解脂耶罗维亚酵母工程菌合成超长链脂肪酸及温度的影响

[背景]解脂耶罗维亚酵母属于产油微生物,大量研究表明该酵母能够高产长链脂肪酸和油脂,但是应用该酵母合成超长链脂肪酸仍待研究。[目的]工程化解脂耶罗维亚酵母合成高值超长链脂肪酸,并研究温度对脂肪酸合成的影响。[方法]合成密码子优化的拟南芥(Arabidopsis thaliana)延长酶基因AtFAE1、非洲芥菜(Brassica tournefortii)延长酶基因BtFAE1和碎米芥属植物Cardamine graeca的延长酶基因CgKCS,分别构建质粒pYLEX1-AtFAE1、pYLEX1-BtFAE1、pYLEX1-CgKCS和pYLEX1-AtFAE1-BtFAE1-CgKCS。以解脂耶罗维亚酵母菌株Po1g为宿主,通过化学法分别转化上述4个质粒,获得工程菌Po1g-AtFAE1、Po1g-BtFAE1、Po1g-CgKCS和Po1g-AtFAE1-BtFAE1-CgKCS,比较评价超长链脂肪酸的合成。在此基础上,过表达内源二酯酰甘油酰基转移酶基因DGAT1(diacylglycerol acyltransferase)提高产油量,并研究温度对生物量、产油、脂肪酸组成的影响。[结果]在解脂耶罗维亚酵母中3个延长酶的延长能力明显不同,AtFAE1主要催化C20:1脂肪酸的合成,BtFAE1更有利于芥酸(C22:1)的合成,而CgKCS能够催化合成神经酸(C24:1),但是三者共表达并未提高神经酸产量。在表达CgKCS基因的菌株中过表达DGAT1基因,细胞油脂含量提高50%。温度实验表明,低温有利于解脂耶罗维亚酵母合成不饱和脂肪酸,反之,高温利于其合成饱和脂肪酸。[结论]脂肪酸延长酶基因CgKCS可直接催化C18:1脂肪酸合成C24:1的超长链脂肪酸,并且通过优化培养温度可提高不饱和脂肪酸的合成。本研究为构建超长链脂肪酸细胞工厂以及发酵优化提供理论和技术参考。     

Temperature and altitude modulate feeding attributes of Mexican beetle,Zygogramma bicolorata Pallister on Parthenium hysterophorus

Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae) is an effective biocontrol agent of Parthenium hysterophorus L. which is an alien invasive herbaceous weed with a pan-tropical distribution. The present study aimed to assess the effects of temperature and altitude on feeding attributes (consumption rate, conversion efficiency and growth rate) of adults from the wild populations of Z. bicolorata inhabiting India and Nepal. Results revealed that adults inhabiting areas of low temperature (24°C ‒ 25°C) and high altitude (415 m ‒1400 m) were large and had higher food consumption rates. In contrast, those inhabiting areas of high temperature (34°C ‒ 36°C) and low altitude (81 m ‒ 229 m) were smaller and had higher food utilization efficiencies. In all the eco-climatic regions, females were larger than males and had higher feeding attributes than their counterparts. Temperature between 27°C and 30°C was found optimal for Z. bicolorata adults to convert and utilize the food biomass to body mass. Above the optimal temperature the feeding attributes decreased. Present results suggest that there exists a possibility for decrease in body size, and thereby weed biocontrol efficiency of Z. bicolorata adults with an increase in temperature due to global climate change.     

