Fertile organs andin situ spores of a matoniaceous fern from the Lower Jurassic of West Hubei

Fossil matoniaceous plants represented byPhlebopteris have been formerly described and illustrated from the Mesozoic of China mainly based on vegetative leaves, and no information is provided on the fine structure of fertile organs. Recent reinvestigation of the Hsiangchi Flora from the Lower Jurassic of West Hubei, China, obtained a rich collection of well preservedPhlebopteris specimens (both sterile and fertile). Studies onPhlebopteris polypodioides Brongniart using LM and SEM reveal not only the structures of sori and sporangia, but also the details ofin situ spores. Comparisons have been made between thein situ spores and other related fossil and extant matoniaceous spores, as well as the dispersed genera.     

An osmundaceous rhizome with sterile and fertile fronds and <Emphasis Type="Italic">in situ</Emphasis> spores from the Jurassic of western Liaoning

A well-preserved permineralized osmundaceous fern was discovered from the Middle Jurassic Tiaojishan Formation of Beipiao, Liaoning Province, China. The rhizome is about 50 cm high and 35–41 cm in diameter and can be attributed to the genus Ashicaulis on the basis of its anatomy. The sterile pinnae are referable to Cladophlebis, the fertile pinnules are of the Todites type with in situ spores of the Osmundacidites type. Osmundaceous rhizomes have been found all over the world including the Jurassic strata of western Liaoning, China. Until now, however, no rhizomes have been reported to be organically connected with leafy organs. Although impressions and compressions of isolated osmundaceous sterile and fertile fronds are common, they were described under separate organ- or morpho-generic names, such as Todites and Cladophlebis. The new discovery provides an opportunity to study an osmundaceous fern in ‘whole plant concept’ to learn more about its morphology and structure, and the paleoclimatic and paleoenvironmental requrirements of this group.     

Discovery of a petrified noeggerathialean strobilus,Discinites sinensis sp. nov. from the Permian of Shizuishan, Ningxia, China

A petrified strobilusDiscinites sinensis sp. nov. is discovered from the Lower Permian of Shizuishan in Ningxia, North China. The epidermal cells of sporophyll are long and narrow, with both ends pointed. Each sporangium is ellipsoid, with the basal part contracted and short, thick sporangiophore-like. The cells of tapetum and sporangial walls are arranged longitudinally. The annulus is not developed. Mega- and microsporangia are similar in shape. They can hardly be distinguished unless they are broken, and the spores are exposed.In situ mega- and microspores are also similar in shape and both of theDeltoidospora type. The new species supports the systematic placement ofDiscinites within Noeggerathiales, and the progymnosperm affinity of the Noeggerathiales.     

对马耳蕨孢子囊早期发育的显微结构研究

利用光镜对对马耳蕨（Polystichum tsus-simense）孢子囊的早期发育进行研究．结论如下：对马耳蕨孢子囊起源于羽状叶的一个表皮细胞,经过横分裂和斜向分裂形成外套层原始细胞和内部细胞,外套层原始细胞分裂分化成孢子囊壁,内部细胞发育形成内外绒毡层和孢子母细胞,孢子母细胞减数分裂形成孢子四分体．在发育过程中,孢子囊腔内先后出现了孢囊被和孢子被．对马耳蕨孢子囊的发育类型属于薄囊蕨型．揭示了对马耳蕨孢子囊的早期发育规律,为蕨类植物的发育生物学和系统进化提供科学依据．     

Atomic force microscopic observation on substructure of pollen exine inCedrus deodara andMetasequoia glyptostroboides

The substructure of pollen exine inCedrus deodara (Roxb.) Loud. andMetasequoia glyptostroboides Hu et Cheng has been examined with an atomic force microscope (AFM). The results indicate that the exine substructure units containing sporopollenin in two species are similar in shape, which are granular, but slightly different in size. InCedrus the substructure unit of pollen exine appears to be 56–99 nm long and 42–74 nm wide, while inMetasequoia it appears to be 81–118 nm long and 43–98 nm wide. It has been observed that the subunits of pollen exine inCedrus arranged tightly to form short-rod-like or spheroidal pollen exine units, several or more than ten of which formed an island-like structure. There are various spaces among these island-like structures which are interconnected to occupy the entire pollen exine. InMetasequoia, the subunits of pollen exine also arranged tightly with a distribution tendency of cluster of 3–10, however, no obvious boundary exists among these clusters. From our results, it is concluded that there is no tendency of helical arrangement for the subunits of pollen exine inCedrus andMetasequoia, and the results support Southworth’ view that subunits of pollen exine are granular shape in lattice structure.     

