Rapid and Sensitive Detection of Xanthomonas fragariae by Simple Alkaline DNA Extraction and the Polymerase Chain Reaction

Methods for DNA preparation from Xanthomonas fragariae in infected or artificially contaminated strawberry plants were compared in diagnostic assays using the polymerase chain reaction (PCR). The bacterium was detected using PCR with primers specific to a region of its hrp gene. Sensitivity of detection was 1.25 ×l 10³ CFU ml^-1 using DNA from bacterial suspensions prepared by an alkali extraction method. This was 10-fold more sensitive than DNA extraction by boiling, and was equal to that in which DNA was prepared by a more involved cetyltrimethylammonium bromide (CTAB) procedure. Sensitivity of detection from artificially contaminated strawberry tissues was 10-fold less than that from cell suspensions. The results indicated that a rapid and simple method of alkali DNA sample preparation is applicable for the sensitive and reliable detection of X. fragariae and possibly other plant pathogenic bacteria.     

Cosmid-derived markers anchoring the bovine genetic map to the physical map

The mapping strategy for the bovine genome described in this paper uses large insert clones as a tool for physical mapping and as a source of highly polymorphic microsatellites for genetic typing, and was one objective of the BovMap Project funded by the European Union (UE). Eight-three cosmid and phage clones were characterized and used to physically anchor the linkage groups defining all the bovine autosomes and the X Chromosome (Chr). By combining physical and genetic mapping, clones described in this paper have led to the identification of the linkage groups corresponding to Chr 9, 12, 16, and 25. In addition, anchored loci from this study were used to orient the linkage groups corresponding to Chr 3, 7, 8, 9, 13, 16, 18, 19, and 28 as identified in previously published maps. Comparison of the estimated size of the physical and linkage maps suggests that the genetic length of the bovine genome may be around 4000 cM. Received: 1 July 1996 / Accepted: 13 September 1996     

Effects of Microplitis croceipes Teratocytes on Host Haemolymph Protein Content and Fat Body Proliferation

Qualitative and quantitative changes in haemolymph proteins in Heliothis virescens were observed in larvae injected with either Microplitis croceipes teratocytes or teratocyte secreted proteins (TSP). Haemolymph protein titres in hosts receiving either 0.5 or 1 larval equivalent (LE) of teratocytes were similar to those of parasitized larvae, whereas a single injection of 4LE of TSP was required to induce a similar response. SDS-PAGE showed that the 82kDa monomer of riboflavin-binding protein and the 74/76kDa monomers of storage proteins were significantly reduced in parasitized larvae and in nonparasitized larvae treated with TSP. Concentrations of a 155kDa monomer (insectacyanin chromoprotein) also were reduced in parasitized larvae and those injected with either teratocytes or TSP. Two monomers (56 and 60kDa) were unique to parasitized larvae. Treated larvae required several days longer than controls to reach a comparable premetamorphic stage (burrowing-digging). Reductions in fat body proliferation similar to those seen in parasitized larvae were observed in larvae treated with either 1LE of teratocytes, or with 2 or 4LEs of TSP. Perivisceral fat body weights from larvae treated with either 0.25 or 0.5LE of teratocytes were significantly reduced, but less so than those which received 1LE. Thus, fat body proliferation in both teratocyte- and TSP-treated larvae was inhibited in a dose-dependent manner. Both light- and transmission electron microscopy observations revealed cytological differences in fat body tissues of larvae injected with either teratocytes or TSP from the condition observed in parasitized larvae and noninjected controls. Gross dissection of periviseral fat body from parasitized, teratocyte-injected and TSP-injected larvae showed tissue much less developed and differing considerably in appearance from controls. Observed differences included reduced size and/or number of lipid bodies and qualitative and quantitative changes in other cytoplasmic organelles.     

Does PBAN play an alternative role of controlling pheromone emission in the cabbage looper moth,Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae)?

