Development of simple sequence repeat (SSR) markers in Eucalyptus from amplified inter-simple sequence repeats (ISSR)

Eucalyptus spp. are widely used in exotic plantations. Since many of these trees are derived from vegetative propagation, the routine identification of clones has become increasingly important. The most widely used molecular based method for fingerprinting these clones is by random amplified polymorphic DNAs (RAPDs). Although this technique is useful, its results are not very repeatable, especially between laboratories. The aim of this study was to develop microsatellite markers that are highly repeatable, and to investigate their value in Eucalyptus fingerprinting. Typically, this process involves the expensive procedure of constructing an enriched genomic library. However, we used an intersimple sequence repeat (ISSR) polymerase chain reaction (PCR)‐based enrichment technique for microsatellite‐rich regions. With this relatively inexpensive method, microsatellite‐rich regions were amplified directly from genomic DNA, after which PCR products were cloned and sequenced. From these microsatellite‐rich sequences, primer sets were constructed to amplify mono‐, di‐, tri‐, hexa‐and nona‐nucleotide repeats. These markers were all inherited in a Mendelian fashion in the progeny of a test cross between two Eucalyptus grandis trees. The primer sets developed were also able to amplify the corresponding microsatellite loci from five different Eucalyptus spp., namely E. grandis, E. nitens, E. globulus, E. camaldulensis and E. urophylla.     

<Emphasis Type="Italic">Fusarium mangiferae</Emphasis> associated with mango malformation in the Sultanate of Oman

Mango malformation, caused by Fusarium mangiferae, represents the most important floral disease of mango. The first symptoms of this disease were noticed in the beginning of 2005 in plantations at Sohar in the Sultanate of Oman. The affected inflorescences were abnormally enlarged and branched with heavy and dried-out panicles. Based on morphology and DNA-sequence data for the genes encoding translation elongation factor 1α and β-tubulin, the pathogen associated with these symptoms was identified as F. mangiferae.     

Effect of Diaporthe RNA virus 1 (DRV1) on growth and pathogenicity of different <Emphasis Type="Italic">Diaporthe</Emphasis> species

A 4.1 kbp positive-strand RNA virus known as Diaporthe RNA virus 1 (DRV1) occurs in hypovirulent, non-sporulating isolates of the fungal pathogen Diaporthe perjuncta. A full-length cDNA clone of DRV1 was developed and RNA transcribed from the cDNA clone used to transfect different Diaporthe spp. The transfected species included three D. ambigua isolates and an unidentified Phomopsis asexual state of a Diaporthe sp. Successful transfections were confirmed using RT-PCR. Although the in vitro-transcribed positive sense single-stranded RNA used for transfection included vector sequences at both ends, the genomes of progeny virus from DRV1-transfected isolates were free of the vector sequences. Transfection resulted in morphological changes in these fungal pathogens. However, the presence of DRV1 did not reduce growth rate in two of the three D. ambigua or the Phomopsis sp. significantly. Pathogenicity studies showed that the transfected isolates have reduced aggresiveness.     

Variation in growth rates and aggressiveness of naturally occurring self‐fertile and self‐sterile isolates of the wilt pathogen Ceratocystis albifundus

Ceratocystis albifundus is the most important fungal pathogen of black wattle (Acacia mearnsii) grown in plantations in southern and eastern Africa. It is a homothallic fungus but also undergoes unidirectional mating type switching. As a result, the ascospore progeny can be either self‐fertile or self‐sterile. The only apparent difference between these mating types is the deletion of the MAT1‐2‐1 gene in self‐sterile isolates. There is some evidence suggesting that self‐sterile isolates grow more slowly than self‐fertile isolates, but this has not been tested rigorously. The aim of this study was to determine whether self‐sterile isolates are less fit by examining growth rate, relative germination rate and pathogenicity. Five self‐sterile isolates were generated from each of five self‐fertile isolates of C. albifundus and these 30 isolates were compared. The results showed that the self‐sterile isolates grew consistently slower and were less pathogenic than the self‐fertile isolates. The germination ratio of self‐fertile to self‐sterile isolates from single ascospores collected from the ascomata of five self‐fertile isolates was on average 7:3. This could be a consequence of the self‐sterile isolates having a lower germination rate. This observation, and the lower growth and pathogenicity levels, suggests that self‐sterile isolates are not likely to compete effectively in nature, raising intriguing questions regarding their role and value to C. albifundus and other fungi having a similar mating system.     

