Nondestructive Raman analysis of polyacetylenes in apiaceae vegetables

Food plants from the Apiaceae family have been found to demonstrate health-promoting properties. Polyacetylenes are bioactive compounds that are considered to contribute substantially to the beneficial properties of Apiaceae plants. This study applied a Raman mapping technique in the investigation of polyacetylene spatial distribution in fresh roots of some Apiaceae species. Fresh root sections were measured directly without any preliminary preparation. For three Apiaceae species, that is, parsnip ( Pastinaca sativa L.), celeriac ( Apium graveolens var. rapaceum L.), and parsley ( Petroselinum crispum ), the presence of polyacetylenes was confirmed due to the detection of strong and well-separated Raman signals of symmetric -C ≡ C-C ≡ C- stretching vibration in the range of 2200-2300 cm(-1). The spectra were used for generation of two-dimensional maps applying the integration and cluster analysis methods. The Raman maps visualized the distribution of total polyacetylenes as well as individual compounds. Heterogeneous and tissue-specific occurrence of polyacetylenes in roots is shown.     

Viability of diploid and triploid larvae of Siberian sturgeon and bester hybrids

The aim of the present study was to produce Acipenser baeri× (Huso huso×Acipenser ruthenus) hybrids in a diploid and triploid state and to study their viability in comparison with the A. baeri from the fish farm stock. A heat shock (37°C) in the 18th minute after fertilization was applied to induce triploidy. The survival rate and the ploidy level of the hybrids obtained were studied. The mortality of triploid hybrids was approximately twice as high as the mortality of diploid hybrids. No significant difference in the survival rate between Siberian sturgeon and their diploid hybrid with bester was noted. Cytogenetic analysis was performed by preparing chromosomes from the gill epithelium. The results showed that all studied fish from the heat‐shocked group were triploid.     

Improving productivity of four fruit species by growth regulators applied once in ultra-low doses to the collar

Summary

Two growth retardants: paclobutrazol (P, Cultar) or 2,3,5-triiodobenzoic acid (TIBA) were applied once alone, or in mixtures (in a range of 5 to 20 mg per tree) to the collar of maiden plum, sour and sweet cherries and apple trees, in early spring of the second year after planting. Plum and apple trees were also treated with benzyladenine (BA) at 10 mg per tree a.i. in a mixture with P or TIBA. Sweet and sour cherry trees were treated with natural phenolic substances: phloridzin (Phi) and quercetin (Que) alone, or in mixtures with P and TIBA. A mid-stem treatment with P and shoot bending were also applied to the plum and apple trees for comparison. Measurements of tree growth and fruiting were made within 4 or 5 years. The reaction of the four species to the treatments varied according to the growth regulator applied. Plum trees responded mostly to TIBA and its mixtures with P. A strong suppression of tree growth and increased fruit productivity, as well as improved fruit quality, were observed. The TIBA application and its mixtures with P were also effective in causing growth reduction of the sweet cherry trees. Treatments with P alone, or mixed with TIBA, were effective in growth limitation of sour cherry trees. Some increase in the reproductive processes was observed only after the TIBA treatment in both species. The apple trees responded to application of mixtures of TIBA with benzyladenine (BA), or with P, and to P alone, with effective growth reduction. But only the P + BA treatment increased significantly the fruiting of apple trees, while the other treatments resulted in crops proportional to the diminished tree sizes. The mid-stem treatment did not affect plum trees but increased growth of apple trees. Shoot bending had no effect on the plum trees but increased fruiting of apple trees. The addition of Phi to half the lower P dose or TIBA, magnified growth suppression in the sweet and sour cherry trees. When applied alone Que caused a small growth inhibition but Phi produced some increase in growth of sour cherry trees. Results obtained show the possibilities of practical applications of growth regulators to the collar. Their reduced doses mixed with natural phenolic substances are equally effective in growth suppression and make fruit production safer and more profitable, especially in plum and sour cherry trees.     

Discrimination between nongenetically modified (Non-GM) and GM plant tissue expressing cysteine-rich polypeptide using FT-raman spectroscopy

Fourier transform (FT)-Raman spectroscopy was applied to the analysis of genetically modified (GM) plant tissue. Transgenic carrot callus and tobacco plants possessing a novel StSn1 gene coding for a cysteine-rich snakin-1 polypeptide were obtained after Agrobacterium-mediated transformation. The presence of the StSn1 gene and its expression were confirmed by polymerase chain reactions using plant DNA and cDNA as templates for the amplification of the transgenes. Raman measurements were taken from lyophilized GM carrot callus tissue, fresh GM tobacco leaves, and from seeds produced by GM tobacco plants as well as from the nontransformed controls. Cluster analysis applied to the obtained spectra allowed clear separation of the GM samples expressing the StSn1 gene and the nontransformed control to distinct groups. Such discrimination was achieved only when wavenumber ranges around 500 cm (-1) were analyzed. The results indicate that discrimination between the GM and non-GM materials was related to S-S stretching vibrations in snakin-1, as it contained six sulfur bridges. Other introduced genes, neomycine phosphotransferase ( nptII) and Chitinase ( chit36), did not cause any detectable changes by Raman spectroscopy in plant tissue. This is the first report on the use of Raman spectroscopy for a nondestructive analysis of GM plant material expressing the gene coding for a cysteine-rich polypeptide.     

