Constitutive nitric oxide synthase (NOS) activities in big-head carp (Aristichthys nobilis)

Nitric oxide synthase (NOS) is an enzyme that catalyzes the formation of nitric oxide (NO), an important biological messenger from L-arginine. There are considerable evidence showing the expression of NOS in mammalian tissues. Information on distribution of NOS activities in various organs and tissues of fish is rare. Non-functional NOS activities were documented in fish semi-quantitatively either by an indirect nicotine-adenine-dinucleotide-phosphate diaphorase (NADPH-d) activity histochemical staining method or by an immunohistochemical method using a cross-reacting antibody to brain NOS. Report on the functional levels of NOS activities in fish is lacking. This report represent the first attempt to document the functional NOS levels in various fish organs and tissues. Constitutive NOS (cNOS) activities in various organs of big-head carp (Aristichthys nobilis) was measured by a chemiluminescence method with a detection limit as low as 10 mol of NO produced. It was found that constitutive NOS activity was highest in the brain, followed by the intestine, stomach, retina, olfactory lobe, swim bladder, skeletal muscle, heart, kidney, ovary and liver. NOS activity could not be detected in the gill filaments. Omission of NADPH in the reaction mixture caused a 57–100% decrease in cNOS activities. However, omission of arginine in the mixture only caused a 56–87% drop in cNOS activities. When compared with cNOS activities documented from other species, a similar pattern of cNOS activities in the various organs and tissues of big-head carp could be seen.     

不规则盐度脉冲对许氏平鲉氨氮、磷排泄的影响

在静水系统中,骤然改变水体盐度,形成盐度脉冲;研究许氏平鲉Sebastodes fuscescens(Houttuyn)经历该脉冲后其氨氮、活性磷酸盐排泄的情况。结果表明,盐度改变后,许氏平鲉的氨氮排泄率比正常盐度(30)下呈现出不同程度的升高趋势;当盐度由30降为25和20时,随着体重增加,氨氮排泄率先增大,后又减小;当盐度由30上升为35时,氨氮排泄率随体重增加而呈下降趋势。盐度改变后,许氏平鲉的活性磷酸盐排泄率随体重增大呈先增大后减小的趋势。在同一体重下,盐度由30降至20时,活性磷酸盐的排泄率最大;盐度由30升至35时,磷的排泄率最小。方差分析表明,盐度脉冲对许氏平鲉氨氮排泄率有显著的影响(P<0.05),对活性磷酸盐排泄率的影响不显著(P>0.05)。该研究对于鱼类养殖的水质管理、养殖场的布设等有重要的指导意义。     

Effect of ABA,arginine and sucrose on protein content of date palm somatic embryos

Abscisic acid (ABA), arginine and sucrose were evaluated for their effects on the morphology, germination rates and protein content of date palm somatic embryos (SE). Different concentrations of these supplements in the culture medium were used. The comparative study of SE length and thickness between treated and untreated SE revealed no differences, except for ABA (20 μM), which increased thickness. A decrease of water content (WC) in favor of an increase in dry weight (DW) was observed in all treated SE, especially with sucrose (90 g l⁻¹) and ABA (20 μM). Only ABA (20 and 4 μM) caused a proliferation rate of the cultures higher than those in the control. Although all the tested compounds increased protein content, ABA (20 μM) was more effective in protein enrichment than arginine and sucrose treatments. The SDS-PAGE protein profiles showed a significant difference between treated and untreated SE. A protein band of 22 kDa, identified as glutelin in a previous work, was accumulated after treatment with 20 μM ABA or 3 mM arginine. These findings may contribute to further understanding of the mechanisms involved in the accumulation of specific storage proteins in several plants.     

Effects of dietary caffeine on growth,body composition,somatic indexes,and cerebral distribution of acetyl‐cholinesterase and nitric oxide synthase in gilthead sea bream (Sparus aurata), reared in winter temperature

This study aims at examining the effect of caffeine administration on growth, feed efficiency, and consumption of sea bream (Sparus aurata), reared in winter temperatures. Moreover, it is questioned whether caffeine has a central action in the brain and its effects are partly mediated via central brain mechanisms. For this, we studied the influences of caffeine treatment on the cerebral pattern of the cholinergic neurotransmission and the novel neuromodulator nitric oxide (NO), by means of acetyl‐cholinesterase (AchE) and nitric oxide synthase (NOS) histochemistry. Five different diets containing 0.0, 0.1, 1.0, 2.0 and 5.0 g caffeine kg^?1 of diet were administrated to five groups of fish. Caffeine adversely affected sea‐bream growth at a concentration higher than 1 g kg^?1 diet and increased feed conversion ratio in the treatments of 2 and 5 g kg^?1 (P < 0.05). The daily consumption of feeds was similar to all groups, indicating that caffeine did not influence diet palatability. AChE‐ and NADPH‐diaphorase histochemistry showed densely labeled cells and fibers mainly in dorsal telencephalon, preoptic, pretectal, hypothalamic areas, optic tectum, reticular formation, cerebellum and motor nuclei. When compared with matched caffeine‐treated animals, no differences in the histochemical pattern and cell densities of cerebral AChE and NADPH‐diaphorase were found.     

