Fine mapping of an Arabidopsis thaliana male sterile mutant EC2-157

An Arabidopsis thaliana male sterile mutant EC2-157 has been isolated using an EMS mutagenesis strategy. Genetic analysis indicated that it was controlled by a single recessive gene called ms157. No pollen grains have been observed in mutant anthers. ms157 Has been mapped to a region of 74kb located in BAC clone T6K22 on chromosome IV using a map-based cloning strategy. As no male sterile genes have been reported in this region, ms157 could be a novel gene related to fertility. The further molecular cloning and functional analysis on this gene should facilitate our understanding of A. thaliana another development. __________ Translated from Journal of Shanghai Normal University (Natural Sciences), 2005, 34(1): 58–63 [译自: 上海师范大学学报 (自然科学版), 2005, 34(1): 58–63]     

Karyotyping of Brassica oleracea L. based on rDNA and Cot -1 DNA fluorescence in situ hybridization

To explore an effective and reliable karyotyping method in Brassica crop plants, Cot-1 DNA was isolated from Brassica oleracea genome, labeled as probe with Biotin-Nick Translation Mix kit, in situ hybridized to mitotic spreads, and where specific fluorescent bands showed on each chromosome pair. 25S and 5S rDNA were labeled as probes with DIG-Nick Translation Mix kit and Biotin-Nick Translation Mix kit, respectively, in situ hybridized to mitotic preparations, where 25S rDNA could be detected on two chromosome pairs and 5S rDNA on only one. Cot-1 DNA contains rDNA and chromosome sites identity between Cot-1 DNA and 25S rDNA was determined by dual-colour fluorescence in situ hybridization. All these showed that the karyotyping technique based on a combination of rDNA and Cot-1 DNA chromosome landmarks is superior to all but one. A more exact karyotype of B. oleracea has been analyzed based on a combination of rDNA sites, Cot-1 DNA fluorescent bands, chromosome lengths and arm ratios. __________ Translated from Journal of Wuhan University (Nat. Sci. Ed.), 2006, 52(2): 230–234 [译自: 武汉大学学报 (理学版)]     

蝴蝶兰花发育的分子生物学研究进展

蝴蝶兰花非常独特且高度进化,如萼片瓣化、瓣片特化为唇瓣、雌雄蕊合生成合蕊柱及子房发育须由授粉启动等,是单子叶植物花发育研究的理想材料。近年来蝴蝶兰花发育分子生物学取得了重要进展。该文就近年来国内外有关蝴蝶兰开花转换及花器官发育相关基因研究以及B类基因与兰花花被的进化发育关系方面的研究进展进行综述。研究表明:MADS基因在蝴蝶兰开花转换及花器官发育过程中起重要作用,推测其中的DEF(DE-FICIENS)-like基因早期经过2轮复制,形成了4类不同的DEF-like基因,进而决定兰花花被属性。蝴蝶兰花发育分子生物学的深入研究,将极大地利于通过基因工程手段提高蝴蝶兰花品质如花色改良及花期调控等,推动分子育种进程。     

A comparison of in vitro anticancerous activity and mechanism of ethanolic extracts from different Ganoderma genus

Five ethanolic extracts from the mycelia of Ganoderma lucidum, G. tsugae, G. oerstedii, G. subamboinense, and G. resinaceum were respectively studied on their anticancerous activities against leukemic HL-60 cell line in vitro. Results showed that all five extracts potently inhibited HL-60 proliferation. The extract from G. lucidum mycelia exerted the highest activity. Annexin V/PI bivariate flow cytometric analysis further revealed that the five extracts significantly induced early apoptosis in HL-60 cells. The results illustrate that not only G. lucidum but also other Ganoderma species can inhibit cancer cells, and their mechanisms are related to induction of apoptosis. __________ Translated from Journal of Shanghai Normal University (Natural Sciences), 2005, 34(2): 77–81 [译自: 上海师范大学学报 (自然科学版), 2005, 34(2): 77–81]     

Analysis of sparteines in the seeds of Ammopiptanthus mongolica

Seven alkaloids were isolated from the seeds of Ammopiptanthus mongolica by thin layer chromatography and silica gel column chromatography, and the chemical structures of five alkaloids, 17-oxosparteine, β-isosparteine, 3α-hydroxysparteine, sparteine, and 3β-hydroxysparteine were identified by ¹H nuclear magnetic resonance (NMR) and electron ionization mass spectrum (EIMS). __________ Translated from Journal of Lanzhou University (Natural Sciences), 2007, 43(2): 43–46 [译自: 兰州大学学报(自然科学版)]     

