Molecular wheat breeding by direct gene transfer

A method for efficient genetic transformation of wheat has been developed using immature embryos as targets for microprojectile-mediated gene transfer and a helium driven particle delivery system. Screening and selection of transgenic cells, somatic embryos and regenerated plants are performed with the gus-gene and the phosphinothricin acetyl transferase (bar) gene coding for Basta-resistance as the selectable marker. On average, one fertile transgenic plant can be obtained from about 100 microprojectile treated, immature embryos. The number of integrated copies of the transferred gene ranges from 1 up to about 10. Stable integrated genes are inherited in most of the transgenic lines in a normal mendelian fashion segregating 3:1 in the F₂. Homozygous, as well as heterozygous, lines have been followed and analysed genetically at the molecular level and up to F₅. Apart from normal stable gene expression, examples have also been found which showed a loss of gene activity or unexpected segregation pattern. For applied aspects, different genes are transferred aiming for improved disease resistance, modification of quality, or other characteristics. First results from these transgenic lines are reported, and problems still existing with the production of stable transgenic wheat lines are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of cowpea (Vigna Unguiculata) feeding on the pancreatic exocrine secretion of pigs

Introduction   Cowpea (Vigna unguiculata) is a valuable protein source that can contribute towards overcoming a predicted protein deficit equivalent to more than 1 000 000 t oilcake in Southern Africa by the year 2000 (N ell et al. 1992). However, like most other legumes, cowpeas contain antinutritional factors (ANFs) (M akinde et al. 1996). It has also been shown that feeding raw cowpea causes reduced growth and histomorphometric changes in the various segments of the intestine (M akinde et al. 1997) attributable to ANFs. The major ANFs in the cowpea are trypsin inhibitors, tannins and phytates (A letor and A ladetimi 1989). Although the protease inhibitors are found in most legumes, only soybean inhibitors have been thoroughly investigated (L iener and K akade 1980), and trypsin inhibitors in other legumes have received little attention.     

甘蓝型油菜(Brassica napus)无花瓣突变体“ Ap-Tengbe ”花瓣性状的遗传规律

本项研究旨在探明甘蓝型油菜突变体Ａｐ－Ｔｅｎｇｂｅ无花瓣性状的遗传规律,‘Ａｐ－Ｔｅｎｇｂｅ’和德国栽培品种‘Ｆａｌｃｏｎ’之间通过正反交产生两个方向的Ｆ１,ＢＣ１－１（Ｆ１和‘Ａｐ－Ｔｅｎｇｅｂｅ’之间）,ＢＣ１－２（Ｆ１和‘Ｆａｌｃｏｎ’之间）以及Ｆ２各遗传群体,根据田间各遗传群体ＰＤｇｒ（花瓣度）的观察和统计,作者提出‘Ａｐ－Ｔｅｎｇｅ’突变体无花瓣性状的遗传由细胞质和两对核隐性基因共同控制     

Water parameters affect anaesthesia induced by eugenol in silver catfish,Rhamdia quelen

The present study investigated the effects of water pH (5.0, 7.0 and 9.0), hardness (0, 20 and 120 mg CaCO₃ L^?1) and temperature (15, 23 and 30 °C) on the induction of sedation and anaesthesia, and subsequent recovery, of silver catfish exposed to eugenol. Moreover, the blood gas tensions (PvO₂ and PvCO₂) and blood pH in silver catfish acclimated to these temperatures were investigated after exposure to eugenol. Water pH, hardness, temperature and fish size affect the efficacy of eugenol in silver catfish, particularly at the lower concentrations tested (20 and 30 mg L^?1). Sedation of this species can be induced at concentrations as low as 20 mg L^?1, but for anaesthesia, a concentration of at least 40 mg L^?1 of eugenol must be used to compensate for the influence of fish size and water quality. Blood gas tension and pH were affected by eugenol anaesthesia, but only in fish acclimated to 30 °C.     

