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.

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.

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.

Suppression of populations of Australian sheep blowfly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae), with a novel blowfly trap

The Australian sheep blowfly, Lucilia cuprina initiates more than 85% of fly strikes on sheep in Australia with an estimated average annual cost of A$280 million to the Australian wool industry. LuciTrap® is a commercially available, selective trap for L. cuprina consisting of a plastic bucket with multiple fly entry cones and a synthetic attractant. The impact of LuciTrap on populations of L. cuprina on sheep properties in five Australian states was evaluated by comparing L. cuprina populations on paired properties with and without LuciTraps over seasons when significant fly populations could be expected. Twenty-four comparisons (trials) were conducted over four years. During times of ‘higher fly density’ (when the 48 h geometric mean of trap catches on the control property was greater than five L. cuprina), the overall geometric mean trap catches for control and trapped properties differed significantly (P<0.001) with mean trap catches of 19.4 and 7.74 L. cuprina respectively. The selectivity of the LuciTrap was confirmed with 59% of all trapped flies being L. cuprina. Chrysomya spp. and Calliphora spp. constituted 9.3% and 1.1% of the catches with a variety of other flies (mainly Sarcophagidae and Muscidae) providing the remainder (31%). L. sericata was only trapped in Tasmania and made up 7.7% of the Lucilia spp. catch in this State. Seventy-two percent of the trapped L. cuprina were female. The deployment of LuciTrap on sheep properties at one trap per 100 sheep from the beginning of the anticipated fly season suppressed the populations of L. cuprina by 60% compared to matched control properties. The LuciTrap is a selective and easy to use fly trap and constitutes an effective, non-insecticidal tool for use in integrated management programs for L. cuprina.

Rice flower production guidelines for growers

Rice flower, an Australian native species, has been cultivated in appreciable quantities as a cut flower crop since 1991. Although the plant can be highly productive, achieving consistently high commercial yields can prove elusive. Rice Flower production guidelines for growers is essential reading for producers who would aspire to grow and market rice flower well. For anyone with an interest in new crop development, this book provides a valuable insight into the intricacies of growing an indigenous species commercially.

Rice flower integrating production and marketing

This book contains guidelines on market-driven production for export markets, with information on how the marketing chain operates and what risks are involved. Using rice flower as an example, the book gives growers strategies to enhance their market performance and improve the profitability of their enterprises. It outlines some practical suggestions for marketing rice flower in Japan, the United States, Taiwan and Hong Kong as well as in Australia, and also provides a draft standard for rice flower for export markets.

Relocation of Intensive Agriculture to Northern Australia: The Case of the Rice Industry

Development of new agricultural industries in northern Australia is seen as a way to provide food security in the face of reduced water availability in existing regions in the south. This report aims to identify some of the possible economic consequences of developing a rice industry in the Burdekin region, while there is a reduction of output in the Riverina. Annual rice production in the Riverina peaked at 1.7 M tonnes, but the long-term outlook, given climate change impacts on that region and government water buy-backs, is more likely to be less than 800,000 tonnes. Growers are highly efficient water users by international standards, but the ability to offset an anticipated reduction in water availability through further efficiency gains is limited. In recent years growers in the Riverina have diversified their farms to a greater extent and secondary production systems include beef, sheep and wheat. Production in north Queensland is in its infancy, but a potentially suitable farming system has been developed by including rice within the sugarcane system without competition and in fact contributing to the production of sugar by increasing yields and controlling weeds. The economic outcomes are estimated a large scale, dynamic, computable general equilibrium (CGE) model of the world economy (Tasman Global), scaled down to regional level. CGE models mimic the workings of the economy through a system of interdependent behavioural and accounting equations which are linked to an input-output database. When an economic shock or change is applied to a model, each of the markets adjusts according to the set of behavioural parameters which are underpinned by economic theory. In this study the model is driven by reducing production in the Riverina in accordance with relationships found between water availability and the production of rice and replacement by other crops and by increasing ride production in the Burdekin. Three scenarios were considered: • Scenario 1: Rice is grown using the fallow period between the last ratoon crop of sugarcane and the new planting. In this scenario there is no competition between rice and sugarcane • Scenario 2: Rice displaces sugarcane production • Scenario 3: Rice is grown on additional land and does not compete with sugarcane. Two time periods were used, 2030 and 2070, which are the conventional time points to consider climate change impacts. Under scenario 1, real economic output declines in the Riverina by $45 million in 2030 and by $139 million in 2070. This is only partially offset by the increased real economic output in the Burdekin of $35 million and $131 million respectively.

