Effects of CO2, temperature and their interaction on the growth, development and yield of cauliflower (Brassica oleracea L. botrytis)

Stands of summer cauliflower were grown within polyethylene-covered tunnels along which a temperature gradient was imposed. Two tunnels were maintained at either normal or elevated CO₂ concentrations. At the last harvest (88 days from transplanting) no interaction between CO₂ and temperature on total biomass was detected. The total dry weight of plants grown at 531 μmol mol⁻¹ CO₂ was 34% greater than those grown at 328 μmol mol⁻¹ CO₂, whereas a 1 °C rise reduced dry weight by 6%. From serial harvests the radiation conversion coefficient was 2.01 g MJ⁻¹ and 1.42 g MJ⁻¹ at 531 μmol mol⁻¹ CO₂and 328 μmol mol⁻¹ CO₂, respectively, but was not greatly affected by differences in temperature. No effect of either CO₂ or temperature on the canopy light extinction coefficient was detected. The rate of progress towards curd initiation increased to a maximum at 15.5 °C, and declined thereafter. Provided the effect of temperature was accounted for, CO₂ enrichment did not affect the time of curd initiation. From serial harvests after curd initiation, the logarithm of curd weight or diameter were negative linear functions of mean temperature from initiation. Increases in curd weight and diameter at 531 compared with 328 μmol mol⁻¹ CO₂ were greater at warmer temperatures (27% at 13 °C compared with 47% at 15 °C, 57 days after initiation). Effects of CO₂ on curd diameter were less than those on curd dry weight because the curd dry matter content was greater at 531 compared with 328 μmol mol⁻¹ CO₂. Thus, the effects of elevated CO₂ concentrations on fresh weight based yield parameters of cauliflower were less than the increase in total dry matter production.     

Preparation of m1AChR-G11 fusion protein and m4AChR-G16 fusion protein and effects of various ligands on them

AIM:To prepare m₁AChR-G₁₁ and m₄AChR-G₁₆ fusion protein in Baculovirus-Sf9 cell system and detect the effects of various muscarinic ligands on the interaction between m₁AChR and G₁₁ and m₄AChR and G₁₆, and screen different kinds of ligands specific for m₁ and m₄. METHODS:To prepare fused DNA of m₁AChR-G₁₁and m₄AChR-G₁₆ in two PCR, then expressed in Sf9 cells and detect the pharmacological function of m₁AChR-G₁₁ fusion protein and m₄AChR-G₁₆ fusion protein by QNB and GTPγS binding experiments; To expore the way of the activation of m₁AChR-G₁₁ and m₄AChR-G₁₆ fusion protein by various ligands includingcetylcholine (ACh), Pilocarpine (Pilo), 4-hydroxy-2-butynyl-1-trimethylammonium-m-chloro-carbanilatechloride (McN-A-343), tetrandrine, pirenzepine (PZ), alcuronium, atropine, R-(+)-hyoscyamine and gallamine by displacement by GDP on GTPγS binding experiments. RESULTS:The expression levels of m₁AChR-G₁₁ and m₄AChR-G₁₆ fusion protein were (45.39±2.62) nmol·g^-1 protein, (47.04±1.58) nmol·g^-1 protein. The affinity of GDP to G₁₁ and G₁₆ partner changed in the presence of different muscarinic ligands. CONCLUSION: The m₁AChR-G₁₁ and m₄AChR-G16 showed the pharmacological specificity to m₁ and m₄ receptor and the efficient signaling of the two partners. Ligands of m₁AChR and m₄AchR mediated different signal transduction by changing the affinity of G₁₁/G₁₆ and GDP. So m₁AChR-G₁₁ fusion protein and m₄AChR-G₁₆ fusion protein can be taken as a tool to screen ligands specific for m₁AChR and m₄AChR.     

Storage conditions and ripening of chiku fruits Achras sapota L.