There is an active process by which sex pheromone reserves of female cabbage looper moths, Trichoplusia ni, are transported to the gland's surface during the nocturnal period of calling. We hypothesized that this mobilization was controlled by a head factor, possibly related to the pheromone biosynthesis activating neuropeptides (PBAN) that in other species stimulate pheromone synthesis. We evaluated the impact of head extracts of T. ni on pheromone emission and glandular content of pheromone. During the photophase injected head extracts stimulated an increased pheromone emission rate in females, but glandular content of pheromone was not affected. Head extracts of H. virescens, a species with known PBAN activity, and synthetic PBAN stimulated an increased pheromone emission rate in T. ni. There was some specificity of the response of female T. ni to PBAN, in that several other unrelated polypeptides did not stimulate this type of response. Previously it had been determined that brain factors do not play a role in stimulating pheromone biosynthesis in T. ni. Our results indicate that there may be additional avenues by which PBAN or related neuropeptides control pheromone emission, including transport of pheromone reserves to the surface of the sex pheromone gland.     

The expression of biologically active human p53 in Leishmania cells: a novel eukaryotic system to produce recombinant proteins.

We have investigated the use of Leishmania cells as a novel eukaryotic expression system for the production of recombinant protein. These cells are easy to maintain, requiring no CO2 incubator or shaker, and can be grown in standard tissue culture media. Leishmania cells can be readily transfected with plasmid DNA by electroporation and transformants selected with antibiotic resistance. Recent studies have shown that it is possible to express foreign genes in Leishmania for the purpose of understanding the biology of this protozoan cell. In the present study we report the use of this system as a means of producing a biologically functional human p53 protein. The conformation of the p53 protein is critical for its ability to bind specific DNA sequences. It is demonstrated that Leishmania-synthesized human p53 is phosphorylated and can bind specifically to its enhancer DNA sequence. These data demonstrate that Leishmania may represent a simple eukaryotic expression system for the production of biologically active recombinant proteins.     

三尖杉叶精油化学成分的研究

三尖杉叶精油化学成分的研究苏应娟,王艇,张宏达（中国科学院武汉植物研究所武汉４３００７４）（中山大学生命科学学院广州５１０２７５）关键词三尖杉,精油,化学分类ＡＳＴＵＤＹＯＮＣＨＥＭＩＣＡＬＣＯＮＳＴＩＴＵＥＮＴＳＯＦＥＳＳＥＮＴＩＡＬＯＩＬＦＲＯＭ...     

高压静电场对绵羊精子存活率的影响

采用不同剂量的高压静电场处理绵羊精液,经分析发现,高压静电场对绵羊精具有激活作用。能提高绵羊精液品质,表现在适当剂量的高压静电场能显著地提高绵羊精子存活率,其中以６００ｋＶ／ｍ剂量处理效果最佳。１００ｋＶ／ｍ和３００ｋＶ／ｍ剂量对精子刺激不足,而９００ｋＶ／ｍ剂量则对精子刺激过程,导致部分精子损伤和死亡,同样达不到预期的效果。     

Preference of a revolving target to a stationary one by the big brown bat, Eptesicus fuscus

Utilizing a three-ramp platform, we studied the detection of a revolving and a stationary target in the presence of background clutter by trained Eptesicus fuscus. During the test, the mean amplitude of echo from either target was always larger than that of the background echoes at the bat-to-target distance of 30, 70 and 100 cm. The amplitude of the echo reflected back from a revolving target was modulated between a maximum and a minimum value. An electric motor was used to revolve a target. The frequency contents of the motor noise were mostly below 1 kHz. While the total percent response of approaching either target is always more than 90% at every bat-to-target distance tested, the bats approach a revolving target more frequently than a stationary one. Echolocation pulses emitted by the bats during the test were recorded and analyzed. The bats shortened their pulse durations and interpulse intervals and lowered the frequency contents as they entered into the crawling phase from the searching phase. Potential interference of background echoes and ambient noise with the performance of the bats is discussed. The preference of a revolving target to a stationary one by the bats is perhaps due to the fact that a revolving target has a higher releasing value than a stationary one does.