欧亚旋覆花提取物对朱砂叶螨生物活性及其体内几种酶活性的影响

植食性害螨是危害农业生产的一个重要生物类群,我国每年用于主要作物的杀螨剂费用约90多亿元(师光禄等,1994)。朱砂叶螨(Tetranychus cinnabarinus)是其中一个重要的种类,可危害粮、棉、油、林木、果树等43科146种植物(李连昌等,     

Differential glutathione S-transferase isozyme activities in rice and early watergrass seedlings

Glutathione S -transferase (GST) isozymes were investigated in one-leaf-stage rice ( Oryza sativa L. cv. Nipponbare) and early watergrass ( Echinochloa oryzicola Vasing) shoots after being induced by treatment with a combination of pretilachlor [2-chloro-2',6'-diethyl-N-(2-propoxyethyl)acetanilide] and fenclorim (4,6-dichloro-2-phenylpyrimidine) using DEAE-Sephacel anion exchange chromatography. Non-treated plants contained GST isozymes that had activity to the following substrates: three isozymes for l-chloro-2,4-dinitrobenzene (CDNB), six isozymes for pretilachlor and three isozymes for fenclorim in rice shoots; and four isozymes for CDNB, three or four isozymes for pretilachlor and two or three isozymes for fenclorim in early watergrass shoots. Glutathione S -transferase isozyme activities of non-treated plants were higher in rice than in early watergrass, especially in the case of GST activity with fenclorim as a substrate. Pretreatment of rice roots with a combination of pretilachlor and fenclorim increased the activity of the constitutively expressed isozymes that exhibited activity with CDNB, pretilachlor and fenclorim. This pretreatment also caused the appearance of one new GST(fenclorim) isozyme. Pretreatment of early watergrass roots with a combination of pretilachlor and fenclorim produced almost no increase in activity of some constitutively expressed isozymes that exhibited activity to CDNB and fenclorim, although it partly increased the peaks to corresponding to pretilachlor. The induction of GST was higher in rice than that in early watergrass. These results indicated that the isozyme pattern and substrate specificity of GST isozymes in rice were different from those in early watergrass. Furthermore, the selectivity of pretilachlor between rice and early watergrass may be related to different constitutively expressed GST(pretilachlor) isozyme activities and the induction of GST(pretilachlor) isozyme activities in the combination treatment.     

Toxic effects of novel organophosphorus insecticide (RPR-V) on certain biochemical parameters of euryhaline fish, Oreochromis mossambicus

The euryhaline fish, Oreochromis mossambicus was exposed to sub-lethal concentration (0.017 mg L⁻¹) of a novel phosphorothionate, 2-butenoic acid-3-(diethoxy phosphinothionyl) ethyl ester (RPR-V) for 30 days and allowed to recover for 7 days. Important biomarker enzymes were assayed in plasma, brain, gill, liver, kidney, and muscle during exposure tenures of day-3, -7, -15, -30, and also at 7 days (withdrawal) after stopping treatment. Acetylcholinesterase (AChE) activities of brain, gill, and muscle were strongly inhibited by 67, 75, and 66%, respectively, on day-30. Exposure (time) dependent increases in alanine aminotransferase (ALAT), and aspartate aminotransferase (ASAT), acid phosphatase (AcP), and alkaline phosphatase (AkP), activities in plasma and kidney; AcP and AkP activities in gill were noticed. However, significant decrease in ALAT, ASAT, AcP, and AkP activities in liver was observed. The depletion of glycogen was observed in liver, brain, and gill tissues, an indication of typical stress related response of the fish with pesticide. A significant increase in lactate dehydrogenase (LDH) activity in gill and brain was observed and decreased in liver and muscle, indicating tissue damage and muscular harm. Depletion of glutathione (GSH) was observed in the above tissues, there by enhancing the lipid peroxidation resulting in cell damage. The induction in hepatic glutathione-S-transferase (GST) levels indicates the protection against the toxicity of xenobiotic-induced lipid peroxidation. There was a significant recovery in all the above biochemical parameters, in all the tissues of fish after a recovery period of 7 days. These results revealed that RPR-V affects the intermediary metabolism of O. mossambicus and the increase of biomarker enzymes in plasma, might be due to the necrosis of liver.     

