Antifungal effect of Penicillium metabolites against some fungi

Microorganisms are increasingly exploited as a source of new biological control agents. Genus Penicillium is a source of novel bioactive molecules which can be used as antifungal agents. The objective of this study was to evaluate the antifungal potential of Penicillium strains. Culture filtrates of two Penicillium species were tested for their antifungal potential by well diffusion assays. Filtrate of Penicillium isolates showed high antifungal effects on mycelial growth of Fusarium oxysporum, Fusarium solani, Macrophomina phaseolina, Aspergillus japonicus var aculeatus and Cladosporium cladosporioides. But Penicillium italicum inhibit the fungal growth from 45 to 68% as compared to Penicillium simplissimum (25–68%). However in case of A. japonicus var aculeatus, Penicillium spp. extracts were equally effective and reduce the colony growth up to 68%. However, P. simplissimum extract was least effective in case of M. phaseolina, where it decreased the colony growth only 25%.     

Rapid assessment of resistance of tissue-cultured water yam (Dioscorea alata) and white guinea yam (Dioscorea rotundata) to anthracnose (Colletotrichum gloeosporioides Penz.)

Yam anthracnose is caused by the pathogen Colletotrichum gloeosporioides Penz. and has been identified as the most important biotic constraint to yam production worldwide. Rapid assessment of the disease is vital to its effective diagnosis and management. In this study, tissue-cultured yam plantlets of five lines of Dioscorea alata and nine of D. rotundata were rapidly assessed for their reactions to two isolates of yam anthracnose. The plantlets, obtained from meristem of the nodal cuttings, were grown for 8?weeks on Murashige and Skoog (MS) basal medium, acclimatised for 3?weeks, hardened for an additional 3?weeks, arranged in screen house in completely randomised design and sprayed with spore inocula prepared from 7?day-old culture of the two strains of Colletotrichum gloeosporioidies Penz. The relative resistance of the different Dioscorea spp. was evaluated using three disease indices – severity at seventh day after inoculation, SD7; area under disease progress curve, AUDPC; and disease severity rate, Rd. A modified rank-sum classification method put TDa 1425 and TDr 2040, with rank sum of 2.0 each, as resistant. TDr 2121, TDr 2287 and TDr 2048 were susceptible with rank sum of 27.50, 25.50 and 24.50, respectively. Dioscorea alata TDa 1425 and Dioscorea rotundata TDr 2040 were recommended in areas endemic with yam anthracnose, and also as parent lines while breeding for resistance to anthracnose.     

Establishment and characterization of an epithelial cell line from thymus of Catla catla (Hamilton, 1822)

A cell line, CTE, derived from catla (Catla catla) thymus has been established by explant method and subcultured for more than 70 passages over a period of 400 days. The cell line has been maintained in L-15 (Leibovitz) medium supplemented with 10% fetal bovine serum. CTE cell line consists of homogeneous population of epithelial-like cells and grows optimally at 28 °C. Karyotype analysis revealed that the modal chromosome number of CTE cells was 50. Partial amplification, sequencing and alignment of fragments of two mitochondrial genes 16S rRNA and COI confirmed that CTE cell line originated from catla. Significant green fluorescent signals were observed when the cell line was transfected with phrGFP II-N mammalian expression vector, indicating its potential utility for transgenic and genetic manipulation studies. The CTE cells showed strong positivity for cytokeratin, indicating that cell line was epithelial in nature. The flow cytometric analysis of cell line revealed a higher number of cells in S-phase at 48 h, suggesting a high growth rate. The extracellular products of Vibrio cholerae MTCC 3904 were toxic to the CTE cells. This cell line was not susceptible to fish betanodavirus, the causative agent of viral nervous necrosis in a large variety of marine fish.     

单头亚菊的组织培养与快速繁殖

以单头亚菊茎段为外植体对其进行组织培养,MS为基本培养基,设置不同激素浓度配比。对实验结果进行观察分析,筛选出合适的配方。启动培养基为Ms＋0．5mg·L-16-BA＋0．01mg·L-1NAA。继代培养基MS＋O．75mg·L-1。6-BA＋0．01mg·L。NAA,可获得较高的增殖率。不定根最适诱导培养基为1／2MS＋O．15mg·L—IBA,生根率达87％以上,组培苗移栽成活率达98％。     

