Comparison of methods for obtaining doubled haploids of carrot

Doubled haploid lines of carrot can be obtained through androgenesis in anther cultures and in isolated microspore cultures. The two methods were compared using three carrot cultivars (‘Kazan F1’, ‘Feria F1’, and ‘Narbonne F1’) at the androgenesis induction stage, during plant regeneration from embryos, and during acclimatization of androgenetic plants as well as their characterization. It was found that cultivar was the main factor affecting the efficiency at each stage of plant production in both anther and isolated microspore cultures. The efficiency of androgenesis in anther cultures of ‘Feria F1’ was considerably higher in comparison with isolated microspore cultures, and more plants were obtained from the embryos of androgenesis-cultured plants. In ‘Kazan F1’ and ‘Narbonne F1’, more acclimatized androgenetic plants were produced from anther cultures. Ploidy assessment of acclimatized plants of ‘Narbonne F1’ showed that the majority of the plants in the population derived from anther cultures had a doubled chromosome (DH) set. On the other hand, the majority of plants obtained from isolated microspore cultures were haploids. When assessing homozygosity, it was found among plants obtained in anther cultures that the percentage of homozygotes for phosphoglucose isomerase (PGI) and aspartate aminotransferase (AAT) depended on the cultivar. In contrast, the majority of plants derived from isolated microspore cultures were homozygous regardless of cultivar

Effect of increased copper ion content in the medium on the regeneration of androgenetic embryos of carrot (Daucus carota L.)

The study was conducted to determine the effect of elevated concentrations of copper in the medium on the regeneration of androgenetic embryos of the carrot cultivar ‘Kazan F1’ obtained in anther cultures and to determine the level of soluble phenols produced in the regenerates under copper stress. Green embryos were laid out on 4 regeneration media based on B5 medium (G a m b o r g et al. 1968) without hormones, containing 0.1 – control, 1, 10, and 100 μM CuSO4×5H2O. The plant material was passaged 3 times, after 4, 9 and 15 weeks. During these passages the emerging structures were examined; they were classified in terms of growth and development in vitro, weighed and counted. The levels of soluble phenols in the freeze-dried regenerates were determined. The elevated concentrations of copper in the regeneration media affected positively the formation of complete plants (rooted rosettes) and secondary embryos during the first 4 weeks of culture. After a longer regeneration time (9, 15 weeks), the elevated concentrations of copper caused negative effects: deformation of rosettes. After 15 weeks, the number of rooted rosettes decreased. The 9-week culture subjected to copper stress brought about an increase in the amounts of soluble phenols. The highest values were recorded in the rosettes treated with 10 μM CuSO4. Prolonged exposure to media containing elevated concentrations of CuSO4 caused a reduction in the accumulation of phenolic compounds in the rosettes

Jednoczesne wykrycie komórek nowotworowych i materiału genetycznego prątków gruźlicy w płynie opłucnowym

Wysięk opłucnowy jest często obserwowanym zjawiskiem w przebiegu chorób układu oddechowego zarówno zapalnych, jak i nowotworowych. Jego przyczyną może być gruźlica, której najczęstszą pozapłucną lokalizacją jest opłucna, a także choroba nowotworowa opłucnej. Potwierdzenie gruźliczej etiologii wysięku opłucnowego bywa trudne, podobnie jak znalezienie pierwotnego ogniska choroby w przypadku wysięku nowotworowego. W części przypadków wykrycie pierwotnego ogniska nowotworu, pomimo zastosowania różnorakich metod diagnostycznych, jest niemożliwe. Poniżej przedstawiono przypadek obustronnego płynu w opłucnej, w którym jednocześnie wykryto komórki raka o nieustalonym punkcie wyjścia oraz stwierdzono dodatni wynik testu genetycznego w kierunku prątków gruźlicy. Pneumonol. Alergol. Pol. 2010; 78, 4: 296-301Pleural effusion is a frequently observed lesion in the course of respiratory diseases such as inflammatory process and cancer metastasis. Its cause may be either tuberculosis (the most common extrapulmonary location is the pleura) and malignant disease of the pleura. Confirmation of tuberculosis is often troublesome. The primary site of cancer may be also difficult to find despite the application of difficult diagnostic methods. Below we present history of 79-year-old female in whom carcinomatous cells and positive result of PCR for Mycobacterium tuberculosis in pleural fluid were discovered simultaneously suggesting the tuberculosis and cancer of unknown primary origin. Pneumonol. Alergol. Pol. 2010; 78, 4: 296-30

Infectious diseases of horseradish (Cochlearia armoracia L.) in Poland.