Effects of salt stress on ion accumulation and antioxidative enzyme activities of Oryza sativa L. and Echinochloa oryzicola Vasing

The effects of salt stress on physiological factors, such as inorganic ion absorption and antioxidative enzyme activities, of rice ( Oryza sativa L. cv. Anapurna) and Echinochloa oryzicola Vasing were investigated. Although having similar morphology, rice and E. oryzicola possessed considerably different salt-tolerance mechanisms. Echinochloa oryzicola was more salt-tolerant than rice. When exposed to salt stress (100 mmol L⁻¹ sodium chloride, NaCl; six days), E. oryzicola had the ability to limit the accumulation of sodium ions (Na⁺), maintained high potassium ion (K⁺) content and had a constantly higher K⁺/Na⁺ ratio than rice. Rice was not effective in limiting Na⁺ absorption but had a higher antioxidative capacity than E. oryzicola . The constitutive activities of superoxide dismutase (SOD) and catalase of rice were three and five times higher than that of E. oryzicola, respectively. Induced activities of SOD, ascorbate peroxidase and glutathione reductase were also higher in rice than in E. oryzicola . The high antioxidative capacity was one of the tolerance mechanisms used by rice to cope with salt stress. Therefore, the salt tolerant-mechanisms are different between the two plants.     

Estimation of the selenium requirement of growing guinea pigs (Cavia porcellus)

The aim of the study was to determine the selenium (Se) requirement of guinea pigs as a species unable to synthesize ascorbic acid. Forty-nine male guinea pigs (average weight 208 ± 3.5 g) were divided into an initial status group and six experimental groups. The animals received a Se deficient Torula yeast based basal diet (<0.02 mg Se and 26 mg α-tocopherol/kg) or a Se addition of 0.05, 0.10, 0.15, 0.20 and 0.25 mg/kg diet as sodium selenate for 10 weeks. There was no significant difference in weight gain (final weight 643 ± 21 g) between the groups and no clinical symptoms of Se deficiency occurred. With the exception of the testes, there was an increasing Se concentration in liver, plasma and haemolysate dependent on supplementation level. Glutathione peroxidase was determined in the plasma and Se dependent glutathione peroxidase (GPx1) in haemolysate, liver, kidney, heart and lung. Thioredoxin reductase (TR) activity was measured in liver, kidney and heart and deiodinase activity in the liver. A phospholipid hydroperoxide reducing activity with Se influence was determined in liver, kidney, heart, testes and brain. With the exception of GPx1 activity in heart and haemolysate and TR activity in the kidney, all enzymes already reached their maximal activity at 0.05 mg Se/kg diet. The activities of GPx1 and TR were used as parameters for broken line analysis and a Se requirement of 0.080 mg Se/kg diet was derived as sufficient for growing guinea pigs adequately supplied with vitamin E.     

柑橘全爪螨不同品系谷胱甘肽S-转移酶基因表达差异分析

为了明确谷胱甘肽S-转移酶（glutathion S-transferases, GSTs）基因与柑橘全爪螨抗性的关系,通过BLAST搜索,对柑橘全爪螨转录组数据库的GST基因进行鉴定,应用MEGA4.0.1软件对GST基因进行同源与进化分析,进一步采用RPKM法对柑橘全爪螨敏感品系和噻螨酮抗性品系GST基因进行表达差异分析。从柑橘全爪螨转录组中获得了30条GST基因,其中11条基因属于Delta家族,10条属于Mu家族,2条属于Omega家族,6条属于Kappa家族,1条属于Zeta家族。基因表达差异分析发现,抗性与敏感品系当中有25条GST基因表达量存在差异,抗性品系中有9条GST基因发生了上调,16条发生了下调,其中上调倍数最高的是GSTd6[log2 Ratio(RS/SS)= 1.0505],由此推断,GST基因的上调可能不是柑橘全爪螨对噻螨酮产生抗性的重要原因。     

甜菜幼苗对干旱胁迫的适应机制

为了研究甜菜幼苗在干旱逆境耐受过程中的应答机制,本研究以‘780016B/12优’为目的材料,通过设置不同浓度（3%、6%,9%以及15%）的PEG6000干旱胁迫来处理甜菜幼苗,并进行恢复生长,来衡量不同干旱胁迫处理间甜菜的相关生理生化指标和目的基因的表达量变化。研究结果表明,逆境胁迫24 h后,随着干旱强度的增加,甜菜幼苗植株的皱缩和萎蔫程度也逐渐严重,并伴随着叶片相对含水量(3.48%~15.86%)以及叶绿素含量(5.60~13.06)的下降;而体内的丙二醛和脯氨酸却大量累积,其中以15% PEG增加的尤为明显,分别高达48.23 μmol/L和201.37 μmol/L左右。逆境恢复后,3% PEG处理的甜菜幼苗的相对含水量(96.94%)、丙二醛含量(17.22 μmol/L)以及植株形态基本恢复到了正常水平;而高浓度PEG胁迫复性后,9% PEG处理的植株的相对含水量虽然恢复了8.48%左右,但丙二醛含量仍旧相对含量高达36.46 μmol/L;而脯氨酸在各组处理中,一直维持在较高水平。qPCR分析表明,干旱胁迫诱导BvGS基因受到了不同程度的诱导表达,分别是处理前的3.85、4.45、5.81、8.20倍左右,即使恢复生长,植株中该基因的表达量虽然有所下降,但仍旧维持在较高水平。因此,可以推测,甜菜幼苗通过调整细胞水势、氧化还原水平、光合作用以及渗透代谢平衡来适应干旱胁迫所带来的失水和质膜过氧化等不良反应,并有可能通过谷胱甘肽的合成来解毒由干旱逆境所带来的氧化伤害。     

