Genetic analysis and molecular characterisation of laboratory and field mutants of Botryotinia fuckeliana (Botrytis cinerea) resistant to QoI fungicides

BACKGROUND: QoI fungicides, inhibitors of mitochondrial respiration, are considered to be at high risk of resistance development. In several phytopathogenic fungi, resistance is caused by mutations (most frequently G143A) in the mitochondrial cytochrome b (cytb) gene. The genetic and molecular basis of QoI resistance were investigated in laboratory and field mutants of Botryotinia fuckeliana (de Bary) Whetz. exhibiting in vitro reduced sensitivity to trifloxystrobin. RESULTS: B. fuckeliana mutants highly resistant to trifloxystrobin were obtained in the laboratory by spontaneous mutations in wild‐type strains, or from naturally infected plants on a medium amended with 1–3 mg L^?1 trifloxystrobin and 2 mM salicylhydroxamic acid, an inhibitor of alternative oxidase. No point mutations were detected, either in the complete nucleotide sequences of the cytb gene or in those of the aox and Rieske protein genes of laboratory mutants, whereas all field mutants carried the G143A mutation in the mitochondrial cytb gene. QoI resistance was always maternally inherited in ascospore progeny of sexual crosses of field mutants with sensitive reference strains. CONCLUSIONS: The G143A mutation in cytb gene is confirmed to be responsible for field resistance to QoIs in B. fuckeliana. Maternal inheritance of resistance to QoIs in progeny of sexual crosses confirmed that it is caused by extranuclear genetic determinants. In laboratory mutants the heteroplasmic state of mutated mitochondria could likely hamper the G143A detection, otherwise other gene(s) underlying different mechanisms of resistance could be involved. Copyright © 2012 Society of Chemical Industry     

植物甜菜碱醛脱氢酶的生物信息学分析

运用生物信息学的方法,对已在GenBank数据库中注册的甜菜、菠菜、山菠菜、玉米、辽宁碱蓬等植物甜菜碱醛脱氢酶(BADH)的氨基酸序列进行分析。结果表明:植物BADH属于稳定蛋白质,含有较丰富的丙氨酸和谷氨酸;不同植物BADH的氨基酸序列具有较高的同源性,含有与酶功能相关的十肽保守序列及半胱氨酸残基;不同植物的BADH具有不同的亚细胞定位;分子进化研究表明,BADH可作为植物遗传分化和分子进化研究的重要依据;氨基酸序列中不存在信号肽;分子中不存在跨膜结构域,可能受蛋白激酶的磷酸化;α?螺旋是多肽链中的主要结构元件;蛋白质保守区域包含典型的醛脱氢酶功能结构域。该研究结果为开展BADH的酶学特性及植物抗逆的分子机理研究提供理论参考。     

日本落叶松 UDPGDH基因的cDNA克隆和表达分析

以日本落叶松杂交Solexa转录组测序获得的数据为依据,经拼接后获得尿苷二磷酸葡萄糖脱氢酶(UDPGDH)原序列,以该序列两端设计引物,在落叶松亲代和子代幼嫩针叶中成功克隆了UDPGDH基因cDNA序列,序列长1 443 bp,编码480个氨基酸。qRT-PCR结果分析表明:UDPGDH基因的表达量在落叶松杂交子代中远远高于亲本,正交子代的表达比反交子代高,说明UDPGDH在形成细胞壁半纤维素前体物过程中起着非常重要的作用,暗示UDPGDH在子代的高表达有利于杂种优势的形成。     

甘草提取物对酒精性肝损伤的防治作用研究

以酒精性肝损伤小鼠为动物模型,探讨甘草提取物对酒精性肝损伤的防治作用.将64只昆明系小白鼠随机分为8组,即正常对照组、治疗组（低、中、高剂量）、预防组（低、中、高剂量）、模型组.模型组以56度白酒16 mL/kg灌胃,预防组和治疗组在酒精灌胃的基础上分别采用低（4.8 g/kg）、中（9.6 g/kg）、高（16 g/kg）不同剂量甘草提取物灌胃,连续14 d,取血液和肝脏样品,测定肝组织谷草转氨酶（GOT）、谷丙转氨酶（GPT）及血清乙醇脱氢酶（ADH）含量,同时采用H.E染色对肝组织进行组织学观察.血样测定结果表明,与模型组比较,低剂量预防组和治疗组GOT降低均不明显,中、高剂量预防组和治疗组GOT降低显著（P＜0.05,P＜0.01）;低剂量治疗组（P＜0.05）和其他给药组（P＜0.01）均能显著降低GPT含量;中、高剂量治疗组（P＜0.05）和低、中剂量预防组（P＜0.05）与高剂量预防组（P＜0.01）均能显著提升ADH含量;组织学观察表明,与模型组比较,中、高剂量治疗组及预防组能有效缓解和恢复肝损伤组织结构.提示甘草提取物对酒精引起的肝损伤具有一定的保护和治疗作用.     

