Multiple Forms of Phospholipase D following Germination and during Leaf Development of Castor Bean

Multiple molecular forms of phospholipase D (PLD; EC 3.1.4.4) were identified and partially characterized in endosperm of germinated seeds and leaves of castor bean (Ricinus communis L. var Hale). The different PLD forms were resolved by nondenaturing polyacrylamide gel electrophoresis, isoelectric focusing, and size-exclusion chromatography. PLD was detected with both a PLD activity assay and immunoblots with PLD-specific antibodies. There were three major forms of PLD, designated types 1, 2, and 3, based on their mobility during nondenaturing polyacrylamide gel electrophoresis. Molecular masses of the PLD variants were estimated at 330, 230, and 270 kD for the types 1, 2, and 3, respectively. Isoelectric points of the native type 1, 2, and 3 PLDs were approximately 6.2, 4.9, and 4.8. Under the in vitro assay conditions used, the three forms of PLD exhibited the same substrate specificity, hydrolyzing phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylglycerol (PG) but not phosphatidylserine (PS) and phosphatidylinositol (PI). The three forms of PLD differed in their substrate preferences, and the order of activities was: PLD 1, PE > PG = PC; PLD 2, PE > PG > PC; PLD 3, PE = PG = PC. The Km values of PLDs 1, 2, and 3 for PC were 1.92, 2.62, and 5.18 mM, respectively. These PLDs were expressed differentially following seed germination and during leaf development. Type 1 was found in the early stages of seedling growth and in young leaves, type 2 was present in all the tissues and growth stages examined, and type 3 was expressed in senescent tissues. The PLDs shifted from largely cytosolic to predominantly membrane-associated forms during leaf development. The present studies demonstrate the structural heterogeneity of plant PLD and growth stage-specific expression of different molecular forms. The possible role for the occurrence of multiple molecular forms of PLD in cellular metabolism is discussed.     

中国盲走螨属一新种和二新纪录：蜱螨亚纲：植绥螨科

本文记述我国植绥螨科盲走螨属Ｔｙｐｈｌｏｄｒｏｍｕｓ一新种和中国二新纪录：鲁盲走螨Ｔ．ｌｕｅｎｓｉｓ ｓｐ．ｎｏｖ．,甲胄盲走螨Ｔ．ｌｏｒｃａｔｕｓ,肥厚盲走螨Ｔ．ｈｉｇｏｅｎｓｉｓ。标本保存于上海复旦大学环境和资源生物系。     

Mathematical models for the swimming pattern of a lamprey

A significant characteristic in a swimming pattern of a lamprey is the generation of a constant phase lag along its body in spite of the wide range of undulation frequencies. In this paper, we discuss a mathematical treatment for coupled oscillators with time-delayed interaction and propose a model for the central pattern generator (CPG) of a lamprey to account for the generation of a constant phase relation, with consideration of the signal conduction time. From this model, it is suggested that the desired phase relation can be produced by long ascending connections from the tail to the neck region of the CPG.     

Hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency: Identification of point mutations in Japanese patients with Lesch-Nyhan syndrome and hereditary gout and their permanent expression in an HPRT-deficient mouse cell line

Two different single nucleotide transitions of hypoxanthine-guanine phosphoribosyltransferase (HPRT) were identified in a Japanese patient with Lesch-Nyhan syndrome (LNS) and a patient with hereditary gout. HPRT enzyme activities in the two patients were severely deficient, but the size and amount of mRNA were normal according to Northern analysis. Entire coding regions of HPRT cDNAs were amplified by PCR and sequenced. A G-to-A substitution at base 208 in exon 3, which predicted glycine 70 to arginine, was detected in the LNS patient (identical mutation with HPRT_Utrecht). A C-to-A substitution at base 73 in exon 2, which predicted proline 25 to threonine, was detected in the gout patient (designated HPRT_Yonago). We transfected normal HPRT cDNA, mutant cDNA with HRPT_Utrecht or mutant cDNA with HPRT_Yonago, respectively, to HPRT-deficient mouse cells and isolated permanent expression cell lines. The HPRT-deficient mouse cells had no detectable HPRT activity and a very low amount of HPRT mRNA. When the HPRT-deficient mouse cells were transfected with normal human cDNA, HPRT enzyme activity increased to 21.8% that of normal mouse cells. The mouse cells transfected with HPRT_Utrecht showed no increase in HPRT activity; however, when the mouse cells were transfected with HPRT_Yonago, the activity increased to 2.4% that of normal activity. The proliferative phenotypes of these cells in HAT medium and in medium containing 6-thioguanine were similar to those of skin fibroblasts from the patients. This series of studies confirmed that each of the two point mutations was responsible for the decreases in HPRT enzyme activity, and the proliferative phenotypes in HAT medium and medium containing 6-thioguanine.     

