胶体金标记法半定量比较植物韧皮部和叶肉组织中细胞壁的纤维素含量
|
摘要
植物细胞壁由多种多糖成分构成,不同类型细胞的细胞壁结构和成分略有不同。本研究利用纤维素酶探针对水稻三种类型细胞壁,包括筛管细胞壁、韧皮部薄壁细胞细胞壁和叶肉细胞细胞壁分别做胶体金标记,半定量比较细胞壁区域内金颗粒的面密度,间接反映三种类型细胞壁的纤维素含量。利用IMARIS软件对金标图像做金颗粒的分割、细胞壁区域面积测算和细胞壁区域内金颗粒的筛选等一系列图像处理技术,可以半自动化对较大批量的图像进行处理,利于统计分析。经过每组30个重复的测算,结果发现筛管细胞壁的纤维素含量显著高于韧皮部薄壁细胞和叶肉细胞。
Plant cell wall is composed of multiple types of polysaccharides,whose structures and compositions are different from each other in different types of cells. In thisstudy,a gold labeling experiment was performed on three types of cell walls( CW) in rice plant,namely CW of sieve element( SE)-SE interface,CW of phloem parenchyma( PP)-PP interface and CW of mesophyll cell( MC)-MC interface. The concentration of cellulose in CW was characterized by the areal density of gold particle which was obtained by semi-quantification studies. Program IMARIS was used to analyze a series of images including semi-automatically segmenting gold particle,CW region,and classifying gold particles. This semi-automatic performance could process theimage in high flux( 30 repeats for each treatment in thisstudy). The results showed that the concentration of cellulose in SE-SE CW was remarkably higher than that in PP-PP CW and in MC-MC CW.
Plant cell wall is composed of multiple types of polysaccharides,whose structures and compositions are different from each other in different types of cells. In thisstudy,a gold labeling experiment was performed on three types of cell walls( CW) in rice plant,namely CW of sieve element( SE)-SE interface,CW of phloem parenchyma( PP)-PP interface and CW of mesophyll cell( MC)-MC interface. The concentration of cellulose in CW was characterized by the areal density of gold particle which was obtained by semi-quantification studies. Program IMARIS was used to analyze a series of images including semi-automatically segmenting gold particle,CW region,and classifying gold particles. This semi-automatic performance could process theimage in high flux( 30 repeats for each treatment in thisstudy). The results showed that the concentration of cellulose in SE-SE CW was remarkably higher than that in PP-PP CW and in MC-MC CW.
引文
[1]POPPER Z,MICHEL G,HERVE C,et al.Evolution and diversity of plant cell walls:from algae to flowering plants[J].Annual Review of Plant Biology,2011,62(1):567-590.
[2]VARNER J E,LIN L S.Plant cell wall architecture[J].Cell,1989,56(2):231-239.
[3]COSGROVE D J.Growth of the plant cell wall[J].Nature Reviews Molecular Cell Biology,2005,6(11):850-861.
[4]van BEL A J,EHLERS K,KNOBLAUCH M.Sieve elements caught in the act[J].Trends in Plant Science,2002,7(3):126-132.
[5]VANBEL A J.The phloem,a miracle of ingenuity[J].Plant,Cell&Environment,2003,26(1):125-149.
[6]HUCKELHOVEN R.Cell wall-associated mechanisms of disease resistance and susceptibility[J].Annual Review of Phytopathology,2007,45(1):101-127.
[7]ZHONG R Q,YE Z H.Secondary cell walls[M].e LS.John Wiley&Sons,Ltd,2009.DOI:10.1002/9780470015902.a0021256.
[8]COSTA G,PLAZANET I.Plant cell wall,a challenge for its characterisation[J].Advances in Biological Chemistry,2016,6(3):70-105.
[9]DAHIYA P,BREWIN N J.Immunogold localization of callose and other cell wall components in pea nodule transfer cells[J].Protoplasma,2000,214(3):210-218.
[10]徐颖,胡东维,娄沂春,等.麦胚凝集素与纤维素酶胶体金探针的混合一步法标记[J].电子显微学报,2000,19(2):179-182.
