Inhibitory Effects of Kojic Acid on Phenoloxidase of Diamondback Moth Plutella xylostella

The effects of kojic acid on phenoloxidase (PO) of Plutella xylostella were investigated, when it had been partially purified by 40% saturated [(NH4)2SO4] and Sephadex G-100 gel filtration. Kojic acid showed inhibitory effects on both monophenolase and o-diphenolase activity of the PO. The inhibitor concentrations leading to 50% (IC50) activity lost were estimated to be 0.07 mmol L^-1 for monophenolase and 1 mmol L^-1 for diphenolase, respectively. Kojic acid can also prolong the lag time of PO for oxidation of L-tyrosine. The inhibition kinetics were analyzed by Lineweaver-Burk plots and kojic acid was found to be a reversible competitive inhibitor with the ki of 0.47 mmol L^-1. The ability of kojic acid to inhibit the enzyme activity may be associated with its ability to chlate copper at the active site. In addition, the iron ion was found to shorten the lag time obviously, but have no important effect on the monophenolase activity.     

Effect of 6-BA and 2, 4-D on Soybean Transformation

Regeneration from cotyledonary nodes and embryonic tips of soybean "Peking" was studied. The disinfectant ways of the mercuric chloride and chlorine gas were used and the concentrations of 6-BA and 2, 4-D were studied in the culture medium. The results showed that the sterilization effect of chlorine gas was better than that of mercuric chloride. The best concentration of 6-BA was 1.0 mg · L-1 and the best concentration of 2, 4-D was 2.0 mg · L-1 in the germinating medium. The number of buds of each explant was 3.56 and 2.98, respectively. The best concentrations of 6-BA and 2, 4-D were 3.0 mg·L-1 and 3.5 mg·L-1 in regeneration of embryonic tips. The best inducing time was 16-20 h and the mean shoots per explant was 2.69 and 2.78, respectively.     

Effect of transferring lignocellulose-degrading bacteria from termite to rumen fluid of sheep on in vitro gas production,fermentation parameters,microbial populations and enzyme activity

The digestive tract of termite(Microcerotermes diversus) contains a variety of lignocellulose-degrading bacteria with exocellulases enzyme activity, not found in the rumen, which could potentially improve fiber degradation in the rumen. The objectives of the current study were to determine the effect of inoculation of rumen fluid(RF) with three species of bacteria isolated from termite digestive tract, Bacillus licheniformis, Ochrobactrum intermedium, and Microbacterium paludicola, on in vitro gas production(IVGP), fermentation parameters, nutrient disappearance, microbial populations, and hydrolytic enzyme activities with fibrous wheat straw(WS) and date leaf(DL) as incubation substrate. Inoculation of RF with either of three termite bacteria increased(P0.05) ammonia-N concentration compared with the control group(free of termite gut bacteria). Termite bacteria inoculation had no effect(P0.05) on gas production characteristics, dry matter, organic matter and neutral detergent fiber disappearance, pH, and concentration and composition of volatile fatty acids. Population of proteolytic bacteria and protozoa, but not cellulolytic bacteria, were increased(P0.05) when RF was inoculated with termite bacteria with both WS and DL substrates. Inoculation of RF with termite bacteria increased protease activity, while activities of carboxymethyl-cellulase, microcrystalline-cellulase, α-amylase and filter paper degrading activity remained unchanged(P0.05). Overall, the results of this study indicated that transferring lignocellulose-degrading bacteria, isolated from digestive tract of termite, to rumen liquid increased protozoa and proteolytic bacteria population and consequently increased protease activity and ammonia-N concentration in vitro, however, no effect on fermentation and fiber degradation parameters were detected. These results suggest that the termite bacteria might be rapidly lysed by the rumen microbes before beneficial effects on the rumen fermentation process could occur.     

Effects of Limiting Amino Acids on Rumen Fermentation and Microbial Community In vitro