Recent emergence of the wheat Lr34 multi-pathogen resistance: insights from haplotype analysis in wheat, rice, sorghum and Aegilops tauschii

Spontaneous sequence changes and the selection of beneficial mutations are driving forces of gene diversification and key factors of evolution. In highly dynamic co-evolutionary processes such as plant-pathogen interactions, the plant's ability to rapidly adapt to newly emerging pathogens is paramount. The hexaploid wheat gene Lr34, which encodes an ATP-binding cassette (ABC) transporter, confers durable field resistance against four fungal diseases. Despite its extensive use in breeding and agriculture, no increase in virulence towards Lr34 has been described over the last century. The wheat genepool contains two predominant Lr34 alleles of which only one confers disease resistance. The two alleles, located on chromosome 7DS, differ by only two exon-polymorphisms. Putatively functional homoeologs and orthologs of Lr34 are found on the B-genome of wheat and in rice and sorghum, but not in maize, barley and Brachypodium. In this study we present a detailed haplotype analysis of homoeologous and orthologous Lr34 genes in genetically and geographically diverse selections of wheat, rice and sorghum accessions. We found that the resistant Lr34 haplotype is unique to the wheat D-genome and is not found in the B-genome of wheat or in rice and sorghum. Furthermore, we only found the susceptible Lr34 allele in a set of 252 Ae. tauschii genotypes, the progenitor of the wheat D-genome. These data provide compelling evidence that the Lr34 multi-pathogen resistance is the result of recent gene diversification occurring after the formation of hexaploid wheat about 8,000 years ago.

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.

Modelling interactions between farm-level structural adjustment and a regional economy: A case of the Australian rice industry

Climate change and on-going water policy reforms will likely contribute to on-farm and regional structural adjustment in Australia. This paper gathers empirical evidence of farm-level structural adjustments and integrates these with a regional equilibrium model to investigate sectoral and regional impacts of climate change and recent water use policy on rice industry. We find strong evidence of adjustments to the farming system, enabled by existing diversity in on-farm production. A further loss of water with additional pressures to adopt less intensive and larger-scale farming, will however reduce the net number of farm businesses, which may affect regional rice production. The results from a regional CGE model show impacts on the regional economy over and above the direct cost of the environmental water, although a net reduction in real economic output and real income is partially offset by gains in rest of the Australia through the reallocation or resources. There is some interest within the industry and from potential new corporate entrants in the relocation of some rice production to the north. However, strong government support would be crucial to implement such relocation.

Responses of Super Rice (Oryza sativa L.) to Different Planting Methods for Grain Yield and Nitrogen-Use Efficiency in the Single Cropping Season

To break the yield ceiling of rice production, a super rice project was developed in 1996 to breed rice varieties with super high yield. A two-year experiment was conducted to evaluate yield and nitrogen (N)-use response of super rice to different planting methods in the single cropping season. A total of 17 rice varieties, including 13 super rice and four non-super checks (CK), were grown under three N levels [0 (N0), 150 (N150), and 225 (N225) kg ha−1] and two planting methods [transplanting (TP) and direct-seeding in wet conditions (WDS)]. Grain yield under WDS (7.69 t ha−1) was generally lower than TP (8.58 t ha−1). However, grain yield under different planting methods was affected by N rates as well as variety groups. In both years, there was no difference in grain yield between super and CK varieties at N150, irrespective of planting methods. However, grain yield difference was dramatic in japonica groups at N225, that is, there was an 11.3% and 14.1% average increase in super rice than in CK varieties in WDS and TP, respectively. This suggests that high N input contributes to narrowing the yield gap in super rice varieties, which also indicates that super rice was bred for high fertility conditions. In the japonica group, more N was accumulated in super rice than in CK at N225, but no difference was found between super and CK varieties at N0 and N150. Similar results were also found for N agronomic efficiency. The results suggest that super rice varieties have an advantage for N-use efficiency when high N is applied. The response of super rice was greater under TP than under WDS. The results suggest that the need to further improve agronomic and other management practices to achieve high yield and N-use efficiency for super rice varieties in WDS.

