Plot size matters: interference from intergenotypic competition in plant phenotyping studies

Genetic and physiological studies often comprise genotypes diverse in vigour, size and flowering time. This can make the phenotyping of complex traits challenging, particularly those associated with canopy development, biomass and yield, as the environment of one genotype can be influenced by a neighbouring genotype. Limited seed and space may encourage field assessment in single, spaced rows or in small, unbordered plots, whereas the convenience of a controlled environment or greenhouse makes pot studies tempting. However, the relevance of such growing conditions to commercial field-grown crops is unclear and often doubtful. Competition for water, light and nutrients necessary for canopy growth will be variable where immediate neighbours are genetically different, particularly under stress conditions, where competition for resources and influence on productivity is greatest. Small hills and rod-rows maximise the potential for intergenotypic competition that is not relevant to a crop’s performance in monocultures. Response to resource availability will typically vary among diverse genotypes to alter genotype ranking and reduce heritability for all growth-related traits, with the possible exception of harvest index. Validation of pot experiments to performance in canopies in the field is essential, whereas the planting of multirow plots and the simple exclusion of plot borders at harvest will increase experimental precision and confidence in genotype performance in target environments.

Competition of sorghum cultivars and densities with Japanese millet (Echinochloa esculenta).

Field studies were conducted at two locations in southern Queensland, Australia during the 2003-2004 and 2004-2005 growing seasons to determine the differential competitiveness of sorghum (Sorghum bicolor L. Moench) cultivars and crop densities against weeds and the sorghum yield loss due to weeds. Weed competition was investigated by growing sorghum in the presence or absence of a model grass weed, Japanese millet (Echinochloa esculenta). The correlation analyses showed that the early growth traits (height, shoot biomass, and daily growth rate of the shoot biomass) of sorghum adversely affected the height, biomass, and seed production of millet, as measured at maturity. "MR Goldrush" and "Bonus MR" were the most competitive cultivars, resulting in reduced weed biomass, weed density, and weed seed production. The density of sorghum also had a significant effect on the crop's ability to compete with millet. When compared to the density of 4.5 plants per m2, sorghum that was planted at 7.5 plants per m2 suppressed the density, biomass, and seed production of millet by 22%, 27% and 38%, respectively. Millet caused a significant yield loss in comparison with the weed-free plots. The combined weed-suppressive effects of the competitive cultivars, such as MR Goldrush, and high crop densities minimized the yield losses from the weeds. These results indicate that sorghum competition against grass weeds can be improved by choosing competitive cultivars and by using a high crop density of > 7.5 plants per m2. These non-chemical options should be included in an integrated weed management program for better weed management, particularly where the control options are limited by the evolution of herbicide resistance.

Gene expression evidence for off-target effects caused by RNA interference-mediated gene silencing of Ubiquitin-63E in the cattle tick Rhipicephalus microplus.

Knowledge of cattle tick (Rhipicephalus (Boophilus) microplus; Acari: Ixodidae) molecular and cellular pathways has been hampered by the lack of an annotated genome. In addition, most of the tick expressed sequence tags (ESTs) available to date consist of similar to 50% unassigned sequences without predicted functions. The most common approach to address this has been the application of RNA interference (RNAi) methods to investigate genes and their pathways. This approach has been widely adopted in tick research despite minimal knowledge of the tick RNAi pathway and double-stranded RNA (dsRNA) uptake mechanisms. A strong knockdown phenotype of adult female ticks had previously been observed using a 594 bp dsRNA targeting the cattle tick homologue for the Drosophila Ubiquitin-63E gene leading to nil or deformed eggs. A NimbleGen cattle tick custom microarray based on the BmiGI.V2 database of R. microplus ESTs was used to evaluate the expression of mRNAs harvested from ticks treated with the tick Ubiquitin-63E 594 bp dsRNA compared with controls. A total of 144 ESTs including TC6372 (Ubiquitin-63E) were down-regulated with 136 ESTs up-regulated following treatment. The results obtained substantiated the knockdown phenotype with ESTs identified as being associated with ubiquitin proteolysis as well as oogenesis, embryogenesis, fatty acid synthesis and stress responses. A bioinformatics analysis was undertaken to predict off-target effects (OTE) resulting from the in silico dicing of the 594 bp Ubiquitin-63E dsRNA which identified 10 down-regulated ESTs (including TC6372) within the list of differentially expressed probes on the microarrays. Subsequent knockdown experiments utilising 196 and 109 bp dsRNAs, and a cocktail of short hairpin RNAs (shRNA) targeting Ubiquitin-63E, demonstrated similar phenotypes for the dsRNAs but nil effect following shRNA treatment. Quantitative reverse transcriptase PCR analysis confirmed differential expression of TC6372 and selected ESTs. Our study demonstrated the minimisation of predicted OTEs in the shorter dsRNA treatments (similar to 100-200 bp) and the usefulness of microarrays to study knockdown phenotypes.