Three goats fitted with cannula were used to provide rumen liquor to investigate the effects of limiting amino acids on rumen fermentation and microbial community in vitro. The removal method was used in the current experiments. Treatments are total essential amino acid （TEAA）, His-removal, Lys-removal, Met-removal, and branch chain amino acid （BCAA）- removal. Results indicated that, pH-value ranged between 5.9 and 6.8, with the highest mean value for the group with BCAA-removal （6.54） in the culture. Concentration of NH3-N ranged between 10.99 to 30.51 mg 100 mL^-1, with the group of TEAA recording the highest average NH3-N concentration （17.85 mg 100 mL^-1）. Yields of microbial protein and limiting degree on microbial growth varied with treatments （P 〈 0.01）, and the lowest accrued in treatment with BCAA-removal （0.1389, 0.1772, and 0.3161 mg mL^-1 for bacteria, protozoa, and mixed microbes, respectively）, compared to the group with TEAA, microbial production of mixed microbes decreased by 44.52%. As for micro-flora, protozoa to bacteria ratio was the lowest for the group with Lys-removal （89.12%）, while the highest for the group with BCAA-removal （127.60%） （P 〈 0.01）. Furthermore, PCR-SSCP analysis revealed that, microbial profile subjected to substrates within bacteria and protozoa groups. It was therefore concluded that, dietary amino acid influenced both rumen fermentation and microbial characteristics.     

The Effect of Yucca schidigera Extract on Ruminal Fermentation and Parameters Traits in Sheep

In a completely randomized block design experiment, 16 ruminally cannulated sheep (40 ± 2.1 kg)fed a 50% concentrate 50% forage diet (DM basis) were given intraruminal doses of powdered Yucca schidigera extract (YSE). Doses of 0 (control), 100, 200, or 300 mg kg-1 diet were given at 8 p.m. and 4 a.m. On 15, 16, and 17 d of the experimental period, ruminal contents were sampled 0, 2, 4, 6, and 8 h after dosing, and blood samples were collected at the end of the experiment (18th and 19th d). Acidity was not affected (P＞0.05) by the addition of YSE. Compared with the control, ruminal propionate concentration was increased by 29.4 and 29.8% (P＞0.05) and the acetic acid concentration was decreased by 15.1 and 19.8% (P＞0.05) at 4 and 6 h after YSE (300 mg kg-1) dosing, respectively. Ruminal ammonia concentration in the first 2 h after feeding was higher (P＜0.05) in the sheep that did not receive YSE (increased by 17.57 mg 100 mL-1) than in those that received 200 mg kg-1 (increased by 6.77 mg 100 mL-1) or 300 mg kg-1 (increased by 6.50 mg 100 mL-1) YSE. Protozoan populations in the rumen were lower (P ＜ 0.05) in the animals that received 300 mg kg-1 of YSE compared with the control. All serum parameters of the four groups were in the normal range and were similar among the treatment groups (P＞0.05), after being fed for 19 d with different doses of YSE. The effect of YSE on ruminal ammonia concentration likely resulted from a decreased concentration of protozoan populations and, presumably, from ammonia binding by YSE.     

Comparsion of the Effects of Succinate and NADH on Postmortem Metmyoglobin Redcutase Activity and Beef Colour Stability

In two experiments, the effects of succinate and NADH（reduced nicotinamide adenine dinucleotide） on metmyoglobin reductase activity and electron transport chain-linked metmyoglobin reduction were investigated and compared. In experiment 1, metmyoglobin（MetMb）, substrate and inhibitors were incubated with mitochondria. Comparsion of the effects of succinate and NADH on MetMb reduction was investigated. The MetMb percentage in sample treated with 8 mol L-1 succinate decreased by about 69% after 3 h incubation, and the effect was inhibited by the addition of 10 mol L-1 electron transfer chain complex II inhibitor malonic acid; the MetMb percentage in samples treated with 2 mol L-1 NADH decreased by 56% and the effect was inhibited by the addition of 0.02 mol L-1 electron transport chain complex I inhibitor rotenone. These results indicated that electron transport chain played an important role in MetMb reduction. Both complex II and complex I take part in the MetMb reduction in mitochondria through different pathways. NADH-MetMb reduction system was less stable than succinateMetMb system. In experiment 2, the beef longissimus dorsi muscle was blended with different concentrations of succinate or NADH. Enhancing patties with higher concentration of succinate or NADH improved colour stability in vacuum packaged samples（P〈0.05）. These results verified that mitochondria electron transport chain is related to the MetMb reduction in meat system.     

Effects of CuSO4 and Uniconazole on Mature Embryo Culture in Japonica Rice

In order to establish the system of high frequency plant regeneration for japonica rice mature embryos, the effects of different concentrations of CuSO4 and uniconazole on in vitro culture of mature embryos were studied using three rice cultivars of Kongyu 131, Longjing 24, and Dongnong 425 as test materials. The results showed that callus induction and differentiation of japonica rice mature embryos were apparently improved on the medium with 10-15 μmol·L-1 CuSO4 and 0.50-1.00 mg·L-1 uniconazole. Induction and differentiation rates of different genotype rice mature embryos displayed different sensitivities to CuSO4 and uniconazole. For the callus induction frequency of three varieties, the optimal concentration of CuSO4 was 15.0 mol·L-1. When the concentration of CuSO4 was 15 μmol·L-1, the plantlet differentiation rates of Kongyu 131 and Dongnong 425 got to the highest, while the concentration of CuSO4 was 10 μmol·L-1 for Longjing 24. For the callus induction and plantlet differentiation rates of Kongyu 131 and Dongnong 425, the ideal concentration of uniconazole was 0.50 mg·L-1 and for Longjing 24 was 1.00 mg·L-1.     

