On the Cytochemistry of Cell Wall Formation in Poplar Trees

Abstract: The ultrastructure of cell walls and the mechanisms of cell wall formation are still not fully understood. The objective of our study was therefore to obtain additional fine structural details on the deposition of cell wall components during the differentiation of xylem cells in hybrid aspen ( Populus tremula L. × P. tremuloides Michx.) we used as a model tree. At the electron microscope level, PATAg staining revealed a successive deposition of polysaccharides with increasing distance from the cambium. Staining with potassium permanganate and UV microspectrophotometry showed that the cell walls were lignified, with some delay to the deposition of polysaccharides. Immunogold labelling of three lignin types in developing cell walls varied with progressive deposition of cell wall layers. Condensed lignin subunits were localized in corners of cells adjacent to the cambium prior to S1 formation, whereas non-condensed lignin subunits became labelled only in later stages - in secondary walls near cell corners and simultaneously with the completion of S1 formation. As S2 polysaccharide deposition progressed, the labelling extended towards the lumen. Labelling of peroxidases revealed their presence in cell corner regions of young xylem cells, still lacking a secondary wall, implying that peroxidases are incorporated into the developing cell wall at early developmental stages. A weak labelling of middle lamella regions and secondary walls could also be seen at later stages. The results are discussed in relation to current knowledge on the succession of polysaccharide and lignin deposition in woody cell walls.     

In situ analysis of lignins in transgenic tobacco reveals a differential impact of individual transformations on the spatial patterns of lignin deposition at the cellular and subcellular levels

Using tobacco transgenic lines altered in the monolignol biosynthetic pathway and which differ in their lignin profiles we have evaluated lignin deposition at the cellular and subcellular levels using several microanalytical techniques. Surprisingly, whereas a Cinnamoyl CoA reductase (CCR) down-regulated line with a strong decrease in lignin content exhibited an overall reduction in lignin deposition in the walls of the different xylem cell types, this reduction was selectively targeted to the fibers in a double transformant (down-regulated for both CCR and Cinnamyl alcohol dehydrogenase (CAD)) displaying a similar degree of global lignin content decrease. Fiber and vessel secondary walls of the transgenic tobacco line homozygous for the ccr antisense gene (CCR.H) down-regulated plants were dramatically destructured, particularly in the S2 sublayer, whereas the deposition of lignins in the S1 sublayer was not significantly modified. In contrast, cell wall organization was slightly altered in xylem cells of the double transformant. The relative distribution of non-condensed and condensed units in lignin, evaluated microscopically with specific antibodies, was differentially affected in the transgenics studied and, in a general way, a drop in non-condensed lignin units (beta- 0-4 interunit linkages) was associated with a loss of cohesion and extensive disorganization of the secondary wall. These results demonstrate that lignification is tightly and independently regulated in individual cell types and cell wall sublayers. They also show that down-regulation of specific genes may induce targeted changes in lignin structure and in spatial deposition patterns of the polymer.     

Biosynthesis of monolignols. Genomic and reverse genetic approaches

The biosynthesis of monolignols is one of the most studied pathways of plant natural product biosynthesis. However, the pathway has recently undergone considerable revision, and it would appear that our understanding of the exact routes for synthesis of the building blocks of lignin and lignans is still not fully understood. Early studies of in vitro enzyme specificity failed to appreciate the catalytic promiscuity of some of the enzymes of the monolignol pathway, and the evolving model of a metabolic grid for monolignol biosynthesis may fail to appreciate the possible extent of metabolic channeling within the pathway. New approaches to the study of monolignol biosynthesis include genomics, advanced cellular imaging techniques, and transgenic manipulation. This article summarizes the use of these approaches to gain a better understanding of the operation of a complex metabolic pathway.     

Cell wall remodeling under salt stress: Insights into changes in polysaccharides,feruloylation, lignification,and phenolic metabolism in maize

Although cell wall polymers play important roles in the tolerance of plants to abiotic stress, the effects of salinity on cell wall composition and metabolism in grasses remain largely unexplored. Here, we conducted an in-depth study of changes in cell wall composition and phenolic metabolism induced upon salinity in maize seedlings and plants. Cell wall characterization revealed that salt stress modulated the deposition of cellulose, matrix polysaccharides and lignin in seedling roots, plant roots and stems. The extraction and analysis of arabinoxylans by size-exclusion chromatography, 2D-NMR spectroscopy and carbohydrate gel electrophoresis showed a reduction of arabinoxylan content in salt-stressed roots. Saponification and mild acid hydrolysis revealed that salinity also reduced the feruloylation of arabinoxylans in roots of seedlings and plants. Determination of lignin content and composition by nitrobenzene oxidation and 2D-NMR confirmed the increased incorporation of syringyl units in lignin of maize roots. Salt stress also induced the expression of genes and the activity of enzymes enrolled in phenylpropanoid biosynthesis. The UHPLC–MS-based metabolite profiling confirmed the modulation of phenolic profiling by salinity and the accumulation of ferulate and its derivatives 3- and 4-O-feruloyl quinate. In conclusion, we present a model for explaining cell wall remodeling in response to salinity.     

平基槭花性别分化的细胞形态学观察

平基槭为杂性花,雄花与两性花同株,本文对其花性别分化过程进行了细胞形态学观察。结果发现,在花性别分化的早期,雄花和两性花的花芽中雌、雄蕊原基均具备,只是在花芽发育到一定时期,雄花的雌蕊原基发生选择性败育,败育发生在大孢子母细胞减数分裂为4个大孢子时期。两性花的雌蕊可以正常膨大结实,雄蕊花药虽然可以形成二核花粉,但不能正常开裂,属于不育雄蕊。初步分析认为,两性花雄蕊花药不能正常开裂与花粉囊壁纤维层木质化程度低有关。     

木质化程度对兔眼蓝浆果不同品种插条扦插生根的影响

在常规扦插条件下,兔眼蓝浆果(Vaccinium ashei Reade)的‘园蓝'('Gardenblue')、‘梯芙蓝'('Tifblue')、‘顶峰'(‘Climax')和‘杰兔'(‘Premier')等4个品种的硬枝插条均不能生根;而春季3月份开始在全光照间歇弥雾条件下扦插6个月后,上述各品种的硬枝插条的生根率分别达到84%、52%、62%和79%. 在全光照间歇弥雾条件下,未木质化的绿枝插条也未能生根;木质化程度不同的绿枝插条均能生根,品种间生根率差异显著,‘园蓝'和‘杰兔' 的生根率显著高于‘梯芙蓝'和‘顶峰';各品种内木质化程度不同的绿枝插条之间的生根率差异显著,木质化程度越低,生根率越高.木质化程度不同的插条还在生根部位、生根数量、插条萌芽和插条基部腐烂等方面存在差异.     

禾本科植物细胞壁的木质化和阿魏酰化

介绍了禾本科植物细胞壁木质化和阿魏酰化的过程及其生理意义,并阐述了木质化和阿魏酰化在细胞壁分子网络形成过程中的作用。