A genetic linkage map in autotetraploid lucerne adapted to northern Australia, and use of the map to identify DNA markers linked to resistance to Phytophthora medicaginis

Phytophthora root rot, caused by Phytophthora medicaginis, is a major limitation to lucerne ( Medicago sativa L.) production in Australia and North America. Quantitative trait loci (QTLs) involved in resistance to P. medicaginis were identified in a lucerne backcross population of 120 individuals. A genetic linkage map was constructed for tetraploid lucerne using 50 RAPD ( randomly amplified polymorphic DNA), 104 AFLP (amplified fragment length polymorphism) markers, and one SSR ( simple sequence repeat or microsatellite) marker, which originated from the resistant parent (W116); 13 markers remain unlinked. The linkage map contains 18 linkage groups covering 2136.5 cM, with an average distance of 15.0 cM between markers. Four of the linkage groups contained only either 2 or 3 markers. Using duplex markers and repulsion phase linkages the map condensed to 7 homology groups and 2 unassigned linkage groups. Three regions located on linkage groups 2, 14, and 18, were identified as associated with root reaction and the QTLs explained 6 - 15% of the phenotypic variation. The research also indicates that different resistance QTLs are involved in conferring resistance in different organs. Two QTLs were identified as associated with disease resistance expressed after inoculation of detached leaves. The marker, W11-2 on group 18, identified as associated with root reaction, contributed 7% of the phenotypic variation in leaf response in our population. This marker appears to be linked to a QTL encoding a resistance factor contributing to both root and leaf reaction. One other QTL, not identified as associated with root reaction, was positioned on group 1 and contributed to 6% of the variation. This genetic linkage map provides an entry point for future molecular-based improvement of lucerne in Australia, and markers linked to the QTLs we have reported should be useful for marker-assisted selection for partial resistance to P. medicaginis in lucerne.

Does the cutting of lucerne (Medicago sativa) encourage the movement of arthropod pests and predators into the adjacent crop?

Lucerne (Medicago sativa) has been suggested as an ideal refuge habitat as part of an integrated pest management (IPM) program because it harbours high numbers of beneficial arthropods. Whether or not cutting of lucerne encourages the movement of these beneficials into adjacent target crops is unknown. Vacuum samples were used to determine the effects of cutting lucerne on arthropod abundance (pests and predators) within lucerne and adjacent soybean (Glycine max) crops. Vacuum-sample collections of arthropods were conducted before and after lucerne cutting on seven occasions in four fields over two seasons. In the lucerne, 10 m by 1 m strips parallel to the crop interface were sampled at 5, 10, 15, 20 and 30 m from the interface. In the soybean, 10 m of row were sampled at the same distances from the crop interface. The abundance of predators in lucerne was reduced immediately after cutting at all distances from the interface. Predator abundance in soybean did not show any change. The cutting of lucerne significantly reduced pest numbers within the lucerne but had little effect on pest abundance in the adjacent soybean. The temporal pattern in pest and predator abundance was very different for each field sampled. Generally, arthropods decreased in abundance after cutting and gradually increased as the lucerne grew back. In soybeans, arthropod numbers fluctuated regardless of the cutting of the lucerne. Cutting of lucerne alone does not guarantee movement of predators into the adjacent target crop. The presence of lucerne fields within a cropping area may have some impact on regional predator populations, and so still be useful for IPM programs, but this has yet to be tested critically.

Identifying and discriminating among lucerne cultivars using plant morphological characters

The use of morphological data obtained from field (plot test) and glasshouse trials to identify and discriminate among four Iranian and two New Zealand lucerne (Medicago sativa L.) cultivars was investigated, following guidelines established by the International Union for the Protection of New Varieties of Plants (UPOV) for cultivar registration and the Organisation for Economic Co-operation and Development (OECD) for seed certification. Data were collected for terminal leaflet length, width and ratio, angle of stem growth, date of first flowering, stem height at first flowering, flower colour, cutting recovery height, and disease scores. None of these characters were sufficient to identify or discriminate among the six cultivars. The results indicate a need to find cost-effective and efficient laboratory techniques to enhance the assessment of distinctness of lucerne cultivars (UPOV) and for determining cultivar purity for lucerne seed certification (OECD).