Correlation between the distribution pattern of virulence genes and virulence of Aeromonas hydrophila strains

Nine strains of Aeromonas hydrophila isolated from diseased fish or soft-shelled tortoise were tested for the presence of three virulence genes including the genes encoding aerolysin, hemolysin, and extracellular serine protease (i.e., aerA, hlyA, and ahpA, respectively). These genes were investigated using polymerase chain reaction (PCR) with specific primers for each gene. And the pathogenicities to Carrassius auratus ibebio of these strains were also assayed. PCR results demonstrated that the distribution patterns of aerA, hlyA, and ahpA were different in these strains. 6/9 of A. hydrophila strains were aerA positive, 8/9 of strains hlyA positive, 7/9 of strains ahpA positive, respectively. However, the assay for pathogenesis showed that two strains (A. hydrophila XS91-4-1 and C2) were strong virulent, two strains (A. hydrophila ST78-3-3 and 58-20-9) avirulent and the rest middle virulent was to the fish. In conclusion, there are significant correlation between the distribution pattern of the three virulence genes and the pathogenicity to Carrassius auratus ibebio. All strong virulent A. hydrophila strains were aerA ⁺ hlyA ⁺ ahpA ⁺ genotype, and all aerA ⁺ hlyA ⁺ ahpA ⁺ strains were virulent. Strains with the genotype of aerA ⁻ hlyA ⁻ ahpA ⁺ have middle pathogenicity. In the present study, we found for the first time that all A. hydrophila isolated from the ahpA positive were virulent to Carrassius auratus ibebio. Additionally, there was a positive correlation between the virulence of A. hydrophila and the presence of aerA and ahpA. __________ Translated from Acta Scientiarum Naturalium Universitatis Sunyatseni, 2006, 45(1): 82–85 [译自: 中山大学学报 (自然科学版)]     

Application of petG-trnP sequence to systematic study of Chinese Cupressus species

Chinese Cupressus L. includes five species. The molecular phylogenetic relationship of the Cupressus species and Chamaecyparis L. were determined by comparing 417–479 bp of chloroplast petG-trnP intergenic spacer sequence. In PAUP^* analysis, Platycladus orientalis was used as the functional out group. By using the maximum likelihood method 1 077 trees were examined and the result showed that one tree had a best score of -Ln=2 232.47. The phylogenetic tree clearly showed that Chamaecyparis nootkatensis was diverged from other Chamaecyparis species. Based on the results, together with evidences from other aspects, we consider that Cupressus funebris and Chamaecyparis nootkatensis should be placed in the genus Cupressus. The use of cpDNA intergenic spacer petG-trnP in Cupressus was also discussed. __________ Translated from Journal of Sichuan University (Natural Science Edition), 2005, 42(5): 1033–1037 [译自: 四川大学学报 (自然科学版) 2005, 42(5): 1033–1037]     

Cloning and expressing a recombinant human tissue inhibitor of metalloproteinase-2 (TIMP-2) in methylotrophic yeast Pichia pastoris and its characterizations

To obtain human tissue inhibitor of metalloproteinase-2 (TIMP-2) cDNA and the secretory expression of TIMP-2 gene in Pichia pastoris, we designed and synthesized a 618 base pairs artificial gene coding for the TIMP-2 with a computer-aided design method using a standard chemical synthesis technique, which was composed of frequently used codons in the highly expressed Pichia pastoris genes. Then the synthetic gene encoding TIMP-2 was checked by means of dideoxynucleotide sequencing. The verified gene of TIMP-2 was cloned to the Escherichia coli-yeast shuttle vector of pPIC9 to construct a recombinant plasmid pPIC9-T2. The plasmid was transformed into GS115 cells of the methylotrophic yeast, Pichia pastoris by electroporation, and we got the expression cell through phenotype selection and induction with methanol. Separation, purification, and bioactivity analysis of the expressed products were performed. __________ Translated from Microbiology, 2006, 33(1): 1–6 [译自: 微生物学通报]     

Chemical compounds and pharmacological effects of Rabdosia excisa

Many kinds of diterpenoids have been isolated from Rabdosia spp. Some of them have anti-microbial effects, counteract inflammation, and inhibit tumor progression activities. We conducted the present study in order to look for bioactive compounds in the medicinal plant Rabdosia excisa. In this study, five compounds were isolated from R. excisa; they were oridonin, isokamebakaurin, oleanolic acid, ursolic acid, and β-sitosterol. In order to identify the function of the extracts, the activity of antibiotics, antioxidation, and immunity test were carried out against these functions. Prospective results were observed in all of the tested items. Translated from Journal of Northeast Normal University (Natural Sciences), 2005, 37(4): 94–98 [译自: 东北师大学报 (自然科学版)]     

