Whole-genome sequencing reveals untapped genetic potential in Africa’s indigenous cereal crop sorghum

Sorghum is a food and feed cereal crop adapted to heat and drought and a staple for 500 million of the world’s poorest people. Its small diploid genome and phenotypic diversity make it an ideal C4 grass model as a complement to C3 rice. Here we present high coverage (16-45 × ) resequenced genomes of 44 sorghum lines representing the primary gene pool and spanning dimensions of geographic origin, end-use and taxonomic group. We also report the first resequenced genome of S. propinquum, identifying 8 M high-quality SNPs, 1.9 M indels and specific gene loss and gain events in S. bicolor. We observe strong racial structure and a complex domestication history involving at least two distinct domestication events. These assembled genomes enable the leveraging of existing cereal functional genomics data against the novel diversity available in sorghum, providing an unmatched resource for the genetic improvement of sorghum and other grass species.

Domestication and the storage starch biosynthesis pathway: Signatures of selection from a whole sorghum genome sequencing strategy

Next-generation sequencing of complete genomes has given researchers unprecedented levels of information to study the multifaceted evolutionary changes that have shaped elite plant germplasm. In conjunction with population genetic analytical techniques and detailed online databases, we can more accurately capture the effects of domestication on entire biological pathways of agronomic importance. In this study, we explore the genetic diversity and signatures of selection in all predicted gene models of the storage starch synthesis pathway of Sorghum bicolor, utilizing a diversity panel containing lines categorized as either ‘Landraces’ or ‘Wild and Weedy’ genotypes. Amongst a total of 114 genes involved in starch synthesis, 71 had at least a single signal of purifying selection and 62 a signal of balancing selection and others a mix of both. This included key genes such as STARCH PHOSPHORYLASE 2 (SbPHO2, under balancing selection), PULLULANASE (SbPUL, under balancing selection) and ADP-glucose pyrophosphorylases (SHRUNKEN2, SbSH2 under purifying selection). Effectively, many genes within the primary starch synthesis pathway had a clear reduction in nucleotide diversity between the Landraces and wild and weedy lines indicating that the ancestral effects of domestication are still clearly identifiable. There was evidence of the positional rate variation within the well-characterized primary starch synthesis pathway of sorghum, particularly in the Landraces, whereby low evolutionary rates upstream and high rates downstream in the metabolic pathway were expected. This observation did not extend to the wild and weedy lines or the minor starch synthesis pathways.

Effect of diet on phosphine toxicity, rate of development and reproduction of the rice weevil Sitophilus oryzae (Linnaeus)

One of the loci responsible for strong phosphine resistance encodes dihydrolipoamide dehydrogenase (DLD). The strong co-incidence of enzyme complexes that contain DLD, and enzymes that require thiamine as a cofactor, motivated us to test whether the thiamine deficiency of polished white rice could influence the efficacy of phosphine fumigation against insect pests of stored grain. Three strains of Sitophilus oryzae (susceptible, weak and strong resistance) were cultured on white rice (thiamine deficient), brown rice or whole wheat. As thiamine is an essential nutrient, we firstly evaluated the effect of white rice on developmental rate and fecundity and found that both were detrimentally affected by this diet. The mean time to reach adult stage for the three strains ranged from 40 to 43 days on brown rice and 50–52 days on white rice. The mean number of offspring for the three strains ranged from 7.7 to 10.3 per female over a three day period on brown rice and 2.1 to 2.6 on white rice. Growth and reproduction on wheat was similar to that on brown rice except that the strongly resistant strain showed a tendency toward reduced fecundity on wheat. The susceptible strain exhibited a modest increase in tolerance to phosphine on white rice as expected if thiamine deficiency could mimic the effect of the dld resistance mutation at the rph2 locus. The strongly resistant strain did not respond to thiamine deficiency, but this was expected as these insects are already strongly resistant. We failed, however, to observe the expected synergistic increase in resistance due to combining thiamine deficiency with the weakly resistant strain. The lack of interaction between thiamine content of the diet and the resistance genotype in determining the phosphine resistance phenotype suggests that the mode of inhibition of the complexes is a critical determinant of resistance.

Population ecology of Heteronyx piceus (Coleoptera: Scarabaeidae) in a peanut/maize cropping system

Large larval populations of the scarabaeid beetle Heteronyx piceus Blanchard that occur under peanuts, but not maize, in the South Burnett region of Australia are the result of a high rate and prolonged period of egg production by females feeding on peanut foliage. Heteronyx piceus is a relatively sedentary species and movement of females between adjacent fields is low. Populations of H. piceus varied markedly with landscape position. High larval populations are more likely (1 in 4 chance) to be encountered on the ‘scrub’ soils in the upper parts of the landscape than in the ‘forest’ soils in the lower half (1 in 20 chance), indicating that soil type/landscape position is a key risk factor in assessing the need for management intervention. The studies indicate that, because of the species' sedentary nature, the most meaningful population entity for management of H. piceus is the individual field, rather than the whole-farm or the region. The implications of this population ecology for management of the pest are discussed in relation to control strategies.

Short-term responses of leaf growth rate to water deficit scale up to whole-plant and crop levels: An integrated modelling approach in maize.

Physiological and genetic studies of leaf growth often focus on short-term responses, leaving a gap to whole-plant models that predict biomass accumulation, transpiration and yield at crop scale. To bridge this gap, we developed a model that combines an existing model of leaf 6 expansion in response to short-term environmental variations with a model coordinating the development of all leaves of a plant. The latter was based on: (1) rates of leaf initiation, appearance and end of elongation measured in field experiments; and (2) the hypothesis of an independence of the growth between leaves. The resulting whole-plant leaf model was integrated into the generic crop model APSIM which provided dynamic feedback of environmental conditions to the leaf model and allowed simulation of crop growth at canopy level. The model was tested in 12 field situations with contrasting temperature, evaporative demand and soil water status. In observed and simulated data, high evaporative demand reduced leaf area at the whole-plant level, and short water deficits affected only leaves developing during the stress, either visible or still hidden in the whorl. The model adequately simulated whole-plant profiles of leaf area with a single set of parameters that applied to the same hybrid in all experiments. It was also suitable to predict biomass accumulation and yield of a similar hybrid grown in different conditions. This model extends to field conditions existing knowledge of the environmental controls of leaf elongation, and can be used to simulate how their genetic controls flow through to yield.

Rice molecular breeding laboratories in the genomics era: Current status and future considerations

Using DNA markers in plant breeding with marker-assisted selection (MAS) could greatly improve the precision and efficiency of selection, leading to the accelerated development of new crop varieties. The numerous examples of MAS in rice have prompted many breeding institutes to establish molecular breeding labs. The last decade has produced an enormous amount of genomics research in rice, including the identification of thousands of QTLs for agronomically important traits, the generation of large amounts of gene expression data, and cloning and characterization of new genes, including the detection of single nucleotide polymorphisms. The pinnacle of genomics research has been the completion and annotation of genome sequences for indica and japonica rice. This information-coupled with the development of new genotyping methodologies and platforms, and the development of bioinformatics databases and software tools-provides even more exciting opportunities for rice molecular breeding in the 21st century. However, the great challenge for molecular breeders is to apply genomics data in actual breeding programs. Here, we review the current status of MAS in rice, current genomics projects and promising new genotyping methodologies, and evaluate the probable impact of genomics research. We also identify critical research areas to "bridge the application gap" between QTL identification and applied breeding that need to be addressed to realize the full potential of MAS, and propose ideas and guidelines for establishing rice molecular breeding labs in the postgenome sequence era to integrate molecular breeding within the context of overall rice breeding and research programs.

Optimising indoor phosphine fumigation of paddy rice bag-stacks under sheeting for control of resistant insects

Three indoor, sheeted bag-stack fumigations of paddy rice using aluminium phosphide were undertaken in Guangdong Province, southern China. We measured the effect of two types of sheeting (polyvinylchloride [PVC] or polyethylene [PE]) and two types of floor sealing (clips or fixing into a slot with a rubber pipe) on phosphine concentration and retention. The aim was to test the feasibility of retaining fumigant at a sufficient concentration for long enough to control known resistant insect pests. Each stack was pressure tested and phosphine concentrations measured daily during the fumigation. Cages of test insects in culture medium, including resistant and susceptible strains, were placed inside each stack and could be observed through the clear sheeting. Highest concentrations for the longest period were obtained in a PVC-covered stack that included a ground sheet and wall sheets sealed to the floor with rubber pipes. A similar PVC-covered stack sealed to the floor with clips instead of pipe did not retain gas as efficiently and required re-dosing. A PE-covered stack, with no ground sheet but also with wall sheets sealed to the floor with pipe, produced an acceptable fumigation. Susceptible Rhyzopertha dominica were controlled in 2 days and the most resistant strain in 15 days. Resistant Cryptolestes ferrugineus survived until day 21. The paddy was still free of insect infestation 7 months later when the bag-stack was opened to mill the rice. Pressure half-lives correlated with gas concentration and retention. Sorption appeared to be a major limiting factor, reducing potential fumigant dosage by about 50%. The trials demonstrated the feasibility of sealing bag-stacks to a standard high enough to control all known resistant strains.

Detecting the attributes of a wheat crop using digital imagery acquired from a low-altitude platform

A low-altitude platform utilising a 1.8-m diameter tethered helium balloon was used to position a multispectral sensor, consisting of two digital cameras, above a fertiliser trial plot where wheat (Triticum spp.) was being grown. Located in Cecil Plains, Queensland, Australia, the plot was a long-term fertiliser trial being conducted by a fertiliser company to monitor the response of crops to various levels of nutrition. The different levels of nutrition were achieved by varying nitrogen application rates between 0 and 120 units of N at 40 unit increments. Each plot had received the same application rate for 10 years. Colour and near-infrared images were acquired that captured the whole 2 ha plot. These images were examined and relationships sought between the captured digital information and the crop parameters imaged at anthesis and the at-harvest quality and quantity parameters. The statistical analysis techniques used were correlation analysis, discriminant analysis and partial least squares regression. A high correlation was found between the image and yield (R2 = 0.91) and a moderate correlation between the image and grain protein content (R2 = 0.66). The utility of the system could be extended by choosing a more mobile platform. This would increase the potential for the system to be used to diagnose the causes of the variability and allow remediation, and/or to segregate the crop at harvest to meet certain quality parameters.

A marker-assisted backcross approach for developing submergence-tolerant rice cultivars

Submergence stress regularly affects 15 million hectares or more of rainfed lowland rice areas in South and Southeast Asia. A major QTL on chromosome 9, Sub1, has provided the opportunity to apply marker assisted backcrossing (MAB) to develop submergence tolerant versions of rice cultivars that are widely grown in the region. In the present study, molecular markers that were tightly linked with Sub1, flanking Sub1, and unlinked to Sub1 were used to apply foreground, recombinant, and background selection, respectively, in backcrosses between a submergence-tolerant donor and the widely grown recurrent parent Swarna. By the BC2F2 generation a submergence tolerant plant was identified that possessed Swarna type simple sequence repeat (SSR) alleles on all fragments analyzed except the tip segment of rice chromosome 9 that possessed the Sub1 locus. A BC3F2 double recombinant plant was identified that was homozygous for all Swarna type alleles except for an approximately 2.3-3.4 Mb region surrounding the Sub1 locus. The results showed that the mega variety Swarna could be efficiently converted to a submergence tolerant variety in three backcross generations, involving a time of two to three years. Polymorphic markers for foreground and recombinant selection were identified for four other mega varieties to develop a wider range of submergence tolerant varieties to meet the needs of farmers in the flood-prone regions. This approach demonstrates the effective use of marker assisted selection for a major QTL in a molecular breeding program.

Using automated supplementation systems to meet growth targets for grazing sheep

Remote drafting technology now available for sheep allows targeted supplementation of individuals within a grazing flock. This paper reports results of three experiments. Experiment 1 examined the weight change of Merino wethers allowed access to either lupin grain or whole cottonseed 0, 1, 2 or 7 days/week for 6 weeks. Experiment 2 examined the weight change of Merino wethers allowed access to either lupins or a sorghum + cottonseed meal (CSM) supplement 0, 2, 4 or 7 days/week for 8 weeks. Experiment 3 investigated the relationship between five allocations of trough space at the supplement self-feeders (5–50 cm/sheep) and the weight change of Merino wethers allowed access to lupins 1 day/week for 8 weeks. In all experiments, the Merino wethers had free access as a single group to drinking water and low quality hay in a large group pen and were allowed access to supplement once per day on their scheduled days of access. No water was available in the areas containing supplement, but one-way flow gates allowed animals to return to the group pen in their own time. There was a linear response in growth rate to increased frequency of access to lupins in Experiments 1 and 2, with each additional day of access increasing liveweight gain by 26 and 21 g/day, respectively. Similarly, the response to the sorghum + CSM supplement was linear, although significantly lower (P < 0.05), at 12 g/day. Providing access to whole cottonseed resulted in no significant change in growth rate compared with the control animals. In Experiment 3, decreasing trough space from 50 to 5 cm/sheep had no effect on sheep liveweight change. It was concluded that the relationships developed here, for growth response to increased frequency of access to lupins or a sorghum + CSM supplement, could be used to indicate the most appropriate frequency of access to supplement, through a remote drafting unit, to achieve sheep weight change targets. Also, that a trough space of 5 cm/sheep appears adequate in this supplementation system.

Rice responses to soil management in a rice-based cropping system in the semi-arid tropics of southern Lombok, Eastern Indonesia

This paper is the first of a series that investigates whether new cropping systems with permanent raised beds (PRBs) or Flat land could be successfully used to increase farmers' incomes from rainfed crops in Lombok in Eastern Indonesia. This paper discusses the rice phase of the cropping system. Low grain yields of dry-seeded rice (Oryza sativa) grown on Flat land on Vertisols in the rainfed region of southern Lombok, Eastern Indonesia, are probably mainly due to (a) erratic rainfall (870-1220 mm/yr), with water often limiting at sensitive growth stages, (b) consistently high temperatures (average maximum - 31 C), and (c) low solar radiation. Farmers are therefore poor, and labour is hard and costly, as all operations are manual. Two replicated field experiments were run at Wakan (annual rainfall = 868 mm) and Kawo (1215 mm) for 3 years (2001/2002 to 2003/2004) on Vertisols in southern Lombok. Dry-seeded rice was grown in 4 treatments with or without manual tillage on (a) PRBs, 1.2 m wide, 200 mm high, separated by furrows 300 mm wide, 200 mill deep, with no rice sown in the well-graded furrows, and (b) well-graded Flat land. Excess surface water was harvested from each treatment and used for irrigation after the vegetative stage of the rice. All operations were manual. There were no differences between treatments in grain yield of rice (mean grain yield = 681 g/m(2)) which could be partly explained by total number of tillers/hill and mean panicle length, but not number of productive tillers/hill, plant height or weight of 1000 grains. When the data from both treatments on PRBs and from both treatments on Flat land, each year at each site were analysed, there were also no differences in grain yield of rice (g/m(2)). When rainfall in the wet season up to harvest was over 1000 mm (Year 2; Wakan, Kawo), or plants were water-stressed during crop establishment (Year 1; Wakan) or during grain-fill (Year 3: Kawo), there were significant differences in grain yield (g/1.5 m(2)) between treatments; generally the grain yield (g/1.5 m(2)) on PRBs with or without tillage was less than that on Flat land with or without tillage. However, when the data from both treatments on PRBs and from both treatments on Flat land, each year at each site, were analysed, the greater grain yield of dry-seeded rice on Flat land (mean yield 1 092 g/1.5 m(2)) than that on PRBs (mean 815 g/1.5 m(2)) was mainly because there were 25% more plants on Flat land. Overall when the data in the 2 outer rows and the 2 inner rows on PRBs were each combined, there was a higher number of productive tillers in the combined outer rows (mean 20.7 tillers/hill) compared with that in the combined inner rows on each PRB (mean 18.2 tillers/hill). However, there were no differences in grain yield between combined rows (mean 142 g/m row). Hence with a gap of 500 mm (the distance between the outer rows of plants on adjacent raised beds), plants did not compensate in grain yield for missing plants in furrows. This suggests that rice (a) also sown in furrows, or (b) sown in 7 rows with narrower row-spacing, or (c) sown in 6 rows with slightly wider row-spacing, and narrower gap between outer rows on adjacent beds, may further increase grain yield (g/1.5 m(2)) in this system of PRBs. The growth and the grain yield (y in g/m(2)) of rainfed rice (with rainfall on-site the only source of water for irrigation) depended mainly on the rainfall (x in mm) in the wet season up to harvest (due either to site or year) with y = 1. 1x -308; r(2) = 0.54; p < 0.005. However, 280 mm (i.e. 32%) of the rainfall was not directly used to produce grain (i.e. when y = 0 g/m(2)). Manual tillage did not affect growth and grain yield of rice (g/m(2); g/1.5 m(2)), either on PRB or on Flat land.

Field evaluation of spinosad as a grain protectant for stored wheat in Australia: efficacy against Rhyzopertha dominica (F.) and fate of residues in whole wheat and milling fractions

The potential of spinosad as a grain protectant for the lesser grain borer, Rhyzopertha dominica, was investigated in a silo-scale trial on wheat stored in Victoria, Australia. Rhyzopertha dominica is a serious pest of stored grain, and its resistance to protectants and the fumigant phosphine is becoming more common. This trial follows earlier laboratory research showing that spinosad may be a useful pest management option for this species. Wheat (300 t) from the 2005 harvest was treated with spinosad 0.96 mg/kg plus chlorpyrifos-methyl 10 mg/kg in March 2006, and samples were collected at intervals during 7.5 month storage to determine efficacy and residues in wheat and milling fractions. Chlorpyrifos-methyl is already registered in Australia for control of several other pest species, and its low potency against R. dominica was confirmed in laboratory-treated wheat. Grain moisture content was stable at about 10%, but grain temperature ranged from 29.3°C in March to 14.0°C in August. Bioassays of all treated wheat samples over 7.5 months resulted in 100% adult mortality after 2 weeks exposure and no live progeny were produced. In addition, no live grain insects were detected during outload sampling after a 9 month storage. Spinosad and chlorpyrifos-methyl residues tended to decline during storage, and residues were higher in the bran layer than in either wholemeal or white flour. This field trial confirmed that spinosad was effective as a grain protectant targeting R. dominica.

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.