‘香槟’月季的组织培养和试管开花诱导

本文对‘香槟’月季（80sachinensis‘Xiangbin’）的组织培养技术和诱导试管开花进行了研究。结果表明：以茎段为外植体能诱导获得无菌苗,适宜的启动培养基为MS＋6-BA1．0mg-L-1＋IBA0．1mg·L-1,幼芽继代增殖的最佳培养基是MS＋6．BA1．0mg·L-1。＋IBA0．1～0．2mg·L-1,诱导生根的适宜培养基为1／2MS＋NAA0．3mg·L-1,生根率达80．0％。诱导试管开花的适宜培养基为MS＋6．BA0．5mg·L-1＋NAA0．1mg·L-1最适宜的诱导试管开花的蔗糖含量是30g·L-1;在三角瓶中培养,试管花可以正常开放,在培养瓶中培养花芽不能正常开放;MS培养基中增加2倍磷的含量,可以提高花芽诱导率,为25．O％;诱导试管开花的最适培养条件为温度21℃,光照强度80～100μmol·m-2．s-1,光照时间16h—d-1。     

THE EUPYRENE-APYRENE DICHOTOMOUS SPERMATOGENESIS OF LEPIDOPTERA. ORGAN CULTURE STUDY ON THE TIMING OF APYRENE COMMITMENT IN THE CODLING MOTH

Lepidopteran spermatogenesis is dichotomous, producing eupyrene (nucleated) and apyrene (anucleated) spermatozoa. The eupyrene precedes the apyrene spermatogenesis. The timing of the switchover from eupyrene to apyrene spermatogenesis was determined by cultivating testes of accurately aged codling moth larvae in a medium containing mammalian serum but neither hemolymph nor insect hormones. In cultures, eupyrene spermatogenesis occurred in testes dissected from either 4th or 5th instar larvae, probably due to macromolecular factor-like activity of the serum of the medium. But apyrene spermatogenesis occurred only in testes explanted during or after the fourth day of the 5th instar larva. It is concluded that: (1) An apyrene spermatogenesis inducing factor (ASIF) becomes active on the fourth day of the 5th instar larva in addition to the already existing macromolecular factor. (2) Primary spermatocytes can develop into either eupyrene or apyrene spermatozoa. (3) The apyrene spermatogenesis commitment and pupal commitment of other tissues coincide about the fourth day of the 5th instar larva.     

Hepatopancreas and ovary are sites of vitellogenin synthesis as determined from partial cDNA encoding of vitellogenin in the marine shrimp,Penaeus vannamei

Summary

The site of yolk protein synthesis in crustaceans has long been a subject of controversy. A portion of the vitellogenin gene structure was reported recently in a freshwater giant prawn (Macrobrachium rosenbergii) and black tiger shrimp (Penaeus monodori), in which the hepatopancreas was confirmed to be the extraovarian site of vitellogenin synthesis. The ovary is also frequently reported to be the site of yolk protein synthesis in penaeid shrimp. The same PCR product was obtained using cDNA from the hepatopancreas or the ovary as a template. The deduced amino acid sequence of Vg in P. vannamei showed high identities of 57% and 78% with those from M. rosenbergii and P. monodon, respectively. The same location of the intron in the sequenced region of genomic DNA was also found between these three species. We therefore concluded that the hepatopancreas and ovary are sites of vitellogenin synthesis in P. vannamei. The partial structure of the vitellogenin gene is further presented.     

Glycosylation: impact,control and improvement during therapeutic protein production

Abstract

The emergence of the biopharmaceutical industry represented a major revolution for modern medicine, through the development of recombinant therapeutic proteins that brought new hope for many patients with previously untreatable diseases. There is a ever-growing demand for these therapeutics that forces a constant technological evolution to increase product yields while simultaneously reducing costs. However, the process changes made for this purpose may also affect the quality of the product, a factor that was initially overlooked but which is now a major focus of concern. Of the many properties determining product quality, glycosylation is regarded as one of the most important, influencing, for example, the biological activity, serum half-life and immunogenicity of the protein. Consequently, monitoring and control of glycosylation is now critical in biopharmaceutical manufacturing and a requirement of regulatory agencies. A rapid evolution is being observed in this context, concerning the influence of glycosylation in the efficacy of different therapeutic proteins, the impact on glycosylation of a diversity of parameters/processes involved in therapeutic protein production, the analytical methodologies employed for glycosylation monitoring and control, as well as strategies that are being explored to use this property to improve therapeutic protein efficacy (glycoengineering). This work reviews the main findings on these subjects, providing an up-to-date source of information to support further studies.