Stay-green alleles individually enhance grain yield in sorghum under drought by modifying canopy development and water uptake patterns

* Stay-green is an integrated drought adaptation trait characterized by a distinct green leaf phenotype during grain filling under terminal drought. We used sorghum (Sorghum bicolor), a repository of drought adaptation mechanisms, to elucidate the physiological and genetic mechanisms underpinning stay-green. * Near-isogenic sorghum lines (cv RTx7000) were characterized in a series of field and managed-environment trials (seven experiments and 14 environments) to determine the influence of four individual stay-green (Stg1–4) quantitative trait loci (QTLs) on canopy development, water use and grain yield under post-anthesis drought. * The Stg QTL decreased tillering and the size of upper leaves, which reduced canopy size at anthesis. This reduction in transpirational leaf area conserved soil water before anthesis for use during grain filling. Increased water uptake during grain filling of Stg near-isogenic lines (NILs) relative to RTx7000 resulted in higher post-anthesis biomass production, grain number and yield. Importantly, there was no consistent yield penalty associated with the Stg QTL in the irrigated control. * These results establish a link between the role of the Stg QTL in modifying canopy development and the subsequent impact on crop water use patterns and grain yield under terminal drought.

Stay-green Drought Adaptation Trait Enhances Sorghum Production in Sub-tropical Australia, Central-Western India and Sub-Saharan Africa

Drought during grain filling is a common challenge for sorghum production in north-eastern Australia, central-western India, and sub-Saharan Africa. We show that the stay-green drought adaptation trait enhances sorghum grain yield under post-anthesis drought in these three regions. A positive relationship between stay-green and yield was generally found in breeding trials in north-eastern Australia that sampled 1668 unique hybrid combinations and 23 environments. Physiological studies in Australia also found that introgressing four individual stay-green (Stg1–4) quantitative trait loci (QTLs) into a senescent background reduced water demand before flowering and hence increased water supply during grain filling, resulting in higher grain yield relative to the senescent control. Studies in India found that various Stg QTLs affected both transpiration and transpiration efficiency, although these effects depended on the interaction between genetic background (S35 and R16) and individual QTLs. The yield variation unexplained by harvest index was related to transpiration efficiency in S35 (R2 = 0.29) and R16 (R2 = 0.72), and was related to total water extracted in S35 (R2 = 0.41) but not in R16. Finally, sixty-eight stay-green enriched lines were evaluated in six countries in sub-Saharan Africa during the 2013/14 season. Analysis of the data from Kenya indicates that stay-green and grain size were positively correlated at two sites: Kiboko (high yielding, r2=0.25) and Masongaleni (low yielding, r2=0.37). Together, these studies suggest that stay-green is a beneficial trait for sorghum production in the semi-arid tropics and is a consequence of traits altering the plant water budget.

Drought adaptation of stay-green sorghum is associated with canopy development, leaf anatomy, root growth, and water uptake

Stay-green sorghum plants exhibit greener leaves and stems during the grain-filling period under water-limited conditions compared with their senescent counterparts, resulting in increased grain yield, grain mass, and lodging resistance. Stay-green has been mapped to a number of key chromosomal regions, including Stg1, Stg2, Stg3, and Stg4, but the functions of these individual quantitative trait loci (QTLs) remain unclear. The objective of this study was to show how positive effects of Stg QTLs on grain yield under drought can be explained as emergent consequences of their effects on temporal and spatial water-use patterns that result from changes in leaf-area dynamics. A set of four Stg near-isogenic lines (NILs) and their recurrent parent were grown in a range of field and semicontrolled experiments in southeast Queensland, Australia. These studies showed that the four Stg QTLs regulate canopy size by: (1) reducing tillering via increased size of lower leaves, (2) constraining the size of the upper leaves; and (3) in some cases, decreasing the number of leaves per culm. In addition, they variously affect leaf anatomy and root growth. The multiple pathways by which Stg QTLs modulate canopy development can result in considerable developmental plasticity. The reduction in canopy size associated with Stg QTLs reduced pre-flowering water demand, thereby increasing water availability during grain filling and, ultimately, grain yield. The generic physiological mechanisms underlying the stay-green trait suggest that similar Stg QTLs could enhance post-anthesis drought adaptation in other major cereals such as maize, wheat, and rice.

