A complex of oxidised chitosan and silver ions grafted to cotton fibres with bacteriostatic properties.

The laccase/TEMPO system was employed to oxidise the C6 primary hydroxyl group on the chitosan (CS) to form a carboxyl group to obtain oxidised chitosan (C-COS). The silver-oxidised chitosan complex(C-COS-Ag) was prepared by reacting C-COS with silver nitrate, then C-COS-Ag and cotton fibres were subjected to a reaction to prepare bacteriostatic fibres. FT-IR and XPS analysis showed that: Ag+ and C-COS were combined in these forms: Ag, [Ag(NH3)2] OH, -COOAg, and Ag2O. C-COS-Ag was combined with cotton fibres by way of ester bonds. The inhibition zone of bacteriostatic fibres was all greater than 11 mm. After 50 washing tests, the bacteriostatic effect of bacteriostatic fibres remained at above 99 %. The amount of silver ions that had migrated from the bacteriostatic fibre was 3.336 mg/kg.

Color evolution of poplar wood chips and its response to lignin and extractives changes in autohydrolysis pretreatment.

Combining chemi-mechanical pulping with autohydrolysis pretreatment is an efficient and value-added utilization approach for lignocellulosic biomass in paper industry. To further promote the utilization of autohydrolyzed biomass in chemi-mechanical pulping, the color evolution of poplar wood chips in autohydrolysis pretreatment and its chromogenic mechanism were investigated by using CIELab color system, FT-IR, NMR and GPC. The results showed that the total color difference ΔE* increased obviously, which were remarkable as the combined hydrolysis factor (CHF) increased. The lignin content led to a more significant influence on the color of poplar wood chips than the extractives. The autohydrolysis pretreatment with a higher CHF accelerated the degradation and subsequent condensation of lignin, resulting in the formation of chromophoric groups, such as Hibbert ketone, quinones and quinone methides. It is of great significance for biomass refinery and paper industry to reveal the color evolution of poplar wood chips caused by autohydrolysis pretreatment from the point of view of chemical components' structure.

Construction of Marker-Free Genetically Modified Maize Using a Heat-Inducible Auto-Excision Vector.

Gene modification is a promising tool for plant breeding, and gradual application from the laboratory to the field. Selectable marker genes (SMG) are required in the transformation process to simplify the identification of transgenic plants; however, it is more desirable to obtain transgenic plants without selection markers. Transgene integration mediated by site-specific recombination (SSR) systems into the dedicated genomic sites has been demonstrated in a few different plant species. Here, we present an auto-elimination vector system that uses a heat-inducible Cre to eliminate the selectable marker from transgenic maize, without the need for repeated transformation or sexual crossing. The vector combines an inducible site-specific recombinase (hsp70::Cre) that allows for the precise elimination of the selectable marker gene egfp upon heating. This marker gene is used for the initial positive selection of transgenic tissue. The egfp also functions as a visual marker to demonstrate the effectiveness of the heat-inducible Cre. A second marker gene for anthocyanin pigmentation (Rsc) is located outside of the region eliminated by Cre and is used for the identification of transgenic offspring in future generations. Using the heat-inducible auto-excision vector, marker-free transgenic maize plants were obtained in a precisely controlled genetic modification process. Genetic and molecular analyses indicated that the inducible auto-excision system was tightly controlled, with highly efficient DNA excision, and provided a highly reliable method to generate marker-free transgenic maize.

Cationic cellulose nanofibers as sustainable flocculant and retention aid for reconstituted tobacco sheet with high performance.

The study that cationic cellulose nanofibers (CCNF) acts as sustainable flocculant and retention aid of precipitated calcium carbonate (PCC) for the preparation of reconstituted tobacco sheet (RTS) was carried out, thanks to the properties of CCNF. In this work, the enhanced flocculation, reflocculation and size properties of PCC flocs induced by CCNF were investigated via a focused beam reflectance measurement (FBRM) system. The physical properties of RTS such as bulk and air permeability etc. were also studied. The results indicated that CCNF could distinctly improve the flocculation and reflocculation properties of PCC with a desirable chord length in the tobacco slurry, and that the PCC retention was also increased with CCNF addition along with a slight decrease of tensile strength of RTS. The mechanisms of flocculation and reflocculation, as well as the reasons of enhanced bulk and air permeability properties ascribed to CCNF were also demonstrated.

Infection of Embryonic Callus with Agrobacterium Enables High-Speed Transformation of Maize.

Several approaches have recently been adopted to improve Agrobacterium-mediated transformation of maize; however, about eight months of in vitro culture are still required to isolate transgenic plants. Furthermore, genetic transformation of maize depends on immature embryos, which greatly increases costs. Here, we report a method that ensures the competency of an embryogenic callus secondary culture under laboratory conditions for Agrobacterium-mediated transformation. Moreover, pretreatment of the cell wall with a mixed lytic enzyme solution prior to Agrobacterium infection, significantly improved transformation efficiency and stability. Average stable transformation efficiency was approximately 30.39%, with peaks of 94.46%. Expression and phenotypic analysis of the Rsc reporter gene were tested in the T0 generation of transgenic plants. Using this system, we successfully regenerated transgenic maize plantlets within three months of the emergence of the embryogenic callus. Additionally, we reduced somaclonal variation accompanying prolonged culture of maize cells in the dedifferentiated state, thus facilitating the molecular breeding of maize.

