昆虫嗅觉相关蛋白的研究进展

昆虫对自然环境中复杂化学信号的识别多依赖于其灵敏的嗅觉系统,选择寄主、觅食、寻找配偶等行为的发生都以嗅觉识别为基础,而完成嗅觉识别还需要多种嗅觉相关蛋白的参与。嗅觉相关蛋白主要包括6种,即气味结合蛋白、化学感受蛋白、气味受体、感觉神经元膜蛋白、离子型受体和气味降解酶。不同种类和性别的昆虫中,嗅觉蛋白的种类、数量和分布各不相同。由于嗅觉蛋白在昆虫识别外界气味分子中的重要作用,国内外近年来对其展开了广泛、深入的研究。本文从几种嗅觉相关蛋白的生化特性、分子结构、生理功能、分布表达部位和研究概况等角度,较详细地综述了近年来国内外昆虫嗅觉相关蛋白的研究进展。     

The effect of food type (formulated diet vs. natural) and fish size on feed utilization in common sole,Solea solea (L.)

This study compares the effect of food type (formulated diet vs. natural food) and fish size on protein and energy utilization efficiencies for growth in common sole, Solea solea (L.). Replicate groups of common sole (mean initial body weight ± SD was 45.7 g ± 2.1 and 111.2 g ± 4.2) received the diets at five (natural feed) or four (formulated diet) feeding levels. The protein utilization efficiency for growth (k_gCP) was higher (P > 0.001) in common sole fed ragworms than in common sole fed the formulated diet (respectively, 0.40 and 0.31). Likewise, the energy utilization efficiency for growth (k_gGE) was higher (P = 0.001) in common sole fed ragworms than in common sole fed the formulated diet (respectively, 0.57 and 0.33). The protein maintenance requirement was not different between food types (P = 0.64) or fish size (P = 0.41) being on average 0.82 g kg^?0.8 day^?1. The energy maintenance requirement was not different between food type (P = 0.390) but differed between fish size (P = 0.036). The gross energy maintenance requirement of small common sole was 35 kJ g^?0.8 day^?1. The gross energy maintenance requirement of large common sole was 25 kJ g^?0.8 day^?1. In conclusion, the low growth of common sole fed formulated diets was related to reduced feed utilization.     

Carbohydrate and crude protein fractions in perennial ryegrass as affected by defoliation frequency and nitrogen application rate

The objective of this study was to evaluate the effects of defoliation frequency (either at two‐ or three‐leaf stage) and nitrogen (N) application rate (0, 75, 150, 300, 450 kg N ha^?1 year^?1) on herbage carbohydrate and crude protein (CP) fractions, and the water‐soluble carbohydrate‐to‐protein ratio (WSC:CP) in perennial ryegrass swards. Crude protein fractions were analysed according to the Cornell carbohydrate and protein system. Carbohydrate fractions were analysed by ultra‐high‐performance liquid chromatography. Sward defoliation at two‐leaf stage increased the total CP, reduced the buffer‐soluble CP fractions and decreased carbohydrate fractions of herbage (P < 0·001). The effect of defoliation frequency was less marked during early spring and autumn (P < 0·001) than for the rest of the seasons. An increase in N application rate was negatively associated with WSC, fructans and neutral detergent fibre (P < 0·001), and positively associated with CP and nitrate (N‐NO₃) contents of herbage. Nitrogen application rate did not affect CP fractions of herbage (P > 0·05). The fluctuations in CP and WSC contents of herbage resulted in lower WSC:CP ratios during early spring and autumn (0·45:1 and 0·75:1 respectively) than in late spring (1·11:1). The herbage WSC:CP ratio was greater (P < 0·001) at the three‐leaf than the two‐leaf defoliation stage and declined as the N application increased in all seasons (P < 0·001). The results of this study indicate that CP and carbohydrate fractions of herbage can be manipulated by sward defoliation frequency and N application rate. The magnitude of these effects, however, may vary with the season.     