Environmental changes reflected by n-alkanes of lake core in Nam Co on the Tibetan Plateau since 8.4 kaB.P.

The n-alkanes are extracted from NMLC-1 core that was drilled in the Nam Co, central Tibet. They are measured by using Gas Chromatography and Mass Spectrometry （GC/MS） for componential and quantitative analyses. According to the constructed depth-age model, the component and concentration of n-alkanes, together with total organic carbon （TOC）, total nitrogen （TN） and carbonate are used to elucidate palaeoenvironmental changes of Nam Co during the past 8.4 ka. The results indicate that Holocene environment performs three stages in the lake area. In the stage of 8.4-6.7 kaB.P., it was warmer while precipitation slightly increased. This stage was ended by an obvious cold/dry event. During 6.7-5.8 kaB.P., temperature increased rapidly and reached its maximum values at about 6.0 kaB.P. The environments were warm/wet optimum for the blooming of terrestrial plants and submerged aquatic plants. After that, temperature decreased continuously and showed the lowest values at about 3.0 kaB.P. From 2.9 kaB.P, to the present, temperature rose again but alternated with cold and warm. The lake area tended to be dry after 1.4 kaB.P. During 600-400 aB.P., the environmental feature was the reflection of ＂Little Ice Age＂.     

High molecular weight (C35 +) n-alkanes of Neogene heavily biodegraded oil in the Qianmiqiao region, North China

With wax content of 1.62%, heavy oil has been produced from the sandstone reservoirs of Neogene Guantao Formation (Ng1^Ⅲ). In the GC and GC-MS RIC profiles of its aliphatic fraction, n-alkanes are totally lost, which shows the result of heavy biodegradedation. However, the remaining trace C₁₃－C₃₆ n-alkanes can be still seen from its m/z 85 mass chromatogram. In addition, a complete series of C₃₅－C₇₃ high molecular weight (HMW) n-alkanes was detected by high-temperature gas chromatography (HTGC). The HMW n-alkane series shows a normal distribution pattern, a major peak at nC₄₃, obvious odd-carbon-number predominance, CP_I37—55 and OEP_45—49 values up to 1.17 and 1.16—1.20 respectively. The present study not only has conformed the strong resistibility of HMW n-alkanes to biodegradation in crude oils as concluded by previous researchers, but also has provided some significant information on source input and maturity for the heavily biodegraded oil in the Qianmiqiao region.     

On the origin of printing in the light of new archaeological discoveries

30 years have pas. since a printeddhārani sūtra scroll was discovered in 1966 in Kyongju. Korea. However. there have been different views about the date and place of its printing and publication in Korea and abroad. Some Korean scholars think that it was translated into Chinese from Sanskrit in 704 in China and printed during 706–751 in the Silla period in Korea. After a further research it is now proved that this sutra was translated into Chinese in 701 and printed in the following year, 702, in Luoyang, during the reign of Empress Wu. This shows that the sutra could not have been printed in the Silla period. The discoveries of theSnddharma pundarik sūtra printed during 690–699 and the Sanskritdharani charm printed around 650–670 in China also show that thedhārani sutra found in Korea is not the earliest one. Both literary records and archaeological findings prove that printing originated in China.     

NMR and IR investigations of ternary inclusion complexes among β-cyclodextrin, rare earth metal ions, and naphthalenediamines

Ternary inclusion complexes β-cyclodextrin (β-CD), rare earth metal ions (YbCl₃, YCl₃), and 1,8-naphthalene- diamine/1,5-naphthalenediamine are synthesized in basic aqueous media, which are characterized via ¹H NMR and IR spectroscopy. The stoichiometric proportion of β-CD:YbCl₃:1,5-naphthalenediamine is 2:1:2, that of β-CD:YCl₃:1,8-naphthalenediamine is 2:1:1, and that of β-CD:YbCl₃:1,8-naphthalenediamine is 2:1:1. The IR spectroscopy of the ternary inclusion complexes in the range of 935–1 000 cm⁻¹ reveals the existence of the coordinate bond M—O or M—N. The possible conformations of the ternary inclusion complexes are depicted. Biography: JIANG Huiming(1972–), male, Associate professor, research direction: molecular complex chemistry, supramolecular chemistry.     

