RNA干扰对烟草NtPPO8基因沉默的效应分析

多酚氧化酶（PPO）是发生酶促褐变的关键酶,PPO活性过高会使烟叶变褐。为明确烟草NtPPO8基因对PPO活性的影响,建立降低NtPPO8基因表达量的病毒诱导的基因沉默（virus induced gene silencing,VIGS）体系,探究使用RNA干扰技术对烟叶NtPPO8基因表达量及PPO活性的影响。以中烟100和K326为材料,分析在移栽后50d进行VIGS载体侵染的植株,并在侵染后10、20、30、40d分别测定NtPPO8基因表达量及PPO活性。结果表明,接种VIGS载体的转化植株,其中在侵染后20d时NtPPO8基因的沉默效率最高,为78.46%;随着侵染时间推移,NtPPO8基因的沉默效率逐渐降低,在接种后40d内有效表达;且转化植株中的病毒载体会随着植株的生长而向上传导,随着接种部位的上移,沉默效率逐渐降低。因此,研究建立VIGS体系能够有效沉默NtPPO8基因的表达,并且有效降低PPO活性,为提高烟叶耐烤性及降低烟叶褐变比例提供了新的思路和方法。     

葡萄多酚含量和多酚氧化酶（PPO）活性与组培苗生根关系的初步研究

研究测定了葡萄属10个种(品种)组培苗的多酚含量和多酚氧化酶(PolyphenolOxidase,PPO)的活性,对其与组培苗的生根指数进行相关性分析,并建立生根指数与多酚含量、多酚氧化酶的多项式回归方程。结果表明:不同种(品种)间多酚含量和PPO活性有显著性差异;多酚含量和PPO活性相关性显著,PPO活性和生根指数不相关,而多酚含量与生根指数呈显著负相关;从回归方程可以看出,PPO活性对生根指数的作用是非线性的,多酚含量对生根指数的作用是线性的,这说明多酚含量对组培苗生根的直接作用很大,PPO活性通过多酚含量间接影响组培苗生根而且直接作用较小。     

超声波辅助提取山药总多酚的研究

以山药为原料,采用超声波辅助提取从山药中提取多酚类物质,通过单因素试验和正交试验得到总多酚的最佳提取条件是:体积分数50%的丙酮溶液、料液比1∶20,提取60 min,提取温度50℃,提取2次、超声功率800 W,多酚提取率为16.97%。     

抑制剂对鲜榨蜜梨汁多酚氧化酶活性的影响

多酚氧化酶(PPO)是酶促褐变的关键酶,以鲜榨蜜梨汁为原料,以邻苯二酚为底物,采用分光光度法测定多酚氧化酶活性,研究氯化钠、柠檬酸、L-半胱氨酸、抗坏血酸和异抗坏血酸5种不同抑制剂对鲜榨蜜梨汁中的多酚氧化酶活性的影响。结果表明,这5种酶抑制剂均能在一定程度上抑制鲜榨蜜梨汁PPO活性,随着其浓度的升高,抑制效应逐渐增强。氯化钠、柠檬酸、L-半胱氨酸、抗坏血酸、异抗坏血酸抑制鲜榨梨汁PPO活性的最佳浓度分别为0.6,0.5 mol/L和2.0,2.0,0.5 mmol/L。     

UV-C irradiation induces defence responses and improves vase-life of cut gerbera flowers