Monensin supplementation of lactating cows fed tropical grasses and cane molasses or grain

Effects of monensin (Mon) on performance of Holstein-Friesian cows fed tropical grasses and cane molasses (M) or cereal grain were examined in three experiments. In experiment I (incomplete 4 x 4 Latin square), three rumen-fistulated cows [188 I I days in milk (DIM)] were fed mixed diets based on rhodes grass (Chloris gayana cv. Callide) bay where M was substituted for wheat grain (W) at rates of 0 (MO), 125 (M 125) or 250 (M250) g/kg dry matter (DM). A fourth diet contained M250 plus 0.02 g Mon/kg DM (M250 + Mon). Substituting M for W tended (P < 0.10) to decrease the ratio of rumen molar proportions of acetate+butyrate (Bu):propionate (Pr) (4.3 versus 3.8 and 4.0 for M0, M125 and M250, respectively). There were no treatment effects (P> 0.10) on intake, organic matter digestibility, milk production or liveweight (LW) change. In experiment 2, 48 cows (173 &PLUSMN; 28.3 DIM) grazing kikuyu (Pennisetum clandestinum cv. common) pastures and supplemented with maize silage and a grain-based concentrate were offered either M (2.6 kg DM/(cow day)) or barley grain (B) (2.7 kg DM/(cow day)). Within each supplement type, half were fed 0 or 320 mg of Mon/(cow day). There were Mon x supplement interactions (Mon x S; P < 0.05) on the rumen molar proportion of Pr and Bu at 15:00 h, with B + Mon having the highest value for Pr (0.259 mmol/mmol) and lowest value for Bu (0.121 mmol/mmol). A Mon x S effect (P < 0.05) on milk fat content was noted with Mon causing a lower value regardless of energy source (31 and 36 g/l versus 40 and 38 g/l for B + Mon, M + Mon, B - Mon and M - Mon, respectively). As a main effect, M as opposed to B, reduced yields of milk (P < 0.05; 16.21/(cow day) versus 18.01/(cow day)) and protein (P < 0.05; 479 g/(cow day) versus 538 g/(cow day)). Monensin reduced milk fat yield (P < 0.05; 669 g/(cow day) versus 562 g/(cow day)), raised milk protein concentration (P < 0.05; 31 g/l versus 29 g/l) and caused LW gain rather than loss (P < 0.05; +0.06 kg/(cow day) versus -0.30 kg/(cow day)). No treatment effects on pasture intake were noted. In experiment 3, 48 cows (91 &PLUSMN; 16.1 DIM) grazing kikuyu pasture and supplemented with grain-based concentrate, sugar cane silage and 2.7 kg DM(cow day) of M were supplemented with either 0 or 320 mg Mon/(cow day). Monensin reduced (P < 0.05) milk fat content (33 g/l versus 30 g/l) and tended (P < 0.10) to reduce milk protein content (29 g/l versus 28 g/l). No effects of Mon on other milk production parameters, LW change or pasture intake were noted. Feeding monensin to mid-lactation Holstein-Friesian cows offered diets based on tropical grasses, and cane molasses or grain, improves rumen fermentation efficiency, thereby improving energy efficiency resulting in higher LW gain. Monensin had no effect on milk yield, but reduced milk fat concentration.

The effect of high temperature on the insecticidal properties of Bt Cotton

Bt transgenic cotton has not shown the same level of resistance to bollworm in China, as in other major Bt cotton growing areas of the world. The objective of this study was to investigate the effects of high temperature on the CryIA insecticidal protein content and nitrogen metabolism, in the leaf of Bt transgenic cotton. The study was undertaken on two transgenic cotton cultivars, one conventional (Xinyang 822) and the other a hybrid (Kumian No. 1), during the 2001 and 2002 growing seasons at the Yangzhou University Farm, Yangzhou, China. In the 2001 study, potted cotton plants were exposed to 37 C for 24 h under glasshouse conditions at three growth stages peak square, peak flowering and peak boll developing periods. Based on the 2001 results, in 2002 the same two cultivars were exposed to the same temperature for 48 h at two growth stages-peak flowering and boll developing periods. The results of the study indicated that the insecticidal protein content of the leaf was not significantly affected by the stress during the square and flowering periods. However, exposure to high temperature for 24h during the boll period reduced the CryIA protein content by approximately 51% in the cultivar Kumian No 1, and 30% in Xinyang 822 in the 2001 study, and by approximately 73 and 63% for 48 h with the same cultivars, respectively, in the 2002 study. Glutamic-pyruvic transaminase (GPT) activity, total free amino acid and soluble protein content, and the activity of protease in the leaf, showed relatively little change in response to high temperature in the flowering period. However, exposure to high temperature in the boll period resulted in the following changes - a reduction of GPT activity, a sharp increase in free amino acid content, a significant decrease in soluble protein content, and significant increases in the activity of protease. The results suggest that high temperature may result in the degradation of soluble protein in the leaf, with a resulting decline in the level of the toxin CryIA. It is believed that this may be the cause of the reduced efficacy of Bt cotton in growing conditions in China, where temperatures during the boll period often reach 36-40&DEG; C. © 2004 Elsevier B.V All rights reserved.