Predicting Additive and Non-additive Genetic Effects from Trials Where Traits Are Affected by Interplot Competition

There are two key types of selection in a plant breeding program, namely selection of hybrids for potential commercial use and the selection of parents for use in future breeding. Oakey et al. (in Theoretical and Applied Genetics 113, 809-819, 2006) showed how both of these aims could be achieved using pedigree information in a mixed model analysis in order to partition genetic effects into additive and non-additive effects. Their approach was developed for field trial data subject to spatial variation. In this paper we extend the approach for data from trials subject to interplot competition. We show how the approach may be used to obtain predictions of pure stand additive and non-additive effects. We develop the methodology in the context of a single field trial using an example from an Australian sorghum breeding program.

Can competition with pasture be used to manipulate bellyache bush (Jatropha gossypiifolia L.) population biology?

Bellyache bush (Jatropha gossypiifolia L.) is an invasive weed that poses economic and environmental problems in northern Australia. Competition between pasture and bellyache bush was examined in North Queensland using combinations of five pasture treatments (uncut (control); cut as low, medium, and high pasture; and no pasture) and four bellyache bush densities (0, 2, 6 and 12plantsm(-2)) in a buffel grass (Cenchrus ciliaris L.) dominated pasture. The pasture treatments were applied approximately once per year but no treatments were applied directly to the bellyache bush plants. Measurements of bellyache bush flowering, seed formation, and mortality were undertaken over a 9-year period, along with monitoring the pasture basal cover and plant species diversity. Maximum flowering rates of bellyache bush occurred after 9 years (97%) in plots containing no pasture, with the lowest rates of 9% in uncut control plots. Earliest flowering (322 days after planting) and seed formation (411 days) also occurred in plots with no pasture compared with all other pasture treatments (range 1314-1393 days for seed formation to occur). No seeds were produced in uncut plots. At the end of 9 years, mortality rates of bellyache bush plants initially planted averaged 73% for treatments with some pasture compared with 55% under the no-pasture treatment. The percentage of herbaceous plant basal cover in uncut plots was increased 5-fold after 9 years, much greater than the average 2% increase recorded across the low, medium, and high pasture treatments. The number of herbaceous species in uncut plots remained largely unchanged, whereas there was an average reduction of 46% in the cut pasture treatments. Buffel grass remained the species with the greatest basal cover across all cut pasture treatments, followed by sabi grass (Urochloa mosambicensis (Hack.) Dandy) and then red Natal grass (Melinis repens (Willd.) Ziska). These results suggest that grazing strategies that maintain a healthy and competitive pasture layer may contribute to reducing the rate of spread of bellyache bush and complement traditional control techniques such as the use of herbicides.

