外源海藻糖对高温胁迫下肺形侧耳氧化损伤的缓解效应

高温胁迫影响食用菌的品质和产量,由高温引起的氧化胁迫是导致损伤的主要原因.以肺形侧耳(Pleurotus pulmonarius)热敏感菌株CCMSSC 00494和耐热菌株CCMSSC 00499为试验材料,研究了外源海藻糖对其菌丝体高温胁迫下氧化损伤的缓解效应.结果表明,高温(40℃)胁迫48 h,热敏感菌株中硫代巴比妥酸反应物(TBARS)含量(代表氧化损伤程度)高于耐热菌株.外源海藻糖处理可以显著降低高温胁迫下菌丝体内O2-产生速率、H2O2含量、脂氧合酶(LOX)活性和TBARS含量水平,缓解高温胁迫所引发的氧化损伤.另外,外源海藻糖对超氧化物歧化酶(SOD)活性有保护和提高的作用,对过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性有抑制作用,对过氧化物酶(POD)活性影响不明显.外源海藻糖对热敏感菌株的保护效应高于耐热菌株.     

Potassium fertilization improves apple fruit (Malus domestica Borkh. Cv. Fuji) development by regulating trehalose metabolism

Potassium (K) fertilization and antioxidant enzymes both positively influence plant growth and development. However, it is not known whether K treatment improves fruit development via increasing soluble sugar. In this study, K-treated apple (Malus domestica Borkh.) fruit was harvested from 60 to 150 days after full bloom and was analyzed for ROS production and trehalose metabolism. The results show that K fertilization improved fruit firmness, increased growth according to several parameters, increased soluble sugar content, and decreased ROS production. The ascorbate metabolic pathway more effectively reduced ROS production than catalase and peroxidase (POD) did under K treatment. Trehalose-treated fruit also showed higher activity of ascorbate-related enzymes (DHAR, GR, and APX) compared with non-treated fruit. The changes of antioxidant enzyme activity in trehalose-treated fruit corresponded to those in K-treated fruit. Moreover, trehalase (TREH) activity in fruit was notably reduced by K treatment. This demonstrates that K influences ROS production via regulating trehalose content and TREH activity in fruit. This study provides new insight into the K mechanism which improves fruit development, including fruit firmness and size.     

谷氨酸棒杆菌麦芽寡糖基海藻糖合成酶基因的克隆与表达

采用PCR方法从谷氨酸棒杆菌（Corynebacterium glutamicum）基因组中分离得到2.4 kb的麦芽寡糖基     

Analytical validation of a portable human Accu-Chek glucometer in honeybee hemolymph

Glucose and trehalose are the main energy sources used by honeybees (Apis mellifera) for daily activities. However, there is no validated point-of-care method to reliably measure both sugars. We performed an analytical validation of a portable human glucometer (Accu-Chek; Roche) for glucose measurement in honeybee hemolymph compared to a reference method (GluCH, UniCel DxC 600; Beckman Coulter). We used 30 pooled hemolymph samples collected from the antennae of anesthetized honeybees and diluted 1:4 in 0.9% saline. We evaluated dilution linearity, spike recovery, and inter- and intra-assay imprecision. Glucose concentration was measured over time (2 h, 4 h, 8 h, 12 h, 1 d, 2 d, 3 d, 7 d, 21 d, 28 d) at various storage temperature (25°C, 4°C, −20°C, −80°C). The trehalose concentration was measured indirectly by trehalase hydrolyzation. Glucose concentrations measured by both instruments had a strong correlation (0.985, p < 0.0001) and a bias of −7.33 mmol/L (±1.96SD: 13.70 to −28.36), with linear agreement at <20 mmol/L (physiologic value: 100 mmol/L). The accuracy of the glucometer decreased at >20 mmol/L. Recovery of 115–130% of diluted spikes indicated good specificity. Inter- and intra-assay imprecision were 2.50% and 2.21%, respectively. Glucose concentrations fluctuated in stored samples dependent on time and temperature; however, glucose concentrations were constant with storage at −80°C for ≥28 d. The Accu-Chek glucometer is an adequate instrument to measure honeybee glucose concentration in hemolymph diluted with 0.9% NaCl, with good accuracy and precision at <20 mmol/L. Hemolymph storage at −80°C is suitable for long-term conservation of glucose.     

外源海藻糖对家蚕生长和茧丝品质的影响

[目的]探讨外源海藻糖对家蚕的生长和茧丝品质的影响。[方法]通过对家蚕品种菁松×皓月进行添食试验,研究添加海藻糖对家蚕的生长和茧丝品质的影响。[结果]添加海藻糖后家蚕上蔟更为整齐,茧丝纤度降低,而生存率、体重、4龄结茧率、普茧率、解舒率、粒茧丝长和解舒丝长都增加。综合评价结果表明,添加20 mmol/L海藻糖的家蚕成绩最优。[结论]外源性海藻糖对提高家蚕生长和茧丝品质具有积极作用。     