Ecological significance of rice (Oryza sativa) planting density and nitrogen rates in managing the growth and competitive ability of itchgrass (Rottboellia cochinchinensis) in direct-seeded rice systems

Current understanding is that high planting density has the potential to suppress weeds and crop-weed interactions can be exploited by adjusting fertilizer rates. We hypothesized that (a) high planting density can be used to suppress Rottboellia cochinchinensis growth and (b) rice competitiveness against this weed can be enhanced by increasing nitrogen (N) rates. We tested these hypotheses by growing R. cochinchinensis alone and in competition with four rice planting densities (0, 100, 200, and 400 plants m-2) at four N rates (0, 50, 100, and 150 kg ha-1). At 56 days after sowing (DAS), R. cochinchinensis plant height decreased by 27-50 %, tiller number by 55-76 %, leaf number by 68-84 %, leaf area by 70-83 %, leaf biomass by 26-90 %, and inflorescence biomass by 60-84 %, with rice densities ranging from 100 to 400 plants m-2. All these parameters increased with an increase in N rate. Without the addition of N, R. cochinchinensis plants were 174 % taller than rice; whereas, with added N, they were 233 % taller. Added N favored more weed biomass production relative to rice. R. cochinchinensis grew taller than rice (at all N rates) to avoid shade, which suggests that it is a "shade-avoiding" plant. R. cochinchinensis showed this ability to reduce the effect of rice interference through increased leaf weight ratio, specific stem length, and decreased root-shoot weight ratio. This weed is more responsive to N fertilizer than rice. Therefore, farmers should give special consideration to the application timing of N fertilizer when more N-responsive weeds are present in their field. Results suggest that the growth and seed production of R. cochinchinensis can be decreased considerably by increasing rice density to 400 plants m-2. There is a need to integrate different weed control measures to achieve complete control of this noxious weed.

The plasticity of NBS resistance genes in sorghum is driven by multiple evolutionary processes

Background Increased disease resistance is a key target of cereal breeding programs, with disease outbreaks continuing to threaten global food production, particularly in Africa. Of the disease resistance gene families, the nucleotide-binding site plus leucine-rich repeat (NBS-LRR) family is the most prevalent and ancient and is also one of the largest gene families known in plants. The sequence diversity in NBS-encoding genes was explored in sorghum, a critical food staple in Africa, with comparisons to rice and maize and with comparisons to fungal pathogen resistance QTL. Results In sorghum, NBS-encoding genes had significantly higher diversity in comparison to non NBS-encoding genes and were significantly enriched in regions of the genome under purifying and balancing selection, both through domestication and improvement. Ancestral genes, pre-dating species divergence, were more abundant in regions with signatures of selection than in regions not under selection. Sorghum NBS-encoding genes were also significantly enriched in the regions of the genome containing fungal pathogen disease resistance QTL; with the diversity of the NBS-encoding genes influenced by the type of co-locating biotic stress resistance QTL. Conclusions NBS-encoding genes are under strong selection pressure in sorghum, through the contrasting evolutionary processes of purifying and balancing selection. Such contrasting evolutionary processes have impacted ancestral genes more than species-specific genes. Fungal disease resistance hot-spots in the genome, with resistance against multiple pathogens, provides further insight into the mechanisms that cereals use in the “arms race” with rapidly evolving pathogens in addition to providing plant breeders with selection targets for fast-tracking the development of high performing varieties with more durable pathogen resistance.