Chiku fruits (Achras sapota L.) were found to be climacteric with the respiratory peak occurring at the same time, or 1–2 days after peak ethylene production. The optimum storage temperature was near 20° C. Storage life at this temperature could be increased by removing ethylene (C₂H₄) and adding 5–10% (v/v) carbon dioxide (CO₂) to the storage atmosphere. Extreme relative humidities or high concentrations of CO₂ impaired the quality of stored fruits. Ripening was not affected by treatment with oxygen, C₂H₄ or indoleacetic acid. Ascorbic acid and glucose levels increased with ripening but ascorbic acid decreased when the fruit became over ripe. Recommended storage conditions are about 20° C with 5–10% CO₂ coupled with the complete removal of C₂H₄ from the storage atmosphere. Short-term holding of the fruit at lower temperatures is also possible.     

The genetic susceptibility of HLA-DRB1 alleles in esophageal neoplasm of Hubei Han Chinese

AIM: To probe into the genetic susceptibility of HLA-DRB1 alleles to esophageal neoplasm in Hubei Han Chinese. METHODS: HLA-DRB1 gene polymorphism in 42 patients with esophageal neoplasm and 136 normal control subjects was studied by PCR and sequence. RESULTS: Allele frequency of HLA-DRB1 *0901 allele was significantly higher in esophageal cancer patients than those in normal controls(0.2500 vs 0.1397, P =0.028; the odds ratiO².053; etiologic fraction 0.1282).There were no association between the rested HLA-DRB1 alleles with patients. CONCLUSION: Individuals carrying HLA-DRB1 *0901 may be susceptible to esophagealo carcinoma, its nucleotide sepuence approachs to the corresponded allele sequence(exoN²)published in GenBank.     

番茄细菌性斑点病菌、溃疡病菌、青枯病菌和疮痂病菌的四重PCR检测方法

针对引起番茄细菌性病害的细菌性斑点病菌（Pseudomonas syringae pv. tomato,Pst）、溃疡病菌（Clavibacter michiganensis subsp. michiganensis,Cmm）、青枯病菌（Ralstonia solanacearum,Rs）以及疮痂病菌（Xanthomonas campestris pv. vesicatoria,Xcv）,建立了四重PCR检测技术,为病害的快速、准确诊断提供技术支持。根据gap1基因设计Pst特异性引物BW-F/BW-R,经PCR条件优化,扩增出了375 bp的特异性片段。将设计的引物与已报道的3种细菌特异性引物组合,通过设定不同的退火温度、引物浓度、循环次数以及延伸时间,探索影响四重PCR扩增的因素,优化了其反应体系。四重PCR反应体系中的引物对BW-F/BW-R、Fan1/Fan2、RS-1-F/RS-3-R和XCVF/XCVR可分别扩增出细菌性斑点病菌、溃疡病菌、青枯病菌和疮痂病菌长度为375、146、716和517 bp的特异性目的片段,反应体系退火温度为57.1 ℃,4对引物的终浓度分别为0.24、0.16、0.16和0.08 μmol ? L-1,延伸时间45 s,35个循环。该四重PCR反应体系可快速检测田间番茄发病植株中的细菌性斑点病菌、溃疡病菌、青枯病菌和疮痂病菌,灵敏度达到10-1 ng ? μL-1。     

Measurement of soil compaction tolerance of Lagestromia speciosa (L.) Pers. using chlorophyll fluorescence

Trees planted along roadsides and on public recreation areas are subjected to environmental stresses such as poor soil, air pollution and heat. However, very little information is available on the trees’ tolerance to the various stress factors faced in an urban environment in Malaysia, such as soil compaction. The effects of soil compaction on a range of chlorophyll fluorescence parameters (F₀,F_m,F_v/F_m) in foliar tissue of Lagestromia speciosa, a widely planted Malaysian street tree, were examined. Results showed that soil compaction was between 170 and 315 MPa in the study areas. Soil compaction above 180 MPa affected tree form and reduced chlorophyll fluorescence. It is concluded that chlorophyll fluorescence offers a rapid screening technique for assessing soil compaction tolerance of L. speciosa.     

