昆虫感受气味物质的分子机制研究进展

摘要：昆虫的嗅觉识别过程是非常复杂的，多种蛋白参与了这一过程，这些蛋白包括气味结合蛋白、气味降解酶以及气味受体等。综述了气味结合蛋白、气味降解酶、气味受体以及化学电信号的转化和传导等方面的最新研究进展。     

一个棉铃虫触角特异表达基因cDNA片段的克隆

摘要：昆虫触角嗅觉感器的主要功能是识别空气中的挥发性气味分子，它们直接与外界环境接触，外界环境中许多分子对昆虫细胞具有毒性，昆虫触角中必定存在某种机制分解这些毒性物质以及多余的气味刺激物。利用差异显示PCR的方法分离得到了3条棉铃虫(Helicoverpa armigera )触角专一性的mRNA片段，在GenBank中进行同源搜索表明，克隆A4-1与昆虫谷胱甘肽-S-转移酶基因家族具有很高的同源性。Northern杂交表明，克隆A4-1在棉铃虫触角中专一性表达，并且在雄蛾触角中的表达量明显比雌虫中高，这与棉铃虫性外激素结合蛋白基因的表达相似。根据该基因在昆虫触角中的表达情况以及已经报道的谷胱甘肽-S-转移酶在昆虫中所起的作用，推测该基因与性外激素的分解有关，同时可能参与触角中毒性分子的分解。     

拟南芥snc2-1D通过WRKY70激活受体类蛋白介导的免疫反应

植物在长期的自然进化过程中形成了两种抵御病原菌的策略方法:一种是由植物细胞膜受体感受病原菌相关分子模式激活的免疫反应(PTI),主要的识别受体包括FLS2、EFR和CERK1等;另一种是抗性蛋白特异性识别某些病原菌的效应蛋白激活免疫反应(ETI),这类的抗性蛋白统称为R蛋白。研究表明,植物体内主要存在三类R蛋白,最大的一类是包含核苷酸结合位点     

麦长管蚜嗅觉相关蛋白Gqα基因的克隆及真核表达

根据已报道无脊椎动物嗅觉相关蛋白Gqα的氨基酸序列保守区设计简并引物,以麦长管蚜(Sitobion avenae)有翅成蚜cDNA为模板,结合RT-PCR及RACE技术扩增编码麦长管蚜Gqα蛋白的cDNA序列,其cDNA序列开放阅读框长为 1 062 bp,编码352个氨基酸,预测蛋白分子量为40.8 kD,与GenBank多个物种Gqα cDNA序列及氨基酸序列均有很高相似性(≥82.17%)(GenBank登录号为EF638906),并具有Gα亚基q类蛋白的典型特征。利用TOPO及Gateway技术,通过体外重组反应,构建苜蓿丫纹夜蛾核型多角体杆状病毒(Autographa californica nuclear polyhedrosis virus, AcMNPV)重组病毒。将N端融合了V5-His6标记的V5-His6Gqα序列整合到AcMNPV病毒基因组DNA中, 得到具有独立转染活性的重组杆状病毒。转染粉纹夜蛾(Trichoplusia ni)离体细胞系Tn-5B1-4(Tn细胞),进行Gqα蛋白的真核表达。SDS-PAGE检测到预期分子量约42 kD的蛋白带,Western blot检测到表达产物。表明含有6个组氨酸标签及V5抗体结合位点的Gqα融合蛋白V5-His6Gqα在昆虫离体细胞系中成功地表达。     

基于iTRAQ技术的不同耐热性水稻花药差异蛋白分析

为从蛋白质水平揭示耐热与热敏感水稻花药响应高温胁迫的分子机制,本研究以耐热水稻品系996和热敏感水稻品系4628为材料,采用iTRAQ技术对高温和适温条件下水稻花药进行差异蛋白质组学分析。结果表明,在996高温-996适温、4628高温-4628适温、996高温-4628高温和996适温-4628适温4个比较组中,共筛选到957个差异表达蛋白,其中上调表达蛋白398个,覆盖率达20%以上的蛋白占鉴定总蛋白的50.96%。GO分析显示,抽穗开花期花药差异蛋白主要集中于代谢过程和细胞过程,在细胞组成方面主要分布于胞内部分、细胞器和细胞,分子功能主要涉及催化活性、绑定等。花药差异蛋白通路分析显示,来源于bZIP和DOF家族的2个转录因子差异表达,DOF家族基因上调,bZIP家族基因下调;18个HSPs基因中大部分表达上调;与植物激素和信号传导相关基因大部分表达下调;10个活性氧(ROS)相关基因中5个表现为上调;与次生代谢相关基因大部分下调。本研究结果为揭示水稻花药高温胁迫的应答分子调控机制提供了一定的理论基础。     