Phylogeography of the green turtle, Chelonia mydas, in the Southwest Indian Ocean

Patterns of mitochondrial DNA (mtDNA) variation were used to analyse the population genetic structure of southwestern Indian Ocean green turtle (Chelonia mydas) populations. Analysis of sequence variation over 396 bp of the mtDNA control region revealed seven haplotypes among 288 individuals from 10 nesting sites in the Southwest Indian Ocean. This is the first time that Atlantic Ocean haplotypes have been recorded among any Indo-Pacific nesting populations. Previous studies indicated that the Cape of Good Hope was a major biogeographical barrier between the Atlantic and Indian Oceans because evidence for gene flow in the last 1.5 million years has yet to emerge. This study, by sampling localities adjacent to this barrier, demonstrates that recent gene flow has occurred from the Atlantic Ocean into the Indian Ocean via the Cape of Good Hope. We also found compelling genetic evidence that green turtles nesting at the rookeries of the South Mozambique Channel (SMC) and those nesting in the North Mozambique Channel (NMC) belong to separate genetic stocks. Furthermore, the SMC could be subdivided in two different genetic stocks, one in Europa and the other one in Juan de Nova. We suggest that this particular genetic pattern along the Mozambique Channel is attributable to a recent colonization from the Atlantic Ocean and is maintained by oceanic conditions in the northern and southern Mozambique Channel that influence early stages in the green turtle life cycle.

The genetic structure of Australasian green turtles (Chelonia mydas): Exploring the geographical scale of genetic exchange

Ecological and genetic studies of marine turtles generally support the hypothesis of natal homing, but leave open the question of the geographical scale of genetic exchange and the capacity of turtles to shift breeding sites. Here we combine analyses of mitochondrial DNA (mtDNA) variation and recapture data to assess the geographical scale of individual breeding populations and the distribution of such populations through Australasia. We conducted multiscale assessments of mtDNA variation among 714 samples from 27 green turtle rookeries and of adult female dispersal among nesting sites in eastern Australia. Many of these rookeries are on shelves that were flooded by rising sea levels less than 10 000 years (c. 450 generations) ago. Analyses of sequence variation among the mtDNA control region revealed 25 haplotypes, and their frequency distributions indicated 17 genetically distinct breeding stocks (Management Units) consisting either of individual rookeries or groups of rookeries in general that are separated by more than 500 km. The population structure inferred from mtDNA was consistent with the scale of movements observed in long-term mark-recapture studies of east Australian rookeries. Phylogenetic analysis of the haplotypes revealed five clades with significant partitioning of sequence diversity (Φ = 68.4) between Pacific Ocean and Southeast Asian/Indian Ocean rookeries. Isolation by distance was indicated for rookeries separated by up to 2000 km but explained only 12% of the genetic structure. The emerging general picture is one of dynamic population structure influenced by the capacity of females to relocate among proximal breeding sites, although this may be conditional on large population sizes as existed historically across this region.

Developmental and physiological traits associated with high yield and stay-green phenotype in wheat

Water availability is a key limiting factor in wheat production in the northern grain belt of Australia. Varieties with improved adaptation to such conditions are actively sought. The CIMMYT wheat line SeriM82 has shown a significant yield advantage in multi-environment screening trials in this region. The objective of this study was to identify the physiological basis of the adaptive traits underpinning this advantage. Six detailed experiments were conducted to compare the growth, development, and yield of SeriM82 with that of the adapted cultivar, Hartog. The experiments were undertaken in field environments that represented the range of moisture availability conditions commonly encountered by winter crops grown on the deep Vertosol soils of this region. The yield of SeriM82 was 6-28% greater than that of Hartog, and SeriM82 exhibited a stay-green phenotype by maintaining green leaf area longer during the grain-filling period in all environments where yield was significantly greater than Hartog. However, where the availability of deep soil moisture was limited, SeriM82 failed to exhibit significantly greater yield or to express the stay-green phenotype. Thus, the stay-green phenotype was closely associated with the yield advantage of SeriM82. SeriM82 also exhibited higher mean grain mass than Hartog in all environments. It is suggested that small differences in water use before anthesis, or greater water extraction from depth after anthesis, could underlie the stay-green phenotype. The inability of SeriM82 to exhibit stay-green and higher yield where deep soil moisture was depleted indicates that extraction of deep soil moisture is important.

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.