Effect of Akt/GSK-3β/Snail signaling pathway on EMT in A549/DDP cells mediated by TGF-β1

AIM: To observe the expression of Akt/GSK-3β/Snail signaling pathway-related molecules in cisplatin-resistant cell line A549/DDP mediated by transforming growth factor-β₁ (TGF-β₁), and to explore the association of Akt/GSK-3β/Snail signaling pathway with epithelial-mesenchymal transition (EMT). METHODS: The A549/DDP cells were divided into TGF-β₁ (+) group, TGF-β₁ (-) group and LY294002 group. The morphological changes of A549/DDP cells treated with TGF-β₁ were observed under microscope. The protein expression of E-cadherin and N-cadherin was determined by the methods of immumofluorescence and Western blot. The protein levels of Akt, p-Akt, GSK-3β, p-GSK-3β^Ser9 and Snail were also detected by Western blot. RESULTS: The A549/DDP cells in TGF-β₁ (+) group were dispersive, showed a spindle-like shape and developed pseudopodia. This transformation was conformed to classic EMT markers. Compared with TGF-β₁ (-) group, the protein expression of E-cadherin in TGF-β₁ (+) group was significantly decreased (P<0.05), and N-cadherin was significantly increased (P<0.05). The protein levels of p-Akt, p-GSK-3β^Ser9 and Snail were also significantly increased (P<0.05). Compared with TGF-β₁ (+) group, the protein levels of p-Akt, p-GSK-3β^Ser9 and Snail were significantly decreased in LY294002 group (P<0.05). No difference of Akt and GSK-3β expression between TGF-β₁ (-) group and TGF-β₁ (+) group was observed. CONCLUSION: The mechanism of EMT in A549/DDP cells might be related to Akt/GSK-3β/Snail signaling pathway activated by TGF-β₁.     

USP14 regulates H2O2 induced oxidative stress in H9c2 cells

AIM:To evaluate the effect of inhibiting ubiquitin-specific protease 14(USPl4) activity on oxidative stress induced by H₂O₂ of H9c2 cells.METHODS:The H9c2 cells were incubated with H₂O₂ at 25 μmol/L for 2 h to establish the oxidative stress injury model.The cells were divided into control group,H₂O₂ group,IU1 group (25 μmol/L or 50 μmol/L) and IU1+H₂O₂ group.The H9c2 cells activity was measured by MTS assay.The level of intracellular reactive oxygen species (ROS) and cell survival rate were analyzed by flow cytometry assay.The changes of the mitogen-activated protein kinase (MAPK) family related proteins were detected by Western blot.RESULTS:Compared with control group,the cell activity and the viability rate in H₂O₂ group were decreased (P<0.05),while the intracellular ROS,the protein levels of Bax/Bcl-2,P53,p-ERK1/2,p-JNK and p-P38 were increased (P<0.05).Compared with H₂O₂ group,the cell activity and the viability rate of the H9c2 cells in IU1+H₂O₂ group were increased (P<0.05),while the intracellular ROS,the protein levels of Bax/Bcl-2,P53,p-ERK1/2,p-JNK and p-P38 were decreased (P<0.05).CONCLUSION:Inhibition of USPl4 activity reduces the oxidative stress injury of the H9c2 cells.The mechanism may be related to inhibition of the MAPK signaling and down-regulation of apoptosis related proteins.     

Genetic diversity and relationships of lotus (Nelumbo) cultivars based on allozyme and ISSR markers

