Insulin-like growth factor signalling and its significance as a biomarker in fish and shellfish research

The insulin-like growth factor signalling system comprises insulin-like growth factors, insulin-like growth factor receptors and insulin-like growth factor-binding proteins. Along with the growth hormones, insulin-like growth factor signalling is very pivotal in the growth and development of all vertebrates. In fishes, insulin-like growth factors play an important role in osmoregulation, besides the neuroendocrine regulation of growth. Insulin-like growth factor concentration in plasma can assess the growth in fishes and shellfishes and therefore widely applied in nutritional research as an indicator to evaluate the performance of selected nutrients. The present review summarizes the role of insulin-like growth factor signalling in fishes and shellfishes, its significance in aquaculture and in evaluating growth, reproduction and development, and discusses the utility of this system as biomarkers for early indication of growth in aquaculture.

     

Evaluation of acoustic wave propagation velocities in the ocular lens and vitreous tissues of pigs, dogs, and rabbits

OBJECTIVE: To evaluate propagation velocity of acoustic waves through the lens and vitreous body of pigs, dogs, and rabbits and determine whether there were associations between acoustic wave speed and age, temperature, and time after enucleation. SAMPLE POPULATION: 9 pig, 40 dog, and 20 rabbit lenses and 16 pig, 17 dog, and 23 rabbit vitreous bodies. PROCEDURE: Acoustic wave velocities through the ocular structures were measured by use of the substitution technique. RESULTS: Mean sound wave velocities in lenses of pigs, dogs, and rabbits were 1,681, 1,707, and 1,731 m/s, respectively, at 36 degrees C. Mean sound wave velocities in the vitreous body of pigs, dogs, and rabbits were 1,535, 1,535, and 1,534 m/s, respectively, at 38 degrees C. The sound wave speed through the vitreous humor, but not the lens, increased linearly with temperature. An association between wave speed and age was observed in the rabbit tissues. Time after enucleation did not affect the velocity of sound in the lens or vitreous body. The sound wave speed conversion factors for lenses, calculated with respect to human ocular tissue at 36 degrees C, were 1.024, 1.040, and 1.055 for pig, dog, and rabbit lenses, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Conversion factors for the speed of sound through lens tissues are needed to avoid underestimation of the thickness of the lens and axial length of the eye in dogs during comparative A-mode ultrasound examinations. These findings are important for accurate calculation of intraocular lens power required to achieve emmetropia in veterinary patients after surgical lens extraction.     

Rapid screening of selected European winter wheat varieties and segregating populations for the Glu-D1d allele using PCR

Forty-two winter wheat varieties and 193 F₂ and BC₁F₂ seeds were screened for Glu-D1d allele encoding the HMW glutenin subunits 5 + 10 using polymerase chain reaction (PCR). The segregating populations originated from crosses involving wheat parents with good and poor bread-making quality. A clear PCR product of 450 bp, representing 1Dx5 of the Glu-D1d allele was identified in 24 varieties and 111 hybrid seeds. Four different Glu-D1 alleles: a (2 + 12), b (3 + 12), c (4 + 12) and d (5 + 10) were detected using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Only genotypes possessing Glu-D1d gave a positive PCR signal, hexaploid triticale and 4 × wheat lacking Glu-D1 locus gave a negative signal. The efficiency of PCR selection for bread-making quality in early generations using half seed is discussed.     

An efficient <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation protocol for black pepper (<Emphasis Type="Italic">Piper nigrum</Emphasis> L.) using embryogenic mass as explant

A protocol was developed for an efficient Agrobactertium-mediated transformation of black pepper plants through somatic embryogenesis. Embryogenic mass derived from primary somatic embryos that were obtained from the micropylar region of mature germinating seeds of black pepper was found to be the ideal target tissue for transformation. Genetic fidelity test of embryogenic mass-derived plantlets by RAPD using 23 random primers revealed no genetic variation among the progenies and the parent plant. Among the antibiotics used for selection of transformants, cefotaxime at 100 μg mL⁻¹ was found to be optimum to control Agrobacterium besides its ability to promote somatic embryo proliferation. In the case of kanamycin, a step-wise increase in concentration from 25 to 50 and then to 100 μg mL⁻¹ were found to be optimum. Embryogenic mass co-cultivated with Agrobacterium carrying the β-glucuronidase (GUS) reporter gene were cultured on plant growth regulator-free Schenk and Hildebrandt (SH) medium and transformants were selected in selection medium containing cefotaxime and step-wise increase in kanamycin concentration. The transient GUS gene expression was determined histochemically. Transformants that survived in the selection medium were hardened in the greenhouse. An average of nine hardened putative plantlets was obtained per gram of embryogenic mass. The presence of transgene in these plantlets was assayed by PCR, dot blot, and Southern blot hybridization. Results presented demonstrated for the first time an efficient transformation and regeneration of black pepper without the use of growth regulators. This simple efficient procedure would allow transformation of black pepper with genes of desirable characters.     