The study of inhibitory effects and mechanism of carboxylate chitooligomer on melanin, prepared by laccase/TEMPO system.

A carboxylate chitooligomer (C-COS) containing carboxyl groups attached to chitooligomer (COS) molecules has been prepared by laccase/2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) system, which is a green-chemistry method. Several experiments were designed to evaluate inhibition effects on melanin and mechanisms of C-COS. The results indicated that C-COS exhibited more distinct anti-melanogenic effects compared to COS. C-COS inhibits melanin production with tyrosine (Tyr) and DOPA as the substrate of melanin formation, and the inhibition rates are, respectively, 89.07% and 84.45%, which reach 1.4-2 times those of COS. UV-vis spectroscopy was used to elucidate the interaction mechanism between C-COS and tyrosinase (TYR). It is C-COS chelating with metal Cu ions in tyrosinase (TYR) that decreases the enzyme activity. Half-maximal inhibitory concentrations (IC50) of C-COS were calculated as 13.49 and 4.07 mg/mL for monophenolase (cresolase) and diphenolase (catecholase), respectively.

The hydration mechanism and hydrogen bonding structure of 6-carboxylate chitooligosaccharides superabsorbent material prepared by laccase/TEMPO oxidation system.

6-carboxylate chitooligosaccharides (6-CCOS), as a superabsorbent material, were prepared by way of the laccase/TEMPO oxidation system. It exhibited a higher moisture-absorption ability and stronger affinity for water. To understand the real reasons for this, the hydrogen bonding structure of 6-CCOS and the hydration mechanism of the molecule were investigated using infrared (IR), differential scanning calorimetry (DSC), and nuclear magnetic resonance (NMR). It was found that the introduction of a strongly hydrophilic carboxylate ion on the C6 site of chitooligosaccharides molecule was conducive to the enhancement of the interaction between polysaccharide polymers and water molecules. The most important was the formation of hydrogen bonds connected between carboxylate ion and residual water. In addition, the hydration mechanism of 6-CCOS molecules was presumed to be that more water molecules from outside were incorporated into the already embedded water molecules within the polymer. The whole molecule was woven into a huge water-polymer network structure through intermolecular hydrated hydrogen bonds.

Nickel(ii)-catalyzed tandem C(sp2)-H bond activation and annulation of arenes with gem-dibromoalkenes.

A nickel(ii)/silver(i)-catalyzed tandem C(sp2)-H activation and intramolecular annulation of arenes with dibromoalkenes has been successfully achieved, which offers an efficient approach to the 3-methyleneisoindolin-1-one scaffold. Attractive features of this system include its low cost, ease of operation, and its ability to access a wide range of isoindolinones.

Molecular structural differences between maize leaf and endosperm starches.

The morphology, whole molecular size distribution and chain-length distribution of maize leaf starch have been characterized and compared to its endosperm starch, to better understand differences between leaf and endosperm starch structure, and the relationship with the functions of starch in these organs. Leaf starch is found to have amylopectin with much shorter chains (virtually none with a degree of polymerization, DP, above 70) than the endosperm amylopectin, which has significant numbers of chains with DP up to ∼120, and has much smaller molecular size (and is present at a much lower amount) than endosperm starch. It is postulated that these pronounced differences arise from the distinct starch synthesis pathways in these organs, and are consistent with the starches' distinct botanical functions: short-term storage requiring relatively rapid degradation for leaf starch, and high crystallinity and high energy density requiring slow degradation for endosperm starch.

Oxidation of primary hydroxyl groups in chitooligomer by a laccase-TEMPO system and physico-chemical characterisation of oxidation products.

The aim of this study was to investigate the oxidation of chitooligomer by a laccase-TEMPO system which had not previously been examined. Chitooligomer was treated with laccase and TEMPO in order to evaluate the potential of a laccase-TEMPO system to improve the moisture absorption, moisture retention, and antioxidant abilities of chitooligomer. Chitooligomer was prepared by degradation of high molecular weight chitosan with hydrogen peroxide followed by oxidation using a laccase-TEMPO system. (13)C NMR and carboxylate ion content detection results indicated that the laccase-TEMPO system could selectively oxidise the C6 hydroxyl group of the chitooligomer into carboxyl group; molecular weight distribution changes suggest that the structure of the oxidised product had changed and the molecular size and molecular weight decreased with the molecules in aqueous solution having a compact structure. Oxidation of chitooligomer by a laccase-TEMPO system resulted in a significant improvement in the moisture absorption, moisture retention and antioxidant abilities. The oxidised product has potential application values in the pharmaceutical and cosmetics industries.

Photo-assisted proton exchange and chemical etching on Fe-doped lithium niobate crystals.