Potential to increase yield in lucerne (Medicago sativa subsp sativa) through introgression of Medicago sativa subsp falcata into Australian adapted material

The effect of interspecific heterosis in crosses between Medicago sativa subsp. sativa and M. sativa subsp. falcata was assessed. Three sativa and 3 falcata plants were crossed in a diallel design. Progeny dry matter yield and natural plant height were assessed in a replicated field experiment at Gatton, Queensland. Yield data were analysed using the method of residual maximum likelihood (REML) and Griffing's model 1. There were significant differences between the reciprocal, general combining ability (GCA), and specific combining ability (SCA) effects. As expected, S-1 populations were lower yielding than their respective intraspecific cross and falcata x falcata crosses were significantly lower yielding than sativa x sativa crosses. Some of the interspecific crosses indicated substantial SCA effects, yielding at least as well as the best sativa x sativa crosses. We have demonstrated the potential usefulness of unselected M. sativa subsp. falcata as a heterotic group in the improvement of yield in northern Australian adapted lucerne material, and discuss how it could be incorporated into future breeding to overcome the yield stagnation currently being experienced in Australian programs.

Properties of a unique mutant of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus that exhibits a partial many polyhedra and few polyhedra phenotype on extended serial passaging in suspension cell cultures

Serial passaging of wild-type Helicoverpa armigera, single-nucleocapsid (HaSNPV) in H. zea (HzAMI) illsect Cell Cultures results ill rapid selection for the few polyhedra (FP) phenotype. A unique HaSNPV mutant (ppC19) was isolated through plaque purification that exhibited a partial many polyhedra (MP) and FP phenotype. Oil serial passaging in suspension cell cultures, ppC19 produced fivefold more polyhedra than a typical FP mutant (FP8AS) but threefold less polyhedra than the wild-type virus. Most importantly, the polyhedra of ppC19 exhibited MP-like virion occlusion. Furthermore, ppC19 produced the same amount of budded virus (BV) as the FP mutant, which was fivefold higher than that of the wild-type virus. This selective advantage was likely to explain its relative stability in polyhedra production for six passages when compared with the wild-type Virus. However, subsequent passaging of ppC19 resulted in a steel) decline in both BV and polyhedra yields, which was also experienced by FP8AS and the wild-type virus Lit high passage numbers. Genomic deoxyribonueleic Licid profiling of the latter suggested that defective interfering particles (DIPS) were implicated in this phenomenon and represented another Undesirable mutation during serial passaging of HaSNPV Hence, a strategy to isolate HaSNPV Clones that exhibited MP-like polyhedra production but FP-like BV production, coupled with low multiplicities of infection during scale-up to avoid accumulation of DIPS, could prove commerically invaluable.

DNA markers linked to yield, yield components, and morphological traits in autotetraploid lucerne (Medicago sativa L.)

We have mapped and identifed DNA markers linked to morphology, yield, and yield components of lucerne, using a backcross population derived from winter-active parents. The high-yielding and recurrent parent, D, produced individual markers that accounted for up to 18% of total yield over 6 harvests, at Gatton, south-eastern Queensland. The same marker, AC/TT8, was consistently identified at each individual harvest, and in individual harvests accounted for up to 26% of the phenotypic variation for yield. This marker was located in linkage group 2 of the D map, and several other markers positively associated with yield were consistently identified in this linkage group. Similarly, markers negatively associated with yield were consistently identified in the W116 map, W116 being the low-yielding parent. Highly significant positive correlations were observed between total yield and yield for harvests 1-6, and between total yield and stem length, tiller number, leaf yield/plant, leaf yield/5 stems, stem yield/plant, and stem yield/5 stems. Highly significant QTL were located for all these characters as well as for leaf shape and pubescence.