Genetic diversity and genetic relationships of lotus (Nelumbo Adanson) cultivars were evaluated using allozyme and ISSR markers. The samples used covered 11 accessions of possible hybrids between Nelumbo nucifera and Nelumbo lutea and 92 accessions of N. nucifera including 69 flower lotus, 13 rhizome lotus, 5 seed lotus and 5 wild lotus. For allozyme studies, a total of 31 alleles at 23 loci of 18 enzyme systems were detected of which 5 (21.7%) loci Aat, Idh, Mdh-2, Pgd, Sod were polymorphic. The loci of Aat and Idh included two alleles, Mdh-2, Pgd and Sod included three alleles. Eighteen genotypes were detected with the 13 alleles of the 5 polymorphic loci. The parameters of average allele number, observed heterozygosity, expected heterozygosity and Shannon information index of 92 N. nucifera samples were 1.35 ± 0.71, 0.06 ± 0.21, 0.05 ± 0.14, 0.10 ± 023, respectively. Thirteen ISSR primers generated 93 loci, of which 37.63% were polymorphic across all samples. The percentage of polymorphic loci, average allele number, expected heterozygosity and Shannon information index of 92 N. nucifera samples were 26.67%, 1.30 ± 0.46, 0.10 ± 0.18 and 0.15 ± 0.25, respectively for the ISSR data. The ‘Bottleneck effect’ and rapid propagation of clones after the ice ages may explain the low genetic diversity of lotus. The dendrograms based on ISSR and allozymes were not congruent. Based on the ISSR data, the 103 samples were divided into the N. nucifera group (Group I), and the group containing inter-specific hybrids between N. nucifera and N. lutea (Group II). The flower lotus, rhizome lotus, and seed lotus each has multiple sources of origin. Plant size, a criterion commonly used in the classification of cultivars of lotus, is not correlated with genetic variation. Flower color is correlated with the cultivar classification to some degree, but its variation is complex in the hybrids.     

In vitro propagation of Symphytum species

Methods are described for the in vitro propagation of Symphytum × uplandicum Nyman from bud, root and stem explants. Highest shoot numbers were produced from root explants > 4 mm in diameter, cultured vertically with their distal cut surface on the medium. The most suitable medium for shoot production was Murashige and Skoog's (MS) with 0.3 mg l⁻¹ 6-benzylaminopurine (BAP). These shoots developed roots on MS medium without hormones and were successfully transplanted into pots. Subculturing shoots onto MS medium containing BAP, kinetin (K), 6-λ,λ-(dimethylallylamino)-purine or gibberellic acid (GA₃) at 0.5, 1.0, 5.0, 10.0 or 30.0 mg l⁻¹ failed to stimulate outgrowth of axillary buds in culture. In vitro propagation from root explants was also achieved with S. asperum Lepech., S. officinale L. and S. × uplandicum cultivars ‘Bocking 1’, ‘Bocking 2’, ‘Bocking 4’ and ‘Bocking 17’, but not with S. bohemicum, S. grandiflorum DC, S. tuberosum L. or S. × uplandicum ‘Bocking 7’ and ‘Variegatum’.     

Effect of senegenin on H2O2-induced damage in hippocampal neurons of SD rats

AIM: To observe the effect of senegenin (Sen) on hippocampal neuron injuries induced by H₂O₂.METHODS: Hippocampal neurons were isolated from neonatal SD rats. The primarily cultured neurons were divided into control group, H₂O₂ group, Sen group and Sen+H₂O₂ group. The cell viability, the content of malondialdehyde(MDA) and the activity of superoxide dismutase(SOD) in the neurons were detected after treated with Sen. The morphological changes of nucleus of the neurons were observed by Hoechst 33258 staining. The mRNA expression of bcl-2 and bax was quantified by real-time PCR. The protein levels of Bcl-2 and bax were measured by Western blotting. The activity of caspase-3 was also assayed.RESULTS: Compared with H₂O₂ group, the levels of antioxidative enzyme were increased in Sen+H₂O₂ group (P<0.05). In addition, mRNA expression of bcl-2 increased and that of bax decreased (P<0.05) in Sen+H₂O₂ group. Moreover, Sen increased the protein level of Bcl-2, and reduced the protein level of Bax and the activity of caspase-3 in the neurons exposed to H₂O₂ (P<0.05).CONCLUSION: The protective effect of Sen on hippocampal neurons with H₂O₂ -induced injury may be involved in the mechanisms of     

Down-regulation of miR-153-3p attenuates H2O2-induced H9C2 cell injury by promoting Nrf2 expression