Plant hosts of the phytoplasmas and rickettsia-like-organisms associated with strawberry lethal yellows and green petal diseases

Candidatus Phytoplasma australiense (Ca. P. australiense) is associated with the plant diseases strawberry lethal yellows (SLY), strawberry green petal (SGP), papaya dieback (PDB), Australian grapevine yellows (AGY) and Phormium yellow leaf (PYL; New Zealand). Strawberry lethal yellows disease is also associated with a rickettsia-like-organism (RLO) or infrequently with the tomato big bud (TBB) phytoplasma, the latter being associated with a wide range of plant diseases throughout Australia. In contrast, the RLO has been identified only in association with SLY disease, and Ca. P. australiense has been detected only in a limited number of plant host species. The aim of this study was to identify plant hosts that are possible reservoirs of Ca. P. australiense and the SLY RLO. Thirty-one plant species from south-east Queensland were observed with disease between 2001 and 2003 and, of these, 18 species tested positive using phytoplasma-specific primers. The RLO was detected in diseased Jacksonia scoparia and Modiola caroliniana samples collected at Stanthorpe. The TBB phytoplasma was detected in 16 different plant species and Ca. P. australiense Australian grapevine yellows strain was detected in six species. The TBB phytoplasma was detected in plants collected at Nambour, Stanthorpe, Warwick and Brisbane. Ca. P. australiense was detected in plants collected at Nambour, Stanthorpe, Gatton and Allora. All four phytoplasmas were detected in diseased Gomphocarpus physocarpus plants collected at Toowoomba, Allora, Nambour and Gatton. These results indicated that the vector(s) of Ca. P. australiense are distributed throughout south-east Queensland and the diversity of phytoplasmas detected in G. physocarpus suggests it is a feeding source for phytoplasma insect vectors or it has a broad susceptibility to a range of phytoplasmas.

Prothoracic Gland Semiochemicals of Green Lacewings.

Adult chrysopids have paired prothoracic glands (PG) that are thought to produce defensive secretions (allomones). We analyzed PG extracts of the following green lacewings from North and South America, Australia, and China: Ceraeochrysa cubana (Brazil); Chrysopa (= Co.) oculata, Co. nigricornis, Co. incompleta, Co. quadripunctata (USA), and Co. septempunctata (China); Chrysoperla (= Cl.) rufilabris (USA) and Cl. sp. (Brazil); Plesiochrysa ramburi and Mallada spp. (Australia). PG secretions are characteristic for species within a genus, except for Chrysopa spp. (Z)-4-Tridecene is ubiquitous, but (Z,Z)-4,7-tridecadiene is a major PG constituent in some Chrysopa spp. and in P. ramburi. Earlier reports that Co. oculata and Co. nigricornis produce 1-tridecene were shown to be in error. Chrysopa PG secretions are distinguished by the presence or absence of N-3-methylbutylacetamide, plus skatole (3-methylindole). Skatole is also identified for the first time from the Plesiochrysa and Ceraeochrysa. The PG secretion in Plesiochrysa ramburi is characterized by the presence of (Z)-4-undecene instead of (Z)-4-tridecene, and N-3-methylbutylpropanamide instead of the acetamide, resembling the PG secretions of Chrysopa nigricornis, Co. septempunctata and Co. incompleta. The chemotaxonomic value of PG semiochemicals is discussed, including evidence for subgroups within the genus Chrysopa as it now stands.

Collation, analysis and research of thermal benefits of green life in the urban landscapes

This project reviewed international research conducted on the possible role of plants in alleviating high temperatures in our living spaces. The literature review served to identify the work that has already been carried out in the area and to highlight the gaps to be filled by experimental research. A pilot study then investigated the thermal properties of six of the most common landscaping materials. This project clearly shows that plants can play a significant role in modifying the thermal conditions of urban environments. Tall trees can shade nearby buildings and allow for reductions in cooling costs. In addition to basic shading, the dispersal of heat via the plant’s natural transpiration stream has long been recognised as an important component of the urban energy balance. It has been shown that urban temperatures can be up to 7°C higher than nearby rural areas, illustrating the impact of plants on their environment. These benefits argue against the idea of removing plants from landscapes in order to save on water in times of drought. Similarly, the idea of switching to artificial turf is questionable, since artificial turf still requires watering and can reach temperatures that far exceed the safe range for players. While vegetation offers evaporative cooling, non-vegetative, impervious surfaces such as concrete do not, and can therefore cause greater surface and soil temperatures. In addition, the higher temperatures associated with these impervious surfaces can negatively affect the growth of plants in surrounding areas. Permeable surfaces, such as mulches, have better insulating properties and can prevent excessive heating of the soil. However, they can also lead to an increase in reflected longwave radiation, causing the leaves of plants to close their water-conducting pores and reducing the beneficial cooling effects of transpiration. The results show that the energy balance of our surroundings is complicated and that all components of a landscape will have an impact on thermal conditions.