A novel method for thermodynamic study on binding of copper ion with Alzheimer’s amyliod β peptide

The interaction of Cu²⁺ with the first 16 residues of the Alzheimer's amyliod β peptide, Aβ (1-16), was studied by employing isothermal titration calorimetry at pH 7.2 and 37℃ in aqueous solution. The Gholamreza Rezaei Behbehani (GRB) solvation model was used to reproduce the enthalpies of Cu²⁺+ Aβ(1-16) interaction over the whole Cu²⁺ concentrations. The binding parameters recovered from the solvation model were attributed to the structural change of Aβ (1-16) due to the metal ion interaction. It was found that there is a set of two identical and non interacting binding sites for Cu²⁺ ions. The molar enthalpy of binding is ΔH=27.895 kJ/mol. The association binding constants are 1.895 μm^-1 and 1.891 μm^-1 for the first and second binding sites respectively.     

Evolutionary mode of metallothioneins inferred from Cd-MT genes of Tetrahymena

FromTetrahymena thermophila (strain BF5), the coding region ofCd-MT gene was cloned and sequenced, and identified as MTTI isoform. A serial duplicate structure is discovered in its amino acid sequence, which separates the coding region into three parts (Part 1: 7–61: Part 2: 64–118; Part 3: 122–162). The alignments among them and comparison with the corresponding parts of MT1 isoform suggest that MT1 and MTT1 isoforms both come from the same ancient gene that is homologous to Part 1, and Cd-MTs ofTetrahymena are aroused by such ancient gene duplication. The prediction of secondary structures and the analysis of the disulfide-bonding state of cysteine show that there are a lot of differences between MT1 and MTT1 isoforms, which maybe relate to their function mechanism. Foundation item: Supported by KSCX of Chinese Academy of Sciences Grant (SW-102) and the Allocation from the Earmaarked Grant for Hong Kong. Biography: WAN Mingliang (1980-). male, Ph. D. candidate, research direction: molecular ecotoxicology of protozoan.     

Regenerating plants from cryopreserved adventitious buds of haploids in rice

A large number of adventitious buds were induced fromin vitro cultured young inflorescences of haploids of rice. Having been subcultured on solidified subculture media at 26°C for 7 days, the adventitious buds were loaded into 1.8 mL plastic cryotubes with cryoprotectant and kept on ice for 45–60 min. After cooled at a rate of 1.0°C/min down to −40°C, the samples, were kept in liquid nitrogen. The adventitious buds which have been cryopreserved for about 30 days were thawed rapidly in 38–40°C water and then plated on solidified MS medium containing 3% sucrose, 0.5 mg/L NAA and 2.0 mg/L kinetin. After plated, 23%–32% of adventitious buds resumed growth and 15%–22% regenerated plantlets. The results of this work indicated that the adventitious buds derived fromin vitro cultured young inflorescences is a critical factor for the success and subculturing adventitious buds on MS medium containing 3% sucrose and 4% sorbitol or 20% potato extract is essential to the procedure. The effective cryoprotectant is 10% DMSO (dimethyl sulfoximide)+0.5 mol/L sorbitol. Supported by the Natural Science Foundation of Hubei Province Zhang Zhihong: born in 1963, Lecturer     

Molecular cloning of a fulllength cDNA for ECBP21 from Angelica dahurica

ECBP21 is an extracellular calmodulin-binding protein which was first detected and purified from extracellular extracts of suspension-cultured cells of Angelica dahurica. The purified protein was electroblotted onto PVDF membrane and the amino acid sequences from 1 to 20 were determined. Using degenerate oligonucleotides of the sequence, a full-length cDNA coding for ECBP21 was isolated by a combination of RT-PCR and 5′-RACE cloning. The cDNA contains 947 nucleotides and codes for a precursor protein of 216 amino acids. The N-terminal 1–25 amino acid sequence is a predicted signal peptide and the other 26–216 amino acid sequence is a mature peptide. The 26–45 amino acid sequence shows identity with the N-terminal amino acid sequence of purified ECBP21 from Angelica dahurica. The fragment of encoding the mature protein was cloned into pET-28b(+) and transformed into E. coli BL21(DE3). A protein with relative molecular mass 21 ku was expressed in E. coli. Using a biotinylated-CaM gel overlay technique, the expression protein was tested for its ability to bind CaM. The results indicated that the expression protein is a Ca²⁺-dependent CaM-binding protein. Thus, these results further defined the cDNA clone for ECBP21. This work laid a foundation for elucidating biological functions of ECBP21 by using molecular biological means.     