Rapid salinity changes affect the survival and physiology of a penaeid prawn: Implications of flood events on recruitment to the fishery

Estuaries act as nurseries for many penaeid prawns, but these habitats are highly susceptible to salinity decline through flooding. The rate of salinity decline and duration of exposure to non‐optimal salinity may affect survival and subsequent recruitment of prawns to the fishery. This study aimed to determine the effect of salinity fluctuations observed in local estuaries during flood events using a novel dilution approach. Mortality of juvenile Melicertus plebejus (Hess) was assessed after 24 hr exposure to 24 rates of salinity decline ranging from 0.01% to 20% per hr. After the salinity decline, prawns were held at the final salinities for five days before again assessing mortality as well as aerobic metabolic rate and prawn water content. Salinity decline from 36 to ~0.8 led to 50% mortality, but continued exposure to low salinity for five days increased mortality at this salinity to 99% and shifted the 50% mortality point to salinity ~5. Aerobic metabolic rate and water content data suggested the cause of mortality due to exposure to salinities < 5 was osmoregulatory failure. Rapid salinity declines over 24 hr and sustained low salinity due to flooding could compromise the survival of juvenile prawns, potentially reducing recruitment to the fishery.     

Iridovirus infections in finfish – critical review with emphasis on ranaviruses

Viruses in three genera of the family Iridoviridae (iridoviruses) affect finfish. Ranaviruses and megalocytiviruses are recently emerged pathogens. Both cause severe systemic disease, occur globally and affect a diversity of hosts. In contrast, lymphocystiviruses cause superficial lesions and rarely cause economic loss. The ranavirus epizootic haematopoietic necrosis virus (EHNV) from Australia was the first iridovirus to cause epizootic mortality in finfish. Like other ranaviruses, it lacks host specificity. A distinct but closely related virus, European catfish virus, occurs in finfish in Europe, while very similar ranaviruses occur in amphibians in Europe, Asia, Australia, North America and South America. These viruses can be distinguished from one another by conserved differences in the sequence of the major capsid protein gene, which informs policies of the World Organisation for Animal Health to minimize transboundary spread of these agents. However, limited epidemiological information and variations in disease expression create difficulties for design of sampling strategies for surveillance. There is still uncertainty surrounding the taxonomy of some putative ranaviruses such as Singapore grouper iridovirus and Santee‐Cooper ranavirus, both of which cause serious disease in fish, and confusion continues with diseases caused by megalocytiviruses. In this review, aspects of the agents and diseases caused by ranaviruses are contrasted with those due to megalocytiviruses to promote accurate diagnosis and characterization of the agents responsible. Ranavirus epizootics in amphibians are also discussed because of possible links with finfish and common anthropogenic mechanisms of spread. The source of the global epizootic of disease caused by systemic iridoviruses in finfish and amphibians is uncertain, but three possibilities are discussed: trade in food fish, trade in ornamental fish, reptiles and amphibians and emergence from unknown reservoir hosts associated with environmental change.     

Dietary inclusion of detoxified <Emphasis Type="Italic">Jatropha curcas</Emphasis> kernel meal: effects on growth performance and metabolic efficiency in common carp, <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.

Jatropha curcas is a multipurpose and drought-resistant shrub or small tree widespread all over the tropics and subtropics. Its seeds are rich in oil, and the Jatropha kernel meal obtained after oil extraction is rich in protein. However, presence of toxic and antinutritional constituents restricts its use in fish feed. Jatropha kernel meal was detoxified. Common carp, Cyprinus carpio, fingerlings (15; av. body mass 10.9 ± 0.65 g) were randomly distributed in three groups with five replicates. A 6-week feeding experiment was conducted in a respirometer system to evaluate the growth performance, nutrient utilisation and energy budget. Fish were fed isonitrogenous diets (38% crude protein): control diet (C _ontrol group) containing fish meal (FM) protein based protein and two other diets replacing 75% FM protein with detoxified Jatropha kernel meal (DJKM, J _atropha group) and soybean meal (SBM, S _oybean group). At the end of the experiment, body mass gain, metabolic growth rate, protein efficiency ratio, protein productive value, energy retention, efficiency of metabolised energy for growth and efficiency of energy retention were determined. These parameters were high and statistically similar for C _ontrol and J _atropha groups and significantly lower (P < 0.05) for S _oybean group. Whereas a reverse trend was observed for energy expenditure per g protein retained in fish body. No significant differences were found in heat released, gross energy uptake, metabolised energy intake, metabolisability, energy expenditure, energy expenditure per g protein fed and apparently unmetabolised energy. Conclusively, common carp–fed plant protein (DJKM and SBM) and FM protein–based diets exhibited equal average metabolic rate.     