Genetic improvement of lucerne for anthracnose (Colletotrichum trifolii) resistance

Anthracnose, caused by Colletotrichum trifolii, is one of the most serious diseases influencing lucerne persistence and productivity in eastern Australia. The disease is largely controlled by plant resistance; however, new pathotypes of C. trifolii have developed in Australia, seriously limiting the productive life of susceptible cultivars. This paper describes an incompletely recessive and quantitatively inherited resistance to C. trifolii identified in a clone (W116) from cv. Sequel. S-1, F-1, F-2 and backcross populations of W116 and D (highly susceptible clone) were studied for their reaction to C. trifolii race 1. Resistance was found to be quantitatively inherited, and quantitative trait loci associated with resistance and susceptibility were identified in a backcross population (D x W116) x D using random amplified polymorphic DNA and amplified fragment length polymorphic markers. A multi-locus region on linkage group 4 was found to contribute significantly to the resistance phenotype. The application of DNA markers to allow exploitation of this quantitatively inherited resistance in lucerne breeding is discussed.

Impacts of feeding system and season on milk composition and Cheddar cheese yield in a subtropical environment

Milk obtained from cows on 2 subtropical dairy feeding systems were compared for their suitability for Cheddar cheese manufacture. Cheeses were made in a small-scale cheesemaking plant capable of making 2 blocks ( about 2 kg each) of Cheddar cheese concurrently. Its repeatability was tested over 10 separate cheesemaking days with no significant differences being found between the 2 vats in cheesemaking parameters or cheese characteristics. In the feeding trial, 16 pairs of Holstein - Friesian cows were used in 2 feeding systems (M1, rain-grown tropical grass pastures and oats; and M5, a feedlot, based on maize/barley silage and lucerne hay) over 2 seasons ( spring and autumn corresponding to early and late lactation, respectively). Total dry matter, crude protein (kg/cow. day) and metabolisable energy (MJ/cow.day) intakes were 17, 2.7, and 187 for M1 and 24, 4, 260 for M5, respectively. M5 cows produced higher milk yields and milk with higher protein and casein levels than the M1 cows, but the total solids and fat levels were similar (P > 0.05) for both M1 and M5 cows. The yield and yield efficiency of cheese produced from the 2 feeding systems were also not significantly different. The results suggest that intensive tropical pasture systems can produce milk suitable for Cheddar cheese manufacture when cows are supplemented with a high energy concentrate. Season and stage of lactation had a much greater effect than feeding system on milk and cheesemaking characteristics with autumn ( late lactation) milk having higher protein and fat contents and producing higher cheese yields.

Strategies to obtain stable transgenic plants from non-embryogenic lines: complementation of the nn(1) mutation of the NORK gene in Medicago sativa MN1008

Strategies to introduce genes into non-embryogenic plants for complementation of a mutation are described and tested on tetraploid alfalfa (Medicago sativa). Genes conditioning embryogenic potential, a mutant phenotype, and a gene to complement the mutation can be combined using several different crossing and selection steps. In the successful strategy used here, the M. sativa genotype MnNC-1008(NN) carrying the recessive non-nodulating mutant allele nn(1) was crossed with the highly embryogenic alfalfa line Regen S and embryogenic hybrid individuals were identified from the F1 progeny. After transformation of these hybrids with the wild-type gene (NORK), an F2 generation segregating for the mutation and transgene were produced. Plants homozygous for the mutant allele and carrying the wild-type NORK transgene could form root nodules after inoculation with Sinorhizobium meliloti demonstrating successful complementation of the nn(1) mutation.

In vitro production of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus

Naturally occurring insect viruses are a promising means of intentionally causing disease in insects but they do not compete successfully with synthetic chemicals in the commercial marketplace. Furthermore, their use for pest control is still restricted. One factor preventing the development of baculoviruses as effective biopesticides is concern over the production issue. In vitro instability during propagation of these viruses in suspension cells is the major limitation to the in vitro production ofbaculoviruses in cell cultures. In this study, an isolated baculovirus (HaSNPV) was cultivated using serial passaging in a suspension cell culture. The results show a reduction in the occlusion body production during six passages, due to the passage effect. However the purification of an HaSNPV clone suggested better stability. A simple method used in this work for the serial passaging of this virus is discussed.