稀释液中添加海藻糖对猪冷冻精液质量参数和活性氧的影响

本试验旨在探讨海藻糖对猪精子冷冻效果以及精子内活性氧(ROS)的含量影响。试验分对照组和4个海藻糖处理组(0.025,0.05,0.1和0.2 mol/L)。精子冷却后,添加0.1 mol/L海藻糖处理组精子活力显著(P<0.05)高于对照组,而精子内和稀释液中各组间ROS发生量都没有显著的差异。精子冷冻解冻后,添加0.05 mol/L海藻糖处理组的精子活力显著(P<0.05)高于对照组;添加0.05 mol/L和0.1 mol/L海藻糖处理组的精子生存率显著(P<0.05)高于对照组;添加0.1 mol/L海藻糖处理组顶体完整的精子百分比显著(P<0.05)高于对照组;4个海藻糖处理组膨胀精子百分比都显著(P<0.05)高于对照组;添加0.025 mol/L海藻糖处理组ROS水平显著(P<0.05)低于对照组。结果表明:海藻糖对精子冷冻保存是有益的,且能抑制精子内ROS的发生,但它的作用机制有待于进一步的研究。     

两个褐飞虱海藻糖转运蛋白基因的结构及调控海藻糖代谢功能

【目的】海藻糖是昆虫中的主要血糖物质,在昆虫的发育及生理活动中发挥重要功能。其中,海藻糖转运蛋白(Tret)在将海藻糖从其生成组织(例如脂肪体)运输到其消耗组织的过程中起着重要作用。本研究通过分析褐飞虱(Nilaparvata lugens)两条Tret1的序列结构,进一步抑制Nl Tret1的表达,探讨这两个Nl Tret1在褐飞虱体内的生物学功能。【方法】以褐飞虱两条Tret1序列为研究对象,利用生物信息学技术分析其蛋白结构以及与其他昆虫之间的同源性。采用RNAi(RNA interference)技术将合成的外源ds RNA(double-stranded RNA)注射到实验室饲养褐飞虱种群体内,抑制其体内Nl Tret1的表达,分别在注射48 h后取材,抽取总RNA并用反转录试剂盒合成第一链c DNA,采用q RT-PCR(quantitative real-time PCR)技术检测ds Nl Tret1的干扰效果以及RNAi后褐飞虱体内海藻糖代谢通路中相关基因的表达,最后测定葡萄糖、海藻糖和糖原含量以及海藻糖酶活性。【结果】生物信息学分析表明,Nl Tret1-like X...     

水生栖热菌FL-03海藻糖合酶的分离纯化及特性研究

将从水生栖热菌FL-03中获得的海藻糖合酶粗酶液经硫酸铵分级沉淀、DEAE Sepharose CL-6B离子交换层析S、ephacryl S-200HR凝胶过滤层析后,纯化68.48倍,产率为18.4%。研究表明:该酶分子量约为101 kD,最适反应温度为60℃,最适pH为7.0,在40~80℃、pH 6.0~9.0范围内具有较高的反应活性和稳定性。Fe3 对该酶具有一定的激活作用,Cu2 、Tris、Zn2 对该酶具有较强的抑制作用。该酶具有高度的底物特异性,只能专一地将麦芽糖转化为海藻糖。在40℃4、8 h条件下,麦芽糖转化为海藻糖的转化率最大可达到79%。     

外源海藻糖调节西瓜细胞渗透胁迫抗性的研究

为探究外源海藻糖对渗透胁迫下西瓜细胞生长的影响,本研究以西瓜悬浮培养细胞为试材,测定外源海藻糖预处理后,渗透胁迫对西瓜悬浮培养细胞生长量、细胞外pH、细胞内ROS（活性氧簇）相对含量和微管骨架的影响。结果表明：渗透胁迫下西瓜细胞的生长量明显受到抑制,并且渗透胁迫可诱导西瓜悬浮培养细胞质外体碱化,ROS迸发,细胞微管骨架发生解聚;外源添加海藻糖可在一定程度上缓解渗透胁迫对西瓜悬浮细胞生长量的抑制作用,调节pH、ROS表达水平、并维持微管骨架结构的完整性。以上研究结果表明渗透胁迫下,海藻糖对西瓜细胞具有维持细胞生长、保护亚细胞结构并调节抗性反应的功能。     

海藻糖对盐胁迫下两个沙棘品种逆境生理的影响

摘要： 通过研究海藻糖对沙棘逆境生理指标的影响规律,探索海藻糖的抗盐生理机制,为生产中应用海藻糖缓解沙棘 盐胁迫提供理论依据 。试验在盆栽条件下,设置空白对照、盐胁迫 + 不同浓度的海藻糖(0、0.2、0.4、0.6 mmol·L-1 )共计 5个处理,3次重复 。结果表明:0.4 mmol·L-1 的海藻糖可以显著提高盐胁迫下两个沙棘品种叶片内的叶绿素、游离脯氨 酸含量,降低 MDA含量;海藻糖会提高盐胁迫下两个品种沙棘叶片内海藻糖-6-磷酸磷酸酶(TPP)活性,提高可溶性糖和可 溶性蛋白含量;0.4mmol·L-1 的海藻糖对提高 TPP活性效果优于0.2mmol·L-1和 0.6mmol·L-1处理,有利于可溶性糖的 积累,缓解盐胁迫对沙棘的危害 。海藻糖对 TPP活性的调控作用可能是其缓解沙棘盐胁迫的生理机制,0.4 mmol·L-1 为适 宜的喷施浓度。