Geographical variation of skull morphology and its functional significances in the red-cheeked squirrel

Skulls of the red-cheeked squirrel (Dremomys rufigenis) from various geographical locations: Malaysia (peninsular area), Vietnam (south district)-Laos, and Thailand (north district) were osteometrically examined. The skull size of the squirrels in the southern (Malaysia) population was fundamentally larger than that in the northern (Vietnam, Laos and Thailand) populations. The proportion indices indicated that the splanchnocranium was relatively longer in the Malaysia population, and that the interorbital space was narrower in Vietnam-Laos, and Thailand populations. We suggest that the long nose and laterally-oriented orbits in the skull may be better adapted for terrestrial-insectivorous life in the Malaysia population and the binocular sense facilitated by rostrally-oriented eyes contributes to the arboreal-fruit eating behavior in the two northern populations. The Malaysia population was clearly distinguished from the other populations by the principal component analysis. We suggest that the geographical barrier of the Isthmus of Kra influences the morphological variation of the skull among the squirrel populations.     

核桃试管嫩茎生根的形态结构及激素调控研究

 以核桃品种‘新早丰’试管嫩茎为试材, 对其诱导生根过程中的形态结构及相关的生长素(IAA) 和脱落酸(ABA) 变化进行了研究。证实诱导生根过程中核桃嫩茎不定根原基发生于形成层, 特别是髓射线正对的形成层部分; 根原基起始分化期为诱导第6 天左右, 伸长期是第10天; 如果12 d之后仍放在诱导培养基中, 生根率下降, 并且出现茎基愈伤化、茎尖变黑和叶片脱落等现象; 若生根诱导10 d后转入无植物生长调节剂培养基, 培养5 d左右可见根尖突出表皮, 根系发育正常; 同时与不定根形态发生相应的内源IAA和ABA的变化是根原基的发生期和伸长期, 内源IAA出现高峰, 内源ABA呈上升趋势,IAA /ABA值在根原基的发生前为最大, 随后降低。本研究不仅从形态结构证实了二步生根法的合理性, 而且从生理学角度阐述了不定根发生的IAA /ABA调控机制。     

Comparative functional anatomy of hindlimb muscles and bones with reference to aquatic adaptation of the sea otter

Although the sea otter (Enhydra lutris) is a complete aquatic species, spending its entire life in the ocean, it has been considered morphologically to be a semi-aquatic animal. This study aimed to clarify the unique hindlimb morphology and functional adaptations of E. lutris in comparison to other Mustelidae species. We compared muscle mass and bone measurements of five Mustelidae species: the sea otter, Eurasian river otter (Lutra lutra), American mink (Neovison vison), Japanese weasel (Mustela itatsi) and Siberian weasel (M. sibirica). In comparison with the other 4 species, E. lutris possessed significantly larger gluteus, popliteus and peroneus muscles, but smaller adductor and ischiopubic muscles. The popliteus muscle may act as a medial rotator of the crus, and the peroneus muscle may act as an abductor of the fifth toe and/or the pronator of the foot. The bundles of the gluteus superficialis muscle of E. lutris were fused with those of the tensor fasciae latae muscle and gluteofemoralis muscles, and they may play a role in femur abduction. These results suggest that E. lutris uses the abducted femur, medially rotated crus, eversion of the ankle and abducted fifth digit or extended interdigital web as a powerful propulsion generator. Therefore, we conclude that E. lutris is a complete aquatic animal, possessing differences in the proportions of the hindlimb muscles compared with those in other semi-aquatic and terrestrial mustelids.     

拟南芥未知基因At018融合蛋白原核表达条件的优化及纯化

本课题组在前期的研究中利用Al Cl3胁迫筛选碱茅全长c DNAs酵母表达文库获得到一个与铝毒害相关的未知基因(基因编号为018,用put018表示),该基因与拟南芥中未知基因(基因号为At5g57345,本研究中用At018表示)有较高的同源性。为了获得At018的纯化蛋白,本研究将拟南芥At018基因克隆后构建原核表达载体p GEX-6p-3-At018,再将其转入大肠杆菌BL21(DE3)菌株,利用异丙基-β-D-硫代半乳糖苷(IPTG)进行诱导表达。同时运用传统方法优化诱导条件,以提高重组融合蛋白的表达效率。SDS-PAGE分析结果表明,30℃下0.1 mmol/L IPTG的条件下诱导3 h后,GST-At018融合蛋白的表达量最大,蛋白分子质量与预测值相符,该蛋白主要以可溶性形式存在;接着利用Glutathione Sepharose4 Fast Flow亲核层析树脂纯化最终获得GST-At018融合蛋白。本研究可为进一步进行At018的功能解析提供基础。     