Virucidal effects of various agents—including protease—against koi herpesvirus and viral haemorrhagic septicaemia virus

In a search for alternative, environmentally friendly and effective disinfecting agents, a commercially available protease—Neutrase^®—was tested in this work for inactivation of koi herpesvirus (KHV) and of viral haemorrhagic septicaemia virus (VHSV). For comparison, the stability of these viral pathogens in similar configurations at various pH values and concentrations of peracetic acid or quicklime, typically used for disinfection, was tested. Therefore, virus suspensions were incubated with various concentrations of different agents for 24 hr and the titre of the remaining infectious particles was determined by virus titration. Furthermore, the treatment of both viruses, with the agents at concentrations that were previously appointed as effective, was also examined in the presence of solid material (quartz sand). All procedures investigated in this study, including the protease treatment, were able to reduce the titre of KHV and VHSV below the detection limit of the titration. Although further studies are necessary, this is the first report of the application of a protease for the inactivation of the selected fish pathogens, demonstrating the great potential of the latter for disinfection.     

Genetic diversity in cultivated yam bean (<Emphasis Type="Italic">Pachyrhizus</Emphasis> spp.) evaluated through multivariate analysis of morphological and agronomic traits

Yam bean [Pachyrhizus DC.] is a legume genus of the subtribe Glycininae with three root crop species [P. erosus (L.) Urban, P. tuberosus (Lam.) Spreng., and P. ahipa (Wedd.) Parodi]. Two of the four cultivar groups found in P. tuberosus were studied: the roots of ‘Ashipa’ cultivars with low root dry matter (DM) content similar to P. erosus and P. ahipa are traditionally consumed raw as fruits, whereas ‘Chuin’ cultivars with high root DM content are cooked and consumed like manioc roots. Interspecific hybrids between yam bean species are generally completely fertile. This study examines the genetic diversity of the three crop species, their potentials for breeding and the identification of useful traits to differentiate among yam bean genotypes and accessions. In total, 34 entries (genotypes and accessions) were grown during 2000?2001 at two locations in Benin, West Africa, and 75 morphological and agronomical traits, encompassing 50 quantitative and 25 qualitative characters were measured. Diversity between entries was analyzed using principal component analysis, cluster analysis, multivariate analysis of variance and discriminant function analysis. Furthermore, phenotypic variation within and among species was investigated. Intra- and interspecific phenotypic diversity was quantified using the Shannon–Weaver diversity index. A character discard was tested by variance component estimations and multiple regression analysis. Quantitative trait variation ranged from 0.81 (for total harvest index) to 49.35% (for no. of storage roots per plant). Interspecific phenotypic variation was higher than intraspecific for quantitative traits in contrast to qualitative characters. Phenotypic variation was higher in overall for quantitative than qualitative traits. In general, intraspecific phenotypic variation ranged from 0.00 to 82.61%, and from 0.00 to 80.03% for quantitative and qualitative traits, respectively. Interspecific phenotypic variation ranged from 0.00 to 95.02%, and 0.00?81.58% for the two trait types, respectively. The Shannon–Weaver diversity index (H′) was in general high and over 0.80 for most of the trait. Diversity within P. tuberosus was higher than within P. erosus and P. ahipa. Across the 50 quantitative and 25 qualitative traits, the Shannon–Weaver diversity index of intra- and interspecific variation was around 0.83 and 0.51, respectively and was lower for qualitative than for quantitative traits. Monomorphism was observed in eight qualitative traits and one quantitative character. The first, second and third principal components explained, respectively, 39.1, 21.3 and 8.3% of the total variation in all traits. Pachyrhizus erosus, P. ahipa, and P. tuberosus (‘Chuin’ and ‘Ashipa’) were clearly separated from each other by these analyses. Multivariate analysis of variance indicates significant differences between Pachyrhizus species for all individual or grouped traits. Discriminant function analysis revealed that the first two discriminant functions were almost significant. Biases due to unbalanced sample size used per species were small. Within each species a similar amount of diversity was observed and was determinable to 70% by only ten traits. We conclude that the cultivated yam bean species represent distinct genepools and each exhibits similarly large amounts of genetic diversity.