Rep-PCR Based Genomic Fingerprinting of Isolates of Leptosphaeria maculans from Poland

Leptosphaeria maculans, the ascomycete fungus which causes blackleg disease of oilseed rape, has been considered for a long time as a single species divided into aggressive and non-aggressive pathogenicity groups which differ in their economic importance. However, the development of accurate biochemical and molecular characterisation methods has demonstrated that the world-wide L. maculans population actually comprises at least two species. The aim of this research was to assess the ability of rep (repetitive element based)-PCR genomic fingerprinting methods, initially developed for bacterial identification, to characterise a collection of 90 isolates of L. maculans from Poland, in comparison with reference isolates from the IBCN (International Blackleg of Crucifers Network) collection. REP (repetitive extragenic palindromic)-, ERIC (enterobacterial repetitive intergenic consensus)-, and BOX primers for rep-PCR genomic fingerprinting, and primers derived from LMR1, a L. maculans specific repeated element, were tested. Rep-PCR and LMR1-based analyses were able to discriminate the different components of the species complex and to evaluate the genetic diversity within each member of the complex. These analyses suggested that Polish populations of L. maculans mainly belong to the non-aggressive species, rather than the aggressive species which is prevalent in Western Europe, Canada and Australia.     

Inostemma melicerta Walker, 1835 (Hym., Platygastridae), ein neuer Parasit vonResseliella piceae Seitner, 1906 (Dipt., Cecidomyiidae) in Weißtannensamen

A hitherto unknown parasite of theResseliella piceae larvae, the pest of fir-seeds (Abies alba Mill.), was found and identified asInostemma melicerta. The material originated from the Piwniczna Management Unit in Beskid S?decki Mts. in southern Poland.     

A follow-up study of neurologic and radiographic findings in working German Shepherd Dogs with and without degenerative lumbosacral stenosis

OBJECTIVE: To identify radiographic abnormalities associated with degenerative lumbosacral stenosis (DLSS) in German Shepherd Dogs (GSDs) and determine whether specific radiographic abnormalities could be used to identify dogs at risk of developing DLSS. DESIGN: Cohort study. ANIMALS: 33 GSDs working as police dogs. PROCEDURES: Results of physical, neurologic, and orthopedic examinations were used to identify dogs with DLSS. Survey radiography of the lumbosacral junction was performed, and radiographs were compared with radiographs obtained 3 years earlier. RESULTS: DLSS was diagnosed in 15 of the 33 (45%) dogs. Thirteen of the 15 dogs with DLSS and 14 of the 18 dogs without DLSS had radiographic abnormalities of the lumbosacral junction. Twenty-two (67%) dogs were able to perform unrestricted duties, including 3 dogs with suspected DLSS. Six (18%) dogs had been excluded from active duty during the period of surveillance because of DLSS. Significant progression in specific clinical and radiographic signs was detected, but multiple logistic regression analysis did not identify any radiographic signs that could be used to predict the development of DLSS. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that survey radiography cannot be used to predict development of DLSS in working GSDs.     

利用油菜花瓣实验检测油菜菌核病

摘要：油菜菌核病是由Sclerotinia sclerotiorum (Lib) de Bary引起的,是世界范围内造成损失最严重的病害之一。该病害主要通过感染了核盘菌子囊孢子的花瓣落在植株茎、叶上来侵染植株。本实验采用添加了两种不同PH指示剂的改良型Steadman’s培养基检测被子囊孢子感染的花瓣,样品采自合肥的两块不同试验田,在油菜花期（3月下旬～4月中旬）每天采摘新鲜花瓣在室温下培养,结果表明整个花期,除1块田1天没有检测出外,其余每天都有被子囊孢子感染的花瓣出现（感染子囊孢子的花瓣接种后培养基变色）。油菜初花期被子囊孢子感染的花瓣较少,而盛花期和终花期被感染的花瓣较多,第一块田被感染的花瓣数在两种培养基上出现的百分率分别为32.0％和40.2％,第二块田分别为27.3％和35.7％。第一块田油菜菌核病的发病率和病指分别为2.7％和1.8,第二块田分别为1.5％和0.4。子囊孢子感染的花瓣数与后期油菜菌核病的发病率呈显著正相关（r=0.829,r0.05=0.811）。研究被感染的花瓣数可以帮助决定最适宜的病害防治时期。     

Does 2-hydroxyflutamide Inhibit Apoptosis in Porcine Granulosa Cells? - An In Vitro Study

In mammalian ovaries, the majority of follicles are lost before ovulation by atresia. This degenerative process is initiated or caused by granulosa cell apoptosis. To reveal the androgen-dependent mechanism of selective follicular atresia, the culture model system for agonism and antagonism of the androgen receptor has been established. We examined the influence of an androgen receptor antagonist, 2-hydroxyflutamide (2-Hf), on the incidence of apoptosis in cultured porcine granulosa cells. They were incubated (6 and 12-h) in the presence of testosterone (T, 10(-7)M), 2-Hf (1.7×10(-4) M) or both T and 2-Hf (T+2-Hf), and then analyzed by flow cytometry with fluorescein labelled annexin V. To better imitate in vivo conditions, the intact porcine follicles (6-8 mm in diameter) have been incubated in an organ culture system with the addition of the same factors. Sections obtained from follicles fixed after culture were stained with hematoxylin and eosin, and the presence of apoptosis-related DNA strand breaks was evaluated by the TUNEL method. Estradiol and progesterone concentrations in the culture media were measured by radioimmunoassays. The addition of T or 2-Hf to the culture media caused an increase in the number of apoptotic granulosa cells, while treatment with T+2-Hf decreased it in both in vitro and organotypic models. Follicles cultured with the addition of T or 2-Hf exhibited morphological changes indicating follicular atresia. Granulosal estradiol secretion was considerably stimulated by T+2-Hf. The highest increase in follicular estradiol secretion was observed after the anti-androgen addition. In both granulosal and follicular cultures, the production of progesterone declined in the presence of T or 2-Hf but increased after their simultaneous addition. In conclusion, androgen receptor antagonist 2-Hf attenuates induction of granulosa cell apoptosis in the presence of a high T level. The nature of this protective mechanism as yet is unknown and requires further research.