Discrimination between Oryza malampuzhaensis Krish. et Chand. and Oryza officinalis Wall ex Watt based on RAPD markers and morphological traits

Oryza malampuzhaensis, one of the tetraploid taxa of the genusOryza, is geographically restricted to Western Ghats of South India. Its taxonomic status is not well established and is generally treated as a tetraploid race of O. officinalis. Sixty-three morphological traits and 262 random amplified polymorphic DNA (RAPD) markers generated by 23 random decamer primers were used to assess the genetic relationship between O. malampuzhaensis and O. officinalis. Pair wise comparisons based on both RAPDs and morphological traits revealed 60% genetic distance between the two taxa and was significantly higher (p <0.01) than the corresponding intra-specific distances. Cluster and principal component analysis (PCA) of genetic distance estimations based on RAPDs and morphological traits clearly differentiated the two taxa. High frequency of discrete O. malampuzhaensis specific RAPDs (21%) and the significantly higher (p < 0.05) mean number of amplification products per individual in O. malampuzhaensis observed in the study reflect its allopolyploid nature. Low genetic diversity within O. malampuzhaensis revealed by RAPD analysis indicates the recent origin of this taxa. The RAPD analysis further revealed the possibility that the putative ‘C’ genomeprogenitor of O. malampuzhaensis is a close relative of O. officinalis. In addition, amplification products diagnostic to O. malampuzhaensis were identified. The results of the present study clearly demonstrated that the O. malampuzhaensis is a distinct entity, and support the recent conclusion that O. malampuzhaensis has diverged enough to deserve species status. This revised version was published online in August 2006 with corrections to the Cover Date.     

Evaluation of random amplified polymorphic DNA (RAPD) markers in Hevea brasiliensis

The applicability of random amplified polymorphic DNA (RAPD) markers in the cultivated rubber tree, Hevea, was evaluated using 43 decamer oligonucleotide primers in a set of 24 clones selected in different South-East Asian countries. A total of 220 0.35–3.5 kb DNA fragments were amplified, of which 111 were polymorphic. Of these, 80 fragments (RAPD markers) which were repeatable and clearly scorable across all genotypes were used to estimate genetic distances among the clones tested. The estimated genetic distances ranged from 0.05 (RRII 308 and PB 5/51) to 0.75 (RRIC 100 and SCATC 88–13). A mean genetic distance of 0.5 indicates a rather high genetic variability among the tested clones. As expected, because of the breeding history of Hevea, UPGMA cluster analysis and Principal Coordinate Analysis (PCoA) indicated the absence of a distinct geographical grouping. The possible application of RAPD markers for clone identification and also for analysis of genetic relationships among Hevea clones is discussed.     

Impact of Fertility Variation on Gene Diversity and Drift in two Clonal Seed Orchards of Teak (Tectona grandis Linn. f.)

Two 25 year old teak clonal seed orchards comprising 15 (CSO-I) and 20 clones (CSO-II), respectively, selected mostly from moist forests of Western Ghats (latitude 10° N) in southern India, were evaluated for fertility, offspring diversity, and genetic drift. The orchards differed in fertility of clones as well as flower and fruit production per ramet. Fertility was highly skewed in CSO-II, where one clone (originating from higher latitude −17° N, in Eastern Ghats of peninsular India) produced 55% of the fruits and 68% of the flowers in the orchard, in contrast to a similar contribution from four most fertile clones in CSO-I. Fertility variation, measured as `sibling coefficient' (1.7 in CSO-I and 8.3 in CSO-II), was high in CSO-II resulting in high coancestry and low effective population size (3 times lower than CSO-I) in the seed crop. In CSO-I, 58% of the clones contributed effectively to seed production compared to only 12% effective contribution resulting in eight times higher genetic drift in CSO-II. Placing limits on how much seed can be collected per clone might be useful in restricting over representation of highly reproductive clones thereby increasing genetic diversity in the seed crop.