Improved production of doubled haploids of winter and spring triticale hybrids via combination of colchicine treatments on anthers and regenerated plants

Double haploids (DH), obtained during androgenesis in vitro or by genome diploidisation in regenerated haploids, are one type of basic materials used in triticale breeding programmes. The aim of this study was to improve DH production by a combination of colchicine treatment methods on a sample of five winter and five spring triticale hybrids. Colchicine was applied in vitro either in the C17 medium to induce embryo-like structures (ELS) or in the 190-2 medium for green plant (GP) development. Regenerants which remained haploid were immersed in a colchicine solution either when placed on the medium prior to transferring to soil or when growing in pots, followed by the application or absence of cooling. Colchicine treatment during anther culture affected neither ELS nor GP development, but significantly increased the number of DH plants in comparison to spontaneous chromosome doubling. The highest efficiency was recorded when colchicine was applied in the induction medium (55%) versus the regeneration medium (44.5%) or no colchicine treatment (30%). The effectiveness of chromosome duplication in haploid plants ranged from 32 to 64.5% and it was the highest for the treatment on the medium followed by cooling. Individual hybrids differed regarding their capability of regeneration and chromosome doubling, which were consistent only to a low or moderate extent. However, taken together, winter and spring hybrids did not differ significantly. Combined colchicine application resulted in a high yield of DH production, 82.6% for all triticale hybrids, and can provide a considerable number of fertile DH lines for triticale breeding programmes.     

Rooting affects the photosystem II activity: in vitro and ex vitro studies on energy hybrid sorrel

Rumex tianschanicus × Rumex patientia is a high-biomass-yielding plant suitable for fuel and biogas production. The protocol of the hybrid sorrel micropropagation was used to study the changes in the photosystem II (PSII) activity as well as to analyse the ultrastructure of the chloroplasts. The lowest effective PSII quantum yield [Y(II)] and an apparent electron transport rate of PSII [ETR(II)] were observed for adventitious shoots that had been regenerated in vitro, before rooting. These fluorescence parameters were higher and similar for both the leaves of the same adventitious shoots that had been rooted under in vitro conditions and for the shoots that had been acclimated and grown in ex vitro conditions. The analysis indicated that the PSII activity strongly depends on the formation of properly functioning roots and that in vitro or ex vitro culture conditions are, at least to some degree, less important. TEM analysis revealed that chloroplasts from plants rooted in vitro were sufficiently mature and acclimatization processes have less impact on their development. This is the first report concerning the analysis of PSII activity and the ultrastructure of the chloroplasts at all of the stages of micropropagation, i.e. adventitious shoot formation in vitro, rooting in vitro and acclimation to ex vitro conditions. It strongly indicated that rooting under in vitro conditions, rather than the acclimation to ex vitro conditions, plays a key role in the development of a completely functional photosynthetic apparatus in hybrid sorrel.     

Epidemiological assessment of the risk of canine mast cell tumours based on the Kiupel two-grade malignancy classification

Background

The degree of differentiation of mast cell tumours (MCTs) is the most important feature and reflects the morphological characteristics and metastatic potential of the tumour and its likely response to treatment and the prognosis. The aim of this study was to epidemiologically analyse the risk of MCT development in dogs according to breed, age, sex, size and anatomical location of the tumour using the Kiupel grading system. The analysis involved 492 dogs selected based on a histopathological assessment of 2763 canine skin tumours. A logistic regression analysis was performed to determine the odds ratios (ORs) with 95% confidence intervals.

Results

Mast cell tumours accounted for 17.8% of all diagnosed canine skin tumours. The highest risk of high-grade MCTs was noted in the Shar-Pei (OR 28.18, P?<?0.001) and Weimaraner (OR 6.45, P?=?0.023). The highest risk of low-grade MCTs was determined in the Boxer (OR 6.72, P?<?0.001), and Pug (OR 6.13, P?=?0.027). The scrotum (OR 31.72, P?<?0.001), inguinal area (OR 17.69, P?<?0.001) and axilla (OR 6.30, P?<?0.001) had the highest risk of high-grade MCTs. The risk of high-grade MCTs increased with age and peaked in the oldest dogs, aged 11–16 years (OR 9.55, P?<?0.001). A higher risk of low-grade tumours was noted in younger dogs (aged 4–6 years) (OR 8.54, P?<?0.001) and females (OR 1.43, P?=?0.001). Statistical analysis further revealed a higher risk of both low (OR 3.47, P?<?0.001) and high-grade MCTs (OR 1.71, P?=?0.006) in medium-sized dogs.

Conclusions

This study demonstrated relationships between Kiupel grading system and phenotypic traits, age and location of canine MCTs confirming the complex biological nature of this tumour.

     

Hearing impairment caused by mutations in two different genes responsible for nonsyndromic and syndromic hearing loss within a single family

Usher syndrome is rare genetic disorder impairing two human senses, hearing and vision, with the characteristic late onset of vision loss. This syndrome is divided into three types. In all cases, the vision loss is postlingual, while loss of hearing is usually prelingual. The vestibular functions may also be disturbed in Usher type 1 and sometimes in type 3. Vestibular areflexia is helpful in making a proper diagnosis of the syndrome, but, often, the syndrome is misdiagnosed as a nonsyndromic hearing loss. Here, we present a Polish family with hearing loss, which was clinically classified as nonsyndromic. After excluding mutations in the DFNB1 locus, we implemented the next-generation sequencing method and revealed that hearing loss was syndromic and mutations in the USH2A gene indicate Usher syndrome. This research highlights the importance of molecular analysis in establishing a clinical diagnosis of congenital hearing loss.     

Molecular Typing by Pulsed-Field Gel Electrophoresis of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> from Root Voles

The root voles intestinal strains of Bacillus thuringiensis, were characterised by pulsed-field gel electrophoresis (PFGE). For 14 isolates, three pulsotypes were found, with the use of SmaI or NotI as restriction enzymes. Strains in each pulsotypes presented identical DNA patterns, indicating that the population structure of B. thuringiensis from root voles is clonal. The similarities in banding patterns were estimated at 56% and 33% for SmaI and NotI digests, respectively. The strains under study differed significantly in the size of their entire genome, which varied between 2.4 and 4.2 Mb. No significant differences were detected among the isolates subjected to biochemical properties determined by API tests. Present study showed that genomic diversity is a common feature of B. thuringiensis originating from one ecological niche. PFGE appears to be a useful technique for use in studies on the spread of B. thuringiensis in the environment. Received: 14 May 2002 / Accepted: 21 June 2002     

Parasitic microfungi of the Tatra Mountains: 5. <Emphasis Type="Italic">Plasmopara</Emphasis> representatives on the species of <Emphasis Type="Italic">Geranium</Emphasis>

A list of species and the distribution of the members of Plasmopara (Chromista, Peronosporales) on Geranium spp. in the Tatra National Park (Western Carpathians) biospheric reserve are given. Three fungal species parasitizing 3 plant species were recorded in the area. Plasmopara praetermissa is a species new to Poland and Slovakia.