An in vivo and in silico analysis of novel variation in TMBIM6 gene affecting cardiopulmonary traits of Indian goats

Transmembrane Bax Inhibitor Motif-containing 6 (TMBIM6) gene acts as calcium leak channel and negatively regulates autophagy and autophagosome formation. The TMBIM6 gene was amplified and searched for variation in three different goat populations (i.e. Black Bengal, Ganjam and Raighar) of Odisha state of the India. The result indicated two substitutions i.e. 55th position (C55T) and 95th position (C95A) in the amplified region of the gene resulting in change of amino acids (Leu > Phe and Thr > Asn). The identified SNPs were combined to form haplotypes and animals were grouped accordingly. Structural analysis showed minor changes (5%) in between mutant and wild TMBIM6 protein structures. However, any functional variation could not be identified with respect to the calcium ligand and open pore state. But an alteration of calcium binding site was found. The binding interaction of calcium with the TMBIM6 protein was hydrophobic in nature in closed state whereas hydrophilic in open pore stage. The stress releasing function was the result of calcium leakage controlled by amino acids coded by exon 4 and exon 5 regions of TMBIM6 gene. The effect of breed and haplotype on cardiopulmonary traits was studied. The data on cardiopulmonary traits of body i.e. rectal temperature, skin temperature, heart rate and respiration rate were recorded when ambient temperature usually remained the highest. The statistical analysis showed, significant difference in rectal temperature, skin temperature and respiration rate among these goat populations. The haplotypes (CC and TA) were found to have a significant (P < 0.05) effect on rectal temperature, skin temperature and respiration rate. However, any such significant effect could not be identified in recorded heart rate. The objective of the present study to identify the genetic variations in TMBIM6 gene having significant effect on cardiopulmonary traits which can be further uses as the molecular markers to improve heat tolerance mechanism in goats.     

Temperature preference of juvenile lumpfish (Cyclopterus lumpus) originating from the southern and northern parts of Norway

Fish are ectothermic animals and have body temperatures close to that of the water they inhabit. They can still control their body temperatures by selecting habitats with temperatures that maximize their growth, feed conversion and wellbeing. Lumpfish, Cyclopterus lumpus, is widely distributed in the North Atlantic Ocean and therefore exposed to variable water temperatures. Lumpfish is extensively used as cleanerfish in salmon farming in Norway and exposed to a wide temperature range along the north-south axis of the Norwegian coastline. But, if these temperature ranges correspond to the preference temperatures of lumpfish is not known. If lumpfish has adapted to regional temperatures along the Norwegian coast, differences in preference temperature for fish from different regions should be evident. In a selective breeding perspective, different selection lines for preference temperature would then be useful for further development of lumpfish as a cleanerfish.We subjected lumpfish juveniles weighing 154–426g originated from northern (Group North – GN) and southern (Group South – GS) Norway to a temperature preference test, using an electronic shuttle box system. The system allowed the fish to control the water temperature by moving between two chambers, and thereby choosing its preferred temperature in the range from 5 to 16 °C. We started the temperature at 7.8 ± 1.37 °C for GN and 7.58 ± 1.34 °C for GS, but all the fish except four (two each from GN and GS) chose lower temperatures (5.03–7.6 °C) in the first 18 h and stayed closer to that temperature during the next 30 h. Based on the results, GN and GS lumpfish preferred 6.92 ± 1.8 and 6.2 ± 1.2, respectively, and there was no significant difference between the groups. Neither was there any significant difference in growth rates (SGR) between the two groups. Based on our results, we suggest that lumpfish from any geographical origin along the Norwegian coast can be used anywhere in Norway. It follows that lumpfish from a single selection line could be used at any salmon farm in Norway independent of its location.     

The highest kingdom of Anolis: Thermal biology of the Andean lizard Anolis heterodermus (Squamata: Dactyloidae) over an elevational gradient in the Eastern Cordillera of Colombia

Vertebrate ectotherms may deal with changes of environmental temperatures by behavioral and/or physiological mechanisms. Reptiles inhabiting tropical highlands face extreme fluctuating daily temperatures, and extreme values and intervals of fluctuations vary with altitude. Anolis heterodermus occurs between 1800 m to 3750 m elevation in the tropical Andes, and is the Anolis species found at the highest altitude known. We evaluated which strategies populations from elevations of 2200 m, 2650 m and 3400 m use to cope with environmental temperatures. We measured body, preferred, critical maximum and minimum temperatures, and sprint speed at different body temperatures of individuals, as well as operative temperatures. Anolis heterodermus exhibits behavioral adjustments in response to changes in environmental temperatures across altitudes. Likewise, physiological traits exhibit intrapopulation variations, but they are similar among populations, tended to the “static” side of the evolution of thermal traits spectrum. The thermoregulatory behavioral strategy in this species is extremely plastic, and lizards adjust even to fluctuating environmental conditions from day to day. Unlike other Anolis species, at low thermal quality of the habitat, lizards are thermoconformers, particularly at the highest altitudes, where cloudy days can intensify this strategy even more. Our study reveals that the pattern of strategies for dealing with thermal ambient variations and their relation to extinction risks in the tropics that are caused by global warming is perhaps more complex for lizards than previously thought.     