Cloning and expressing DBT (dibenzothiophene) monooxygenase gene (dszC) from Rhodococcus sp. DS-3 in Escherichia coli

Dibenzothiophene (DBT) monooxygenase (DszC) catalysis, the first and also the key step in the microbial DBT desulfurization, is the conversion of DBT to DBT sulfone (DBTO₂). In this study, dszC of a DBT-desulfurizing bacterium Rhodococcus sp. DS-3 was cloned by PCR. The sequence cloned was 99% homologous to Rhodococcus erythropolis IGTS8 that was reported in the Genebank. The gene dszC could be overexpressed effectively after being inserted into plasmid pET28a and transformed into E. coli BL21 strain. The expression amount of DszC was about 20% of total supernatant at low temperature. The soluble DszC in the supernatant was purified by Ni²⁺ chelating His-Tag resin column and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to electronics purity. Only one band was detected by Western-blotting, which is for the antibody released in mouse against purified DszC in the expression product of BL21 (DE3, paC5) and Rhodococcus sp. DS-3. The activity of purified DszC was 0.36 U. DszC can utilize the organic compound such as DBT and methyl-DBT, but not DBT derivates such as DBF, which has no sulfur or inorganic sulfur. __________ Translated from Acta Scientiarum Naturalium Universitatis Nankaiensis, 2005, 38(6): 1–6 [译自: 南开大学学报 (自然科学版), 2005, 38(6): 1–6]     

Cloning and expression of the gene encoding (R)-specific carbonyl reductase from Candida parapsilosis CCTCC M203011

The gene which encodes (R)-specific carbonyl reductase (rCR) from Candida parapsilosis CCTCC M203011 was cloned, sequenced and compared with genes from the GenBank. The results indicated that rCR gene was 1011 bp, encoding a protein of 336 amino acids with a molecular weight of 35.9 kDa, and its nucleotide sequence showed 99% similarity to those of other members of the alcohol dehydrogenase superfamily. The rCR gene could express in recombinant strain Escherichia coli JM109, and the expression plasmid could produce (R)-1-pheny-1,2-ethanediol (100% e.e., 80.14% yield) from β-hydroxyacetophenone without any additive to regenerate NAD⁺ from NADH. __________ Translated from Microbiology, 2006, 33(4): 112–118 [译自: 微生物学通报]     

Biological characteristics of wild Cycas fairylakea population in Guangdong Province, China

There are five wild populations of Cycas fairylakea in Guangdong Province, China, three of which are newly found. A study of the biological characteristics of C. fairylakea populations showed that this species had a narrow colonization area within 300 hm², and an island pattern of distribution. Because of the overexploitation, urbanization, environment pollution, plant diseases, and insect pests, the wild populations and individuals of C. fairylakea decreased markedly in the past decades. All five populations have an opposite pyramid age structure, few coning plants, few seed production, and low level of seed germination rate or sterility. In conclusion, C. fairylakea in Guangdong Province was threatened seriously and an urgent need to take effective efforts to protect the plants and habitats in its location sites was required. __________ Translated from Acta Scientiarum Naturalium Universitatis Sunyatseni, 2005, 44(6): 97–100 [译自: 中山大学学报 (自然科学版), 2006, 44(6): 97–100]     

Breeding and application of Jiafuzhan, a new elite early indica rice cultivar in China

After 20 years of dedicated research, Jiafuzhan has been successfully developed under the new technologies in breeding high-quality early indica rice cultivars. Its rice quality has almost reached the A-level Editable Rice of Agriculture Department of China, and its average production reaches 400–500 kg/(666.7 m²). This new cultivar also has other characteristics such as enhanced resistance of blast and fallen, steady productivity, and strong adaptability. Jiafuzhan has been put into production of over 11.4 × 10⁴ hm² in Fujian Province and has been introduced and extended in other Provinces like Jiangxi, Guangdong, and Guangxi, China. The successes of breeding Jiafuzhan is a solution to the existing perennial problems in the rice industry, such as poor grain quality of big-grain rice and early indica rice, low productivity, and poor blast resistance of elite rice. __________ Translated from Journal of Xiamen University (Natural Science), 2006, 45(1): 114–119 [译自: 厦门大学学报 (自然科学版)]     

