Changes in enzymatic and structural properties of grass carp fast skeletal myosin induced by the laboratory-conditioned thermal acclimation and seasonal acclimatization

ABSTRACT:   Myosins were prepared from fast skeletal muscles of grass carp thermally acclimated to 10, 20 and 30°C in the laboratory as well as from those seasonally acclimatized and collected in January (winter) 2003 and May (spring), August (summer) and November (autumn) 2002. The maximal initial velocities ( V _max) of actin-activated Mg²⁺-ATPase activity for myosins from the 10°C-acclimated and winter grass carp were 1.7–1.8-fold as high as those from the 30°C-acclimated and summer fish. The inactivation rate constant ( K _D) of Ca²⁺-ATPase for myosin from the 10°C-acclimated grass carp was three to fourfold higher than those for myosins from the fish acclimated to 20°C and 30°C, whereas myosin from winter grass carp was about sevenfold as high as that for myosin from summer fish. Myosins from spring and autumn fish showed K _D values comparable to those of the fish acclimated to 30°C and 10°C, respectively. In differential scanning calorimetry analysis, the transition temperature ( T _m) was observed near 38°C and 45–46°C with most myosins. However, the lowest T _m at 32–33°C was given as one of the major endotherms in myosins from the 10°C-acclimated, autumn and winter fish. These responses of grass carp to changed environmental temperatures were almost similar to those for common carp reported previously.     

Qualitative and quantitative strategies of thermal adaptation of grass carp (Ctenopharyngodon idella) cytoplasmic malate dehydrogenases

Malate dehydrogenase isozymes of grass carp,Ctenopharyngodon idella, were identified by mitochondrial preparation and thermal denaturation. The structural and kinetic characteristics of chromatographically separated thermostable and thermolabile cMDHs were different in (1) half-life at 42°C, 10 min and 24 h, respectively, (2) optimal substrate, oxaloacetate and malate, concentrations, and (3) the apparent Michaelis-Menten constants of NADH and oxaloacetate.Total MDH activity in white muscle of 11°C-acclimated fish was about twice that of the 30°C-acclimated group. In addition, the ratio of the thermostable to thermolabile cMDH activity in white muscle of 30°C-acclimated fish was significantly higher than that of 11°C-acclimated fish. These results suggest that temperature acclimation can induce temperature compensation in MDH activity and differential expression of thermostable and thermolabile cMDH isozymes in freshwater fish.     

Comparison of apparent digestibility coefficient among replicates and different stocking density in red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis>

Two consecutive feeding trials were conducted to know: (i) how the apparent digestibility coefficient (ADC) varies among replicates on different days and during the course of the experiment (Trial 1); and (ii) the effect of stocking density (e.g. 7.17, 5.56, and 3.92 kg fish/t water) on the ADC of nutrients and energy in red sea bream (Trial 2). In both trials, 0.5% Cr₂O₃ was used as an inert marker. In Trial 1, there were no significant differences in ADC within the replicates although the values on different days varied significantly during the second week. The ADC of all parameters in the third and fourth weeks was significantly higher than in the second week (P<0.05). Although statistically insignificant, the lower stocking densities (3.92 and 5.56 kg fish/t water) showed better growth performance and superior ADC of protein, lipid, and energy than the higher stocking density in Trial 2. The results indicated that it may be better to acclimate fish to the diet for at least three weeks to obtain more reliable data by using a single tank for each treatment for digestibility experiments. Results also indicated that growth and digestibility performance can be stimulated by controlling stocking density.     

柽柳冷适应蛋白基因

冷适应蛋白(CAP)是指生物体受到寒冷刺激后,诱导表达的一类特异蛋白质,由冷诱导有关的基因在冷诱导过程中启动转录和翻译的。低温条件下,冷适应蛋白表达量在持续不断增加,对低温条件下蛋白质合成的调节以及mRNA的折叠起重要作用。是一类重要的与细胞耐寒相关基因。我们从构建的紫杆柽柳(Tamarixandrossowii)cDNA文库中分离得到了冷适应蛋白基因片段,并应用RACE技术获得其全长cDNA序列,该基因cDNA序列长度为1176bp,其中开放读码框(ORF)从49到663碱基位置,长度为615bp,编码204个氨基酸组成的多肽,预测其蛋白质分子量为23kD,预测其理论等电点为9.23。该基因已经在GenBank上注册,其基因序列登录号AY587773,氨基酸序列登录号为AAT01418。该基因的克隆为植物分子育种提供了新的基因资源。     

Effects of Thermal and Salt Treatments during Imbibition on Germination and Seedling Growth of Sorghum at 42/19 °C

This investigation tested the influence of thermal treatment of seeds at various stages of imbibition on the germination percentage, speed and growth response of seeds and seedlings. Seeds of sorghum (Sorghum bicolor L Moench) were treated in 2, 4 or 6 g NaCl l^?1 solutions and exposed for 2 h to 45 °C during the first, second or third days of imbibition. Thereafter, the seeds were dried and germinated at 42/19 °C (day/night temperature). Salt treatments did not improve the final germination percentage but increased germination speed over untreated seeds. The higher the salt concentration used, the greater the dry weights of plumules and radicles. Thermal treatment on the third day of imbibition yielded higher germination percentages than untreated seeds, while thermal treatment on the second day gave faster germination. Both the second and third day thermal treatments gave superior germination indices and higher plumule‐to‐radicle ratios. It is concluded that thermal treatment may assist in acclimating seeds to heat stress.     