The magnitude of physical exercise-induced hyperthermia is associated with changes in the intestinal permeability and expression of tight junction genes in rats

We investigated whether the magnitude of exercise-induced hyperthermia influences intestinal permeability and tight junction gene expression. Twenty-nine male Wistar rats were divided into four groups: rest at 24 °C and exercise at 13 °C, 24 °C or 31 °C. The exercise consisted of a 90-min treadmill run at 15 m/min, and different ambient temperatures were used to produce distinct levels of exercise-induced hyperthermia. Before the experimental trials, the rats were treated by gavage with diethylenetriaminepentaacetic acid labeled with technetium-99 metastable as a radioactive probe. The rats' core body temperature (T_CORE) was measured by telemetry. Immediately after the trials, the rats were euthanized, and the intestinal permeability was assessed by measuring the radioactivity of blood samples. The mRNA levels of occludin and zonula occludens-1 (ZO-1) genes were determined in duodenum samples. Exercise at 24 °C increased T_CORE to values close to 39 °C, without changing permeability compared with the resting trial at the same environment. Meanwhile, rats’ T_CORE exceeded 40 °C during exercise at 31 °C, leading to greater permeability relative to those observed after exercise in the other ambient temperatures (e.g., 0.0037%/g at 31 °C vs. 0.0005%/g at 13 °C; data expressed as medians; p < 0.05). Likewise, the rats exercised at 31 °C exhibited higher mRNA levels of ZO-1 and occludin genes than the rats exercised at 24 °C or 13 °C. The changes in permeability and gene expression were positively and significantly associated with the magnitude of hyperthermia. We conclude that marked hyperthermia caused by exercise in the warmer environment increases intestinal permeability and mRNA levels of tight junction genes.     

高强度聚焦超声换能器温度场的数值仿真

相控阵高强度聚焦超声换能器可以通过换能器上不同阵元发射超声波的时间不同来实现变焦、多焦点。该论文应用Westervelt方程的近似式,结合Pennes热传导方程,以人体乳房为例,FDTD(finite difference time domain)仿真对比研究平面阵列相控聚焦换能器与曲面阵列相控聚焦换能器形成温度场的特性,同时数值仿真研究不同占空比的正弦激励函数、不同治疗频率、声强对曲面阵列相控聚焦换能器超声温度场的影响。研究结果表明曲面阵列相控聚焦换能器能有效地减少皮肤处的温升,对皮肤的伤害较小;对于曲面阵列相控聚焦换能器,不同占空比的正弦激励函数形成的可治疗区域(60℃以上)大小差别不大,但最高温度不同;随着频率升高,形成的可治疗区域体积减小;随着输入声强的增大,可治疗区域变大,但焦距不变。     

洱海螺旋鱼腥藻生长生理特性的初步研究

近年来,洱海正处于中营养水平向富营养湖泊的过渡阶段,蓝藻水华也频繁发生。本文作者在洱海大规模水华暴发期间,分离、纯化了水华优势种螺旋鱼腥藻,并对其生长生理特性进行了初步研究,以期为探讨洱海鱼腥藻水华发生的环境影响因素提供基础的参考资料。实验结果表明,氮含量1.5mmol/L、磷含量12µmol/L、光照强度30E/ m·s、pH值8－10及温度25℃时,螺旋鱼腥藻生长状况最好,生物量及相对生长速率较高。不同氮、磷浓度下的氮磷代谢活性表明：氮浓度在0－0.36mmol/L时,硝酸还原酶活性随着氮浓度的增加而增强;氮浓度在0.36－6 mmol/L时,酶活性由其生长状况决定,生长越好,酶活性越高。碱性磷酸酶受磷影响较大,随着磷浓度的增加其活性逐渐减弱,磷充足时,氮对其活性并无显著影响。此外,洱海螺旋鱼腥藻可在低的氮、磷浓度下生长,这与其氮磷代谢活性的调节作用有关。     