鹿衔草化学成分的研究：羟基肾叶鹿蹄草甙的结构鉴定

从鹿衔草（Ｐｙｒｏｌａ ｃａｌｌｉａｎｔｈａ Ｈ．Ａｎｄｒｅｓ）分离得到两个化合物。经光谱分析,确定其中１个化合物为新化合物,命名为羟基肾叶鹿蹄草甙（１, ｈｙｄｒｏｘｙｌｒｅｎｉｆｏｌｉｎ）,另一化合物为儿茶素（３）。     

黑龙江依兰早第三纪植物群的古气候分析

对黑龙江依兰煤矿煤层间的大量植物叶痕化石研究表明：依兰植物群有蕨类植物２ 种,裸子植物１０ 种,被子植物５８ 种,分属３４ 科４６属。植物群可分为两个植物组合：一个是下煤层上顶板的矿页岩层化石的组合,称Ａ 段组合,时代为早始新世。植物种类丰富,含有较多常绿阔叶成分,属北亚热带的常绿阔叶和落叶阔叶、针阔叶混生林。通过植物叶相特征分析,其全缘叶比例为３８％ 。用气候诺模图得出其古气候为年均温１３．２℃,年温差２０℃;另一个植物组合是煤系地层之上,即上煤层顶部的油页岩层中的Ｂ段化石组合,时代为早渐新世。植物以落叶成分为主,属暖温带落叶阔叶林和针阔叶混生林。全缘叶比例为３０％ 。古气候年均温为１１℃,年温差２５℃。表明植物区系组成完全不同,显示出气候随时代发生了演变,而使区系逐渐发展到今日的寒温带气候和植被     

向日葵幼苗环旋运动的三维轨迹

采用光学显微镜标尺和垂线原理制作的三维空间点测定仪,对向日葵（Ｈｅｌｉａｎｔｈｕｓ ａｎｎｕｕｓＬ．）幼苗的环旋运动进行了连续测量。结果表明：向日葵环旋运动的轨迹有椭圆型、摆动型和不规则型;同一植株在不同生长阶段所表现的环旋运动轨迹不一定相同,同一株龄的不同个体也不一定具有相同的运动轨迹;运动的方向有左旋和右旋,圆周运动光源可以显著地改变运动方向;从三维角度看,在整个下胚轴生长阶段,环旋运动的振幅存在一个由小变大再由大变小的变化规律     

非同位素PCR－单链构象多态性技术的建立和应用

ＰＣＲ－单链构象多态性技术问世以来,成为研究基因突变的工具,特别是在分子肿瘤学研究中,广泛应用于癌基因,抑癌基因突变的研究,常规ＰＣＲ－ＳＳＣＰ采用同位素标记ＰＣＲ产物,测序板电泳分离突变,在操作和费用上有种种局限,文章建立了一种非同位素ＰＣＲ－ＳＳＣＰ技术;通过不对称ＰＣＲ获得单链,普通ＰＡＧＥ分离,经银染检出突变,用这种方法,还研究了四株鼻咽癌细胞株ＣＮＥ１,ＣＮＥ２,ＨＫ１和ＳＵＮＥ１中肿瘤     

Abscisic acid and mefluidide in the regulation of cold hardiness development in Solanum tuberosum L. potato

Plants of Solanum tuberosum L. potato do not cold acclimate when exposed to low temperature such as 5°C, day/night. When ABA (45 M) was added to the culture medium, stem-cultured plantlets of S. tuberosum, cv. Red Pontiac, either grown at 20°C/15°C, day/night, or at 5°C, increased in cold hardiness from –2°C (killing temperature) to –4.5°C. The increase in cold hardiness could be inhibited in both temperature regimes if cycloheximide (70 M) was added to the culture medium at the inception of ABA treatment. Cycloheximide did not inhibit cold hardiness development, however, when it was added to the culture medium 3 days after ABA treatment.When pot-grown plants were foliar sprayed with mefluidide (50 M), ABA content increased from 10 nmol to 30 nmol g^–1 dry weight and plants increased in cold hardiness from –2°C to about –3.5°C. The increases in free ABA and cold hardiness occurred only in plants grown at 20°C/15°C; neither ABA nor cold hardiness increased in plants grown at 5°C.The results suggest that an increase in ABA and a subsequent de novo synthesis of proteins are required for the development of cold hardiness in S. tuberosum regardless of temperature regime, and that the inability to synthesize ABA at low temperature, rather than protein synthesis, appears to be the reason why S. tuberosum does not cold acclimate.     

Increased shikonin production by hairy roots of Lithospermum erythrorhizon in two phase bubble column reactor

Summary Plant hairy root cultures of Lithospermum erythrorhizon were carried out to produce shikonin derivatives by employing in situ extraction with n-hexadecane in a shake flask and a bubble column bioreactor. Over 95 % shikonin produced was recovered in the n-hexadecane layer. In flask cultures the maximum concentration of shikonin with n-hexadecane extraction was 3 times higher than that obtained without extraction. In the two phase bubble column reactor, 572.6 mg/L of shikonin and 15.6 g/L of dry cell mass were obtained after 54 days. Shikonin was produced at a constant level of 10.6 mg/L day during this period.