AIM To study the effect of microRNA-153-3p (miR-153-3p) knock-down on oxidative injury of H9C2 cells induced by H₂O₂ and its specific mechanism. METHODS The oxidative stress injury of H9C2 cell model was induced by H₂O₂, and then the cell viability and the expression of miR-153-3p were detected by MTT assay and RT-qPCR, respectively. The effects of miR-153-3p knock-down on the H9C2 cell injury under oxidative stress were studied by RNA interference technology. The targets of miR-153-3p were identified by Western blot and dual-luciferase reporter assay. RESULTS MTT assay showed that the viability of H9C2 cells was decreased with the increase in H₂O₂ concentration (P<0.05). The results of RT-qPCR showed that the expression of miR-153-3p was increased with the increase in H₂O₂ concentration (P<0.05). Knock-down of miR-153-3p increased the viability of H9C2 cells under oxidative stress, decreased the cell apoptosis and the content of malondialdehyde (MDA), and increased the activity of superoxide dismutase (SOD). The expression of nuclear factor E2-related factor 2（Nrf2） and antioxidant response element（ARE） activity were increased with the increase in H₂O₂ concentration (P<0.01). TargetScan analysis and dual-luciferase reporter assay showed that Nrf2 was one of the potential target genes of miR-153-3p. The results of Western blot further showed that over-expression of miR-153-3p inhibited the expression of Nrf2 (P<0.01), while down-regulation of miR-153-3p increased the expression of Nrf2 (P<0.01). Dual interference with Nrf2 and miR-153-3p significantly reduced H9C2 cell viability, promoted the apoptosis, increased MDA content, and decreased SOD activity in the presence of H₂O₂ (P<0.01). CONCLUSION Inhibition of miR-153-3p expression attenuates the injury of H9C2 cells induced by H₂O₂ through up-regulating Nrf2/ARE signaling pathway.     

Rose (Rosa hybrida L.) EST-derived microsatellite markers and their transferability to strawberry (Fragaria spp.)

The ‘Genome database for Rosaceae (GDR)’ provides a large collection of expressed sequence tags (ESTs) harboring simple sequence repeats (SSRs) from several Rosaceae genera, including Rosa (rose). Primer pairs flanking SSR were designed for 312 unique Rosa ESTs based on GDR database. Eight rose (Rosa hybrida L.) genotypes were tested for PCR amplification, and 287 (92%) of the primer pairs generated allele-specific PCR bands that were readily scored. From 183 (63.7%) primer pairs that evidenced polymorphic alleles among the eight rose cultivars, 20 pairs evidencing EST sequence homology to known gene functions and high levels of polymorphism were selected and utilized for DNA fingerprinting and genetic diversity assessments of 47 rose hybrids. A total of 202 polymorphic bands were scored and generated unique fingerprints for each rose hybrid. The Nei–Li genetic similarity coefficients among 1081 pair-wise comparisons of 47 cultivars exhibited a broad range of genetic variations from 0.30 (‘Grand King’ and ‘Carnival’) to 0.99 (‘First Red’ and ‘Red Champ’). UPGMA cluster analysis divided 47 hybrids into five major groups and two sub-groups. The cross-species transferability of 273 EST-SSR primer pairs was evaluated using four genotypes of the strawberry, a genus member of the Rosaceae family. PCRs on the DNA samples of strawberry were successful for 165 primer pairs; among these, 123 pairs amplified 243 polymorphic bands. As surrogates of the marker transfer, the phenetic relationship among the four strawberry genotypes was evaluated. Genetic similarity coefficients varied from 0.78 (‘Maehyang’ and ‘Janghyee’) to 0.64 (‘Janghyee’ and ‘Pragana’). The results of cluster analysis showed that the three octaploid strawberry cultivars were quite similar, whereas the diploid ‘Pragana’ was related distantly at the genomic DNA level. The EST-SSR markers developed in the present study can be efficiently utilized for genetic diversity studies in Rosaceae.     

