超高效液相色谱-串联质谱法测定猪牛组织中甲基盐霉素残留量

建立超高效液相色谱-串联质谱法测定猪牛组织中甲基盐霉素残留量。猪、牛组织中残留的甲基盐霉素,经乙腈溶液提取,硅胶固相萃取柱净化,超高效液相色谱-串联质谱法测定,外标法定量。甲基盐霉素在5 ng/mL～250 ng/mL的浓度范围内呈现良好的线性关系,甲基盐霉素在猪、牛肌肉、肝脏、肾脏、脂肪中的检测限均为2.5μg/kg,定量限均为5.0μg/kg,猪牛肌肉、肝脏、肾脏、脂肪中5.0 ng/g～100 ng/g添加浓度范围内的回收率均值为71.9%～88.4%,批内批间RSD值均<15%。     

UASB工艺处理高浓度Vc生产废水的试验研究

采用UASB反应器对高浓度Vc生产废水进行了实验室处理研究。试验结果表明:Vc废水的厌氧处理运行性能因水质不同而存在很大差异。转化母液的容积负荷在8kg COD.m-3d-1以下时,去除率基本稳定在65%以上。当负荷继续提高时,去除率会急剧下降并保持在50%左右。精制母液和提取母液的容积负荷在10kgCOD.m-3d-1时去除率仍能稳定在70%以上。这些表明当两套反应系统的容积负荷控制在一定范围时,UASB反应器可以有效去除大部分的有机污染物。同时也说明转化母液废水中存在相当数量的难生物降解化合物。GC/MS水质分析结果表明,其中含有大量10C以上的长直链或双苯环等结构的难生物降解物。系统中厌氧颗粒污泥的产甲烷活性随着反应器中污泥驯化的进行和负荷的增加而上升。     

Evidence of diadromy in the French Polynesian Kuhlia malo (Teleostei: Percoidei) inferred from otolith microchemistry analysis

Abstract – Strontium/Calcium (Sr/Ca) and Barium/Calcium (Ba/Ca) ratios were measured in 60 otoliths of the French Polynesian flagtail Kuhlia malo. Both elemental ratios were needed to correctly distinguish residence in marine, brackish and fresh water. High Sr/Ca and low Ba/Ca around the nucleus of all otoliths provided evidence of marine residency during the early life of all individuals. At about 0.5 mm from the core, Ba/Ca increased rapidly and remained high in all otoliths. High and unstable Sr/Ca ratios were attributed to estuarine residency, whereas low and stable Sr/Ca ratios were attributed to freshwater residency. These chemical signatures inferred that following recruitment to rivers, some fish remained in the estuary, while others moved upstream. Some individuals shared their time between fresh and brackish waters later in life. The findings of this study have important implications for the protection of nearshore, brackish and freshwater environments for the conservation of this French Polynesian diadromous species.     

W/O微乳液体系中SA和MS清除·O_2~-的研究

以水溶液体系作为对照,利用分光光度法研究了W/O型微乳液体系中水杨酸（SA）和水杨酸甲酯（MS）清除超氧阴离子自由基（·O2-）的性能.结果发现,在水浴时间为20min、pH值为8.20和邻苯三酚加入量适宜的情况下,W/O微乳液体系中SA、MS对·O2-的清除率分别可达72.92%、81.47%.微小的pH值变化会引起邻苯三酚自氧化速率的显著变化,电解质溶液对其也有明显的影响,但对SA、MS清除·O2-的能力影响不大.     