草鱼鳃组织蛋白双向电泳技术的建立及优化

为建立并优化草鱼鳃蛋白质组双向电泳的技术体系,实验将草鱼鳃经裂解处理后,通过固相p H梯度胶条进行一向等电聚焦和SDS聚丙烯酰胺凝胶垂直电泳进行蛋白分离,对不同的裂解液配方、蛋白纯化方法、IPG胶条的分离范围、上样量和染色方法等进行了探索和优化,并应用lmage Master 2D Platinum 7.0软件对电泳图谱进行了分析。结果表明:采用裂解液中添加DTT来取代Tris和TBP、选择分离范围为pH4-7的IPG胶条和150μg的主动水化上样,电泳结束后对凝胶进行硝酸银染色,获得了覆盖范围广、低背景、高分辨率、适合差异蛋白分析的双向电泳参考图谱。     

Effects of dietary protein and lipid levels on growth performance,fatty acid composition and antioxidant‐related gene expressions in juvenile loach Misgurnus anguillicaudatus

Dojo loach (Misgurnus anguillicaudatus) is one of the most important cultured freshwater fish in several East Asian countries. However, a little information is available in its nutritional requirements. Thus, this study was conducted to evaluate the effects of feeding varying levels of dietary protein and lipid on growth, fatty acid composition and antioxidant‐related gene expressions in juvenile loach. Six practical diets at three levels of protein (30%, 40% and 50%) and two levels of lipid (6% and 12%) were fed to loach juveniles (initial weight 0.40 g) in triplicated groups (20 fish per replicated) for a period of 8 weeks. Results showed that regardless of lipid level, body weight gain of fish was significantly increased with incremental dietary protein level. Meanwhile, feed conversion ratio was significantly decreased by dietary protein levels, and the lowest value was observed in fish fed dietary protein levels of 50%, regardless of dietary lipid level. Moreover, the percentage of 22:6n‐3 in viscera was significantly increased by different protein levels. The expression level of catalase was significantly increased with incremental dietary protein level with both lipid levels. Meanwhile, the expression level of hepatic nuclear factor erythroid 2‐related factor 2 (Nrf2) was downregulated with incremental dietary protein level with 6% of lipid level, but the expression was upregulated with incremental dietary protein level with 12% of lipid level. In conclusion, these data suggested that 6% lipid and 50% protein in diet was optimal for loach during early development stage.     

Calibration and validation of optical chlorophyll‐measuring devices for use in predicting crude protein concentration in tropical grass herbage

The objective of this experiment was to evaluate the Fieldscout CM 1000 NDVI and Yara N‐Tester as easy‐to‐use and cost‐effective tools for predicting foliar chlorophylls (a, b and total) and crude protein (CP) concentrations in herbage from three tropical grass species. Optical chlorophyll measurements were taken at three stages (4, 8 and 12 weeks) of regrowth maturity in Guinea grass (Panicum maximum) and Mulato II (Brachiaria hybrid) and at 6 and 12 weeks maturity in Paspalum spp (Paspalum atratum). Grass samples were harvested subsequent to optical measurements for laboratory analysis to determine CP and solvent‐extractable chlorophylls (a, b and total) concentrations. Optical chlorophyll measurements and CP concentrations were highly correlated (Yara N‐Tester: r² = 0·77–0·89; Fieldscout CM 1000 NDVI: r² = 0·52–0·84). Crude protein prediction models from the Yara N‐Tester and Fieldscout CM 1000 NDVI accounted for 70–89% and 44–73% CP variability, respectively, in Mulato II and Guinea grass. The Yara N‐tester produced more accurate and reliable CP estimates based on very high concordance correlation coefficient [CCC (0·73–0·91)] and low rMSPE, mean and regression bias. It is concluded that the Yara N‐Tester produces more accurate and reliable CP estimates of tropical pastures.     

Effects of the dietary protein to energy ratio on growth,feed utilization and body composition in Macrobrachium nipponense

Oriental river prawn (Macrobrachium nipponense) has been widely cultured in Asian countries. However, its nutritional studies are very limited. In the present 8‐week study, we investigated the effects of dietary protein to energy ratio (P/E ratio) on the growth, feed utilization and body composition in juvenile M. nipponense (initial weight 0.302 ±0.03 g). Two‐factor experiment was designed and nine semi‐purified diets were formulated to contain three lipid levels (20, 80 and 140 g kg^?1) and three protein levels (330, 380 and 430 g kg^?1), producing P/E ratios from 16.5 to 23.4 mg KJ^?1 protein. The results indicated that the growth, survival rate and protein efficiency were dose dependently improved by the increased dietary lipid, but not dietary protein content. Increased dietary lipid content and/or protein content increased lipid accumulation in whole body, hepatopancreas and muscle, but did not change the feed intake and hepatopancreas weight. In conclusion, our present study indicated that M. nipponense is a species with relatively high‐energy requirement. It could utilize dietary lipid content up to 140 g kg^?1, while the dietary protein with more than 330 g kg^?1 would not promote growth and protein efficiency. Taken together, 330 g kg^?1 dietary protein and 140 g kg^?1 dietary lipid level with P/E ratio 16.49 could be optimum for M. nipponense.     