香蕉中一个新的S-腺苷甲硫氨酸合成酶基因的克隆及采后表达分析

利用抑制消减文库从香蕉中分离到一个cDNA片断,结合RACE技术获得该基因的全长序列。该全长cDNA共含1 285个碱基,通过Blastx同源性分析结果显示它与香蕉的一个S-adenosyl-L-methionine synthase(SAMS)基因(GenBank序列号为AF004317)具有82%的碱基同源性,编码的氨基酸顺序有93%同源性,但在cDNA的5’和3’非编码区同源性较低。设计特异引物对此基因进行RT-PCR分析表明,正常成熟的香蕉果实,采后当天表达量较高,随后略有降低,至采后12 d又达到一个相对较高值后迅速下降;高锰酸钾处理的果实,整个成熟期该基因均表现一个比较高的表达量;乙烯利处理的果实,采后表达量明显下降且处在一个比较稳定的水平。     

香草兰花芽分化期蛋白质及碳水化合物变化研究

以墨西哥香草兰花芽分化期花芽、混合花芽、叶芽及其功能叶为研究对象,研究香草兰花芽分化期蛋白质及碳水化合物变化及其差异,结果表明：香草兰叶芽蛋白质含量持续下降,花芽和混合花芽的蛋白质含量在花序分化初期上升至顶峰而后下降,说明香草兰花芽在花序分化前需要累积大量蛋白质。叶芽的可溶性糖、蔗糖含量均无显著变化,花芽和混合花芽均存在明显的上升和下降趋势;在整个花芽分化期,花芽可溶性糖和蔗糖含量均高于叶芽。叶芽的淀粉含量呈升高趋势,花芽和混合花芽的淀粉含量呈先上升后下降趋势。在整个花芽分化期过程中,花芽、混合花芽功能叶C/N比均高于叶芽功能叶,且花芽功能叶和叶芽功能叶间差异达显著水平。     

外源ABA对苗期低温下冬小麦蔗糖含量及其关键酶基因表达的影响

为了探讨外源ABA对苗期低温下冬小麦植株蔗糖代谢的调节作用,以抗寒性强的品种东农冬麦1号和抗寒性弱的品种济麦22为试验材料,在三叶期分别于不同温度下经ABA处理48 h后取叶片和地下茎,研究外源ABA对苗期低温下冬小麦的蔗糖含量及蔗糖代谢相关酶基因表达的影响。结果表明,外源ABA处理使低温下抗寒性强的东农冬麦1号中积累了更多的蔗糖,尤其是叶片,而在济麦22中则抑制了蔗糖的积累。4℃以上温度时,外源ABA促进了东农冬麦1号叶片和地下茎中UGP在蔗糖合成中的作用;在4℃以下低温时,外源ABA则促进了UGP在蔗糖分解中的作用。4℃以上温度时,外源ABA提高了东农冬麦1号叶片中Ta SPS基因和地下茎中Ta SAInv基因的表达,而抑制了济麦22各器官中Ta SPS基因和Ta SAInv基因的表达。此外,4℃以上温度时,外源ABA抑制了两个小麦品种各器官中Ta SS基因的表达。表明抗寒性强的东农冬麦1号对外源ABA可能更加敏感,其蔗糖合成能力的提高将有利于冬小麦植株抵御低温,进而维持植株的存活。     

小麦籽粒淀粉合成酶基因表达与酶活性特征的研究

为研究小麦籽粒淀粉合成酶基因表达与酶活性的特征,以普通小麦品种秦麦11(粒重36.942 mg/粒,淀粉含量61.02%)和中优9507(粒重50.636 mg/粒,淀粉含量68.36%)为材料,采用实时荧光定量RT-PCR方法,对灌浆期籽粒淀粉合成酶相关基因的表达进行了研究,并对其表达量与相应酶活性的相关性做了分析.结果表明,腺苷二磷酸葡萄糖焦磷酸化酶基因(AGPase)、束缚态淀粉合成酶基因(GBSSI)、可溶性淀粉合成酶基因(SSS)、淀粉分支酶基因(SBE)表达均呈单峰曲线变化,在花后3 d 内检测不到其表达,花后4～6 d 这些基因开始表达.AGPase和SSS在花后12 d左右表达量达到高峰,GBSSI和SBE在花后15 d左右的表达量最大.自花后18 d开始,各基因的表达量均显著下降.中优9507在表达高峰期(即花后第12、15、18 d)的酶基因相对表达量均显著高于秦麦11,且差异均达显著水平.整个灌浆期间中优9507的四种淀粉合成酶活性高于秦麦11,尤其在灌浆中期差异更显著.相关分析表明,AGPase、SSS、SBE基因在花后15 d的相对表达量与相应酶活性间呈极显著正相关,而GBSS基因的相对表达量与GBSS酶活性间相关不显著,暗示AGPase、SSS、SBE基因在籽粒淀粉合成过程中属于转录水平调控,而GBSSI基因属于转录后调控.     

金花茶松散型愈伤组织诱导与悬浮细胞系建立

采用组织培养及显微观察技术,以金花茶植株茎段、叶片、花药及离体培养的金花茶体细胞胚为试验材料,对松散型愈伤组织的获得及悬浮细胞系的建立进行了研究。结果表明:在黑暗条件下,以金花茶体细胞胚切块为外植体,在MS+KT 0.5 mg/L+NAA 8.0 mg/L+硝酸银5mg/L+蔗糖30 g/L培养基上诱导出愈伤组织后,将其转接到该诱导培养基上连续培养3个月,最后从培养物中挑选状态良好的松散型愈伤组织在分别含有肌醇与硝酸银的2种培养基上交替培养,可以获得金花茶松散型愈伤组织;方差分析表明:蔗糖添加量、肌醇添加量及硝酸银添加量均对金花茶悬浮培养液细胞密度有显著影响且后两者分别与光照条件存在相互作用;将松散型愈伤组织在分别添加100 mg/L肌醇与2.5 mg/L硝酸银的液体增殖培养基上交替培养,可以建立并保持金花茶悬浮细胞系。该研究结果可为金花茶大规模细胞培养提供科学依据。