity of vacuole
5. The position effect determined the direction of cell division so the differentiation of the stem tip protomeristem was not the same in different areas.
Tunica and corpus zone are the cell center for differentiation and the fundamental for seedling keeping sustainable growing.
Cell at peripheral meristem zone kept a high frequency in the bearing period, which was the most active and closely related to the seedling morphorgenes is.
The pith meristem was responsible for bearing pith and enhancing the growth of seedling.
6. The ATPase mainly located in cell nuclear and mitochondrial while but ATPase activity was high both on the eel I nuclear and mitochondria of procambium. The cell membrane showed very high activity because the cell division began which enhanced the metabolism of materials and energy among eel Is.
There was primary meristem on the stem tips of sugarcane. The activity of ATPase located on this kind of cell was much higher than those located on the neighborhood cells. This i nd i cated that these cells is active in cell division and physiological metabolism.
The ATPase activity on the cell membrane, vacuole membrane, plasmadesmus and intercellular space is higher than those on the sugarcane nodes.
本实验通过对6个不同甘蔗品种(粤糖91／976、月。粤糖86／368、桂糖11号、新台糖16号、美国CP80和农林8号)茎尖生长点采用压片，石蜡连续切片，半薄切片，超薄切片和酶组织化学定位等方法进行研究，取得主要结果如下：
1)甘蔗一天内(白天)不同时间都存在细胞分裂，大部分品种均在10:00出现细胞分裂高峰期，12:00～14:00是细胞分裂的低谷。早熟品种细胞分裂高峰期出现的时期早一些，晚熟品种则晚一些。甘蔗茎径和各生长时期细胞分裂指数呈正相关。
2)甘蔗生长锥原生分生组织具有明显的细胞—组织学区域
甘蔗茎尖生长锥原生分生组织可以划分为四个区域：原套原始细胞区、原体原始细胞区、髓分生区以及周缘分生区。不同区域之间细胞核及细胞的长、宽、面积、灰度值、液泡化程度指标差异显著，这些指标是甘蔗茎尖生长锥可以分区的重要依据。
从ATP酶的分布看，生长锥不同区域细胞ATP酶活性分布不一样，周缘分生区细胞ATP酶分布密度最大，其次到原体原始细胞区再到原套原始细胞区，髓分生区ATP酶分布相对较少。这也是甘蔗茎尖分区的重要依据。
3)周缘分生区细胞在甘蔗整个生育期都维持比较高的分裂频率，
是由原始细胞产生的苗端最活跃的、直接参加到苗的形态形成的分生
组织。
的甘蔗茎尖不同区域细胞ATP酶活性分布特征不同
原生分生组织ATP酶的活性主要分布在细胞核和线粒体上，而细
胞膜和小的液泡膜内则呈负反应。原形成层细胞的细胞核和线粒体上
ATP酶活性都比较强，细胞膜也显出了很高的活性。
甘蔗茎尖具有初生们粗分生组织，这些组织细胞的细胞核、细胞
膜、线粒体、胞间连丝上ATP酶活性都明显比周围的细胞高。
甘蔗节上的细胞ATP酶的活性比节间的高，这与甘蔗茎的增粗有
密切的关系。
Sugarcane is one of the most important economic crops in Guangxi. It is very meaningful to study the thickening mechanism by studying on the structure of protomerism, growth and differentiation, which formed at the time histomorpho I ogy.
In this experiment six sugarcane varieties: Yuetang91/976, Yangtang86/368, GT11, TT16, CP80, Nonglin 8 were used. The pressed disc, paraffin, semi slice, ultramicrotomy and Enzyme histology chemical locat i on methods were applied in the experiment. The results were followings:
1. Cell division took place in anytime and at 10 AM cell it reached the climax for most sugarcane varieties.
2. The breadth of growing tip was positively relative to the sugarcane stem.
3. In the view of cell morphology and the location of ATP Enzyme, the growing prick of sugarcane had significant cell-histology areas, which were tunica -primary cell zone (the growing tip had only one layer of tunica), corpus-primary cell zone, peripheral meristem zone and pith zone.
4. The indexes such as, cell nuclear in the growing tips, the length, breadth, areas and intensity of vacuole
5. The position effect determined the direction of cell division so the differentiation of the stem tip protomeristem was not the same in different areas.
Tunica and corpus zone are the cell center for differentiation and the fundamental for seedling keeping sustainable growing.
Cell at peripheral meristem zone kept a high frequency in the bearing period, which was the most active and closely related to the seedling morphorgenes is.
The pith meristem was responsible for bearing pith and enhancing the growth of seedling.
6. The ATPase mainly located in cell nuclear and mitochondrial while but ATPase activity was high both on the eel I nuclear and mitochondria of procambium. The cell membrane showed very high activity because the cell division began which enhanced the metabolism of materials and energy among eel Is.