菜豆根瘤菌Rhizobium etli CFN42全基因组含信号肽分泌蛋白的鉴定与分析

利用菜豆根瘤菌Rhizobium etli CFN42全基因组测序结果,结合计算机技术和生物信息学分析方法,对菜豆根瘤菌蛋白质组中的分泌蛋白进行预测和分析,以便更深入地了解共生固氮的分子机制。经过分析发现,在该根瘤菌5 963个蛋白质中,具有信号肽的分泌蛋白332个,占总数的5.4%主要涉及底物转运、物质代谢和信号转导等方面。在有功能描述的186个分泌蛋白中,识别了Ⅰ、Ⅱ、Ⅲ、Ⅳ型分泌系统和丝氨酸蛋白酶,这些蛋白质可能参与了根瘤菌对宿主植物的侵染,是两者分子对话的关键参与者;在假定蛋白中,通过结构域分析,发现了SH3b、BA14K和EFh结构域,含有这些结构域的分泌蛋白可能参与了根瘤信号转导途径。在根瘤菌共生结瘤过程中,分泌蛋白发挥了重要作用。     

基于DSP的苹果气味检测装置设计与实践

依据划分苹果等级的气味指标,由气味传感器阵列和DSP构成电子鼻系统采集苹果气味信号进行处理,建立苹果气味识别模型。研究气味传感器阵列的组成形式以及其采集到的数据,设计了传感器与DSP的接口数据采集电路以及视频显示接口电路。     

常规稻与杂交稻谷的仿生电子鼻分类识别

气味是进行稻谷品种及其品质识别的重要方法之一,作为一种基于仿生嗅觉的机器检测方法,仿生电子鼻在水稻品种的分类识别中具有较好的应用前景。常规稻与杂交稻在食味品质等方面存在一定的差异,为了解应用电子鼻进行常规稻谷与杂交稻谷识别的可行性,采用PEN3电子鼻对同季同地域收获的3种常规稻(中香1号、湘晚13、瑶平香)和3种杂交稻(伍丰优T025、品36、优优122)稻谷样品的气味信息进行了采集和分析。首先通过过载分析(Loadings)法分析了电子鼻检测稻谷气体挥发物时的各传感器贡献率,分别针对基于特征值的提取和稻谷气味检测对电子鼻传感器阵列中的传感器进行了优选,阐明了稻谷气体挥发物检测中应以对硫化物、氮氧化合物、芳香成分和有机硫化物敏感的传感器为主。随后,分别采用主成分分析法(principal component analysis,PCA)、线性判别法(linear discriminant analysis,LDA)和BP神经网络对6种不同稻谷之间、常规稻与杂交稻之间的分类识别进行了研究。结果表明,PCA分析法与LDA分析法在对6种不同稻谷之间的分类以及常规稻与杂交稻之间的分类中均未取得理想的效果,存在部分样本数据点重叠或样本数据点较近的情况,在实际应用中易发生混淆;而BP神经网络在对6种不同稻谷之间的分类中对测试集的识别正确率分别达到了90%,在常规稻与杂交稻之间的分类识别中对测试集的识别正确率达到了96.7%。上述试验验证了电子鼻用于常规稻与杂交稻稻谷分类识别的有效性,为常规稻与杂交稻的快速、无损分类识别提供了一种新的方法。     

吹扫式仿生嗅觉检测装置的设计与性能试验

为研究仿生嗅觉检测装置的检测性能,该文研制了一种吹扫式仿生嗅觉检测装置,选取传感器阵列灵敏度、响应时间和恢复时间作为装置性能指标,分析了传感器加热电压、待检气体湿度以及流量等控制参数对嗅觉检测性能的影响,并进一步验证优化控制参数的仿生嗅觉检测装置对气味检测的可行性和检测性能。试验结果表明:传感器阵列灵敏度随加热电压增大而增大,其响应和恢复时间随加热电压增大而减小,加热电压为5.0 V时,装置性能较佳,其传感器阵列灵敏度分布范围为2.260~4.823,响应和恢复时间分布范围分别为46~53 s、44~70 s;增加气体湿度会使传感器阵列灵敏度减小,同时也会延长响应和恢复时间,气体相对湿度为30%时,装置性能较佳;传感器阵列灵敏度随载气流量的增加先递增后减小,其响应和恢复时间随流量的增加先减小后递增,流量为100 m L/min时,装置性能较佳,其传感器阵列灵敏度分布范围为2.853~7.559,响应和恢复时间分布范围分别为35~50 s、30~50 s;在优化控制参数下,待检气体体积分数范围控制在0.002%~0.020%时,装置检测灵敏度较高,范围为3 577.1~6 700.7;线性特性和重复性较好,决定系数范围为0.901~0.997,变异系数范围为0.832%~9.696%,能满足仿生嗅觉气味检测的要求,可为后续开展仿生嗅觉的应用性研究提供数据参考与技术支撑。     

