Large Scale Structural Analysis of cDNAs in the Model Legume, Lotus japonicus

Lotus japonicus possesses certain characteristics suited to molecular genetic and genomic analyses and has been adopted as a model species in the study of legume plants. To make a catalogue of genes expressed in L. japonicus and understand biological processes specific to legume plants, large scale EST analyses have been performed. To date, more than 26,000 EST sequences of L. japonicus have been deposited in the public databases. These sequences were developed by five laboratories using different organs. In this review, information obtained from two EST projects carried out in Japan is presented. Some 7137 non-redundant EST groups from young plants and 718 groups from flower buds were generated. A similarity search revealed that homologues of nodulin genes in other legume plants, as well as genes related to secondary metabolism, seed development and the reproductive process, were included in the EST collection, indicating the usefulness of the EST clones in the study of biological phenomena distinctive to legume plant species. Received 23 August 2000/ Accepted in revised form 22 September 2000     

麦冬的配糖体成分

从川麦冬（Ｏｐｈｉｏｐｏｇｏｎ ｊａｐｏｎｉｃｕｓ）根部乙醇提取物中分离得到四个已知化合物,经波谱分析确定其结构分别为：龙脑７－Ｏ－β－Ｄ－吡喃葡萄糖甙（１）,龙脑７－Ｏ－〔β－Ｄ－呋喃芹糖（１→６）〕－β－Ｄ吡喃葡萄糖甙（２）,４－烯丙基－１,２－苯本酚１－Ｏ－〔α－Ｌ－吡喃鼠李糖（１→６）〕－β－Ｄ－吡喃葡萄糖甙（３）,５－烯－１β,３β,１６β,２２Ｓ－胆甾四醇１－Ｏ－α－Ｌ－吡喃鼠李糖１６     

海水酸化及升温对刺参生长及能量收支的影响

以东亚浅海生态系统中的关键种——刺参(Apostichopus japonicus)为实验对象,研究了CO₂驱动的海水酸化及升温对其生长及能量收支的影响。实验设置对照组(大连近海水温, pCO₂ 400μatm)、升温组(大连近海水温+3℃, pCO₂ 400μatm)、酸化组(大连近海水温, pCO₂ 1100μatm)和酸化升温组(大连近海水温+3℃,pCO₂ 1100μatm)。结果表明:与对照组相比,温度升高3℃对刺参的生长无显著影响;酸化组刺参的特定生长率最低,较对照组降低0.19%/d,个体体重的变异系数最大;酸化升温组刺参的终末体重和特定生长率与对照组相较无显著差异,但其摄食率和排粪率均显著高于对照组。升温组和酸化组的刺参能量的分配模式与对照组相比未发生明显改变,但酸化升温组刺参的能量分配模式发生显著变化,其粪便能所占摄食能的比例显著升高。研究表明,海水酸化抑制了刺参的生长但未改变其能量的分配,生长的降低主要取决于摄食减少;而海水酸化与温度升高的共同作用可能会通过...     

海参微卫星DNA的筛选与克隆的初步分析

微卫星DNA(microsatelliteDNA)广泛存在于真核生物的基因组中。由于其具有突变频率快、多态性丰富、呈共显性遗传、通用性等特点,已成为近年来被广泛应用的分子遗传标记。对刺参基因组DNA的提取及其微卫星的筛选与克隆进行了初步分析。     

Development and characterization of cell lines from heart, liver, spleen and head kidney of sea perch Lateolabrax japonicus