Preparation of gfp-bcl-XL-contained recombinant adenovirus vector by the homologous recombination in bacteria

AIM: To prepare gfp-bcl-X_L-contained recombinant adenovirus(rAd-gfp-bcl-X_L).METHODS: Bcl-X_L gene was amplified from pEGFP-C₃-bcl-X_L, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-X_L. Then pAdTrack-CMV-bcl-X_L was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-X_L and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-X_L into 293 cells. PCR test indicated that the recombinant Ad contained bcl-X_L gene. The titer of the purified rAd-gfp-bcl-X_L was 6.5×10¹² PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-X_L. This affords a good gene transfer vector for the gene therapy in human’s diseases.     

Lipoxin A4 reduces lipopolysaccharide-induced expression of cyclooxygenase 2 and prostaglandin E2 in human bronchial epithelial cells

AIM: To explore the effects of lipoxin A₄ on the expression of cyclooxygenase 2 (COX-2) in human bronchial epithelial cells (HBECs). METHODS: HBECs were incubated with various concentrations (0.1, 1 and 10 mg/L) of lipopolysaccharide(LPS) for 9 h, or 1 mg/L LPS for different time (3 h, 6 h and 9 h). The levels of COX-2 mRNA in HBECs and prostaglandin E₂ (PGE₂) in the culture supernatant were measured. In addition, the HBECs were exposed to lipoxin A₄ at concentration of 0, 100 and 400 μmol/L after stimulated with LPS at concentration of 1 mg/L for 9 h, and the supernatant of the culture cells was collected for determining the content of PGE₂ by ELISA. The cells were also harvested, and the mRNA and protein levels of COX-2 were analyzed by RT-PCR and Western blotting, respectively. RESULTS: LPS increased the mRNA expression of COX-2 and production of PGE₂ in a dose and time dependent manners in HBECs. Induction of COX-2 mRNA and protein by LPS were inhibited by lipoxin A₄ in a dose-dependent manner. Lipoxin A₄ also significantly decreased LPS-induced production of PGE₂. CONCLUSION: Lipoxin A₄ down-regulates LPS-induced expression of COX-2 and consequently inhibits the production of PGE₂ in HBECs.     

Physico-chemical and antioxidant properties of cornelian cherry fruits (Cornus mas L.) grown in Turkey

Cornus mas L. is a naturally growing dogwood species in Anatolia. In present study, physical, chemical and antioxidant properties of cornelian cherry fruits were studied. The fruit weight was in the range of 0.39–1.03 g, fruit length 14.24–22.20 mm, fruit width 9.59–13.21 mm, flesh/seed ratio 1.34 to 6.72. Hunter L values of the samples ranged between 10.82 and 19.69, and a value was between +6.25 and +15.59, and b value was between +3.46 and +6.64. In addition to the levels of dry matter, soluble solids, pH, total acidity, total sugar content, reduced sugar content, unreduced sugar content, ascorbic acid, total anthocyanin and total phenolics were within the range of 15.88–28.19%, 12.50–21.00%, 3.11–3.53, 1.10–2.53%, 76.80–154.00 g kg⁻¹, 52.80–120.00 g kg⁻¹, 0.00–32.30 g kg⁻¹, 0.16–0.88 mg g⁻¹, 1.12–2.92 mg g⁻¹ and 2.81–5.79 mg g⁻¹, respectively. On the other hand, ferric reducing antioxidant power (FRAP) and EC₅₀ values were between 16.21 mmol g⁻¹ and 94.43 mmol g⁻¹, 0.29–0.69 mg mL⁻¹. Anthocyanin extracts of the fruits were analysed by high-performance liquid chromatography (HPLC) with UV–vis detection. Pelargonidin 3-glucoside was the main pigment found in cornelian cherry fruits.     

Changes during the respiratory climacteric in ripening plantain fruits

Fruits of a Ghanaian plantain (Musa, AAB group) of the “French” type, cultivar ‘Apem’, were used to study changes before and after commencement of the climacteric at 25°C. Fruits exhibited an extended preclimacteric period during which C₂H₄ production was undetectable, then a well defined maximum in C₂H₄ production prior to a 10-fold increase in CO₂ production. Maximum C₂H₄ production was much higher than in dessert banana cultivars (Musa, AAA and AAAA groups). During ripening, the high initial content of alcohol-insoluble solids was almost completely converted to sugars, accompanied by rapid peel colouration and pulp softening.     

