Control of Brennandania lambi (Acari: Pygmephoroidea) by freezing: evaluation of its efficacy and effects on mushroom growth and yield

Laboratory and field studies were conducted to evaluate freezing as a control measure against Brennandania lambi (Krczal) which infests cultivated mushrooms (Agaricus bisporus) in Shanghai primarily through contaminated spawn. Laboratory experiments showed that exposure of contaminated spawn at -10° C for 24 h killed all stages of B. lambi. These mites probably died due to the freezing of body tissues during sustained exposure at -10° C for 24 h. because the supercooling points for the egg, active larva, quiescent larva, non-gravid adult and gravid adult of B. lambi were respectively -11.34, -10.96, -11.43, -7.65 and -11.20° C, whereas the freezing points were respectively -6.68, -6.84. -6.56, -3.96 and -6.75 °C. Semi-field experiments showed that compost inoculated with B. lambi-infested spawn that had been exposed at -10 °C for 24 h had no mite infestation during spawn running and thus mycelium growth was normal. Laboratory and field experiments during 1989 to spring 1990 showed that freezing spawn had no effect on mushroom yield. Further field experiments in two farms during fall 1990 showed no effect of freezing spawn on yield in one farm and increased yield in the other farm. A field experiment during 1991 to 1992 also indicated that freezing spawn -10 °C for 24 h to 34 h had no effect on mushroom yield.     

The genetic characterisation of novel multi-addition doubled haploid lines derived from triticale x wheat hybrids

Two novel 46-chromosome doubled haploid lines, W66 and M17, derived from separate hexaploid triticale x bread wheat crosses, were characterised using cytological and biochemical markers. Both lines were shown to be relatively stable cytologically, over 11 and 8 generations of selfing, respectively. By examining mitotic and meiotic chromosomes, the stabilities of the two lines were shown to be similar with frequencies of 2n=46 in 74.2–85.5% of cells. However, over selfed generations, the rye chromosomes were shown to have lost some of their heterochromatin, which made it difficult to establish their continued presence using cytological techniques, such as C-banding alone. Cytological evidence from pairing studies, C-banding, and fluorescence in-situ hybridization, showed that both M17 and W66 are wheat/rye multi-addition lines with rye chromosome constitutions of 1R+6R, and 1R+4R, respectively. These conclusions were confirmed by isozyme and storage-protein analysis.     

In vitro survival of murine morulae after quick freezing in the presence of chemically defined macromolecules and different cryoprotectants

We studied the ability of frozen-thawed mouse morulae to develop in vitro when the cryoprotectant proteins were substituted with one of the following nonorganic macromolecules: polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), and ficoll. We also determined how these agents interacted with 3 different cryoprotectants: glycerol (GLY), propylene glycol (PG), and ethylene glycol (EG). The influence of both of the above factors was measured on the basis of post-thaw morphological appearance, the percentage of development to the expanded blastocyst stage and the total cell count. Morulae (n=950) were collected from superovulated mice. Those classified as good or excellent were distributed among the 12 different freezing solutions, obtained by combining the 3 cryoprotectants with the 4 macromolecules (the 3 mentioned above, plus a control of 5% fetal calf serum) in phosphate buffered saline (PBS). Embryos frozen in PVA, PVP and ficoll tended to be a little difficult to recover from the straws. Development to the expanded blastocyst stage was significantly lower (P<0.05) in propylene glycol (43.6%) than in ethylene glycol (79.5%) or in glycerol (76.1%). Polyvinyl alcohol provided a higher survival rate when combined with glycerol (90.3) or ethylene glycol (95.0), but when it was combined with propylene glycol, only 56.5% of embryos survived after thawing. A positive interaction was observed between glycerol and PVA and between ethylene glycol and PVA or ficoll. The results indicate that fetal serum could be successfully substituted for any of the 3 chemically defined macromolecules. However, our findings also suggest that the use of PG as a cryoprotectant should be avoided when mouse morulae are frozen using the quick freezing method.     

Map-based cloning in crop plants: tomato as a model system II. Isolation and characterization of a set of overlapping yeast artificial chromosomes encompassing the jointless locus

Constructs carrying the entire or part of the tobacco nitrate reductase cDNA (NIA) cloned between the promoter and terminator sequences of the 35S RNA of the cauliflower mosaic virus were introduced into tobacco, in an attempt to improve nitrate assimilation. Several transgenic plants that had elevated NIA mRNA and nitrate reductase (NR) activity were obtained. In addition, a few plants that exhibited a chlorotic phenotype characteristic of NR-deficient mutants were also obtained. One of these plants contained no NIA mRNA, no NR activity and accumulated nitrate. This phenotype was therefore assumed to result from co-suppression of 35S-NIA transgenes and host NIA genes. NR-deficient plants were also found among the progeny of transformants overexpressing NIA mRNA. Genetic analyses indicated that these NR-deficient plants were homozygous for the 35S-NIA transgene, although not all homozygous plants were deficient for NR. The ratio of normal to NR-deficient plants in the progeny of homozygous plants remained constant at each generation, irrespective of the state of expression of the NIA genes (active or inactive) in the previous generation. This ratio also remained unchanged when field trials were performed in two areas of France: Versailles and Bergerac. The analysis of homozygous plants revealed that co-suppression was reversible at some stage of sexual reproduction. Indeed, host genes and transgenes reactivated at each generation, and co-suppression always appeared after a lag period of normal growth, suggesting that the phenomenon is developmentaly regulated. We observed that the triggering of cosuppression was delayed when plants were initially grown under limited light and/or watered with limited nitrate supply (light and nitrate both being required for the expression of the host NIA genes). However, this delay did not affect the final ratio between normal and NR-deficient plants after transfer to nitrate-fertilized fields. Independent transformants exhibited either different co-suppression ratios or no co-suppression at all, irrespective of the transgene copy number, suggesting that genomic sequences surrounding the transgene might play a role in determining co-suppression.     