Biological characteristics and thermal requirements of a Brazilian strain of the parasitoid Trichogramma pretiosum reared on eggs of Pseudoplusia includens and Anticarsia gemmatalis

A new strain of the parasitoid Trichogramma pretiosum, was collected in Rio Verde County, State of Goiás, Central Brazil, and designated as T. pretiosum RV. This strain was then found to be the most effective one among several different strains of T. pretiosum tested in a parasitoid selection assay. Therefore, its biological characteristics and thermal requirements were studied, aiming at allowing its multiplication under controlled environmental conditions in the laboratory. The parasitoid was reared on eggs of Pseudoplusia includens and Anticarsia gemmatalis at different constant temperatures within an 18–32 °C temperature range. The number of annual generations of the parasitoid was also estimated at those temperatures. Results have shown that T. pretiosum RV developmental time, from egg to adult, was influenced by all temperatures tested within the range, varying from 6.8 to 20.3 days and 6.0 to 17.0 days on eggs of P. includens and A. gemmatalis, respectively. The emergence of T. pretiosum RV from eggs of A. gemmatalis was higher than 94% at all temperatures tested. When this variable was evaluated on eggs of P. includens, however, the figures were higher than that within the 18–30 °C range (more than 98%), and were also statistically higher than the emergence observed at 32 °C (90.2%). The sex ratio of the parasitoids emerged from eggs of A. gemmatalis decreased from 0.55 to 0.29 at 18–32 °C, respectively. However, for those emerged from eggs of P. includens, the sex ratio was similar (0.73, 0.72 and 0.71) at 20, 28 and 32 °C, respectively. The lower temperature threshold (Tb) and thermal constant (K) were 10.65 °C and 151.25 degree-days when the parasitoid was reared on eggs of P. includens; and 11.64 °C and 127.60 degree-days when reared on eggs of A. gemmatalis. The number of generations per month increased from 1.45 to 4.23 and from 1.49 to 4.79 when the parasitoid was reared on eggs of P. includens and A. gemmatalis, respectively, following the increases in the temperature.     

Tackling aquatic invasions: risks and opportunities for the aquarium fish industry

The aquarium trade is an important and rapidly growing vector for introduced species in the United States. We examined this vector by surveying pet stores in the San Francisco Bay–Delta region to compile a list of aquarium fish species commonly stocked. We identified which of these species might be able to survive in the Bay–Delta, and investigated store representatives’ knowledge and attitudes about biological invasions. A restrictive analysis using conservative estimates of fish temperature tolerances and environmental conditions found that the local aquarium trade includes 5 fish species that can survive in a temperate system such as the Bay–Delta. Under more inclusive parameters, up to 27 fish species met the criteria for survival in the Bay–Delta. We further explored these results by comparing potential invader incidence between different types of stores. In the more restrictive analysis, three national retail chains stocked significantly more potentially invasive species than independent aquarium stores, but there was no difference in the more inclusive analysis. A significantly higher percentage of fish taxa were easily identifiable and well-labeled in chain stores than in independent stores. Most aquarium store representatives indicated willingness to take action to reduce the threat of trade-related introductions, although chain store employees were more willing to assign responsibility for reducing this threat to the aquarium industry than were independent store employees. Management efforts for this vector should focus on (a) improving labeling and identification of fish species in stores, (b) expanding the often spotty data on fish physiological tolerances, especially for saltwater species, (c) educating customers and store employees about the risks posed by pet release, and (d) providing better options for responsible disposal of unwanted fish. Electronic Supplementary Material   The online version of this article (doi:) contains supplementary material, which is available to authorized users.