接头连接介导的PCR 扩增马铃薯抗晚疫病基因侧翼序列研究

利用接头连接介导的PCR 技术，以前期NBS profiling 试验获得的1 条马铃薯抗晚疫病基因
片段为基础，设计巢式基因特异性引物，扩增获得两侧序列。结果表明：基因组DNA 经DraⅠ酶切后，
在已有536 bp 片段的左侧和右侧均得到序列延伸，右侧扩增得到2 301 bp，左侧扩增得到820 bp，去除
边界序列后，向左延伸789 bp，向右延伸2 273 bp，拼接后共长3 443 bp。采用GENSCAN 进行基因预测，
发现包含内含子在内的晚疫病抗性基因全长2 413 bp，在该预测基因 的UTR 区域设计特异性引物，在马
铃薯抗病和感病的基因型中扩增包含完整基因在内的2 571 bp 序列，发现该特异候选基因与晚疫病抗性相
连锁。     

The effect of human complement C5b~9 complex on nitric oxide synthesis in glomerular mesangial cells of rats

AIM:To study the effect of human complement C_5b~9 complex on nitric oxide(NO) synthesis of glomerular mesangial cells (MC). METHODS: First, the human complement C_5b~9 complexes were isolated and glomerular MC of rats were cultured. Second, the MC were stimulated with C_5b~9 complex and changes of metabolism products of NO(NO₃^- and NO₂) in MC culture supernatant at 6,24 and 48 hours after C₊5b~9 stimulating were detected. Moreover, cGMP levels in cultured MC were also measured. RESULTS: NO₃^-／NO₂^- contents from culture supernatant and cGMP levels in MC were increased parallelly after C_5b~9 complex stimulation. Further, NO synthesis was inhibited by L-N^G-nitro-arginine-methylester(L-NAME). CONCLUSION: NO synthesis of rat glomerular MC was incerased by human complement C_5b~9 stimulation.     

Genetic characterisation of oak seedlings, epicormic, crown and micropropagated shoots from mature trees by RAPD and microsatellite PCR

DNA was isolated from seedlings of Quercus robur, collected from a single provenance, and from epicormic, crown shoots and in vitro shoots from a single tree of Q. petraea using a CTAB method of extraction. DNA was obtained in sufficient quantity and purity, from 13 out of 30 seedlings, and from all isolations from epicormic and in vitro shoots (2.5–10.0 μg/g fresh/ weight). Smearing was minimised at a primer concentration of 0.12 μM with Taq polymerase at 0.5 unit/reaction. Nine primers produced 142 bands, 28 of which were polymorphic. A similarity index showed that 11 seedlings were closely related with high coefficients (0.85–0.90), but each could be identified from another using only 9 primers (OPA-02 and -05, OPG-04 and -05, OPE-01, -02, -03, -08, -09). DNA was isolated from crown, epicormic and in vitro leaves originating from a single 150-yr old tree of Q. petraea and analysed by randomly amplified polymorphic DNA (RAPD) and microsatellites. With each primer, a characteristic RAPD pattern was obtained, and it was common to all six epicormic shoots derived from different parts of a single branch of this tree; also to the shoots from the crown of the same tree with OPE1 OPA-05, OPA-08, OPA-01, OPA-02, OPA-04, OPA-05, OPG-02, OPG-10, OPE-12. Similarly, the RAPD pattern obtained from shoot cultures in vitro, derived from individual nodes of epicormic shoots produced by six different branch segments, were uniform for each of 15 primers. This work was repeated using microsatellite PCR. Three microsatellite loci AG16, AG 1/2 and AG 1/5 were amplified by PCR. It showed a uniformity of these microsatellite loci in shoots from the crown of the tree, and from epicormic shoots cultures derived from six different sections of branch.