We report the photo-assisted proton exchange and chemical etching on Fe-doped LiNbO(3) crystals. Selective proton exchange and chemical etching are realized through the 455nm-laser irradiation on the crystal surface in pyrophosphoric acid. Optical microscopy and Micro-IR spectroscopy analysis show that the hydrogen incorporation is confined spatially by the laser irradiation. Moreover, under the laser irradiation, + z surface is found to be more easily etched than -z surface. This unexpected etching anisotropy is attributed to the photogalvanic effect of the crystal.

The 5' stem-loop and its role in mRNA stability in maize S cytoplasmic male sterility.

The co-transcribed orf355-orf77 region of the mitochondrial genome is associated with S cytoplasmic male sterility (CMS-S) in maize; the amounts of its 1.6- and 2.8-kb transcripts were previously shown to be greatly reduced in fertility-restored microspores relative to the amounts in sterile plants. To investigate the mechanism underlying this reduction, detailed analysis of the 5' and 3' termini of these transcripts was conducted. Using 3' RACE analysis, the polyadenylation sites of the 1.6- and 2.8-kb transcripts were mapped adjacent to a 3' stem-loop, which may play an important role in stabilizing their 3' ends. No difference was found between the polyadenylation sites in sterile and fertility-restored microspores that could account for the differences in orf355-orf77 transcript levels. The 5' terminus of the 1.6-kb transcript was further studied by primer extension; the result revealed that there was a deletion of nine nucleotides only in fertility-restored microspores, and that this deletion eliminated a 5' stem-loop sequence. We propose that the elimination of the 5' stem-loop in the fertility-restored microspores could be the cause of the degradation of the 1.6-kb transcript. Because the 2.8-kb transcript can be cleaved to generate the 1.6-kb transcript, the amount of the 2.8-kb transcript is also reduced in fertility-restored microspores.

Expression analysis of orf77 and R region in mitochondrial DNA of S-type CMS maize.

It is generally recognized that S type of CMS in maize associated with the recombination region R in mitochondria. Complicated DNA recombination and changes of transcripts of R are observed in several subgroups of S cytoplasm. R region includes two open reading frames (orf355 and orf77) and they are all chemeric. Among the sequence of orf77, there are three stretches similar to atp9 of mitochondria. Many different S cytoplasms are found in maize in China and researches on them show that similar R region and sequence are also found in the mitochondrial DNA in Tangxu, Shuang and J (cytoplasm. Northern analysis of Tangxu, J cytoplasm with Probe R and orf77 indicates that orf77 co-transcribes with R region and the nuclear restorer gene Rf3 affects their expression. In all tested S cytoplasm plants with the genotype of s-rf3rf3, both probe orf77 and R can detect six transcripts of 2.8, 1.6, 1.1, 0.9, 0.7 and 0.4 kb. In the presence of the nuclear restorer-of-fertility gene Rf3, previous transcripts of 2.8 kb and 1.6 kb are disappeared, but the other four transcripts are not changed. In T and C cytoplasm of CMS, only four transcripts of 1.1, 0.9, 0.7 and 0.4 kb appear when hybrid to probe R. Further Northern analysis of probe atp9 proved that all the four transcripts of 1.1, 0.9, 0.7 and 0.4 kb were actually transcripts of gene atp9, so only the transcripts of 2.8 kb and 1.6 kb are particular to R region of S. In addition, atp9, atp6 and cox II each has the same transcription pattern in tassel with different genotypes respectively. It can be included that R region and orf77 are the most important candidate gene for S-CMS.

[Construction of physical map and polymorphism analysis of mtDNA region R from Mo17CMS-J of maize].

Total DNA from twenty-six CMS lines of maize under Mo17, 77 and W23 nuclear background were used for PCR amplification, including N, T, C, S four groups of cytoplasms. The primers was prefabricated according the sequence of R region published by Zebala (1997). Through these amplifications, mitochondrial DNA fragments were obtained from maize total DNA. Generally the results in one group are identical. And they are different from the others. The amplified fragments were sequenced and also give us much more information about the structure of mitochondrial genes that may lead to CMS. In order to isolate and identify the CMS genes, we developed a new platform to construct physical map of chromosome DNA by means of restriction enzyme double-digestion. The elongation of contigs is based on Southern hybridization. Having retrieved DNA from agarose gel after electrophoresis by beta-agarase, we labelled it with 32P-dCTP as a probe. We detected the positive clones in the gene library. Two contigs were revealed. And a restriction map covering 40 kb was constructed, including R region.

[Progress of molecular biology of CMS in maize].

In the paper,we have summarized the molecular biological accomplishment acquired and accepted by most of maize researchers on CMS of maize. A brief review of current molecular biological progress of CMS of maize are displayed in the paper. These progresses concern in the positioning,cloning and maker-assisted selection of nucleic genes associated with fertility,expression and cloning of cytoplasmic genes associated with male sterility. In order to elucidate the molecular mechanism of CMS of maize, the areas about cloning and expression profiling of male sterile nucleic genes,and functional genomics of mitochondria,and interaction cytoplasmic genes with nucleic genes will need to be researched in the future.