Red Algal Bromophenols as Glucose 6-Phosphate Dehydrogenase Inhibitors

Five bromophenols isolated from three Rhodomelaceae algae (Laurencia nipponica, Polysiphonia morrowii, Odonthalia corymbifera) showed inhibitory effects against glucose 6-phosphate dehydrogenase (G6PD). Among them, the symmetric bromophenol dimer (5) showed the highest inhibitory activity against G6PD.     

Characterization of biochar and their influence on microbial activities and potassium availability in an acid soil

Application of biochar to soil has increased considerably during recent years because of its effectiveness as a soil amendment causing beneficial effects on soil health. However, the effects have been reported to vary and depend upon types of feedstock and pyrolysis conditions during biochar production. Therefore, characterization of biochar is extremely important for its efficient utilization as a soil amendment. In the present study, biochar was prepared from agro-industrial by-products (rice husk and sugarcane bagasse) and weeds (Parthenium and Lantana) under similar pyrolysis conditions. Lantana biochar (LBC) showed the highest pH (10.4) while the lowest value (8.5) being recorded in rice husk biochar (RHBC). The energy-dispersive X-ray spectroscopy (EDS) analysis indicated that LBC and Parthenium biochar (PBC) were superior with respect to potassium (K) content than sugarcane bagasse biochar (SBBC) and RHBC. The Fourier-Transform Infrared Spectroscopy (FTIR) study exhibited the existence of different functional groups in biochar. All the biochar treated soils showed significantly higher microbial activities with different degrees. Application of LBC and PBC at 4.50 g kg^?1 soil significantly increased K availability in soil. Lantana biochar and PBC amended the soil at 9 g kg^?1 significantly increased the soil pH thus makes these biochar as potential liming materials.     

黑穗醋栗果实生长发育过程中抗坏血酸含量 及相关酶活性的变化

【目的】 研究不同品种不同生长时期黑穗醋栗果实内抗坏血酸（AsA）含量及其合成代谢过程中相关酶活性的变化,以明确果实生长发育过程中AsA含量与代谢合成相关酶之间的关系,为全面揭示黑穗醋栗果实AsA积累规律提供理论依据。【方法】 以3个不同黑穗醋栗品种（‘亚德’‘布劳德’和‘黑丰’）为试材,测定果实在幼果期、膨大期、半转色期、转色期和成熟期时还原型抗坏血酸（AsA）、氧化型抗坏血酸（DHA）、还原型谷胱甘肽（GSH）和氧化型谷胱甘肽（GSSG）含量以及AsA合成与代谢相关酶活性。【结果】 不同品种黑穗醋栗果实大小、AsA含量以及AsA相关代谢物水平存在明显的多样性。其中‘亚德’单果重最大,为1.97 g。果实生长发育过程中,总抗坏血酸（T-AsA）和AsA含量在3个品种中变化趋势一致,均在幼果期含量最高,其中‘亚德’幼果期果实中AsA含量最高,为83.17 μmol·g ^-1 FW,随着果实的生长迅速下降,在成熟期降至21.28 μmol·g ^-1 FW;3个品种果实中GSH和T-GSH含量随着果实发育呈升高趋势,但不同品种升高时期或增加幅度不同;GSSG含量在不同品种间存在较大差异,成熟果中‘黑丰’含量最低,为0.008 μmol·g ^-1 FW,仅为‘亚德’的10.2%。AsA-GSH循环再生代谢中,脱氢抗坏血酸还原酶（DHAR）和单脱氢抗坏血酸还原酶（MDHAR）活性在果实膨大期达到最高,成熟期降至最低,其中‘布劳德’果实中DHAR和MDHAR活性略高于‘亚德’和‘黑丰’;谷胱甘肽还原酶（GR）活性在幼果期最高,‘亚德’幼果期果实中GR活性最高（0.06 μmol·min ^-1·g ^-1 FW）,之后随着果实的生长发育不断下降,抗坏血酸过氧化物酶（APX）活性变化与之相似;L-半乳糖途径的关键酶L-半乳糖-1,4-内酯脱氢酶（GalLDH）活性随着果实生长发育的变化趋势与AsA含量变化相一致,且‘亚德’果实中GalLDH活性在幼果期和成熟期均高于其他两个品种。通过相关性分析发现,GalLDH与T-AsA、AsA、DHA、DHAR和MDHAR呈现极显著正相关关系,相关系数可达0.91以上,即果实中GalLDH活性越高,果实中AsA含量也越高;DHAR和MDHAR与T-AsA、AsA间也存在极显著正相关关系,而APX与T-GSH、GSH间相关性较强。【结论】 黑穗醋栗幼果期果实中AsA含量最高,且品种间差异显著;GalLDH、MDHAR和DHAR可能是黑穗醋栗果实中AsA合成代谢的关键酶,黑穗醋栗果实中AsA含量积累主要取决于GalLDH活性,说明合成途径起着更关键的作用,而AsA-GSH循环再生途径相关酶对AsA合成也有一定的贡献,黑穗醋栗高AsA含量的积累是由合成途径与循环途径共同作用的结果。