Effects of dietary reduced glutathione on the growth and antioxidant capacity of juvenile Atlantic salmon (Salmo salar)

To investigate the effects of dietary reduced glutathione (GSH) on the growth performance and antioxidant capacity of juvenile Atlantic salmon (Salmo salar), 396 juvenile fish with initial body weight of 143.07 ± 6.56 g were randomly distributed into four groups fed four diets with graded supplementation levels of GSH (0, 100, 200 and 400 mg/kg diet) for 83 days. The results showed that the appropriate GSH supplementation (100 and 200 mg/kg diet) significantly increased the growth performance, activities and gene mRNA expression levels of glutathione peroxidase (GPx) and glutathione transferase (GST), and the content of GSH and total antioxidant capacity (TAOC), whereas it significantly decreased activities and gene mRNA expression levels of superoxide dismutase (SOD) and catalase (CAT), and the content of malondialdehyde (MDA; p < 0.05). However, the excess dietary GSH (400 mg/kg diet) had an adverse effect on the all above indexes. Interestingly, the dietary GSH had the opposite effect on GSH‐related antioxidant enzymes (GPx and GST) and other antioxidant enzymes (SOD and CAT). The results showed that the diet with 200 mg/kg GSH supplementation was optimal for the juvenile Atlantic salmon, which had a measured GSH content of 209.54 mg/kg.     

向日葵3个谷胱甘肽-S-转移酶GST基因的克隆及表达模式分析

从向日葵转录组测序结果中获得了3个对盐胁迫有响应的谷胱甘肽-S-转移酶（GST,EC2.5.1.18）GST基因,构建了系统进化树并进行分析,得知这3个基因属于Tau型GST,并将它们命名为HaGSTU26（HanXRQChr04g0127901）、HaGSTU8（HanXRQChr06g0177581）、HaGSTU27（HanXRQChr10g0316331）,然后以向日葵Sk02R为试验材料,克隆这3个基因,并进行了不同组织和不同胁迫条件下的表达分析。序列分析表明：HaGSTU26基因组为1674bp,CDS（编码蛋白序列）长666bp,编码221个氨基酸,由2个外显子和1个内含子组成;HaGSTU8基因组为2271bp,CDS长654bp,编码217个氨基酸,由2个外显子和1个内含子组成; HaGSTU27基因组为663bp,CDS长663bp,编码220个氨基酸,由1个外显子组成。实时荧光定量PCR分析表明：向日葵HaGSTU26、HaGSTU8、HaGSTU27基因在不同组织（根、幼叶、成熟叶、茎、幼茎、苞叶）中表达量不同,其中,HaGSTU26基因和HaGSTU27基因在根中表达量最高,而HaGSTU8基因在苞叶中表达量最高,但这3个基因均在成熟茎中的表达量最低。在不同胁迫条件下,测定这3个基因在向日葵幼苗中的表达量,结果表明在盐及ABA胁迫下,基因表达量均随着处理时间的增加而呈现先增加后下降的趋势。其中,在盐胁迫下,HaGSTU26基因在12 h后相对表达量最高,HaGSTU8及HaGSTU27基因表达量在3h后相对表达量最高。在ABA胁迫后,HaGSTU26及HaGSTU27基因表达量在12 h后相对表达量最高,HaGSTU8在24 h后相对表达量最高。在冷胁迫下,HaGSTU26和HaGSTU27基因上调表达,它们分别在3、24 h后相对表达量最高,HaGSTU8基因下调表达,其相对表达量随处理时间的延长呈现逐渐减少的趋势。在热胁迫条件下,这3个基因的相对表达量随着胁迫时间的延长而增加,均在24 h后表达量最高。以上结果说明这3个基因对不同非生物胁迫（盐、ABA、冷、热胁迫）均有响应。