Serum concentrations of thyroxine, 3,5,3'-triiodothyronine, thyrotropin, and prolactin in dogs before and after thyrotropin-releasing hormone administration

Serum concentrations of thyrotropin (TSH), prolactin, thyroxine, and 3,5,3'-triiodothyronine in 15 euthyroid dogs and 5 thyroidectomized and propylthiouracil-treated dogs after thyrotropin-releasing hormone (TRH) administration were measured. Although thyroidectomized and propylthiouracil-treated dogs had higher (P less than 0.01) base-line concentrations of TSH in serum than did euthyroid dogs, concentrations of TSH after TRH administration varied at 7.5, 15, and 30 minutes with 14 of 45 samples obtained from healthy dogs having lower TSH concentrations than before TRH challenge. Similarly, concentrations of 3,5,3'-triiodothyronine in the serum of euthyroid dogs 4 hours after TRH administration were similar (P less than 0.05) to concentrations before TRH challenge. Although the mean concentration of thyroxine in serum was elevated (P less than 0.05) 4 hours after administration of TRH to euthyroid animals, as compared with base-line levels, the individual response was variable with concentrations not changing or decreasing in 4 dogs. Therefore, the TRH challenge test as performed in the current investigation was of limited value in evaluating canine pituitary gland function. Although mean concentrations of TSH in serum were higher (P less than 0.05) in euthyroid dogs after TRH administration, the response was too variable among individual animals for accurate evaluation of pituitary gland function. Concentrations of prolactin in the sera of dogs after TRH administration, confirmed previous reports that exogenously administered TRH results in prolactin release from the canine pituitary and indicated that the TRH used was biologically potent.     

Circumcostal gastropexy for preventing recurrence of gastric dilatation-volvulus in the dog: an evaluation of 30 cases

Follow-up evaluation (mean, 13.7 months) was obtained in 30 dogs with gastric dilatation-volvulus that were surgically treated with circumcostal gastropexy. Evaluation was performed by questionnaire (29 dogs), radiographic contrast studies (23 dogs), and necropsy (6 dogs). The gastropexy was thought to be intact in all 23 dogs examined radiographically. Necropsy of 6 dogs dying from causes unrelated to gastric dilatation-volvulus (mean of 12.7 months after surgery) revealed an intact gastropexy site in each dog. Only 1 (3.3%) dog had clinical recurrence of gastric dilatation after surgery. Five additional dogs dying postoperatively were necropsied and had intact gastropexy sites.     

Corticimorbus sinomyrti gen. et sp. nov. (Cryphonectriaceae) pathogenic to native Rhodomyrtus tomentosa (Myrtaceae) in South China

The Cryphonectriaceae includes many important tree pathogens, especially of the Myrtales. Disease surveys on Myrtales in South China revealed a stem canker disease on native Rhodomyrtus tomentosa (Myrtaceae, Myrtales) trees in the proximity of Eucalyptus plantations in GuangXi Province and in a natural forestry area in the Hong Kong Region. Fruiting structures with typical characteristics of the Cryphonectriaceae were observed on the canker surfaces. The fungus was identified based on DNA sequence comparisons and morphological features, and its pathogenicity was tested on R. tomentosa under field conditions. DNA sequence comparisons for the partial large subunit, partial internal transcribed spacer nuclear ribosomal DNA, and two regions of the β‐tubulin gene showed that the fungus represents a previously undescribed genus and species in the Cryphonectriaceae. Based on morphology, the fungus is most similar to species in the genus Chrysoporthe, but can be distinguished from this and the other genera of Cryphonectriaceae. The fungus provided with the name Corticimorbus sinomyrti gen. et sp. nov., is described. Field inoculations showed that it is highly pathogenic to R. tomentosa trees, with the ability to kill inoculated branches within 4 weeks. Pathogenicity tests also showed that C. sinomyrti is pathogenic to a tested Eucalyptus clone. Pathogenicity on both native R. tomentosa and non‐native Eucalyptus suggests that this fungus should be monitored carefully to limit its possible spread to commercially grown Eucalyptus in South China.     

Presence of Leptin and Its Receptor in the Hypothalamus,Uterus and Ovaries of Swine Females Culled with Distinct Ovarian Statuses and Parities

Leptin acts on energy metabolism, affecting reproductive functions through activation of its receptors in the brain and in reproductive organs. This study compared the presence of leptin and its receptor (ObR‐b) in hypothalamus neurons, endometrial glands and oocytes of culled swine females across ovarian statuses and parities. Immunohistochemistry was done in samples of uterus, ovaries and hypothalamus from 28 culled females, using polyclonal antibodies antileptin and ObR‐b. Immunolabelling was compared for sows categorized by parity at culling (0, 1, 2–4 and <4) and ovarian status (luteal and follicular phases of the oestrous cycle and with cysts). Immunolabelling for leptin and ObR‐b in neurons and oocytes was weaker in females with cysts (p < 0.05) than in those at the follicular phase. In endometrial glands, leptin immunolabelling was less intense in females with cysts (p < 0.05), but immunolabelling for ObR‐b was similar across ovarian statuses (p > 0.05). In sows culled with 2–4 parities, leptin immunolabelling in neurons and endometrial glands was more intense than in nulliparous females (p < 0.05). In comparison with sows culled at greater parities, ObR‐b immunolabelling for nulliparous females was less intense in endometrial glands and in oocytes (p < 0.05), but more intense in neurons (p < 0.05). Thus, in swine, the presence of leptin and ObR‐b varies across parities and is more intense in the uterus, ovaries and hypothalamus of females that were cycling before culling than in those having cystic ovaries.