Effect of immunopotentiator of the Chinese herbal medicine on the quantity of T lymphocyte in chicken

In order to study the mechanism of immunopotentiator, the quantity of T lymphocyte was observed. Total 240 1-day chikens were divided into 3 groups randomly: one contrast group and two groups being drunk immunopotentiator according to the concentration 10 mL·L-1 and 5 mL·L-1 lasting for 48 d. The number of T lymphocyte in blood was measured by E-rosette when the chikens were at 12-, 24-, 36-, and 48-day. The results showed that the percent of T lymphocyte in the trial group was obviously higher than that of the contrast, and the 10 mL·L-1 group was higher than that of the 5 mL·L-1 group; the quantity distribution of T lymphocyte in intestinal mucosa lymphoid tissue of 14-day, 21-day chiken was surveyed in the contrast group and the 10 mL·L-1 group with the method of routine histology-slices and ANAE, the results showed that the quantity of T lymphocyte in the 10 mL·L-1 group was significantly higher than that of the contrast, which indicated that immunopotentiator increased markedly the quantity of T lymphocyte, and the 10 mL·L-1 group was higher than that of the 5 mL·L-1 group.     

Dynamic Changes in the Calcium Content of Several Apple Cultivars During the Growing Season

Dynamic changes in calcium content were investigated in eight apple cultivars. The results showed that the calcium concentration in leaves and shoots increased with fruit development. The cultivars displayed only a small difference in the calcium concentration during the early stage of development, the difference became very significant at the late stage of development, especially in shoots. In shoots, for example, calcium content was highest in Starkrimson （19 638.6 mg kg^-1） and lowest in Fuji （8 751.3 mg kg^-1）. Calcium concentration was highest in young fruits and was found to decrease with the growth of fruit, and was characterized by a dramatic drop at the rapid expansion stage. There was a significant difference among cultivars. Young Starkrimson fruits contained the highest calcium concentration of 506.52 mg kg^-1 among cultivars tested, followed by Pink Lady and Fuji. The calcium concentration in mature fruits from high to low is as follows： Starkrimson, Sansa, Pink Lady, Senshu, Gala, Fuji, Red General and New Century. In this study, it was found that eight cultivars continuously assimilated calcium during the whole growing season, especially at the young stage when fruit took up 35-46% of total calcium. The calcium content in fruitlets was low in all cultivars; in the expansion stage, there was rapid absorption of about 30% of total calcium, whereas in the ripening fruit, content of calcium was reduced. The calcium accumulation increased with fruit growth in stalk, similar to that in fruit.     

番木瓜种子中异硫氰酸苄酯（BITC）的抑癌试验

利用MTT法检测番木瓜种子中的异硫氰酸苄酯(BITC)在体外对肝癌细胞等6种常见癌细胞的生长抑制率。结果表明,番木瓜种子提取物中BITC的浓度达到5μmol.L-1(相当于0.745mg.L-1)时,对人肝癌HepG2细胞、人肺癌A549细胞、人乳腺癌MCF-7细胞、人大肠癌HCT-8细胞、人宫颈癌HeLa细胞、人前列腺癌DU-145细胞的生长抑制率均达到70%以上,其中,人肝癌细胞HepG2最敏感,cBITC=2.5μmol.L-1时抑制率可达87%,在cBITC=10μmol.L-1和5.0μmol.L-1时,人大肠癌HCT-8细胞的生长抑制率几乎相同。因此,BITC具有良好的抑癌效果。     