There was primary meristem on the stem tips of sugarcane. The activity of ATPase located on this kind of cell was much higher than those located on the neighborhood cells. This i nd i cated that these cells is active in cell division and physiological metabolism.
The ATPase activity on the cell membrane, vacuole membrane, plasmadesmus and intercellular space is higher than those on the sugarcane nodes.
引文
1 Cecich R A, N R Lersten and J P Miksche. Acytophotometric study of nucleic acids and proteins in the shoot apex of white spruce., Amer. J. Bot., 1972, 59: 442～449
2 Fosket J E and J D Miksche. A histochcrriical study of the seedling shoot apical meristem of till Pinus lambcrlia na, Amer. J, Bot. 1966. 653: 694
3 李扬汉，植物学，上海：上海科学技术出版社，1986，118～122
4 施良，王伏雄，胡玉喜，香榧营养苗端的结构及淀粉动态的研究，植物学报，1988，30(4)；341-346
5 Laux T, Mayer K F X, Berger J, Jorgens G. The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Development, 1996, 122: 87-96
6 Endrizzi K, Moussian B, Haecker A, Levin J Z, Laux T. The SHOOT MERISTEMLESS gene is required for maintenance of undifferentiated cells in Arabidopsis shoot and floral meristems and acts at a different regulatory level than the meristem genes WUSCHEL and ZWILLE. Plant J, 1996, 10:967-979
7 Long J A, Moan E I, Medford J l, Barton M K. A member of the KNOTTED class of homeodomain proteins encoded by the STM gene of Arabidopsis. Nature, 1996, 379: 66-69
8 Clark S E, Jacobsen S E, Levin J Z, Meyerowitz E M. The CLAVATA and SHOOT MERISTEMLESS loci competitively regulate meristem activity in Arabidopsis, Development, 1996, 122:1567～1575
9 Laux T, Schoof H, Maintaining the shoot meristem-the role of Clavata 1. Trends Plant Sci, 1997, 2:325-363
10 Williams R W, Wilson J M, Meyerowitz E M. A possible role for kinase-associated protein phosphatase in the Arabidopsis CLAVATAI signaling pathway. Proc Natl acad Sci USA, 1997, 94:1067-1072
11 Aida M, Ishida T, Fukaki H, Fujisawa H, Fujisaka M. Genes involved in organ separation in Arabidopsis: an analysis of the cup-shaped cotyleden mutant, Plant Cell, 1997, 9:841—857
12 Souer E, Van Houwelingen A, Kloos D, Mol J, Koes R. The no apical meristem gene of Petunia is required for pattern formation in embryos and flowers and is expressed at meristem and primodia boundaried. Cell, 1996, 85:159～170