Cloning and characterization of PhPI9 involved in floral development from Phalaenopsis Orchid

In the attempt to discover new genes involved in the floral development in monoeotyledonousin species,we have cloned and characterized the homologous PISTALLATA-like (PI-like) gone from Phalaenopsis hybrid cultivar named PhPI9 (Phalaenopsis PI STILLATA # 9).The eDNA of PhPI9 has a fragment of 834 bp and has 60% identity with the PISTILATA from Arabidopsis.The deduced amino acid sequence of PhPI9 had the typical PI-motif.It also formed a subelade with other monoeot PI-type genes in phylogenetie analysis.Southern analysis showed that PhPI9 was present in the Phalaenopsis orchid genome as a single copy.Furthermore,it was expressed only in the lip of the Phalaenopsis flower and no expression was detected in vegetative organs.Thus,as a B-function MADS-box gone,PhP19 specifies floral organ identity in orchids.     

Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid

Plant genomes appear to exploit the process of gene duplication as a primary means of acquiring biochemical and developmental flexibility. The best example is the gene encoding chalcone synthase (CHS, EC2.3.1.74), the first committed step in flavonoid biosynthesis. In this study, we examined the molecular evolution of three CHS family members of Phalaenopsis including a novel chs gene (phchs5), which is slowly evolved. The inferred phylogeny of the chs genes of Phalaenopsis with other two orchid plants, Bromoheadia finlaysoniana and Dendrobium hybrid, suggested that gene duplication and divergence have occurred before divergence of these three genera. Relatively quantitative RT-PCR analysis identified expression patterns of these three chs genes in different floral tissues at different developmental stages. Phchs5 was the most abundantly expressed chs gene in floral organs and it was specifically transcribed in petal and lip at the stages when anthocyanin accumulated (stage1–4). Phchs3 and phchs4 were expressed at much lower levels than phchs5. Phchs3 was expressed in pigmented tissue (including lip, petal and sepal) at middle stages (stages 2–4) and in colorless reproductive tissue at late stage (stage 5). Phchs4 was only expressed in petal at earlier stages (stage 1–3) and in lip at middle stage (stage 4). These results present new data on differentiation of gene expression among duplicate copies of chs genes in Phalaenopsis.     

Cloning, expression and characterization of L-cysteine desulfhydrase gene from Pseudomonas sp. TS1138

L-cysteine desulfhydrase (CD) plays an important role in L-cysteine decomposition. To identify the CD gene in Pseudomonas sp. TS1138 and investigate its effect on the L-cysteine biosynthetic pathway, the CD gene was cloned from Pseudomonas sp. TS1138 by polymerase chain reaction (PCR) method. The nucleotide sequence of CD gene was determined to be 1,215 bp, and its homology with other sequences encoding CD was analyzed. Then the CD gene was subcloned into pET-21a(+) vector and expressed in Escherichia coli (E. coli) by isopropyl-β-D-thiogalactopyranoside (IPTG) inducement. The recombinant CD was purified by Ni-NTA His-Bind resin, and its activity was identified by the CD activity staining. The enzymatic properties of the recombinant CD were characterized and its critical role involved in the L-cysteine biosynthetic pathway was also discussed. __________ Translated from Microbiology, 2006, 33(4): 21–26 [译自: 微生物学通报]     

Proteomic approach for acute-phase proteins of hemolymph and muscles in Scylla serrata challenged by a pathogenic bacterium

Acute-phase response is documented to be a significant mechanism of innate immunity in vertebrates and invertebrates. In this study, proteomic methodologies were applied for different protein expressions in hemolymph of Scylla serrata challenged by Vibrio parahaemolyticus after immunization, and in muscles of the crabs separately challenged by V. parahaemolyticus, V. anguillarum and Aeromonas hydrophila. Up-regulated cryptocyanin is documented in the hemolymph and up-regulated calexcitin, wingless (fragment) and tachykinin-related peptide in the muscle as acute-phase proteins. All the four altered proteins were responsible for bacterial stress, but cryptocyanin seemed to be a memory response protein against the challenge by a live bacterium after immunization of the live cells. These up-regulated proteins can be indicative of an understanding of immunity of a crab. __________ Translated from Journal of Xiamen University (Natural Science), 2005, 44(4): 559–562, 44(Sup.): 191–194 [译自: 厦门大学学报(自然科学版), 2005, 44(4): 559–562, 44(增刊): 191–194]