Poland is an important horseradish grower in Europe, therefore the problems concerning its cultivation and diseases are considered vital. Horseradish diseases can be classified as non-infectious and fungal, bacterial or virus origin. The most important fungal disease is white-rust caused by Albugo candida, which can be responsible for up to 50% loss of yield, in the case of heavy infection. Bacterial diseases are not very important now. Turnip mosaic virus (TuMV), arabis mosaic virus (AMV) and tomato black ring virus (TBRV) were found so far on horseradish in Poland. The most important is TuMV, which was found in almost 100% collected horseradish samples and caused yield loss about 40%. The virus TuMV belongs to the Potyvirus genus (VC57.0.1) from the Potyviridae family (VC57), which is the largest family of plant viruses. It infects many economically important plants. Attempts to eradicate TuMV by means of thermotherapy proved ineffective. At the Research Institute of Vegetable Crops, methods of obtaining TuMV-free horseradish plants (together with methods of its fast multiplication) have been developed in in vitro cultures

Influence of Polyamines on Red Beet (<i>Beta vulgaris</i> L. ssp. <i>vulgaris</i>) Gynogenesis

The influence of polyamines (PAs), putrescine (Put) and spermidine (Spd) on the efficiency of gynogenesis in ovule cultures of red beet (syn. beetroot) (Beta vulgaris L. vulgaris) cultivar “Czerwona Kula” and breeding accessions no. 3/2010 and no. 7/2008 was investigated. The effect of Put on the process of plant regeneration from gynogenetic embryos was studied. The response to the applied PAs was strongly dependent on the genotype. In “Czerwona Kula”, an increase in the number of obtained embryos was achieved by using each of the two PAs in the B5 medium. The effect of Spd was stronger. Put added to the regeneration medium at the concentration of 0.5 mg L−1 increased the number of obtained plants. All shoots placed on the rooting medium supplemented with 160 mg L−1 Put formed roots. The distribution of ploidy and homozygosity of gynogenetic plants depended on the genotype. Of the tested genotypes, the highest number of haploid plants, 68%, was obtained in red beet “Czerwona Kula”. The highest percentage of homozygotes, 69% for the glucose phosphate isomerase (GPI, E.C.5.3.1.9) isoenzyme and 100% for the aspartate aminotransferase (AAT, E.C.2.6.1.1) isoenzyme, was obtained in the population of gynogenetic plants of cultivar “Czerwona Kula”

Development of embryoids by microspore and anther cultures of red beet (Beta vulgaris L. subsp. vulgaris)

So far there is no information about receiving red beet androgenic embryos by androgenesis. Several factors were tested which affected this process: starch accumulation in microspores, correlation between bud length and microsporogenesis course, induction and regeneration medium composition. Ploidy level of obtained regenerants were evaluated. Treating anthers with α-amylase or watering donor plants with gibberellin increased number of obtained androgenic embryos. The highest percentage (80%) of microspores at uninuclear stage appeared in buds with 1.3-1.5 mm. The B5 medium with 100 g·L-1 sucrose and 0.1 mg·L-1 2,4-D (2, 4-dichlorophenoxyacetic acid) proved to be better for inducing androgenesis than MS medium supplemented with 0.2 mg·L-1 BAP (6-benzylaminopurine) and 0.5 mg·L-1 IAA (indole-3-acetic acid). First androgenic embryos were placed on B5 medium without plant growth regulators and then on MS medium containing 0.2 mg·L-1 BAP and 1 mg·L-1 NAA (α-naphthaleneacetic acid). Androgenic embryos died on B5 regeneration medium without plant growth regulators. On MS medium first shoots and callus with and without roots were obtained. Rosettes withered during following passages whereas callus tissue developed further. The quantity of DNA in this tissue equivalent to 4X chromosomes

Simultaneous Detection of Tumour Cells and Positive Genetic Test Results for Mycobacterium tuberculosis in Pleural Effusion

Pleural effusion is a frequently observed lesion in the course of respiratory diseases such as inflammatory process and cancer metastasis. Its cause may be either tuberculosis (the most common extrapulmonary location is the pleura) and malignant disease of the pleura. Confirmation of tuberculosis is often troublesome. The primary site of cancer may be als difficult to find despite the application of difficult diagnostic methods. Below we present history of 79-year old female in whom carcinomatous cells and positive result of PCR for Mycobacterium tuberculosis in pleural fluid were discovered simultaneously suggesting the tuberculosis and cancer of unknown primary origin