Expanding the biomass resource:sustainable oil production via fast pyrolysis of low input high diversity biomass and the potential integration of thermochemical and biological conversion routes

Waste biomass is generated during the conservation management of semi-natural habitats, and represents an unused resource and potential bioenergy feedstock that does not compete with food production. Thermogravimetric analysis was used to characterise a representative range of biomass generated during conservation management in Wales. Of the biomass types assessed, those dominated by rush (Juncus effuses) and bracken (Pteridium aquilinum) exhibited the highest and lowest volatile compositions respectively and were selected for bench scale conversion via fast pyrolysis. Each biomass type was ensiled and a sub-sample of silage was washed and pressed. Demineralization of conservation biomass through washing and pressing was associated with higher oil yields following fast pyrolysis. The oil yields were within the published range established for the dedicated energy crops miscanthus and willow. In order to examine the potential a multiple output energy system was developed with gross power production estimates following valorisation of the press fluid, char and oil. If used in multi fuel industrial burners the char and oil alone would displace 3.9 × 105 tonnes per year of No. 2 light oil using Welsh biomass from conservation management. Bioenergy and product development using these feedstocks could simultaneously support biodiversity management and displace fossil fuels, thereby reducing GHG emissions. Gross power generation predictions show good potential.

Desempenho agronômico, bromatológico e estabilidade fenotípica de sorgo silageiro em Uberlândia - MG

Sorghum (Sorghum bicolor (L.) Moench) is a good alternative to be used as silage, especially in places with water scarcity and high temperatures, due to their morphological and physiological characteristics. The appropriate management, as the ideal seeding time, interferes both productivity and the quality of silage. The work was conducted with the objective of evaluating the agronomic and bromatological performance of varieties and hybrids of silage sorghum and their phenotypic stability in two seasons, season and off-season, in the city of Uberlândia, Minas Gerais. The experiments were performed at Capim Branco Experimental Farm of Federal University of Uberlândia (UFU), located in the referred city. There were two sowing dates in the same experimental area, off-season (March to June 2014) and season (November 2014 to March 2015), and the varieties and hybrids were evaluated in both situations. The design was a randomized block with 25 treatments (hybrids and varieties of sorghum) and three replications. Agronomical and bromatological data were subjected to an analysis of variance; averages were grouped by Scott-Knott test at 5% of probability, through Genes computer program; and to estimate the stability, it was opted for Annicchiarico method. The flowering of cultivars, dry matter productivity, plant height, Acid Detergent Fiber (ADF), Neutral Detergent Fiber (NDF) and Crude Protein (CP) are affected by the environment and the variety. Regarding productivity and quality of the fiber, SF11 variety was superior, independent of the rated environment. In relation to the performance stability of dry matter, the varieties SF15, SF11, SF25, PROG 134 IPA, 1141572, 1141570 and 1141562 were highlighted. For the stability of the quality of fibers (FDA and FDN), the variety 1141562 stood out. The environment reduces the expression of characters “days of flowering”, “plant height” and “productivity of dry matter of hybrids”. From the 25 hybrids analyzed for productivity and stability of dry matter performance, seven were highlighted, regardless of the rated environment: Volumax commercial hybrid and experiments 12F39006, 12F39007, 12F37014, 12F39014, 12F38009 and 12F02006.

Catabolic flexibility of mammalian-associated lactobacilli

Metabolic flexibility may be generally defined as “the capacity for the organism to adapt fuel oxidation to fuel availability”. The metabolic diversification strategies used by individual bacteria vary greatly from the use of novel or acquired enzymes to the use of plasmid-localised genes and transporters. In this review, we describe the ability of lactobacilli to utilise a variety of carbon sources from their current or new environments in order to grow and survive. The genus Lactobacillus now includes more than 150 species, many with adaptive capabilities, broad metabolic capacity and species/strain variance. They are therefore, an informative example of a cell factory capable of adapting to new niches with differing nutritional landscapes. Indeed, lactobacilli naturally colonise and grow in a wide variety of environmental niches which include the roots and foliage of plants, silage, various fermented foods and beverages, the human vagina and the mammalian gastrointestinal tract (GIT; including the mouth, stomach, small intestine and large intestine). Here we primarily describe the metabolic flexibility of some lactobacilli isolated from the mammalian gastrointestinal tract, and we also describe some of the food-associated species with a proven ability to adapt to the GIT. As examples this review concentrates on the following species - Lb. plantarum, Lb. acidophilus, Lb. ruminis, Lb. salivarius, Lb. reuteri and Lb. sakei, to highlight the diversity and inter-relationships between the catabolic nature of species within the genus.