Microcalorimetric study on the antibiotic activity of clarithromycin and erythromycin

By using LKB-2277 Bioactivity Monitoring System, the heat effect changes in the process of inhibitory action of clarithromycin and erythromycin onEscherichia coli at 37°C were determined. Quantitative analysis showed that relationship between antibiotic concentrationc and rate contantk ofEscherichia coli growth, and half inhibitory ratio concentration IC₅₀: clarithromycin:k=0. 030 03–1. 1736×10⁻³ c, 8. 45 mg ·L⁻¹; erythromycin:k=0.031 08–8.4657×10⁻⁴ c, 14. 45 mg·L⁻¹. As a result of the microcalorimetry experiments, it not only indicated that antibacterial activity of clarithromycin was stronger than that of erythromycin, but also reported the changeable features of thermodynamics of the bacterial cell in biological, biochemical and metabolic process under different drug action. Foundation item: Supported by Natinal Natural Science Fundation of China (29973030), Natural Science Fundation of Hubei Province (98J052) and Post-doctoral Science Fundation of China Biography, SHEN Xue-song (1956-), Associate professor Research direction: biothermochemistry.     

Transformation of plant young proembryos by electroporation

It is first reported that plant young proembryos expressed exogenous reporter genes by electroporation. Young proembryos with 8–32 cells and globular proembryos with 250–400 cells could be isolated by enzymatic maceration combined with microdissection. After electroporation withGUS orGFP genes, the proembryos were cultured for 1–2 d in KM8p medium. At the field strength of electroporation 500–1500 V/cm, blue reaction of GUS or green fluorescence of GFP could be observed in the proembryos. The highest transient expression frequency of young proembryos (2.2%) was obtained at the field strength of 750 V/cm, whereas the highest frequency of globular proembryos (5.9%) was obtained at the field strength of 1 250 V/cm. Taking the proportion of transformed cells in the whole cells of proembryos as efficient transformation frequency, the efficient transformation frequency of the young proembryos was 7 times that of the globular proembryos.     

The role of β18-β19 loop structure in insecticidal activity of Cry1Ac toxin from Bacillus thuringiensis

The β18-β19 loop in domain Ⅲ of Cry1Ac toxin is unique among Bacillus thuringlensis Cry proteins. In this study, the role of the loop structure in insecticidal activity of Cry1Ac toxin was investigated. Alanine scanning mutations within the loop were initially generated and most mutants were over-expressed and reduced toxicity at different degrees, except mutant N546A that showed almost 2 times enhanced toxicity against Helicoverpa armigera larvae. Further mutagenic analysis of N546 revealed that a charged amino acid in this position would cause very unfavorable influence on insecticidal activity. In addition, the deletion of N546 led to protein instability because of destruction of the loop integrity. Besides, mutant W544F was much more toxic than W544Y, indicating that hydrophobic nature of the position was important for maintaining the stability and activity of Cry1Ac protein. These findings are the first biological evidence for a structural function of β18-β19 loop in insecticidal activity of the Cry1Ac toxin.     

Evaluation and characterization of an endosperm-specific sbeⅡa promoter in wheat

Starch,the main component of the wheat grain,is the product of a complex biochemical pathway. The sbeⅡα gene plays a key role in controlling the synthesis of starch, in particular, the biosynthesis of amylopectin,in maturing wheat grain.To investigate its regulatory mechanisms and endosperm-specific expression pattern, the sbeⅡα promoter (3094 bp in length) was cloned using APCR and sequenced.The effect of a series of deletions was studied using a GUS transient assay system. Results showed that the 3094 bp sequence (sbe.g construct) exhibited full stable promoting activity and that the activities of 5′ or 3′ deletions reduced levels of GUS expression. Some constructs with internal deletions showed only weak activity, however,sbe.e, with a deletion from -1579—--1210 bp resulted in higher levels of expression than the full-length promoter sequence, sbe.g. This indicates that motifs such as the -300 bp element, G-box and/or P-box act as positive elements and are necessary in determining the promoter‘s endosperm-specific pattern and that negative repressor elements or motifs may also be present within the -1579—-1210 bp sequence. The age of wheate ndosperm tissue used in the GUS-transient assay system is shown to be of significant importance.     

Preparation of exine-detached pollen inNicotiana tabacum

A method of preparing exine-detached pollen inNicotiana tabacum was established. Anthers containing early-middle binucleate pollen were cold-pretreated at 4–6°C for 7–14 days, and were suspended in 0.3 mol/L sucrose solution for 2 days. During this process, the exine of most pollen grains dehisced. Then they were transferred into an enzyme solution containing 1% cellulase, 1% pectinase, 0.1% pectolyase, 1 mol/L mannitol, 0.3 mol/L sorbitol, 0.5% potassium dextran sulphate and K3 medium macro elements. After 15–20 min enzymatic maceration, the exine was detached resulting in the release of exine-detached pollen. Factors affecting preparation of exine-detached pollen were investigated, including cold-pretreatment, osmoticum concentration and enzymes used. Xia Huijun: born in Oct. 1963, Ph.D. Current research interest is in plant reproductive biology Supported by the National Natural Science Foundation of China