UV-C (λ = 254 nm) irradiation was effective in reducing Botrytis cinerea floret specking (i.e., lesion development) and maintaining a better postharvest quality of cut gerbera flowers. A range of UV-C doses (0.5-10.0 kJ m⁻²) was tested on ‘Ice cream’ and ‘Ecco’ gerbera flowers to activate germicidal and inducible defence mechanisms. Irradiation of B. cinerea cultures with 0.5, 1.0, 2.5 and 5.0 kJ m⁻² UV-C resulted in up to a 10-fold reduction of conidial germination percentages and significant (P < 0.05) delay of mycelium growth, compared to the non-irradiated control cultures. Moreover, lesion diameters on gerbera florets inoculated with UV-C irradiated B. cinerea cultures were reduced by up to 70%, suggesting that UV-C had a negative effect on the pathogenic strength of the fungi. Lesion diameters on florets of UV-C irradiated gerberas were reduced by up to 55% giving evidence that defence responses in the host tissue were induced. Concentration of total phenolics seemed to be unaffected by 0.5 kJ m⁻² UV-C treatment in both cultivars, but polyphenol oxidase (PPO) activity increased and remained higher compared to the non-irradiated control flowers throughout the 48 h storage period at 20 °C. The increase of PPO suggests that this enzyme might play an important role in host defence mechanisms that suppressed B. cinerea floret specking. Gerbera flowers irradiated with 1.0 or 10.0 kJ m⁻² UV-C showed improvement in vase-life by 1.8 and 2.4 d, decrease in stem break percentages by 43 and 29% and delay in stem break incidence by 3.3 and 1.3 d, respectively.     

抑制荸荠中多酚氧化酶活性的研究

通过单因素及多因素试验,探讨抑制荸荠中多酚氧化酶活性的效果。L_9(3~3)正交试验结果表明:柠檬酸预煮液的质量分数0.5%,预煮温度100℃,预煮时间5 min为最优预煮条件。     

基质含水量变化对盆栽‘红富士’苹果果实苯丙氨酸解氨酶活性及多酚含量的影响

为了解基质水分变化对苹果果实中苯丙氨酸解氨酶(PAL)活性及多酚含量的影响,以盆栽‘红富士’苹果为试材,调控基质水分含量,测定不同时期苹果果实中PAL酶活性和多酚含量。结果表明：果皮和果肉中的苯丙氨酸解氨酶活性均呈现先升后降的趋势,但达到高峰的时间有所差异。基质相对含水量分别由50%~55%和65%~70%提高到80%~85%时,果皮PAL酶活性显著高于同时期的其他处理,但果肉中的PAL活性没有差异;果皮和果肉中多酚含量显著低于同时期的其他处理。     

重组葡萄糖氧化酶毕赤酵母高产菌株的高通量快速筛选方法

根据葡萄糖氧化酶的反应原理和毕赤酵母诱导表达的机制,本研究建立了葡萄糖氧化酶毕赤酵母表达菌株筛选方法。首先在MM平板中诱导重组子表达葡萄糖氧化酶,然后添加显色液,阳性酵母菌株周围可见显色圈。经过与传统液体培养筛选方法的比较验证,结果表明,该方法灵敏度高,稳定性和重复性好,筛选结果与传统方法一致。当葡萄糖氧化酶活力介于0–0.1 U/ml的范围内,显色圈的绝对面积与酶活力相关系数可达0.63。该平板筛选方法可以从大量毕赤酵母重组子中快速筛选出表达葡萄糖氧化酶的阳性菌株,并可方便地判断出各菌株之间表达水平的差异。同时,该方法也可为筛选其他重组酶毕赤酵母菌株提供参考。     

Contribution of mineral nutrition to the response of barley seedlings to polyethylene glycol–induced mild water stress

The effect of polyethylene glycol–induced osmotic stress on the activity of nitrate reductase, glutamine synthetase, and glycolate oxidase in leaves of young barley plants grown under two nutrient‐supply regimes was studied. The activity of nitrate reductase gradually decreased after polyethylene glycol (PEG) application, while glutamine synthetase and glycolate oxidase were increased. It is speculated that the enhanced glutamine synthetase and glycolate oxidase activities are due to increased flux of metabolites through the photorespiratory cycle. Prominent increase in concentrations of free proline, reducing sugars, and free amino acids was observed. The possible contribution of these cellular solutes to the process of osmotic adjustment and the role of mineral supply is discussed. It is suggested that low N supply in combination with stress conditions switched the preferred osmolyte type from amino acids (N‐containing) to sugars (C‐containing).