The application of Fourier transform mid-infrared(FTIR) spectroscopy to identify variation in cell wall composition of Setaria italica ecotypes

Cell wall composition in monocotyledonous grasses has been identified as a key area of research for developing better feedstocks for forage and biofuel production.Setaria viridis and its close domesticated relative Setaria italica have been chosen as suitable monocotyledonous models for plants possessing the C4 pathway of photosynthesis including sorghum,maize,sugarcane,switchgrass and Miscanthus×giganteus.Accurate partial least squares regression（PLSR）models to predict S.italica stem composition have been generated,based upon Fourier transform mid-infrared（FTIR）spectra and calibrated with wet chemistry determinations of ground S.italica stem material measured using a modified version of the US National Renewable Energy Laboratory（NREL）acid hydrolysis protocol.The models facilitated a high-throughput screening analysis for glucan,xylan,Klason lignin and acid soluble lignin（ASL）in a collection of 183 natural S.italica variants and clustered them into classes,some possessing unique chemotypes.The predictive models provide a highly efficient screening tool for large scale breeding programs aimed at identifying lines or mutants possessing unique cell wall chemotypes.Genes encoding key catalytic enzymes of the lignin biosynthesis pathway exhibit a high level of conservation with matching expression profiles,measured by RT-q PCR,among accessions of S.italica,which closely mirror profiles observed in the different developmental regions of an elongating internode of S.viridis by RNASeq.

Floristic composition and ecological gradient analyses of the Liakot Forests in the Kalam region of District Swat,Pakistan

Kalam Kohistan is a dry,temperate hilly region in the Hindu Raj Series of the Greater Hindu Kush Mountains with diverse forests.As plant distribution and composition is expression of a range of various environmental variables,ecological and floristic attributes of vegetation in the Laikot Forests were therefore evaluated via consecutive explorative trips.These forests have not been evaluated ecologically before due to harsh climatic conditions and issues of accessibility and the study in hand is one of the first in its nature.Quantitative ecological techniques were used to sample forest vegetation and identify species in quadrats of 2 m×2 m for herbs,5 m×5 m for shrubs and 10 m×10 m for trees.All the data of 195 plant species and environmental factors were analyzed via Two-way Cluster Analysis and Canonical Correspondence Analysis using PCORD and CANOCO software.Among 195 plant species from 63 families were 27 species of Asteraceae,the most-speciose family,followed by Poaceae(20 spp.).The dominant life form was therophyte(86 spp.;44%share),followed by phanerophyte(41 spp.;21%share).For the leaf size spectrum,most prevalent was nanophyll(73 spp.;44%)followed by microphyll(66 spp.,34%)and mesophyll(44 spp.;23%).Calcium carbonate,pH,potassium,sand,silt,organic matter,slope aspect and grazing significantly influenced species composition,distribution and habitat.The floristic variation in the region was diverse due to elevational and aspect gradients and thus devided into three zones.This vegetation zonation can be utilized for forest management,species and habitat conservation.Both in situ and ex situ conservation of threatened plant species may improve their conservation status in future if address properly.

Expression of the C-Terminal Domain of Mammalian TET3 DNA Dioxygenase in Arabidopsis thaliana Induces Heritable Methylation Changes at rDNA Loci

In plants, demethylation of 5-methylcytosine (5 mC) residues is controlled by DNA glycosylases, while in mammals it requires oxidation of 5 mC by TET proteins, a group of Fe(II)/2-oxoglutaratedependent dioxygenases. We analysed the effects of expressing the C-terminal catalytic domain of the human TET3 gene (TET3c) in Arabidopsis thaliana, using an rDNA region as a methylation reporter. In TET3c transformants, epialleles with hypomethylation or hypermethylation patterns can be induced, which is each stably retained in progeny lines even after removal of the TET3c transgene. In TET3c transformants, 5-hydroxymethylcytosine (5 hmC) marks are detected, indicative of the oxidative activity of the transgenic enzyme. 5-formylcytosine (5 fC) is only detectable in TET3c transformants with a DNA glycosylase mutant background suggesting further oxidation of 5 hmC residues to 5 fC by TET3c, and efficient recognition and removal of 5 fC by plant glycosylases. The results suggest that TET3c can be employed to induce heritable locus-specific changes in DNA methylation, and that accumulation of 5 hmC can be used as a marker for TET3c target regions.