13 Poethig R S. Leaf morphogenesis in flowering plants. Plant Cell, 1997, 9. 1077～1087
14 Hareven D, Gutflnger T, Parnis A, Eshed Y, Lifschitz E. The making of a compound leaf:
genetic manipulation of leaf architecture in tomato. Cell, 1996, 84:735-744
15 Laux T, Jurgens G. Embryogenesis. a new start in life. Plant Cell, 1997, 9:989-1000
16 李景原，庞广昌，朱命伟等，玉米茎尖扫描电镜观察及蛋白质电泳初报，河南科学。1996，6，第14卷增刊：14～16
17 李梅兰，曾广文，朱祝军，菜心茎尖nNA甲基化水平及GA、蛋白质含量的变化与花芽分化，浙江大学学报(农业与生命科学版)，2002，28(2)：161-164
18 任桂杰，董合忠，陈永吉等，棉花花芽分化时期茎尖内源激素的变化，西北植物学报，2002，22(2)：32～326
19 Fahn A(吴数明，刘德义译)，植物解剖学，南京：南开大学出版社，1990，366～370
20 K 伊稍著，李正理译，种子植物解剖学，上海：上海科学技术出版社，1982，191～197
21 成小飞 李文钿，秃杉苗端离体培养的形态组织学研究，植物学报，1992，34(12)：919～924
22 胡玉熹，苗端分区漫谈，植物杂志，1983，2；3～5
23 李正理，植物制片技术(第二版)，北京；科学技术出版社，1987
24 林钧安，高锦梁，洪健主编，实用生物电子显微术，沈阳：辽宁科学技术出版社。1989年3月，48～52，189～189
25 孙敬三，钱迎倩主编，植物细胞学研究方法，北京：科学出版社，1987，203～204
26 张泽林，细胞分裂相的决定因素——取材时间，生物学通报1999，34(8)：22
27 李懋学，张学敦，植物染色体研究技术，东北大学出版社，1991，26．
28 Stephens C E. Cytologia.1984, 49:679,
29 黄有总，徐建云等，几个甘蔗新品种的农艺性状比较研究，福建甘蔗，2001，(3)：8～13
30 三味工作室编写，SPSS V 10.0 for window 实用基础教程，北京：北京希望电子出版社，2001，2：98～106；174～209．
31 AC 吉斯，细胞生理学，科学出版社，1984，576～603
32 Foster A S. Structure and growth of the shoot apex in Ginkgo biloba. Bull. Torrcs Rot. Club, 1938: 65: 531-556.
33 Gifford Sr E M and G E Corson. The shoot apex in seed plants. Lot. Rev., 1971, 37: 143-229
34 Johnson M A. The shoot apex in Gymnosperms. Phytomorphology, 1951, 1:188-204
35 Madusudanau K N, S Nandakumar. Starch localization in shoot apt during ontogeny of pincapple. Phytomorphology, 1984, 34:135-139
36 Pillai S K, B Chacko. Anatomical and histochemical studies of the shoot apex of' Cedrus cleo clara. Phyzomorpho/ogy, 1978, 26: 275-283
37 Riding R T, E M Gifford, Jr., Histochemical changes occurring at the seedling shoot apex
of Pinus radiata. Can. J. Bot., 1973, 51: 501-512
38 Hall J L, A histochemical study of adenosine triphosphatase in young root tips. Planta, 1969, 89:254-265
39 Sexton B. and J Sutcliffe, Some observations on the characteristics. and distribution of adenosine triphosphatase in young roots of Pisum sativuin. Ann. Bot., 1969, 33:689-694
40 Shifrin N, L Levine, Cytocbemieal localization of adenosine triphosphatase in plant root meristem. J. Cell Sci., 1968, 3: 423-436
41 简令成，孙龙华，孙德兰，根尖分生区、伸长区和根毛区细胞内ATP酶活性的超微结构定位，植物学报，1982，24(5)：408-411
42 田国伟，申家恒，小麦胚珠在受精过程中ATP酶的超微细胞化学定位，植物学报，1993，35(5)：329—336
43 Joshi P A, Steward J MeD. Ultrastructural localization of ATPase activity In cotton fiber during elongation. Proto—plasma, 1988, 143:1—10
44 何才平，杨弘远，金鱼草胚珠中ATP酶活性的超微细胞化学定位，植物学报，1991，33(2)：85—90
45 彭时尧，庄伟健，刘利华，甘蔗叶不同部位ATP酶活性细胞化学定位，植物学报，1989，31(1)：24～28
46 刘林，范树国，张再君，西瓜柱头乳突细胞分泌活动期间ATP酶活性超微结构定位，植物学通报，2000，17(2)：179～184
47 Gilder J, Cronshaw J, Adenesine triphophatase in the phloem of Cueurbita, Planta, 1973, 110:189～204
48 第二军医大学电镜室，复旦大学生物系电镜室主编，细胞超微结构及功能[附电子显微镜技术]，上海：上海科学技术出版社出版，1981，83～93
2 Fosket J E and J D Miksche. A histochcrriical study of the seedling shoot apical meristem of till Pinus lambcrlia na, Amer. J, Bot. 1966. 653: 694
3 李扬汉，植物学，上海：上海科学技术出版社，1986，118～122
4 施良，王伏雄，胡玉喜，香榧营养苗端的结构及淀粉动态的研究，植物学报，1988，30(4)；341-346
5 Laux T, Mayer K F X, Berger J, Jorgens G. The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Development, 1996, 122: 87-96
6 Endrizzi K, Moussian B, Haecker A, Levin J Z, Laux T. The SHOOT MERISTEMLESS gene is required for maintenance of undifferentiated cells in Arabidopsis shoot and floral meristems and acts at a different regulatory level than the meristem genes WUSCHEL and ZWILLE. Plant J, 1996, 10:967-979