Reproductive interference between honey bee species in artificial sympatry

Reproductive isolation between closely related species is often incomplete. The Western honey bee, Apis mellifera, and the Eastern hive bee, A. cerana have been allopatric for millions of years, but are nonetheless similar in morphology and behaviour. During the last century the two species were brought into contact anthropogenically, providing potential opportunities for interspecific matings. Hybrids between A. mellifera and A. cerana are inviable, so natural interspecific matings are of concern because they may reduce the viability of A. cerana and A. mellifera populations – two of the world's most important pollinators. We examined the mating behaviour of A. mellifera and A. cerana queens and drones from Caoba Basin, China and Cairns, Australia. Drone mating flight times overlap in both areas. Analysis of the spermathecal contents of queens with species-specific genetic markers indicated that in Caoba Basin, 14% of A. mellifera queens mated with at least one A. cerana male, but we detected no A. cerana queens that had mated with A. mellifera males. Similarly, in Cairns, no A. cerana queens carried A. mellifera sperm, but one third of A. mellifera queens had mated with at least one A. cerana male. No hybrid embryos were detected in eggs laid by interspecifically-mated A. mellifera queens in either location. However A. mellifera queens artificially inseminated with A. cerana sperm produced inviable hybrid eggs, or unfertilised drones. This suggests that reproductive interference will impact the viability of honey bee populations wherever A. cerana and A. mellifera are in contact. This article is protected by copyright. All rights reserved.

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.

Interference and management of parthenium: The world's most important invasive weed

Parthenium (Parthenium hysterophorus L.) is one of the most aggressive herbaceous weeds of the Asteraceae family. It is widely distributed, almost across the world and has become the most important invasive weed. Comprehensive information on interference and control of this devastating species is required to facilitate better management decisions. A broad review on the interference and management of this weed is presented here. Inspite of its non-tropical origin, parthenium grows quite successfully under a wide range of environmental conditions. It is spreading rapidly in Australia, Western Africa, Asia, and Caribbean countries, and has become a serious weed of pastures, wastelands, roadsides, railwaysides, water courses, and agricultural crops. The infestations of parthenium have been reported to reduce grain and forage yields by 40–90%. The spread of parthenium has been attributed to its allelopathic activity, strong competitiveness for soil moisture and nutrients, and its capability to exploit natural biodiversity. Allelochemicals released from parthenium has been reported to decrease germination and growth of agronomic crops, vegetables, trees, and many other weed species. Growth promoting effects of parthenium extracts at low concentrations have also been reported in certain crops. Many pre- and post-emergence herbicides have been evaluated for the control of parthenium in cropped and non-cropped areas. The most effective herbicides are clomazone, metribuzin, atrazine, glyphosate, metsulfuron methyl, butachlor, bentazone, dicamba, and metsulfuron methyl. Extracts, residues, and essential oils of many allelopathic herbs (Cassia, Amaranthus, and Xanthium species), grasses (Imperata and Desmostachya species), and trees (Eucalyptus, Azadirachta, Mangifera species, etc.) have demonstrated inhibitory activities on seed germination and seedling growth of parthenium. Metabolites of several fungi, e.g., Fusarium oxysporun and Fusarium monilifonne, exhibit bioherbicidal activity against seeds and seedlings of this weed. Intercropping, displacement by competitive plant species like Cassia species, bisset bluegrass, florgen blugress, buffelgrass, along with the use of biological control agents like Mexican beetle, seed-feeding and stem-boring weevils, stem-galling and leaf-mining moth, and sap-feeding plant hopper, have been reported as possible strategies for the management of parthenium. An appropriate integration of these approaches could help minimize spread of parthenium and provide sustainable weed management with reduced environmental concerns.

The critical period for weed control in dry-seeded rice

The critical crop-weed competition period in a dry-seeded rice system is an important consideration in formulating weed management strategies. Field experiments were conducted in the summer seasons of 2012 and 2013 at the Punjab Agricultural University, Ludhiana, India, to determine the extent of yield loss in two different rice cultivars (PR 114 and PR 115) with different periods of weed interference. Twelve weed control timings were used to identify critical periods of weed competition in dry-seeded rice. PR 114, a long-duration rice cultivar (145 d) having slower initial growth than PR 115 (125 d), was more prone to yield losses. In both years, 100% yield loss was observed where weeds were not controlled throughout the season. In weed-free plots, the grain yield of PR 114 was 6.39-6.80 t ha-1, for PR 115, it was 6.49-6.87 t ha-1. Gompertz and logistic equations fitted to yield data in response to increasing periods of weed control and weed interference showed that, PR 114 had longer critical periods than PR 115. Critical weed-free periods to achieve 95% of weed-free yield for PR 114 was longer than for PR 115 by 31 days in 2012 and 26 days in 2013. Weed infestation also influenced the duration of critical periods. Higher weed pressure in 2012 than in 2013 increased the duration of the critical period of crop-weed competition in that year. The identification of critical crop-weed competition periods for different cultivars will facilitate improved decision-making regarding the timing of weed control and the adoption of cultivars having high weed-suppressing abilities. This will also contribute to the development of integrated weed management in dry-seeded rice systems.