[11]XIE L,LU M F,SONG X J,et al.Structure and components of the globular and filamentous viroplasms induced by rice black-streaked dwarf virus[J].Micron,2017,98:12-23.
[12]SCHOLANDER P F,BRADSTREET E D,HEMMINGSEN E A,et al.Sap pressure in vascular plants[J].Science,1965,148(3668):339-346.
[13]LUCAS W J,GROOVER A,LICHTENBERGER R,et al.The plant vascular system:evolution,development and functions F[J].Journal of Integrative Plant Biology,2013,55(4):95.
[14]李云琴,王华,谢礼,等.免疫胶体金标记定位昆诺藜中的胼胝质和胼胝质降解酶[J].电子显微学报,2014,33(6):558-563.
[15]张梦婕,丁琦,贺新强,等.低温对毛果杨韧皮部筛分子胼胝质沉积的影响[J].电子显微学报,2015,34(2):121-125.
[16]AMICO F D,SKARMOUTSOU E.Quantifying immunogold labelling in transmission electron microscopy[J].Journal of Microscopy,2008,230(1):9-15.
[17]LUCOCQ J.Quantitation of gold labelling and antigens in immunolabelled ultrathin sections[J].Journal of Anatomy,1994,184(1):1-13.
[2]VARNER J E,LIN L S.Plant cell wall architecture[J].Cell,1989,56(2):231-239.
[3]COSGROVE D J.Growth of the plant cell wall[J].Nature Reviews Molecular Cell Biology,2005,6(11):850-861.
[4]van BEL A J,EHLERS K,KNOBLAUCH M.Sieve elements caught in the act[J].Trends in Plant Science,2002,7(3):126-132.
[5]VANBEL A J.The phloem,a miracle of ingenuity[J].Plant,Cell&Environment,2003,26(1):125-149.
[6]HUCKELHOVEN R.Cell wall-associated mechanisms of disease resistance and susceptibility[J].Annual Review of Phytopathology,2007,45(1):101-127.
[7]ZHONG R Q,YE Z H.Secondary cell walls[M].e LS.John Wiley&Sons,Ltd,2009.DOI:10.1002/9780470015902.a0021256.
[8]COSTA G,PLAZANET I.Plant cell wall,a challenge for its characterisation[J].Advances in Biological Chemistry,2016,6(3):70-105.
[9]DAHIYA P,BREWIN N J.Immunogold localization of callose and other cell wall components in pea nodule transfer cells[J].Protoplasma,2000,214(3):210-218.
[10]徐颖,胡东维,娄沂春,等.麦胚凝集素与纤维素酶胶体金探针的混合一步法标记[J].电子显微学报,2000,19(2):179-182.
[11]XIE L,LU M F,SONG X J,et al.Structure and components of the globular and filamentous viroplasms induced by rice black-streaked dwarf virus[J].Micron,2017,98:12-23.
[12]SCHOLANDER P F,BRADSTREET E D,HEMMINGSEN E A,et al.Sap pressure in vascular plants[J].Science,1965,148(3668):339-346.
[13]LUCAS W J,GROOVER A,LICHTENBERGER R,et al.The plant vascular system:evolution,development and functions F[J].Journal of Integrative Plant Biology,2013,55(4):95.
[14]李云琴,王华,谢礼,等.免疫胶体金标记定位昆诺藜中的胼胝质和胼胝质降解酶[J].电子显微学报,2014,33(6):558-563.
[15]张梦婕,丁琦,贺新强,等.低温对毛果杨韧皮部筛分子胼胝质沉积的影响[J].电子显微学报,2015,34(2):121-125.
[16]AMICO F D,SKARMOUTSOU E.Quantifying immunogold labelling in transmission electron microscopy[J].Journal of Microscopy,2008,230(1):9-15.
[17]LUCOCQ J.Quantitation of gold labelling and antigens in immunolabelled ultrathin sections[J].Journal of Anatomy,1994,184(1):1-13.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |