Alkaloid production by isolates of the sorghum ergot pathogen (Claviceps africana) from Australia and other countries

Isolates of Claviceps africana from Australia, Africa, Asia, and America were tested for the production of dihydroergosine (DHES), and its biogenic precursors dihydroelymoclavine (DHEL) and festuclavine (FEST), in culture. Several growth media were evaluated to optimise alkaloid production with little success. The best of these involved 2-stage culturing on high-sucrose substrate. Australian C. africana isolates varied widely and inconsistently in alkaloid production, with DHES concentrations in mycelium ranging from: <0.1 to 9 mg DHES/kg; <0.1 to 1.6 mg DHEL/kg; and <0.1 to 0.4 mg FEST/kg. In a separate experiment using similar culturing techniques, DHES was produced by 2 of 3 Australian isolates, 1 of 3 USA isolates, 1 of 4 Indian isolates, the sole Puerto Rican isolate, the sole Japanese isolate, but not the sole South African isolate. In this experiment, DHES concentrations detected in mycelium of Australian isolates (0.1-1.0 mg DHES/kg) were of similar magnitude to isolates from other countries (0.2-1.8 mg DHES/kg). Three C. africana isolates, including one that produced only traces of alkaloid in culture after 8 weeks, were inoculated onto panicles of sterile male sorghum plants. After 8 weeks, all 3 isolates produced 10-19 mg DHES/kg in the panicles, demonstrating that the growing plant favoured more consistent alkaloid production than culture medium.

Effect of feeding sorghum ergot (Claviceps africana) to sows during mid-lactation on plasma prolactin and litter performance

Diets containing 3% sorghum ergot (16 mg alkaloids/kg, including 14 mg dihydroergosine/kg) were fed to 12 sows from 14 days post-farrowing until weaning 14 days later, and their performance was compared with that of 10 control sows. Ergot-fed sows displayed a smaller weight loss during lactation of 24 kg/head vs. 29 kg/head in control sows (p > 0.05) despite feed consumption being less (61 kg/head total feed intake vs. 73 kg/head by control sows; p < 0.05). Ergot-fed sows had poorer weight gain of litters over the 14-day period (16.6 kg/litter vs. 28.3 kg/litter for controls; p < 0.05) despite an increase in consumption of creep feed by the piglets from the ergot-fed sows (1.9 kg/litter compared with 1.1 kg/litter by the control; p > 0.05). Sow plasma prolactin was reduced with ergot feeding after 7 days to 4.8 μg/l compared with 15.1 μg/l in the control sows (p < 0.01) and then at weaning was 4.9 μg/l compared with 8.0 μg/l (p < 0.01) in the control sows. Two sows fed ergot ceased lactation early, and the above sow feed intakes, body weight losses with litter weight gains and creep consumption indirectly indicate an ergot effect on milk production.

DArT markers: Diversity analyses and mapping in Sorghum bicolor

The sequential nature of gel-based marker systems entails low throughput and high costs per assay. Commonly used marker systems such as SSR and SNP are also dependent on sequence information. These limitations result in high cost per data point and significantly limit the capacity of breeding programs to obtain sufficient return on investment to justify the routine use of marker-assisted breeding for many traits and particularly quantitative traits. Diversity Arrays Technology (DArT™) is a cost effective hybridisation-based marker technology that offers a high multiplexing level while being independent of sequence information. This technology offers sorghum breeding programs an alternative approach to whole-genome profiling. We report on the development, application, mapping and utility of DArT™ markers for sorghum germplasm. Results: A genotyping array was developed representing approximately 12,000 genomic clones using PstI+BanII complexity with a subset of clones obtained through the suppression subtractive hybridisation (SSH) method. The genotyping array was used to analyse a diverse set of sorghum genotypes and screening a Recombinant Inbred Lines (RIL) mapping population. Over 500 markers detected variation among 90 accessions used in a diversity analysis. Cluster analysis discriminated well between all 90 genotypes. To confirm that the sorghum DArT markers behave in a Mendelian manner, we constructed a genetic linkage map for a cross between R931945-2-2 and IS 8525 integrating DArT and other marker types. In total, 596 markers could be placed on the integrated linkage map, which spanned 1431.6 cM. The genetic linkage map had an average marker density of 1/2.39 cM, with an average DArT marker density of 1/3.9 cM. Conclusion: We have successfully developed DArT markers for Sorghum bicolor and have demonstrated that DArT provides high quality markers that can be used for diversity analyses and to construct medium-density genetic linkage maps. The high number of DArT markers generated in a single assay not only provides a precise estimate of genetic relationships among genotypes, but also their even distribution over the genome offers real advantages for a range of molecular breeding and genomics applications.

Graze to grain - measuring and modelling the effects of grazed pasture leys on soil nitrogen and sorghum yield on a Vertosol soil in the Australian subtropics

Highly productive sown pasture systems can result in high growth rates of beef cattle and lead to increases in soil nitrogen and the production of subsequent crops. The nitrogen dynamics and growth of grain sorghum following grazed annual legume leys or a grass pasture were investigated in a no-till system in the South Burnett district of Queensland. Two years of the tropical legumes Macrotyloma daltonii and Vigna trilobata (both self regenerating annual legumes) and Lablab purpureus (a resown annual legume) resulted in soil nitrate N (0-0.9 m depth), at sorghum sowing, ranging from 35 to 86 kg/ha compared with 4 kg/ha after pure grass pastures. Average grain sorghum production in the 4 cropping seasons following the grazed legume leys ranged from 2651 to 4012 kg/ha. Following the grass pasture, grain sorghum production in the first and second year was < 1900 kg/ha and by the third year grain yield was comparable to the legume systems. Simulation studies utilising the farming systems model APSIM indicated that the soil N and water dynamics following 2-year ley phases could be closely represented over 4 years and the prediction of sorghum growth during this time was reasonable. In simulated unfertilised sorghum crops grown from 1954 to 2004, grain yield did not exceed 1500 kg/ha in 50% of seasons following a grass pasture, while following 2-year legume leys, grain exceeded 3000 kg/ha in 80% of seasons. It was concluded that mixed farming systems that utilise short term legume-based pastures for beef production in rotation with crop production enterprises can be highly productive.

Sorghum stay-green QTL individually reduce post-flowering drought-induced leaf senescence

Sorghum is an important source of food, feed, and biofuel, especially in the semi-arid tropics because this cereal is well adapted to harsh, drought-prone environments. Post-flowering drought adaptation in sorghum is associated with the stay-green phenotype. Alleles that contribute to this complex trait have been mapped to four major QTL, Stg1-Stg4, using a population derived from BTx642 and RTx7000. Near-isogenic RTx7000 lines containing BTx642 DNA spanning one or more of the four stay-green QTL were constructed. The size and location of BTx642 DNA regions in each RTx7000 NIL were analysed using 62 DNA markers spanning the four stay-green QTL. RTx7000 NILs were identified that contained BTx642 DNA completely or partially spanning Stg1, Stg2, Stg3, or Stg4. NILs were also identified that contained sub-portions of each QTL and various combinations of the four major stay-green QTL. Physiological analysis of four RTx7000 NILs containing only Stg1, Stg2, Stg3, or Stg4 showed that BTx642 alleles in each of these loci could contribute to the stay-green phenotype. RTx7000 NILs containing BTx642 DNA corresponding to Stg2 retained more green leaf area at maturity under terminal drought conditions than RTx7000 or the other RTx7000 NILs. Under post-anthesis water deficit, a trend for delayed onset of leaf senescence compared with RTx7000 was also exhibited by the Stg2, Stg3, and Stg4 NILs, while significantly lower rates of leaf senescence in relation to RTx7000 were displayed by all of the Stg NILs to varying degrees, but particularly by the Stg2 NIL. Greener leaves at anthesis relative to RTx7000, indicated by higher SPAD values, were exhibited by the Stg1 and Stg4 NILs. The RTx7000 NILs created in this study provide the starting point for in-depth analysis of stay-green physiology, interaction among stay-green QTL and map-based cloning of the genes that underlie this trait.

Modelling environmental effects on phenology and canopy development of diverse sorghum genotypes

The ability to predict phenology and canopy development is critical in crop models used for simulating likely consequences of alternative crop management and cultivar choice strategies. Here we quantify and contrast the temperature and photoperiod responses for phenology and canopy development of a diverse range of elite Indian and Australian sorghum genotypes (hybrid and landrace). Detailed field experiments were undertaken in Australia and India using a range of genotypes, sowing dates, and photoperiod extension treatments. Measurements of timing of developmental stages and leaf appearance were taken. The generality of photo-thermal approaches to modelling phenological and canopy development was tested. Environmental and genotypic effects on rate of progression from emergence to floral initiation (E-FI) were explained well using a multiplicative model, which combined the intrinsic development rate (Ropt), with responses to temperature and photoperiod. Differences in Ropt and extent of the photoperiod response explained most genotypic effects. Average leaf initiation rate (LIR), leaf appearance rate and duration of the phase from anthesis to physiological maturity differed among genotypes. The association of total leaf number (TLN) with photoperiod found for all genotypes could not be fully explained by effects on development and LIRs. While a putative effect of photoperiod on LIR would explain the observations, other possible confounding factors, such as air-soil temperature differential and the nature of model structure were considered and discussed. This study found a generally robust predictive capacity of photo-thermal development models across diverse ranges of both genotypes and environments. Hence, they remain the most appropriate models for simulation analysis of genotype-by-management scenarios in environments varying broadly in temperature and photoperiod.

Seasonal incidence of Stenodiplosis sorghicola (Coquillett) (Diptera: Cecidomyiidae) and its parasitoids on Sorghum halepense (L.) Pers. in south-eastern Queensland, Australia

To quantify the role of Johnson grass, Sorghum halepense, in the population dynamics of the sorghum midge, Stenodiplosis sorghicola, patterns of flowering of Johnson grass and infestation by sorghum midge were studied in two different climatic environments in the Lockyer Valley and on the Darling Downs in south-eastern Queensland for 3 years. Parasitism levels of S. sorghicola were also recorded. In the Lockyer Valley, Johnson grass panicles were produced throughout the year but on the Darling Downs none were produced between June and September. In both areas, most panicle production occurred between November and March and infestation by S. sorghicola was the greatest during this period. The parasitism levels were between 20% and 50%. After emergence from winter diapause, one to two generations of S. sorghicola developed on S. halepense before commercial grain sorghum crops were available for infestation. Parasitoids recorded were: Aprostocetus diplosidis, Eupelmus australiensis and two species of Tetrastichus. Relationships between sorghum midge population growth rate and various environmental and population variables were investigated. Population size had a significant negative effect (P < 0.0001) on population growth rate. Mortality due to parasitism showed a significant positive density response (P < 0.0001). Temperature, rainfall, open pan evaporation, degree-days and host availability showed no significant effect on population growth rate. Given the phenology of sorghum production in south-eastern Queensland, Johnson grass provides an important bridging host, sustaining one to two generations of sorghum midge. Critical studies relating population change and build-up in sorghum to sorghum midge populations in Johnson grass are yet to be performed.

Genetic erosion and changes in distribution of sorghum (Sorghum bicolor L. (Moench)) landraces in north-eastern Ethiopia

Ethiopia is believed to be the centre of origin and domestication for sorghum, where sorghum remains one of the main staple crops. Loss of biodiversity is occurring at an alarming rate in Ethiopia and crops, including sorghum, have long been recognized as vulnerable to genetic erosion. A major collection of sorghum germplasm was made in 1973 by Gebrekidan and Ejeta from north-eastern Ethiopia. A new collection of landraces was made in 2003, and these were field evaluated at Sirinka in 2004 along with representative samples from the 1973 collection. Farmer surveys and soil and climate surveys were also performed. Preliminary analysis demonstrated that some important landraces have disappeared either locally or regionally in the past 30 years and many other landraces have become marginalized. Landraces which are less preferred in terms of agronomic value and end use, and introductions, have become increasingly important. Late maturing landraces were found to be particularly vulnerable, with a number disappearing altogether. Farmers have become more risk averse, and factors such as declining soil fertility, more frequent drought and unreliable rainfall, and increased pest infestation have contributed to a change in farmer landrace selection. Data are presented on the variability and unique characters of some of the Ethiopian landraces, and implications for conservation are discussed.

Preparation and in vitro release of drug-loaded microparticles for oral delivery using wholegrain sorghum kafirin protein

Kafirin microparticles have been proposed as an oral nutraceutical and drug delivery system. This study investigates microparticles formed with kafirin extracted from white and raw versus cooked red sorghum grains as an oral delivery system. Targeted delivery to the colon would be beneficial for medication such as prednisolone, which is used in the management of inflammatory bowel disease. Therefore, prednisolone was loaded into microparticles of kafirin from the different sources using phase separation. Differences were observed in the protein content, in vitro protein digestibility, and protein electrophoretic profile of the various sources of sorghum grains, kafirin extracts, and kafirin microparticles. For all of the formulations, the majority of the loaded prednisolone was not released in in vitro conditions simulating the upper gastrointestinal tract, indicating that most of the encapsulated drug could reach the target area of the lower gastrointestinal tract. This suggests that these kafirin microparticles may have potential as a colon-targeted nutraceutical and drug delivery system.

Pre-anthesis ovary development determines genotypic differences in potential kernel weight in sorghum

Kernel weight is an important factor determining grain yield and nutritional quality in sorghum, yet the developmental processes underlying the genotypic differences in potential kernel weight remain unclear. The aim of this study was to determine the stage in development at which genetic effects on potential kernel weight were realized, and to investigate the developmental mechanisms by which potential kernel weight is controlled in sorghum. Kernel development was studied in two field experiments with five genotypes known to differ in kernel weight at maturity. Pre-fertilization floret and ovary development was examined and post-fertilization kernel-filling characteristics were analysed. Large kernels had a higher rate of kernel filling and contained more endosperm cells and starch granules than normal-sized kernels. Genotypic differences in kernel development appeared before stamen primordia initiation in the developing florets, with sessile spikelets of large-seeded genotypes having larger floret apical meristems than normal-seeded genotypes. At anthesis, the ovaries for large-sized kernels were larger in volume, with more cells per layer and more vascular bundles in the ovary wall. Across experiments and genotypes, there was a significant positive correlation between kernel dry weight at maturity and ovary volume at anthesis. Genotypic effects on meristem size, ovary volume, and kernel weight were all consistent with additive genetic control, suggesting that they were causally related. The pre-fertilization genetic control of kernel weight probably operated through the developing pericarp, which is derived from the ovary wall and potentially constrains kernel expansion.

A consensus genetic map of sorghum that integrates multiple component maps and high-throughput Diversity Array Technology (DArT) markers

Background: Sorghum genome mapping based on DNA markers began in the early 1990s and numerous genetic linkage maps of sorghum have been published in the last decade, based initially on RFLP markers with more recent maps including AFLPs and SSRs and very recently, Diversity Array Technology (DArT) markers. It is essential to integrate the rapidly growing body of genetic linkage data produced through DArT with the multiple genetic linkage maps for sorghum generated through other marker technologies. Here, we report on the colinearity of six independent sorghum component maps and on the integration of these component maps into a single reference resource that contains commonly utilized SSRs, AFLPs, and high-throughput DArT markers. Results: The six component maps were constructed using the MultiPoint software. The lengths of the resulting maps varied between 910 and 1528 cM. The order of the 498 markers that segregated in more than one population was highly consistent between the six individual mapping data sets. The framework consensus map was constructed using a "Neighbours" approach and contained 251 integrated bridge markers on the 10 sorghum chromosomes spanning 1355.4 cM with an average density of one marker every 5.4 cM, and were used for the projection of the remaining markers. In total, the sorghum consensus map consisted of a total of 1997 markers mapped to 2029 unique loci ( 1190 DArT loci and 839 other loci) spanning 1603.5 cM and with an average marker density of 1 marker/0.79 cM. In addition, 35 multicopy markers were identified. On average, each chromosome on the consensus map contained 203 markers of which 58.6% were DArT markers. Non-random patterns of DNA marker distribution were observed, with some clear marker-dense regions and some marker-rare regions. Conclusion: The final consensus map has allowed us to map a larger number of markers than possible in any individual map, to obtain a more complete coverage of the sorghum genome and to fill a number of gaps on individual maps. In addition to overall general consistency of marker order across individual component maps, good agreement in overall distances between common marker pairs across the component maps used in this study was determined, using a difference ratio calculation. The obtained consensus map can be used as a reference resource for genetic studies in different genetic backgrounds, in addition to providing a framework for transferring genetic information between different marker technologies and for integrating DArT markers with other genomic resources. DArT markers represent an affordable, high throughput marker system with great utility in molecular breeding programs, especially in crops such as sorghum where SNP arrays are not publicly available.

Growth and carcass characteristics of cast-for-age Merino ewes fed sorghum-based feedlot diets

Grain feeding low bodyweight, cast-for-age (CFA) sheep from pastoral areas of eastern Australia at the end of the growing season can enable critical carcass weight grades to be achieved and thus yield better economic returns. The aim of this work was to compare growth and carcass characteristics for CFA Merino ewes consuming either simple diets based on whole sorghum grain or commercial feed pellets. The experiment also compared various sources of additional nitrogen (N) for inclusion in sorghum diets and evaluated several introductory regimes. Seventeen ewes were killed initially to provide baseline carcass data and the remaining 301 ewes were gradually introduced to the concentrate diets over 14 days before being fed concentrates and wheaten hay ad libitum for 33 or 68 days. Concentrate treatments were: (i) commercial feed pellets, (ii) sorghum mix (SM; whole sorghum grain, limestone, salt and molasses) + urea and ammonium sulfate (SMU), (iii) SMU + whole cottonseed at 286 g/kg of concentrate dry matter (DM), (iv) SM + cottonseed meal at 139 g/kg of concentrate DM, (v) SMU + virginiamycin (20 mg/kg of concentrate) for the first 21 days of feeding, and (vi) whole cottonseed gradually replaced by SMU over the first 14 days of feeding. The target carcass weight of 18 kg was achieved after only 33 days on feed for the pellets and the SM + cottonseed meal diet. All other whole grain sorghum diets required between 33 and 68 days on feed to achieve the target carcass weight. Concentrates based on whole sorghum grain generally produced significantly (P < 0.05) lower carcass weight and fat score than pellets and this may have been linked to the significantly (P < 0.05) higher faecal starch concentrations for ewes consuming sorghum-based diets (270 v. 72 g/kg DM on day 51 of feeding for sorghum-based diets and pellets, respectively). Source of N in whole grain sorghum rations and special introductory regimes had no significant (P > 0.05) effects on carcass weight or fat score of ewes with the exception of carcass weight for SMU + whole cottonseed being significantly lower than SM + cottonseed meal at day 33. Ewes finished on all diets produced acceptable carcasses although muscle pH was high in all ewe carcasses (average 5.8 and 5.7 at 33 and 68 days, respectively). There were no significant (P > 0.05) differences between diets in concentrate DM intake, rumen fluid pH, meat colour score, fat colour score, eye muscle area, meat pH or meat temperature.

Concentrates based on sorghum grain provide a basis for a finishing system for crossbred lambs

In parts of Australia, sorghum grain is a cheaper alternative to other cereal grains but its use and nutritive value in sheep feeding systems is not well understood. The aim of this work was to compare growth and carcass characteristics for crossbred lambs consuming several simple, sorghum-based diets. The treatments were: (1) whole sorghum grain, (2) whole sorghum grain + urea and ammonium sulfate, (3) cracked sorghum grain + urea and ammonium sulfate, (4) expanded sorghum grain + urea and ammonium sulfate, (5) whole sorghum grain + cottonseed meal, and (6) whole sorghum grain + whole cottonseed. Nine lambs were slaughtered initially to provide baseline carcass data and the remaining 339 lambs were gradually introduced to the concentrate diets over 14 days before being fed concentrates and wheaten hay ad libitum for 41, 56 or 76 days. Neither cracking nor expanding whole sorghum grain with added non-protein nitrogen (N) resulted in significantly (P > 0.05) increased final liveweight, growth rates or carcass weights for lambs, or in decreased days on feed to reach 18-kg carcass weight, although carcass fat depth was significantly (P < 0.05) increased compared with the whole sorghum plus non-protein N diet. However, expanding sorghum grain significantly (P < 0.05) reduced faecal starch concentrations compared with whole or cracked sorghum diets with added non-protein N (79 v. 189 g/kg DM after 59 days on feed). Lambs fed whole sorghum grain without an additional N source had significantly (P < 0.05) lower concentrate intake and required significantly (P < 0.05) more days on feed to reach a carcass weight of 18 kg than for all diets containing added N. These lambs also had significantly (P < 0.05) lower carcass weight and fat depth than for lambs consuming whole sorghum plus true protein diets. Substituting sources of true protein (cottonseed meal and whole cottonseed) for non-protein N (urea and ammonium sulfate) did not significantly (P > 0.05) affect concentrate intakes or carcass weights of lambs although carcass fat depth was significantly (P < 0.05) increased and the days to reach 18-kg carcass weight were significantly (P < 0.05) decreased for the whole sorghum plus cottonseed meal diet. In conclusion, processing sorghum grain by cracking or expanding did not significantly improve lamb performance. While providing an additional N source with sorghum grain significantly increased lamb performance, there was no benefit in final carcass weight of lambs from substituting sources of true protein for non-protein N.

Functional dynamics of the nitrogen balance of sorghum. II. Grain filling period.

Maintenance of green leaf area during grain filling can increase grain yield of sorghum grown under terminal water limitation. This 'stay-green' trait has been related to the nitrogen (N) supply-demand balance during grain filling. This study quantifies the N demand of grain and N translocation rates from leaves and stem and explores effects of genotype and N stress on onset and rate of leaf senescence during the grain filling period. Three hybrids differing in potential height were grown at three levels of N supply under well-watered conditions. Vertical profiles of biomass, leaf area, and N% of leaves, stem and grain were measured at regular intervals. Weekly SPAD chlorophyll readings on main shoot leaves were correlated with observed specific leaf nitrogen (SLN) to derive seasonal patterns of leaf N content. For all hybrids, individual grain N demand was sink determined and was initially met through N translocation from the stem and rachis. Only if this was insufficient did leaf N translocation occur. Maximum N translocation rates from leaves and stem were dependent on their N status. However, the supply of N at canopy scale was also related to the amount of leaf area senescing at any one time. This supply-demand framework for N dynamics explained effects of N stress and genotype on the onset and rate of leaf senescence.