Gene Duplication and the Evolution of Plant MADS-box Transcription Factors

Since the first MADS-box transcription factor genes were implicated in the establishment of floral organ identity in a couple of model plants, the size and scope of this gene family has begun to be appreciated in a much wider range of species. Over the course of millions of years the number of MADS-box genes in plants has increased to the point that the Arabidopsis genome contains more than 100. The understanding gained from studying the evolution, regulation and function of multiple MADS-box genes in an increasing set of species, makes this large plant transcription factor gene family an ideal subject to study the processes that lead to an increase in gene number and the selective birth, death and repurposing of its component members. Here we will use examples taken from the MADS-box gene family to review what is known about the factors that influence the loss and retention of genes duplicated in different ways and examine the varied fates of the retained genes and their associated biological outcomes.

The Cooperative Activities of CSLD2, CSLD3, and CSLD5 Are Required for Normal Arabidopsis Development

Glycosyltransferases of the Cellulose Synthase Like D （CS/D） subfamily have been reported to be involved in tip growth and stem development in Arabidopsis. The csld2 and csld3 mutants are root hair defective and the csld5 mutant has reduced stem growth. In this study, we produced double and triple knockout mutants of CSLD2, CSLD3, and CSLD5. Unlike the single mutants and the csld2/csld3 double mutant, the csld2/csld5, csld3/csld5, and csld2/csld3/csld5 mutants were dwarfed and showed severely reduced viability. This demonstrates that the cooperative activities of CSLD2, CSLD3, and CSLD5 are required for normal Arabidopsis development, and that they are involved in important processes besides the specialized role in tip growth. The mutant phenotypes indicate that CSLD2 and CSLD3 have overlapping functions with CSLD5 in early plant development, whereas the CSLD2 and CSLD3 proteins are non-redundant. To determine the biochemical function of CSLD proteins, we used transient expression in tobacco leaves. Microsomes containing heterologously expressed CSLD5 transferred mannose from GDP-mannose onto endogenous acceptors. The same activity was detected when CSLD2 and CSLD3 were coexpressed but not when they were expressed separately. With monosaccharides as exogenous acceptors, microsomal preparations from CSLD5-expressing plants mediated the transfer of mannose from GDP-mannose onto mannose. These results were supported by immunodetection studies that showed reduced levels of a mannan epitope in the cell walls of stem interfascicular fibers and xylem vessels of the csld2/csld3/csld5 mutant.

Cell Wall Biology： Perspectives from Cell Wall Imaging

Polysaccharide-rich plant cell walls are important biomaterials that underpin plant growth, are major repositories for photosynthetically accumulated carbon, and, in addition, impact greatly on the human use of plants. Land plant cell walls contain in the region of a dozen major polysaccharide structures that are mostly encompassed by cellulose, hemicelluloses, and pectic polysaccharides. During the evolution of land plants, polysaccharide diversification appears to have largely involved structural elaboration and diversification within these polysaccharide groups. Cell wall chemistry is well advanced and a current phase of cell wall science is aimed at placing the complex polysaccharide chemistry in cellular contexts and developing a detailed understanding of cell wall biology. Imaging cell wall glycomes is a challenging area but recent developments in the establishment of cell wall molecular probe panels and their use in high throughput procedures are leading to rapid advances in the molecular understanding of the spatial heterogeneity of individual cell walls and also cell wall differences at taxonomic levels. The challenge now is to integrate this knowledge of cell wall heterogeneity with an understanding of the molecular and physiological mechanisms that underpin cell wall properties and functions.

Cell Wall Microstructure Analysis Implicates Hemicellulose Polysaccharides in Cell Adhesion in Tomato Fruit Pericarp Parenchyma

Methods developed to isolate intact cells from both unripe and ripe tomato fruit pericarp parenchyma have allowed the cell biological analysis of polysaccharide epitopes at the surface of separated cells. The LM7 pectic homogalacturonan epitope is a marker of the junctions of adhesion planes and intercellular spaces in parenchyma systems. The LM7 epitope persistently marked the former edge of adhesion planes at the surface of cells separated from unripe and ripened tomato fruit and also from fruits with the Cnr mutation. The LM 11 xylan epitope was associated, in sections, with cell walls lining intercellular space but the epitope was not detected at the surface of isolated cells, being lost during cell isolation. The LM15 xyloglucan epitope was present at the surface of cells isolated from unripe fruit in a pattern reflecting the former edge of cell adhesion planes/intercellular space but with gaps and apparent breaks. An equivalent pattern of LM15 epitope occurrence was revealed at the surface of cells isolated by pectate lyase action but was not present in cells isolated from ripe fruit or from Cnr fruit. In contrast to wild-type cells, the LM5 galactan and LM21 mannan epitopes occurred predominantly in positions reflecting intercellular space in Cnr, suggesting a concerted alteration in cell wall microstructure in response to this mutation. Galactanase and mannanase, along with pectic homogalacturonan-degrading enzymes, were capable of releasing cells from unripe fruit parenchyma. These observations indicate that hemicellulose polymers are present in architectural contexts reflecting cell adhesion and that several cell wall polysaccharide classes are likely to contribute to cell adhesion/cell separation in tomato fruit pericarp parenchyma.

Stem Cell Regulation by Arabidopsis WOX Genes

Gene amplification followed by functional diversification is a major force in evolution. A typical example of this is seen in the WUSCHEL-RELATED HOMEOBOX （WOX） gene family, named after the Arabidopsis stem cell regulator WUSCHEL. Here we analyze functional divergence in the WOX gene family. Members of the WUS clade, except the cambium stem cell regulator WOX4, can substitute for WUS function in shoot and floral stem cell maintenance to different degrees. Stem cell function of WUS requires a canonical WUS-box, essential for interaction with TPL/TPR co-repressors, whereas the repressive EAR domain is dispensable and the acidic domain seems only to be required for female fertility. In contrast to the WUS clade, members of the ancient WOX13 and the WOX9 clades cannot support stem cell maintenance. Although the homeodomains are interchangeable between WUS and WOX9 clade members, a WUS- compatible homeodomain together with canonical WUS-box is not sufficient for stem cell maintenance. Our results suggest that WOX function in shoot and floral meristems of Arabidopsis is restricted to the modern WUS clade, suggesting that stem cell control is a derived function. Yet undiscovered functional domains in addition to the homeodomain and the WUS-box are necessary for this function.

Identification of Quantitative Trait Loci Affecting Hemicellulose Characteristics Based on Cell Wall Composition in a Wild and Cultivated Rice Species

Cell wall hemicellulosic polysaccharides are structurally complex and diverse. Knowledge about the synthesis of cell wall hemicelluloses and their biological roles is limited. Quantitative trait loci （QTL） mapping is a helpful tool for the dissection of complex phenotypes for gene identification. In this study, we exploited the natural variation in cell wall monosaccharide levels between a common wild rice, Yuanj, and an elite indica cultivar, Teqing, and performed QTL map- ping with their introgression lines （ILs）. Chemical analyses conducted on the culms of Yuanj and TeqJng showed that the major alterations are found in glucose and xylose levels, which are correlated with specific hemicellulosic polymers. Gly- cosidic linkage examination revealed that, in Yuanj, an increase in glucose content results from a higher level of mixed linkage I^-glucan （MLG）, whereas a reduction in xylose content reflects a low level of xylan backbone and a varied arabi- noxylan （AX） structure. Seventeen QTLs for monosaccharides have been identified through composition analysis of the culm residues of 95 core ILs. Four major QTLs affecting xylose and glucose levels are responsible for 19 and 21% of the phenotypic variance, respectively. This study provides a unique resource for the genetic dissection of rice cell wall forma- tion and remodeling in the vegetative organs.

除草剂丙酯草醚对大麦的影响和支链氨基酸对其生长的恢复作用（英文）

目的:评价除草剂丙酯草醚对大麦生长的影响,探讨外源添加支链氨基酸对其生长的恢复作用。创新点:从除草剂丙酯草醚作用机理出发,阐明了外源添加支链氨基酸对丙酯草醚作用下大麦各项生理和生长指标的缓解作用,为生长调节物质的进一步开发利用提供实验依据。方法:采用水培和大田实验相结合的方法,研究了除草剂丙酯草醚对大麦生长的影响,并通过进一步测定大麦幼苗各项生理指标和支链氨基酸浓度,表明100mg/L支链氨基酸可显著缓解丙酯草醚对大麦的生理损伤,从而恢复其生长。结论:外源添加支链氨基酸能够缓解除草剂丙酯草醚对大麦生长的抑制作用。

Phyllosticta species associated with freckle disease of banana

The identity of the casual agent of freckle disease of banana was investigated.The pathogen is generally referred to in literature under its teleomorphic name,Guignardia musae,or that of its purported anamorph,Phyllosticta musarum.Based on morphological and molecular data from a global set of banana specimens,several species were found associated with freckle disease.Phyllosticta maculata(from Southeast Asia and Oceania)is introduced as a new name for Guignardia musae,and an epitype is designated from Australia.Phyllosticta musarum(from India and Thailand)is shown to represent a distinct species,and the name is fixed by designation of an epitype from India.Guignardia stevensii is confirmed as distinct species from Hawaii,while Guignardia musicola from northern Thailand is shown to contain different taxa and is regarded as nomen confusum.Phyllosticta cavendishii is described as a new,widely distributed species,appearing primarily on Cavendish,but also on non-Cavendish banana cultivars.