Incidence and antimicrobial resistance of enteropathogens isolated from an integrated aquaculture system

Aims: Biocontrol is an emerging trend aimed at reducing chemical input while increasing plant fitness, productivity and resistance to diseases in sustainable agriculture. An antagonist, pY11T‐3‐1, was herein characterized for potential applications against soil‐borne plant diseases. Methods and Results: In vitro antagonistic assays, the antagonist pY11T‐3‐1 was demonstrated able to obviously reduce the occurrence of the soft rot disease on Pinellia ternata, potato, pepper, tomato, cucumber and eggplant tubers or fruits, with higher prevention (90%) on P. ternata. It showed a broad antagonistic spectrum against 23 tested bacterial and fungal phytopathogens, which were distributed in 14 genus and 17 species. However, it inhibited only two of the seven bacterial nonpathogens. Phenotypic characterizations showed that the antagonist pY11T‐3‐1 was similar to Pseudomonas aeruginosa. Its major fatty acids were 18:1 w7c (22·17%), 16:0 (20·21%), 12:0 2OH (12·45%), 16:1w7c/15 iso2OH (10·95%) and 10:0 3OH (10·79%), which is a different profile from that of Ps. aeruginosa. The 16S rRNA and gyr B gene sequences shared 100 and 99% similarity with Ps. aeruginosa, respectively. The phylogenetic trees showed that it was clustered with Ps. aeruginosa. Conclusions: The antagonist pY11T‐3‐1 was characterized as Ps. aeruginosa with a unique fatty acid profile. Significance and Impact of the Study: With broad antagonistic spectrum and host selectivity, the antagonist pY11T‐3‐1 may provide a more environmental and economical alternative to the control of soil‐borne disease on P. ternata, which needs further investigation.     

生半夏、南星水提物对人胃癌BGC823细胞的侵袭力及HIF-lαmRNA蛋白表达的影响

摘要目的：观察生半夏、南星中药水提物对缺氧环境中人胃癌细胞株BGC823细胞HIF-lα蛋白表达和侵袭力的影响。方法：运用CoCl2（氯化钴）诱导BCG823细胞缺氧,使得细胞中HIF-1α蛋白表达升高．实验组加入生半夏、南星水提物对细胞进行预处理,然后在进行缺氧诱导。通过甲基噻唑基四唑法（MTT）检测细胞活性,使用Transwell检测细胞侵袭能力变化,RT-PCR、Weste-rn blotting分别检测HIF-lαmRNA及蛋白含量及变化。结果：生半夏、南星水提物能抑制人胃癌BGC823细胞的增殖;生半夏、南星水提物均能抑制缺氧诱导胃癌细胞的侵袭力,并且能降低HIF-1αmRNA及蛋白表达。结论：生半夏、南星水提物可抑制人胃癌BGC823细胞的增殖,抑制人胃癌BGC823细胞侵袭力,可能通过降低HIF-1α蛋白表达有关。     

半夏光系统对光照和温度日变化的适应

为探讨半夏(Pinellia ternate)光系统对光照强度和温度日变化的适应机理,该文连续3 d模拟了在同一种变化的光照强度(0～1 600μmol·m~(-2)·s~(-1))日变化下低温(10～18℃)、中温(20～28℃)和高温(28～38℃)的环境条件,测定了光系统Ⅱ(PSⅡ)和光系统I(PSⅠ)的叶绿素荧光参数,通过PSⅡ和PSⅠ光合活性和电子传递能力的变化来研究半夏光合系统对光照强度和温度日变化的适应。结果表明:(1) PSⅡ最小荧光(F_o')和PSⅡ反应中心激发能捕获效率(F_v'/F_m')随光照强度的增加而降低,光照强度的增加是导致光系统的活性降低的主要原因,低温会进一步导致光系统活性的降低;(2)光照强度和温度的增加使PSⅠ受体端热耗散效率[Y(ND)]上升,而PSⅠ供体端热耗散效率[Y(NA)]则降低,光照强度的增加不会导致供体侧较大的激发压,但会使受体侧开始积累较大的激发压,而较低的温度会导致受体侧活性降低,使供体侧积累较高的激发压;(3)高光(光强900μmol·m~(-2)·s~(-1))对半夏的光抑制和光损伤导致了PSⅡ实际光化学量子产量[Y(Ⅱ)]和PSⅠ实际光化学量子产量[Y(I)]的降低,低温进一步加剧了Y(Ⅱ)和Y(I)的降低;(4)在高光下,PSⅠ的电子传递速率ETR(I)的增加启动了环式电子传递(CEF),较高的CEF稳定了高温下的PSⅡ电子传递速率ETR(Ⅱ)的同时也保护PSⅡ免受光的损伤;(5)在3 d的处理中,虽然非光化学猝灭系数(NPQ)随光照强度的增加而上升,但是相对于高温,在低温处理下,半夏较低的NPQ使PSⅡ非调节性能量耗散的量子产量[Y(NO)]一直处于最高水平,表现出明显的光抑制。综上结果表明,低温降低了半夏对高光环境的适应能力,而高温通过增强NPQ,加速CEF的产生,减少光抑制的产生,从而加速光反应的电子传递和维持光反应系统的稳定性。因此,低温胁迫会加剧半夏光系统的损伤,适当提高温度可以增强半夏光反应系统对高光的适应性。     