嗅觉可视化技术及其对四种食醋的识别

不同的食醋散发的香味具有一定的相似性,同时又有各自的特征,怎样快速表达这些特征是一大难点.本研究根据Kenneth S.Suslick教授的思路,筛选出23种卟啉类化合物和7种疏水性pH指示剂,制成一个5x6气体可视化传感器阵列.文章详细介绍了嗅觉可视化系统及其操作步骤,并利用气体可视化传感器阵列对4种传统食醋进行了测试.聚类分析表明,当取相似度为12时,样本被区分为镇江香醋、山西老陈醋、四川麸醋、浙江玫瑰米醋.研究表明嗅觉可视化技术可用于食品气味的可视化分析和图像指纹表达.     

Binding of PF2 lectin from Olneya tesota to gut proteins of Zabrotes subfasciatus larvae associated with the insecticidal mechanism

Zabrotes subfasciatus (Boheman) is the main pest of common beans ( Phaselous vulgaris ). Wild legume seeds from Olneya tesota contain a lectin, PF2, that shows insecticidal activities against this insect. The binding of PF2 to midgut glycoproteins of 20-day-old larvae was evaluated using PF2 affinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the proteins retained on the gel revealed several putative glycoproteins, ranging in mass from 17 to 97 kDa. Subsequent protein digestion and analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) provided amino acid fragments that identified an α-tubulin, cytochrome c oxidase subunit I, an odorant receptor, and a lysozyme from available insect sequence databases. The potential of these proteins to serve as part of the mechanisms involved in the insecticidal activity of PF2 to Z. subfasciatus is discussed.     

Protective roles of plant proteins in conferring tolerance to heat stress

Intensified by global warming, heat stress is one of the major concerns in many farmlands that severely disturb plant productivity. Plant genetic engineering by regulating heat shock proteins, RNA-binding proteins, and molecular chaperones is an efficient strategy for maintaining plant productivity under heat stress conditions. Plant performance and tolerance to heat stress are highly dependent on their balance, interactions, structural, and functional diversities. Thus, this review article is undertaken to discuss the effectiveness of these proteins in conferring tolerance to heat stress and to incentivize their coregulation to achieve better results in breeding programs in future.     

Isolation and identification of two lipid transfer proteins in pomegranate (Punica granatum)

Lipid transfer proteins (LTPs) are a family of low molecular mass (7-9 kDa) polypeptides, the members of which share 35-95% sequence homology. These proteins are widely distributed throughout the plant kingdom and are receiving attention for their biochemical characteristics and biological activity. LTPs are indeed studied in different research fields varying from allergy to food technology, and numerous molecules belonging to this class are progressively being identified and investigated. Proteins from pomegranate juice were fractioned by cation exchange chromatography and analyzed by SDS-PAGE. Two proteins were identified as putative LTPs on the basis of their molecular weights and their electrophoretic behaviors under reducing and nonreducing conditions. Finally, proteins were purified and characterized by mass spectrometry. This analysis confirmed that the two polypeptides are LTPs on the basis of an amino acid sequence common to LTPs from other plant sources and cysteine content. The two proteins, named LTP1a and LTP1b, showed similar molecular masses but different immunological profiles when immunodetected with rabbit antibodies specific for Pru p 3 and human IgE from a patient suffering from pomegranate allergy. The demonstration of the existence of two immunologically unrelated LTPs in pomegranate confirms the variability and the complexity of the plant LTP family. This should be taken into account when the role of these proteins as elicitors of allergies to fruits is investigated and could help to explain the contradictory literature data on pomegranate allergy.     