Effect of high inclusion of rendered animal by‐product ingredients on growth,digestibility and economic performances in climbing perch Anabas testudineus

A 70‐day growth trail was conducted to investigate the effects of inclusion of high levels of meat and bone meal (MBM) and protein concentrate (PC) on growth, digestibility and economic performances of climbing perch, Anabas testudineus. Four isonitrogenous diets were formulated by lowering the level of dietary fishmeal protein at 0 (D₁, control), 70 (D₂), 85 (D₃) and 100% (D₄) with a mixture of MBM and PC protein (1:1). Triplicate groups of 300 fish (mean weight of 0.80 g) stocked in each 40 m² pond and fed the respective test diets. A digestibility trial was conducted after the growth trial in indoor glass aquarium. The result showed that growth parameters were significantly decreased (P < 0.05) with fishmeal replacement levels. However, significant differences were not found in feed conversion ratio and survival of fish. No difference was also found in protein efficiency ratio among D1, D2 and D3. Similar to growth parameters, total fish production was highest in D1, intermediate in D2 and D3; and lowest in D4. Apparent digestibility coefficients of dry matter, protein and lipid were highest (P < 0.05) in D1 and lowest in D4. The economic analysis revealed that the benefit cost ratio was ranked by D₃ (1.81), D₂ (1.71), D₁ (1.66) and D₄ (1.46) respectively. Upon considering the overall performances and unavailability of finite protein sources, it can be concluded that 70–85% fishmeal could be replaced with a mixture of MBM and PC (1:1) in practical diets for climbing perch.     

Impacts of diet on hindgut microbiota and short‐chain fatty acids in grass carp (Ctenopharyngodon idellus)

Diet is known to influence intestinal microbiota in fish, but the specifics of these impacts are still poorly understood. Different protein/fibre ratio diets may result in differing structures and activities of gut microbiota. We examined the hindgut microbiome of grass carp (Ctenopharyngodon idellus) fed three different diets: fish meal (FM, high protein – low fibre), Sudan grass (SG, high fibre – low protein) and compound feed (CF, intermediate). Microbial profiles of fish fed on FM were significantly different from profiles of fish fed CF and SG (F = 18.85, p < .01). Cetobacterium, known to be positively associated with protein digestion, was the dominant microbial group in FM samples (approximately 75.7%), while Lachnospiraceae and Erysipelotrichaceae, thought to be involved in fermentation of plant polysaccharides, were dominant in CF and SG samples (46.8% and 42.9% respectively). Network analyses indicated that the abundance of Lachnospiraceae and Erysipelotrichaceae was in a significantly positive correlation (r = .895, p = .001). Short‐chain fatty acid (SCFA) levels may indicate that the digestibility of diet by microbiota in the grass carp gut decreased from FM to SG (FM>CF>SG). Overall low SCFA levels indicate that hindgut fermentation probably provides a low proportion of energy requirements in grass carp.     

木尔坦棉花曲叶病毒编码蛋白的亚细胞定位分析

 木尔坦棉花曲叶病毒(Cotton leaf curl Multan virus,CLCuMV)是典型的单组分双生病毒,并伴随β卫星分子,是棉花曲叶病的主要病原之一。本研究利用农杆菌介导的瞬时表达系统,将CLCuMV及其卫星分子编码的7个病毒蛋白在本氏烟表皮细胞中表达。通过激光共聚焦显微镜观察发现:V1、C2和C3定位于细胞核;C1和βC1定位在细胞核以及细胞质或细胞膜上,并且在细胞质中形成丝状结构;V2和C4主要定位在细胞质或细胞膜上,细胞核也有微量表达,V2可形成大小不一的颗粒状聚集体结构,C4在细胞膜上可见点状聚集体结构。同时,利用RT-PCR和Western blot对病毒各基因的转录和表达水平进行了分析。CLCuMV编码蛋白的亚细胞定位为蛋白功能的进一步研究提供重要理论依据。