牛奶中吡利霉素残留检测高效液相色谱-串联质谱法研究

建立了牛奶中吡利霉素残留检测的高效液相色谱-串联质谱(HPLC-MS/MS)法。液相色谱条件为:色谱柱为Waters XBridge C18柱(150 mm×2.1 mm,5μm),流动相为乙腈-0.01mol/L乙酸铵溶液(30∶70,V/V),柱温为30℃,流速为0.2 mL/min,进样量为10μL。质谱条件为:电喷雾离子源(ESI ),多反应监测(MRM)方式进行采集;监测离子质荷比(m/z)为411.5>112.0,411.5>363.4。以吡利霉素的同分异构体异吡利霉素作内标,内标法定量。结果表明:吡利霉素的线性范围为20~400 ng/mL,相关系数R2=0.999 9;方法检测限为2 ng/mL,定量限为5ng/mL;从50、100和150 ng/mL三个添加浓度检测结果可以看出,方法的平均回收率为90.4%~104.3%(n=6),批内、批间RSD均小于5%。     

Phenotypic characterisation of Phaeoacremonium and Phaeomoniella strains isolated from grapevines: enzyme production and virulence of extra-cellular filtrate on grapevine calluses

Extra-cellular enzyme production of different Phaeoacremonium spp. and Phaemoniella chlamydospora isolates were used to assay the possibility of inter-specific characterisation. Isolates of Phaeoacremonium aleophilum, Phaeoacremonium angustius, P. viticola and Ph. chlamydospora were grown on solid media and the activities of extra-cellular amylases, lipases, proteases, cellulases, xylanases, laccase, polygalacturonase, pectate lyase, lignin peroxidase, manganese peroxidase, urease and chitinase were assayed. Phaeoacremonium species showed activities of a larger number of enzymes and also enzyme activity was frequently higher suggesting that Phaeoacremonium can be more virulent. To assay if the produced extra-cellular enzymes could reflect the virulence capacity of the two genera, calluses of Vitis vinifera L. (cvs. Baga and Maria Gomes) and of a rootstock (R3309) were inoculated with filtrated culture liquid medium of three isolates of Ph. chlamydospora and one of P. angustius. Filtrates from all strains decreased callus growth and membrane integrity, while soluble protein content of calluses decreased with the strains CAP 054 and 1AS. P. angustius (CAP 054) induced the more severe symptoms in all genotypes. Water content decreased together with an increase of osmolality in both cultivars but not in rootstock suggesting that osmorregulatory capacity is more affected in cultivars. Data show that: (1) Phaeoacremonium and Phaeomoniella genera have different patterns of extra- cellular enzymatic production; (2) these fungi produce extra-cellular compound(s) that induce(s) senescence symptoms in plant cells inhibiting callus proliferation; (3) among the strains tested in plant calluses the most virulent isolate (CAP 054) also produced higher amounts of some extra-cellular enzymes; (5) rootstock calluses were less sensitive to inoculation than grapevine calluses.     

Hybridization and in vitro culture of an orchid hybrid Ascocenda ‘Kangla’

The present study focuses on hybridization program involving two species belonging to two different vandaceous genera, viz., Ascocentrum ampullaceum (Roxb.) Schltr. var. auranticum, a narrow endemic orchid of Manipur and Vanda coerulea Griff., an endangered orchid of Appendix I of CITES (Convention on International Trade in Endangered Species of Wild Fauna and Flora), to synthesize the primary hybrid genus with intermediate and improved characters in the F₁ generation. Observations on the crossability in the present bigeneric cross between V. coerulea and A. auranticum had been achieved with 60% success when V. coerulea was taken as female parent. Murashige and Skoog (MS) basal medium at half-strength was effective for the development of the hybrid seedlings of V. coerulea × A. auranticum followed by Vacin and Went (VW) and Knudson C (KC) media. The best response of seedling growth was observed on MS medium at half-strength supplemented with 2.3 μM kinetin + 0.5 μM α-naphthalene acetic acid with maximum shoot height (2.7 cm), leaf number (4.6) and root number (4.1) after 150 days of inoculation. The survival percentage and growth performance of the seedlings were found to be higher (80% survival) in potting substrate consisting of brick:charcoal in the ratio 2:1 mulched with moss (Sphagnum sp.) than in potting substrate consisting of brick:charcoal:tree fern in the ratio 2:1:1. The first flowering was observed in the hybrid seedlings of V. coerulea × A. auranticum after 2 years of transfer to the ex vitro environment. Morphologically, the flowers differed from that of the parents clearly showing the success of the hybridization experiment. Registration of the hybrid has been made with the Royal Horticultural Society with the nomenclature Ascocenda ‘Kangla’ (No. T128725).     

Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter

Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchina semidecandra were optimized using green fluorescent protein (GFP) as a reporter. The binary vector pCAMBIA1304 harboring the modified green fluorescent protein (mgfp) gene driven by the CaMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period, immersion time, acetosyringone concentration and wounding type. Results obtained were based on the percentage of GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens strain LBA4404 and EHA105 at concentration 1 × 10⁷ cfu ml⁻¹ (OD_600nm 0.8) showed the highest virulence on M. malabathricum and T. semidecandra, respectively. Four days of pre-culture and 2 days of co-cultivation were optimum for M. malabathricum transformation, while 3 days of pre-culture and co-cultivation for T. semidecandra. Result also showed that 60 min of immersion and addition of 200 μM acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T. semidecandra. Mild wounding also significantly increased the efficiency of M. malabathricum transformation.     

不同品种厚皮甜瓜果实成熟过程中挥发性物质成分分析

采用固相微萃取(SPME)与气相色谱-质谱(GC-MS)联用的方法,对品质差异较大的山农黄金1号、Sweet delight(美国)和Takami(日本)厚皮甜瓜果实成熟过程中挥发性物质进行分析鉴定。结果表明,不同品种厚皮甜瓜果实成熟过程中挥发性物质组成和相对含量不同。山农黄金1号、Sweet delight和Takami果实成熟过程中共分离出156种挥发性物质,主要为酯类、醛类和醇类等化合物。山农黄金1号甜瓜果实未成熟时挥发性物质以醛类物质为主;随着果实的发育,近成熟时醇类、醛类物质含量急剧降低,酯类物质含量迅速升高;果实成熟时挥发性物质以酯类物质为主。Sweet delight果实未成熟时挥发性物质以醛类、醇类物质为主;随着果实的发育,醇类物质含量逐渐降低,醛类、酯类物质含量逐渐升高;果实成熟时挥发性物质以醛类物质为主。Takami果实未成熟时挥发性物质以醇类、醛类物质为主;果实成熟时挥发性物质以酯类、醛类物质为主。     

High frequency plantlet regeneration from callus and artificial seed production of rock plant Pogonatherum paniceum (Lam.) Hack. (Poaceae)

Pogonatherum paniceum (Lam.) Hack. is a rock plant with good potential for vegetative recovery on naked lands. A high frequency in vitro regeneration system was developed for P. paniceum. Calli were induced from explants of mature seeds, seedlings, young leaves, and stem segments on Murashige and Skoog (MS) medium supplemented with 1.0 mg L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D), 2.0 mg L⁻¹ α-naphthalene acetic acid (NAA) and 0.2 mg L⁻¹ 6-benzylaminopurine (BAP). High induction rates (59.57%) and regeneration rates (100%) were obtained from mature seed explants; calli were sub-cultured for over 2 years and still retained a high regenerative capacity. One seed explant resulted in 69,997 plants in 1 year. Shoot buds derived from calli were used for encapsulation in liquid MS medium containing 3% sucrose and two different alginate matrices (3% sodium alginate (w/v) + MS medium containing 3% sucrose and 3% sodium alginate + 1% activated carbon (w/v) + MS medium containing 3% sucrose) with a 20-min exposure to 2% CaCl₂ and 0.3% bavistin (w/v). The capsule with 3.0% sodium alginate (w/v) and 1% activated carbon (w/v) showed a higher conversion rate (61.58%) and stronger plantlets under non-aseptic conditions. These systems are useful for the rapid clonal propagation and dissemination of artificial seed material of P. paniceum for eco-recovery.