Water deficit stress,ROS involvement,and plant performance

Crop productivity is impaired by stress factors, biotic or abiotic. The main are pathogens, diseases, insects, cold, heat, salinity, drought, radiation and others. Among these unfavorable conditions, drought is one of the main occurrences and negatively affects crop development. This environmental adversity generally induces the accumulation of reactive oxygen species (ROS). These molecules lead to oxidative stress, and at high levels cause cell effects, like loss of organelle functions, electrolyte leakage, and reduction in metabolic efficiency. High concentration of ROS in cells can still cause molecular damage that include damage in proteins, amino acids, and lipids, and even lead to cell death. To neutralize these damages, plants increase enzymatic antioxidant activity and non-enzymatic antioxidant contents. ROS are essential to life in plants, and at basal levels performs cellular functions, such as signaling and defense responses. Here, we focus on the ROS production, the involvement and damages of these species in water deficit condition, changes in activity of antioxidant enzymes and non-enzymatic antioxidant contents in plants under drought stress. In addition, the signaling reactions of ROS under stress water restriction, changes on yield components of species under water deficit and the antioxidant genes involved in plant responses to stress were also addressed.     

The challenge of conserving grassland insects at the margins of their range in Europe

Insects have experienced higher rates of decline than other popular taxa in the calcareous grasslands of Europe. This paper reviews reasons why insects are so vulnerable to environmental change, particularly at the edges of their range. Case studies of the ant Myrmica sabuleti and the butterflies Polyommatus bellargus and Thymelicus acteon are used to illustrate the mechanisms responsible for this vulnerability and the need for different management regimes to maintain species’ niches in different parts of their range.     

Olfactory cues associated with fungal grazing intensity and secondary metabolite pathway modulate Collembola foraging behaviour

Fungal secondary compounds play an important role for springtail food choice and fitness. Little is known, however, on the role of olfactory cues for Collembola foraging behaviour and whether Collembola can olfactorily perceive volatiles associated with fungal secondary metabolite pathways. We investigated the ability of three species of Collembola (Folsomia candida, Heteromurus nitidus and Supraphorura furcifera) to use olfactory cues for discriminating between fungi of different phylogenetic affiliation (Aspergillus nidulans, Cladosporium cladosporioides, Ascomycota; Laccaria bicolor, Basidiomycota) and toxicity using fungal strains of A. nidulans with reduced secondary metabolite production. Furthermore, we studied if olfactory cues from hyphae injured by grazing affect the foraging behaviour of Collembola. We hypothesized that (i) Collembola are able to olfactorily perceive and respond to fungal species/strains with different secondary metabolite pathways and that (ii) Collembola are able to sense fungal mycelia injured by grazing. Each of the Collembola species studied preferentially oriented their foraging towards fungal strains of A. nidulans with suppressed secondary metabolites, and in particular towards the mutant where the global regulator for secondary metabolites (ΔlaeA) has been silenced. Two of the three Collembola species (H. nitidus and S. furcifera) sensed olfactory cues of previously grazed fungi and moved towards ungrazed fungi, however, the response of S. furcifera was restricted to fungi extensively exposed to grazing (5 days) suggesting that the response varies between Collembola species.Overall, the results support the first and second hypothesis indicating that Collembola (1) are able to olfactorily differentiate fungi of different toxicity and move towards more palatable fungi, and (2) showed a lesser preference for fungi previously exposed to grazing.     

Lectins and protease inhibitors as plant defenses against insects

Plants commonly accumulate lectins and proteinaceous protease inhibitors in their various tissues, sometimes in high concentrations. Much evidence suggests that one of the functions of these proteins is to serve as defenses against insects.     

蛋白质组学在生鲜肉肉色变化机制研究中的应用

肉色是影响消费者购买欲望的最直观因素。宰后生鲜肉肉色的变化机制和肉色改善一直是肉类科学领域研究的热点。肉色体系复杂,其稳定性受动物宰前因素、从肌肉向可食用肉转化过程和宰后商业流通链条件等因素的影响。目前,仍有许多影响途径和作用机制不够清楚。研究者已采用蛋白质组学技术发现了许多与肉色相关的蛋白标志物和可能影响途径。该文基于近年的研究,综述了不同处理条件下与肉色相关的差异蛋白,主要包括结构性蛋白中的肌动蛋白、肌球蛋白和肌联蛋白,以及肌浆蛋白中的伴侣蛋白、热休克蛋白、代谢酶类和氧化还原酶类等,分析了差异蛋白表达对肉色的影响及其表达调控肉色体系的可能机制,为今后肉色变化机制的研究和肉色稳定性的调控提供了思路。