两种季铵盐阳离子表面活性剂对水生生物的毒性效应

为评价两种季铵盐阳离子表面活性剂对水生态的安全性,运用评价化学品对水生生物毒性的标准试验方法,测定了十二烷基二甲基苄基氯化铵(1227)和双八、十烷基季铵盐(C8-10)两种季铵盐阳离子表面活性剂对3种水生生物(斜生栅藻、大型溞和斑马鱼)的毒性。结果表明,在0.05~0.5 mg.L-1的浓度范围内,1227、C8-10对斜生栅藻的毒性表现出明显的剂量-效应关系,随处理浓度的增大抑制效应增强,0.5 mg.L-1C8-10对斜生栅藻具有杀灭作用;同时1227、C8-10与斜生栅藻还存在时间-效应关系,处理后抑制率随时间呈先升高后下降的趋势,72 h时抑制率最高。1227和C8-10对斜生栅藻96 h的EC50分别为0.109、0.103 mg.L-1,对大型溞的48 h LC50分别为1.56×10-2、1.45×10-2mg.L-1,对斑马鱼的96 h LC50分别为2.36、2.19 mg.L-1。C8-10对3种水生生物的毒性均高于1227,两种季铵盐阳离子表面活性剂对大型溞和斑马鱼的致死率(LC50)均随作用时间延长逐渐减小。     

泡桐AFLP反应体系的建立及引物筛选

以豫杂一号泡桐为材料,通过对影响AFLP技术体系的各主要因素的研究,建立了适于泡桐AFLP分析的技术体系.结果表明最佳酶切体系(20μL)为500 ng模板DNA,3 U的Pst 1和Mse I,在37℃下双酶切3 h;20μL最佳连接体系中为酶切产物15 μL,0.25 μmol·L-1Pst I接头,2.5 μmol·L-1 Mse I接头,1 μL 10×T4 Buff-er,2 U T4连接酶,22℃连接18 h;20 μL最佳预扩反应体系中5 μL稀释10倍的连接产物,100 μmol·L-1dNTP,2 U Taq酶,250 μmol.L-1 Pst I和Mse I引物(P+AGT/M+AGT),2 μL 10×PCR Buffer.20 μL最佳选择性扩增反应体系中5 μL稀释20倍预扩增产物,100 μmol·L-1dNTP,2 U Taq酶,350 μmol·L-1 Pst I和Mse I引物(P+AGT/M+AGT),2 μL 10×PCR Buffer.最后,筛选出了97对适宜于泡桐AFLP分析的引物.     

体外氧化μ-钙激活酶对牛肉肌原纤维蛋白降解的影响

在体外,将μ-钙激活酶用不同浓度的氧化剂(0μmol·L-1H2O2+0μmol·L-1Fe2+、300μmol·L-1H2O2+600μmol·L-1 Fe2+、400μmol·L-1 H2O2+800μmol·L-1 Fe2+、500μmol·L-1 H2O2+1 000μmol·L-1 Fe2+)氧化,并用其孵育分离纯化的牛肉肌原纤维蛋白,通过SDS-PAGE和Western blotting分析氧化的μ-钙激活酶对肌原纤维蛋白降解的影响。结果表明:随着氧化剂浓度的升高,μ-钙激活酶降解肌间线蛋白和肌钙蛋白T的能力减弱,同时相对分子质量为30×103肌钙蛋白T降解的产物减少。表明,μ-钙激活酶的氧化抑制其对部分肌原纤维蛋白的降解。     

单壁碳纳米管/玻碳电极同时测定邻苯二酚和对苯二酚的研究

通过在裸玻碳电极表面修饰单壁碳纳米管,制备了单壁碳纳米管/玻碳电极(SWNTs/GC).用循环伏安法研究了邻苯二酚和对苯二酚2种异构体在该电极上的电化学行为.结果表明,该修饰电极对苯二酚的2种异构体表现出了优异的识别能力和电催化性能.对苯二酚和邻苯二酚的氧化峰电位差值为106 mV,这表示2种异构体可以在SWNTs/GC电极上完全分开.基于对苯二酚和邻苯二酚在SWNTs/GC电极上的电化学行为,建立了苯二酚2种异构体同时测定的方法.在最佳的实验条件下,邻苯二酚的氧化峰电流与其浓度在3.0×10-6～8.0×10-5mol·L-1范围内呈线性关系,检出限为1.0×10-6mol·L-1.对苯二酚的氧化峰电流与其浓度在8.0×10-6～1.0×10-4mol·L-1范围内呈线性关系,检出限为2.67×10-6mol·L-1.     