Five cell lines (LJHK, LJS, LJL, LJH-1 and LJH-2) were established from the head kidney, spleen, liver and heart of sea perch Lateolabrax japonicus . The cell lines LJHK, LJS, LJL, LJH-1 and LJH-2 were subcultured 46, 32, 32, 36 and 34 times in minimum essential medium (MEM) supplemented with foetal bovine serum (FBS), sea perch serum and 10 ng ml⁻¹ basic fibroblast growth factor (bFGF). Morphology of primary cultures and subcultures of the five cell lines were observed continuously by microscopy. The suitable temperature for growth was 18 to 30° C for all of these cell lines with the optimum growth at 24° C and a reduced growth rate <18° C. The optimum concentration of FBS was found to be 10% and addition of bFGF to the medium significantly increased the growth rate of the cells. The doubling time of LJS, LJH-1, LJL, LJH-2 and LJHK cells was determined to be 52·7, 54·9, 57, 58·7 and 66 h at a plating density of 1 × 10⁵ cells ml⁻¹ at 24° C, respectively. Chromosome analysis revealed that 42, 48, 38, 43 and 45% cells maintained normal diploid chromosome number (48) in the LJH-1, LJH-2, LJHK, LJL and LJS cell lines, respectively. The LJHK cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. Furthermore, treatment of the LJHK cells with lipopolysaccharide led to increased expression of IL-1β, demonstrating that LJHK cells might be a valuable tool for studying the expression and function of immunomodulatory gene in fishes.     

刺参管足SMART cDNA文库构建及EST分析

以大连广鹿岛野生刺参的管足mRNA为材料,利用SMART cDNA Construction Kit构建了表达型cDNA文库.初始文库的滴度为1.3 × 106PFU/mL,蓝白斑估测量组率合格.扩增后获得96 mL文库,滴度为1.8 × 1010PFU/mL,从扩增文库中随机挑取200个噬菌斑进行PCR检测,电泳检测结果表明重组率大于90％,片段大小在0.25 -2.0 kb之间的插入片段占80.4％,文库构建成功.随机选择122个噬菌斑转化为质拉pTriplEx2并测序,测序成功率83.6％,软件处理后100 bp以上的clean ESTs有72条,测通的为65条,占成功测序样品的63.7％.72条序列经SeqMan软件拼接,获得48条conting/singletons,在线Blast比对发现其中26条具有同源序列并获得注释,另外20条可能为新基因,其功能和结构有待进一步利用生物信息学的方法进行分析和研究.     

秦岭产珠子参叶的达玛烷型皂甙研究(1)

从陕西省秦岭产珠子参(Panax japonicus C.A.Meyer var.major(Burk.)Wu etFeng)的叶中分离到十个新的达玛烷型四环三萜皂甙,经光谱测定和化学降解,其中四个的化学结构分别为珠子参甙(majoroside)F_1(1)、F_2(2)、F_3(3)和F_4(4)。同时,还分离到已知的人参甙(ginsenoside)Rd(5)、Re(6)、Rg_1(7)、Rg_2(8)和F_2(9)。     

花鲈对不同饲料原料的表观消化率

试验以Cr2 O3 为指示物 ,以 70 %基础饲料和 30 %的待测饲料原料组成试验饲料 ,测定花鲈对红鱼粉、豆粕、花生粕、菜籽粕、肉骨粉和棉籽粕中干物质、粗蛋白、粗脂肪、能量和氨基酸的表观消化率。在水温 2 5± 1℃的试验条件下 ,花鲈对鱼粉、豆粕、花生粕的干物质表观消化率均在 6 0 %以上 ;对鱼粉、豆粕、花生粕和菜籽粕的蛋白质、脂肪的表观消化率都在 80 %以上 ;花鲈对能量的表观消化率变化范围在 16 99%— 83 96 %。测得每种原料的 16种氨基酸的表观消化率 :花鲈对原料氨基酸消化率的变化与对蛋白质消化率的变化相一致 ,除了赖氨酸、缬氨酸、蛋氨酸之外 ,花鲈对肉骨粉中氨基酸的利用率最低。花鲈对饼粕类原料中的含硫氨基酸 (如蛋氨酸 )的表观消化率低于其他氨基酸 ,对鱼粉中氨基酸利用率最低的为组氨酸 ,对肉骨粉中氨基酸利用率最低的为苏氨酸。