Cloning of the blasticidin S deaminase gene (BSD) from Aspergillus terreus and its use as a selectable marker for Schizosaccharomyces pombe and Pyricularia oryzae

Aspergillus terreus produces a unique enzyme, blasticidin S deaminase, which catalyzes the deamination of blasticidin S (BS), and in consequence confers high resistance to the antibiotic. A cDNA clone derived from the structural gene for BS deaminase (BSD) was isolated by transforming Escherichia coli with an Aspergillus cDNA expression library and directly selecting for the ability to grow in the presence of the antibiotic. The complete nucleotide sequene of BSD was determined and proved to contain an open reading frame of 393 bp, encoding a polypeptide of 130 amino acids. Comparison of its nulceotide sequence with that of bsr, the BS deaminase gene isolated from Bacillus cereus, indicated no homology and a large difference in codon usage. The activity of BSD expressed in E. coli was easily quantified by an assay based on spectrophotometric recording. The BSD gene was placed in a shuttle vector for Schizosaccharomyces pombe, downstream of the SV40 early region promoter, and this allowed direct selection with BS at high frequency, following transformation into the yeast. The BSD gene was also employed as a selectable marker for Pyricularia oryzae, which could not be transformed to BS resistance by bsr. These results promise that the BSD gene will be useful as a new dominant selectable marker for eukaryotes.     

Identification of non-catalytic conserved regions in xylanases encoded by the xynB and xynD genes of the cellulolytic rumen anaerobe Ruminococcus flavefaciens

xynB is one of at least four genes from the cellulolytic rumen anaerobe Ruminococcus flavefaciens 17 that encode xylanase activity. The xynB gene is predicted to encode a 781-amino acid product starting with a signal peptide, followed by an amino-terminal xylanase domain which is identical at 89% and 78% of residues, respectively, to the amino-terminal xylanase domains of the bifunctional XynD and XynA enzymes from the same organism. Two separate regions within the carboxy-terminal 537 amino acids of XynB also show close similarities with domain B of XynD. These regions show no significant homology with cellulose- or xylan-binding domains from other species, or with any other sequences, and their functions are unknown. In addition a 30 to 32-residue threonine-rich region is present in both XynD and XynB. Codon usage shows a consistent pattern of bias in the three xylanase genes from R. flavefaciens that have been sequenced.     

豇豆单孢锈菌中病毒样颗粒中双链RNA的存在

从北京西郊清华园附近田间豇豆上采集的豇豆单孢锈菌(Uormyces vignal Barcl)夏孢子。萌发后提取双链RNA,电泳分析可测出300—8000碱基对的三组双链RNA。从萌发的孢子中通过差迷离心提取病毒样颗粒,可获得二种类型的病毒样颗粒,一种直径为35—40nm的等轴颗粒。另一种为长短不等的棒状颗粒,用提纯物提取核酸电泳分析与直接从孢子中提取的双链RNA有相同的核酸带,从而证明这些双链RNA存在于病毒样颗粒中。     

十字花科上的链格孢属真菌同工酶凝胶电泳分析

本文报道对从国内外收集到的生于十字花科植物上的3种链格孢属真菌21个菌株进行的10种同工酶的聚丙烯酰胺凝胶电泳分析结果。这10种酶分别是:酯酶(EST)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、多酚氧化酶(PPO)、过氧化氢酶(CAT)、谷氨酸脱氢酶(GDH)、甘露醇脱氢酶(MADH)、乙醇脱氢酶(ADH)、黄嘌呤脱氢酶(XDH)、过氧化物岐化酶(SOD)。对同工酶酶谱资料的聚类分析,在较高的相似性水平上将21个菌株归为3大类群,每一类群所包括的菌株与形态学的鉴定结果完全一致,各菌株的归类关系与其寄主植物、地理来源无关。表明同工酶凝胶电泳在Alternaria种级分类中是一个有用的工具,可以明确地将不同菌株鉴定到种级水平。     

海南省粉褶蕈属的分类研究(Ⅱ)

本文报道了海南省粉褶蕈属[Entoloma(Fr.)Kumm.]的31个种,其中17个为新记录种。并附分种检索表,表中含6个新种。 所有标本保藏于广东省微生物研究所真菌标本室。