桦褐孔菌原生质体制备与再生

采用正交设计方法对桦褐孔菌原生质体最佳制备与再生条件从酶条件、酶解时间、温度等5个方面进行了研究。试验结果表明:其原生质体制备最佳条件是以液体静置培养6d的菌丝体,0.6mol·L-1KCl作为渗透压稳定剂,在30g·L-1溶壁酶作用下,32℃酶解4h,制备率达2.675×107个·mL-1;再生最佳条件是液体静置培养10d的菌丝体,0.6mol·L-1NaCl为渗透压稳定剂,在30g·L-1溶壁酶作用下,32℃酶解2h,再生率为6.83%。     

白刺的原生质体制备与培养

以西伯利亚白刺幼苗子叶为材料,采用L(934)正交试验,比较酶组合、渗稳剂甘露醇浓度、酶解温度和酶解时间对其原生质体制备的影响。试验结果表明,适宜酶解条件为1%纤维素酶OnozukaR-10+0.2%果胶酶Y-23+0.8mol·L-1甘露醇,28℃下酶解3.5h。而且,经过预处理的子叶,其原生质体得率优于未处理的。在MS+2,4-D1mg·L-1+6-BA0.5mg·L-1+0.7mol·L-1甘露醇的固液双层培养基中,原生质体能形成细胞团。     

不同精粗比底物时添加大蒜油对体外瘤胃微生物发酵的影响

体外法研究了底物精粗比(m(干草)∶m(精料))分别为10∶0、7∶3、5∶5、3∶7时,向发酵液中添加大蒜油(0、30、300和3 000 mg.L-1)对瘤胃微生物厌氧发酵24 h的影响。结果显示:24 h产气量随着大蒜油浓度的增加而降低(P<0.05),每隔3 h产气量的动态变化显示大蒜油延缓发酵的进行。大蒜油降低总挥发性脂肪酸(VFA)浓度(P<0.05),但是30 mg.L-1和300 mg.L-1之间没有显著差异(P>0.05);30 mg.L-1和300 mg.L-1大蒜油降低乙酸比例和乙酸/丙酸(P<0.05),增加丙酸和丁酸比例(P<0.05)。3 000 mg.L-1大蒜油增加pH值(P<0.05),而30 mg.L-1和300 mg.L-1时对pH值没有影响(P>0.05)。氨态氮(NH3-N)和菌体粗蛋白浓度从大到小分别依次是对照组、300 mg.L-1、30 mg.L-1、3 000 mg.L-1和30 mg.L-1、对照组、3 000 mg.L-1、300 mg.L-1。相对于其他发酵底物,在高精料时,30 mg.L-1和300 mg.L-1大蒜油的24 h产气量和总VFA浓度的降低较小,而NH3-N浓度、乙酸比例和乙酸/丙酸的降低以及丙酸比例的增加比较明显。结论:大蒜油抑制体外发酵存在剂量依赖效应,具有延缓发酵进程的特点,高精料时中等水平大蒜油对发酵的抑制作用较小,对发酵的优化作用更明显。     

高效液相色谱-荧光检测法同时分析沼液中4种喹诺酮类抗生素

通过对沼液样品预处理过程优化、提取剂与洗脱剂的筛选和检测色谱条件优化,建立了固相萃取-高效液相色谱-荧光检测法分析沼液中4种喹诺酮类抗生素(FQs)的检测方法。该方法选用Na2EDTA溶液作为提取剂,调节样品pH=4,漩涡混匀后经HLB固相萃取柱净化,选用6 m L甲醇为洗脱剂,浓缩后用高效液相色谱-荧光分析法(HPLC-FLD)检测,采用乙腈/0.01 mol·L~(-1)四丁基溴化铵作为流动相,外标法定量。4种FQs在0.002~5.0 mg·L~(-1)浓度范围内呈现良好线性关系,R~2均大于0.990。样品在0.1、0.2、0.5、1.0、2.0、5.0 mg·L~(-1)6个添加水平下的平均回收率在80.4%~105.7%,相对标准偏差为1.0%~5.1%,方法的检出限为0.004~0.01 mg·L~(-1)。应用该方法对南京地区分别以鸡粪、猪粪、牛粪为发酵原料的3个大中型沼气工程的沼液样品进行分析,4种喹诺酮类抗生素均被检出,检测浓度为5~204μg·L~(-1),表明沼液中抗生素污染问题值得关注。     

瓦松快繁体系的构建研究

以瓦松为研究材料, 系统地研究了丛生芽的诱导、增殖及生根培养等,探索瓦松快繁的最佳途径并构建其快繁体系,结果表明: 茎段接种于MS+10 mg·L-16\|BA+02 mg·L-1IBA+20 g·L-1蔗糖培养基上,茎段增殖倍数可达1169,叶片接种于MS+15 mg·L-16\|BA+01 mg·L-1IBA+30 g·L-1蔗糖培养基上,能产生很好的不定芽,其再生芽数达971。在1/2 MS+02 mg·L-1IBA+005 mg·L-1NAA培养基中,生根率达100%,平均每株能产生123条根,植株生长健壮。