过量表达星星草(Puccinellia tenuiflora)液泡型H~+-ATP酶c亚基基因提高转基因酵母的耐盐性

植物液泡型H+-ATP酶(V-ATPase)在次级转运与应对多种逆境胁迫中具有重要作用。本研究克隆并解析了能够生长在盐碱地上的野生星星草V-ATPase c亚基基因(Put VHA-c)的功能。本研究利用荧光蛋白(GFP)标记结合内涵体染色的方法研究了Put VHA-c蛋白在酵母细胞中的定位,并利用转基因酵母分析了Put VHA-c基因在盐胁迫中的作用。共定位实验显示Put VHA-c-GFP融合蛋白主要定位在酵母细胞的内涵体上。Northern杂交和实时定量PCR显示在星星草悬浮细胞中Put VHA-c基因的表达能够被盐胁迫所诱导。研究结果结合过表达Put VHA-c的转基因酵母具有更高的耐盐性的发现,说明Put VHA-c基因参与着盐胁迫的应答。     

Development of monoclonal antibodies (MAbs) to feline interferon (fIFN)-γ as tools to evaluate cellular immune responses to feline infectious peritonitis virus (FIPV)

Feline infectious peritonitis virus (FIPV) can cause a lethal disease in cats, feline infectious peritonitis (FIP). The antibody-dependent enhancement (ADE) of FIPV infection has been recognised in experimentally infected cats, and cellular immunity is considered to play an important role in preventing the onset of FIP. To evaluate the importance of cellular immunity for FIPV infection, monoclonal antibodies (MAbs) against feline interferon (fIFN)-γ were first created to establish fIFN-γ detection systems using the MAbs. Six anti-fIFN-γ MAbs were created. Then, the difference in epitope which those MAbs recognise was demonstrated by competitive enzyme-linked immunosorbent assay (ELISA) and IFN-γ neutralisation tests. Detection systems for fIFN-γ (sandwich ELISA, ELISpot assay, and two-colour flow cytometry) were established using anti-fIFN-γ MAbs that recognise different epitopes. In all tests, fIFN-γ production from peripheral blood mononuclear cells (PBMCs) obtained from cats experimentally infected with an FIPV isolate that did not develop the disease was significantly increased by heat-inactivated FIPV stimulation in comparison with medium alone. Especially, CD8(+)fIFN-γ(+) cells, but not CD4(+)fIFN-γ(+) cells, were increased. In contrast, fIFN-γ production from PBMCs isolated from cats that had developed FIP and specific pathogen-free (SPF) cats was not increased by heat-inactivated FIPV stimulation. These results suggest that cellular immunity plays an important role in preventing the development of FIP. Measurement of fIFN-γ production with the anti-fIFN-γ MAbs created in this study appeared to be useful in evaluating cellular immunity in cats.     

Suppression of Ralstonia solanacearum bacterial wilt disease by an organic hydroponic system

An organic hydroponic system that we developed has potential to control root diseases including bacterial wilt of hydroponically grown tomato. In inoculation tests with Ralstonia solanacearum during tomato plant cultivation in conventional inorganic hydroponics and in our organic system, many of the tomato seedlings in the conventional system wilted and died, but none of the seedlings in the organic hydroponics wilted or developed any symptoms, suggesting that the organic system can suppress this bacterial wilt disease. Interestingly, a rhizosphere biofilm, formed only on roots in the organic hydroponic system, may be responsible for the suppression of the bacterial wilt.     

Anatomical and histological characterization of ileal and jejunal Peyer's patch in lesser mouse deer (Tragulus javanicus)

Tragulidae is a primitive ungulate family within the order Cetartiodactyla, suborder Ruminantia. Domestic ruminants such as cattle, sheep, and goat have two types of Peyer's patches (PP): jejunal and ileal PP, in which there are morphological and functional differences. In this study, lesser mouse deer (Tragulus javanicus) PP was studied by gross anatomical and histological procedures. At the fetal stage, both types of PP were formed in the small intestine. Ileal PP was observed as a single continuous aggregation of lymphatic follicles extending cranially from the ileo-caecal junction. However, jejunal PP was observed as multiple and discrete accumulations of lymphatic follicles. This study showed that the lesser mouse deer has two types of PP in the small intestine. In addition, the anatomical and histological characteristics of jejunal and ileal PP are quite similar to those of other ruminants' jejunal and ileal PP. Further studies are needed to analyze immune function of both PP in lesser mouse deer in order to determine the evolutionary process of Cetartiodactyla.