Comparative studies of the changes in enzymatic activities in hardy and less hardy cultivars of winter wheat in late fall and in winter under snow

Abstract

The synthetic activity of protein and changes in the activity profiles of enzymes during cold acclimation were studied in leaves, stems, and crowns of a hardy winter wheat cultivar ( Triticum aestivum L. cv. Horoshirikomugi) and a less hardy winter wheat cultivar (Triticum aestivum L. cv. Norin 61). The results showed that one of the genetic differences between the two cuitivars is found in the activities of protein synthesis under low temperatures. The profiles of the changes in enzymatic activities showed that the adjustment of enzymatic activities was a tissue-specific event to accommodate environmental changes. The synthesis of the enzymes involved in a peroxide scavenging system such as hexokinase, glucose-6-phosphate dehydrogenase, dehydroascorbate reductase, ascorbate peroxidase, and so on started at the acclimation stage, while the phosphoglucose isomerase, which appears to be a maintenance-type enzyme in the wintering wheat, showed a small increase in the activity at later stages of acclimation period in their stems of both cuitivars. When the field was covered with snow, the catalase activity in the leaves started to decrease abruptly, while catalase activities in the stems and crowns did not exhibit appreciable changes during the life under snow. The life-cycle dependent changes in enzymatic activities appear to occur in late March or early April under snow.     

冷驯化和冷冻条件下不同抗寒性冬小麦品种三个COR基因表达差异的实时荧光定量分析

COR基因是含有CRT/DRE调控元件的一类冷响应基因,在植物抗寒耐寒过程中发挥着重要作用.为了分析不同抗寒性的冬小麦品种中COR基因表达的差异,明确在抗寒中起关键作用的COR基因,本实验以抗寒品种东农冬麦1号和不抗寒品种济麦22为材料,设定一系列低温处理,用实时荧光定量PCR(qRT-PCR)分析低温下Wrab17、Wcor80和Wcs120三个基因的表达差异.结果表明,在所有冷处理中,这三个基因在抗寒品种东农冬麦1号的相对表达量均比不抗寒品种济麦22高,上调表达时间更长,响应时间更短.处理组和对照组中也出现较大差异,在某些处理组中的相对表达量明显更高.其中冷驯化处理8～12 h和冷冻处理4h时Wrab17在东农冬麦1号中分别上调表达11.2、14.7和26倍,同时期同处理的济麦22中此数值分别为2.7、6.2和4.3倍;冷冻处理6h时Wcor80在东农冬麦1号中上调表达6.5倍,在济麦22中则出现小幅下调;冷驯化处理1h后Wcs120在东农冬麦1号中上调8.7倍,在济麦22中未发现上调表达.     

CBF转录因子在提高植物抗逆性中的作用

CBF转录因子在植物胁迫应答途径中发挥十分重要的作用,对CBF转录因子的结构特点、表达调控及其在提高植物抗寒性、抗旱性及抗盐性中的作用的研究进展进行了综述。     

不同低温驯化条件对玉米螟赤眼蜂低温贮藏的影响

低温驯化是昆虫应对外界环境低温的重要生存策略,驯化温度、驯化时间和冷藏时间是低温驯化方法中的关键因子。本文采用正交试验方差分析法检验了不同驯化温度、驯化时间和冷藏时间对用米蛾卵培养的玉米螟赤眼蜂4℃冷藏后的影响,结果显示,驯化温度和驯化时间交互作用对玉米螟赤眼蜂冷藏后的出蜂情况影响显著,当15℃驯化10 d后冷藏,出蜂卵数达到峰值为66.75±3.50,其出蜂率为(64.91±2.04)%;当10℃驯化20 d后冷藏,出蜂卵数为56.75±4.84,出蜂率达到峰值为(75.50±6.78)%。冷藏时间对玉米螟赤眼蜂低温驯化后的出蜂情况影响显著,其出蜂卵数和出蜂率均在冷藏60 d和90 d达到最大,出蜂卵数分别为53.61±3.64、55.39±3.51,出蜂率分别为(66.07±4.17)%、(68.30±4.08)%。研究表明,驯化温度、驯化时间和冷藏时间对低温驯化效果均有一定影响,低温驯化能增强玉米螟赤眼蜂的耐寒性,适于其长期冷藏,本试验中的最佳低温驯化条件为15℃驯化10 d。该研究结果对玉米螟赤眼蜂的低温耐受性研究及低温贮藏技术探讨均具有重要的理论指导意义。