Weed dynamics as influenced by tillage system, sowing time and weed competition duration in dry-seeded rice

Cultural practices alter patterns of crop growth and can modify dynamics of weed-crop competition, and hence need to be investigated to evolve sustainable weed management in dry-seeded rice (DSR). Studies on weed dynamics in DSR sown at different times under two tillage systems were conducted at the Agronomic Research Farm, University of Agriculture, Faisalabad, Pakistan. A commonly grown fine rice cultivar 'Super Basmati' was sown on 15th June and 7th July of 2010 and 2011 under zero-till (ZT) and conventional tillage (CONT) and it was subjected to different durations of weed competition [10, 20, 30, 40, and 50 days after sowing (DAS) and season-long competition]. Weed-free plots were maintained under each tillage system and sowing time for comparison. Grassy weeds were higher under ZT while CONT had higher relative proportion of broad-leaved weeds in terms of density and biomass. Density of sedges was higher by 175% in the crop sown on the 7th July than on the 15th June. Delaying sowing time of DSR from mid June to the first week of July reduced weed density by 69 and 43% but their biomass remained unaffected. Tillage systems had no effect on total weed biomass. Plots subjected to season-long weed competition had mostly grasses while broad-leaved weeds were not observed at harvest. In the second year of study, dominance of grassy weeds was increased under both tillage systems and sowing times. Significantly less biomass (48%) of grassy weeds was observed under CONT than ZT in 2010; however, during 2011, this effect was non-significant. Trianthema portulacastrum and Dactyloctenium aegyptium were the dominant broad-leaved and grassy weeds, respectively. Cyperus rotundus was the dominant sedge weed, especially in the crop sown on the 7th July. Relative yield loss (RYL) ranged from 3 to 13% and 7 to16% when weeds were allowed to compete only for 20 DAS. Under season-long weed competition, RYL ranged from 68 to 77% in 2010 and 74 to80% in 2011. The sowing time of 15th June was effective in minimizing weed proliferation and rectifying yield penalty associated with the 7th July sowing. The results suggest that DSR in Pakistan should preferably be sown on 15th June under CONT systems and weeds must be controlled before 20 DAS to avoid yield losses. Successful adoption of DSR at growers' fields in Pakistan will depend on whether growers can control weeds and prevent shifts in weed population from intractable weeds to more difficult-to-control weeds as a consequence of DSR adoption.

The combined effect of biological control with plant competition on the management of parthenium weed (Parthenium hysterophorus l.)

Parthenium hysterophorus L., (Asteraceae) commonly known as parthenium weed, is a highly invasive plant that has become a problematic weed of pasture lands in Australia and many other countries around the world. For the management of this weed, an integrated approach comprising biological control and plant competition strategies was tested in southern central Queensland. Two competitive pasture plant species (butterfly pea and buffel grass), selected for their high competitive ability, worked successfully with the biological control agent (Epiblema strenuana Walker) to synergistically reduce the biomass of parthenium weed, by between 62 and 69%. In the presence of biological control agent, the corresponding biomass of competitive plants, butterfly pea and buffel grass increased in comparison to when the biological control agent had been excluded, by 15 and 35%, respectively. This suggests that biological control and competitive plants can complement one another to bring about improved management of parthenium weed in Australia. Further, this approach may be adopted in countries where some of the biological control agents are already present including South Africa, Ethiopia, India, Pakistan and Nepal.