7 Long J A, Moan E I, Medford J l, Barton M K. A member of the KNOTTED class of homeodomain proteins encoded by the STM gene of Arabidopsis. Nature, 1996, 379: 66-69
8 Clark S E, Jacobsen S E, Levin J Z, Meyerowitz E M. The CLAVATA and SHOOT MERISTEMLESS loci competitively regulate meristem activity in Arabidopsis, Development, 1996, 122:1567～1575
9 Laux T, Schoof H, Maintaining the shoot meristem-the role of Clavata 1. Trends Plant Sci, 1997, 2:325-363
10 Williams R W, Wilson J M, Meyerowitz E M. A possible role for kinase-associated protein phosphatase in the Arabidopsis CLAVATAI signaling pathway. Proc Natl acad Sci USA, 1997, 94:1067-1072
11 Aida M, Ishida T, Fukaki H, Fujisawa H, Fujisaka M. Genes involved in organ separation in Arabidopsis: an analysis of the cup-shaped cotyleden mutant, Plant Cell, 1997, 9:841—857
12 Souer E, Van Houwelingen A, Kloos D, Mol J, Koes R. The no apical meristem gene of Petunia is required for pattern formation in embryos and flowers and is expressed at meristem and primodia boundaried. Cell, 1996, 85:159～170
13 Poethig R S. Leaf morphogenesis in flowering plants. Plant Cell, 1997, 9. 1077～1087
14 Hareven D, Gutflnger T, Parnis A, Eshed Y, Lifschitz E. The making of a compound leaf:
genetic manipulation of leaf architecture in tomato. Cell, 1996, 84:735-744
15 Laux T, Jurgens G. Embryogenesis. a new start in life. Plant Cell, 1997, 9:989-1000
16 李景原，庞广昌，朱命伟等，玉米茎尖扫描电镜观察及蛋白质电泳初报，河南科学。1996，6，第14卷增刊：14～16
17 李梅兰，曾广文，朱祝军，菜心茎尖nNA甲基化水平及GA、蛋白质含量的变化与花芽分化，浙江大学学报(农业与生命科学版)，2002，28(2)：161-164
18 任桂杰，董合忠，陈永吉等，棉花花芽分化时期茎尖内源激素的变化，西北植物学报，2002，22(2)：32～326
19 Fahn A(吴数明，刘德义译)，植物解剖学，南京：南开大学出版社，1990，366～370
20 K 伊稍著，李正理译，种子植物解剖学，上海：上海科学技术出版社，1982，191～197
21 成小飞 李文钿，秃杉苗端离体培养的形态组织学研究，植物学报，1992，34(12)：919～924
22 胡玉熹，苗端分区漫谈，植物杂志，1983，2；3～5
23 李正理，植物制片技术(第二版)，北京；科学技术出版社，1987
24 林钧安，高锦梁，洪健主编，实用生物电子显微术，沈阳：辽宁科学技术出版社。1989年3月，48～52，189～189
25 孙敬三，钱迎倩主编，植物细胞学研究方法，北京：科学出版社，1987，203～204
26 张泽林，细胞分裂相的决定因素——取材时间，生物学通报1999，34(8)：22
27 李懋学，张学敦，植物染色体研究技术，东北大学出版社，1991，26．
28 Stephens C E. Cytologia.1984, 49:679,
29 黄有总，徐建云等，几个甘蔗新品种的农艺性状比较研究，福建甘蔗，2001，(3)：8～13
30 三味工作室编写，SPSS V 10.0 for window 实用基础教程，北京：北京希望电子出版社，2001，2：98～106；174～209．
31 AC 吉斯，细胞生理学，科学出版社，1984，576～603
32 Foster A S. Structure and growth of the shoot apex in Ginkgo biloba. Bull. Torrcs Rot. Club, 1938: 65: 531-556.
33 Gifford Sr E M and G E Corson. The shoot apex in seed plants. Lot. Rev., 1971, 37: 143-229
34 Johnson M A. The shoot apex in Gymnosperms. Phytomorphology, 1951, 1:188-204
35 Madusudanau K N, S Nandakumar. Starch localization in shoot apt during ontogeny of pincapple. Phytomorphology, 1984, 34:135-139
36 Pillai S K, B Chacko. Anatomical and histochemical studies of the shoot apex of' Cedrus cleo clara. Phyzomorpho/ogy, 1978, 26: 275-283
37 Riding R T, E M Gifford, Jr., Histochemical changes occurring at the seedling shoot apex
of Pinus radiata. Can. J. Bot., 1973, 51: 501-512
38 Hall J L, A histochemical study of adenosine triphosphatase in young root tips. Planta, 1969, 89:254-265
39 Sexton B. and J Sutcliffe, Some observations on the characteristics. and distribution of adenosine triphosphatase in young roots of Pisum sativuin. Ann. Bot., 1969, 33:689-694
40 Shifrin N, L Levine, Cytocbemieal localization of adenosine triphosphatase in plant root meristem. J. Cell Sci., 1968, 3: 423-436
41 简令成，孙龙华，孙德兰，根尖分生区、伸长区和根毛区细胞内ATP酶活性的超微结构定位，植物学报，1982，24(5)：408-411
42 田国伟，申家恒，小麦胚珠在受精过程中ATP酶的超微细胞化学定位，植物学报，1993，35(5)：329—336
43 Joshi P A, Steward J MeD. Ultrastructural localization of ATPase activity In cotton fiber during elongation. Proto—plasma, 1988, 143:1—10
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