猫爪草提取物体外抗肿瘤作用的研究

为探讨猫爪草不同提取物对体外培养的肿瘤细胞的作用,采用系统溶剂法提取猫爪草皂苷及多糖,运用MTT和集落形成实验法观察不同浓度的皂苷及多糖对体外培养的肉瘤S180、艾氏腹水瘤EAC及人乳腺癌细胞株MCF-7的影响。结果发现猫爪草皂苷及多糖对三种肿瘤细胞株的生长和集落形成均有不同程度的影响,皂苷给药量与抑瘤率和集落形成明显地呈正相关关系,而多糖有一最佳浓度。     

Transgenic Tobacco Expressing Pinellia ternata Agglutinin Confers Enhanced Resistance to Aphids

Tobacco leaf discs were transformed with a plasmid, pBIPTA, containing the selectable marker neomycin phosphotransferase gene (nptII) and Pinellia ternata agglutinin gene (pta) via Agrobacterium tumefaciens-mediated transformation. Thirty-two independent transgenic tobacco plants were regenerated. PCR and Southern blot analyses confirmed that the pta gene had integrated into the plant genome and northern blot analysis revealed transgene expression at various levels in transgenic plants. Genetic analysis confirmed Mendelian segregation of the transgene in T1 progeny. Insect bioassays showed that transgenic plants expressing PTA inhibited significantly the growth of peach potato aphid (Myzus persicae Sulzer). This is the first report that transgenic plants expressing pta confer enhanced resistance to aphids. Our study indicates that the pta gene can be used as a supplement to the snowdrop (Galanthus nivalis) lectin gene (gna) in the control of aphids, a sap-sucking insect pest causing significant yield losses of crops.     

Cloning and molecular characterization of a novel lectin gene from Pinellia ternata

The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE-PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was successfully expressed in E. coli BL21 when induced by IPTG. Artificial diet assay revealed that PTA fusion protein had significant levels of resistance against peach potato aphids when incorporated into artificial diet at 0.1% (w/v). The cloning of the pta gene will enable us to further test its effect in depth on aphids by transferring the gene into crop plants.     

半夏属若干变异式样及其演化

通过数量统计和栽培观察,发现半夏属具有子块茎Ｔｎ＾ｍ,子块茎根状茎Ｔｎ＾ｍＲｂ＾ｎ,子块茎根状茎珠芽Ｔａ＾ｍＴｎｚ＾ｎＢｃ＾ｒ和子块茎株芽Ｔａ＾ｍＢｃ＾ｒ４种组合形式的无性繁殖式样。属内经历了从根状茎（Ｒ）揶块茎（Ｔ）到珠芽（Ｂ）和盾形叶到其它叶形的经及其它变化。还讨论了该属的种间进化。     

贵州半夏块茎腐烂病拮抗细菌的筛选与鉴定

【目的】从半夏根际土壤筛选对半夏块茎腐烂病致病菌有拮抗活性的细菌并进行鉴定。【方法】采用稀释分离法和平板对峙法分离拮抗菌,然后根据菌落形态、生理生化特性,结合分子生物学方法进行鉴定。【结果】从半夏根际土壤共分离到228株细菌,其中菌株GZDF2、GZDF3、GZDF4对病原细菌及病原真菌均有拮抗活性,抑菌圈大小达23 mm,并且抑菌谱广。经形态学观察、生理生化特性分析和分子生物学方法,将菌株GZDF2、GZDF3、GZDF4鉴定为短短芽胞杆菌(Brevibacillus brevis)。进一步采用gyr B和rpo B基因对3株短短芽胞杆菌进行聚类分析,结果显示GZDF3菌株与其他2株短短芽胞杆菌存在遗传差异。【结论】半夏根际土壤中分离到的3株短短芽胞杆菌抑菌活性强、抑菌谱广,具有开发成生防剂的潜力。     

掌叶半夏有效提取物对宫颈癌细胞株增殖的抑制作用

目的既往的临床研究证实中药掌叶半夏可有效治疗宫颈癌。在此基础上,课题组观察了中药掌叶半夏有效提取物(Pinellia Extract,PE)对宫颈癌细胞株CaSki和HeLa细胞增殖的作用,先前采用MTT法检测细胞活性以及倒置相差显微镜观察细胞形态的变化,初步显示了PE对CaSki和HeLa细胞的增殖有抑制作用;本文将对PE的增殖抑制作用进行进一步的研究。方法采用扫描电镜和透射电镜观察PE作用后CaSki和HeLa细胞表面及内部超微结构的变化,In-cell Western blot检测不同浓度PE作用15min后CaSki和HeLa细胞磷酸化ERK的表达。结果0.15mg/mlPE作用24h后CaSki和HeLa细胞表面结构及内部结构均发生了显著的变化,提示细胞增殖受到抑制;不同浓度PE作用15min后CaSki和HeLa细胞磷酸化ERK的表达随PE浓度的增加逐渐下降。结论PE对CaSki和HeLa宫颈癌细胞株的增殖有明显抑制作用,表现在细胞表面及内部超微结构的变化;增殖抑制作用之一是阻断ERK的磷酸化。实验结果为进一步研究PE的抗宫颈癌作用机制以及新药的研制开发奠定了一定的理论依据和实验基础。