Cynodon dactylon, Green Couch Grass, Bermuda Grass 'Grand Prix'

‘Grand Prix’ is a selection from a cross between ‘Wintergreen’ and ‘Couch 5’ (also designated C5). ‘Couch 5’ was a selection from an earlier series of crosses by the breeder between ‘Wintergreen’ and a number of Cynodon dactylon accessions, which were collected by the breeder from the Mornington Peninsula area of Victoria between 1986 and 1990. C5 was an experimental breeding line, and was not subsequently reserved as vegetative germplasm. Living material of C5 is no longer in existence. Following the crossing of ‘Couch 5’ and ‘Wintergreen’ in 1998, the resultant seed was germinated on moist blotting paper. Individual seedlings, a total of 150 in number, were planted into 150mm pots and these plants observed during 1998 and 1999. During the summer of 1999-2000, the majority of the seedling plants were culled on the basis of their shoot density, leaf texture, internode length, and colour. In the spring of 2000, the remaining 20 potted seedlings were planted individually into 4m2 plots at the Evergreen Turf farm at Pakenham (Victoria), and allowed to expand fully across these plots. The final selection of Seedling 12 (later designated DN12) in late 2002 was based on shoot density, leaf colour, turf quality, and reduced thatch accumulation as expressed in these plots. Propagation: the original plant has been multiplied through four (4) vegetative expansions prior to PBR application without showing any discernible off types. Breeder: David Nickson, Frankston, VIC. PBR Certificate Number 3133, Application Number 2005/291, granted 12 September 2006.

Cynodon dactylon, Green Couch Grass, Bermuda Grass 'Winter Gem'

‘Winter Gem’ is a selection from a cross between ‘Wintergreen’ and Couch 5 (also designated C5). Couch 5 was a selection from an earlier series of crosses by the breeder between ‘Wintergreen’ and a number of Cynodon dactylon accessions, which were collected by the breeder from the Mornington Peninsula area of Victoria between 1986 and 1990. C5 was an experimental breeding line, and was not subsequently reserved as vegetative germplasm. Living material of C5 is no longer in existence. Following the crossing of Couch 5 and ‘Wintergreen’ in 1998, the resultant seed was germinated on moist blotting paper. Individual seedlings, a total of 150 in number, were planted into 150mm pots and these plants observed during 1998 and 1999. During the summer of 1999-2000, the majority of the seedling plants were culled on the basis of their shoot density, leaf texture, internode length, and colour. In the spring of 2000, the remaining 20 potted seedlings were planted individually into 4m2 plots at the Evergreen Turf farm at Pakenham (Victoria), and allowed to expand fully across these plots. The final selection of Seedling 9 (later designated DN9) in late 2002 was based on shoot density, leaf texture, and retention of winter colour as expressed in these plots. Propagation: The original plant had been multiplied through four (4) vegetative expansions prior to PBR application without showing any discernible off types. Breeder: David Nickson, Frankston, VIC. PBR Certificate Number 3132, Application Number 2005/290, granted 11 September 2006.

Cynodon transvaalensis x Cynodon dactylon, Hybrid Green Couch Grass, Hybrid Bermuda Grass 'P18'

‘P18’ was first produced in 1992 and is a mutant genotype obtained from a hybrid Bermudagrass line believed to be ‘Tifdwarf’, which was grown in a greenhouse owned by H&H Seed Company in Yuma, Arizona. ‘P18’ was selected for its extremely fine leaf texture, its high shoot density under close mowing, its rapid growth rate, and its uniform dark green colour, and was subsequently evaluated for these traits and characteristics. Propagation: vegetative. Breeder: Howard E. Kaewer, Eden Prairie, MN, USA. PBR Application Number 2007/179, Certificate Number 3567, granted 13 August 2007.

Migration of green turtles (Chelonia mydas) from Australasian feeding grounds inferred from genetic analyses

Coastal seagrass habitats in tropical and subtropical regions support aggregations of resident green turtles (Chelonia mydas) from several genetically distinct breeding populations. Migration of individuals to their respective dispersed breeding sites provides a complex pattern of migratory connectivity among nesting and feeding habitats of this species. An understanding of this pattern is important in regions where the persistence of populations is under threat from anthropogenic impacts. The present study uses mitochondrial DNA and mixed-stock analyses to assess the connectivity among seven feeding grounds across the north Australian coast and adjacent areas and 17 genetically distinct breeding populations from the Indo-Pacific region. It was hypothesised that large and geographically proximate breeding populations would dominate at nearby feeding grounds. As expected, each sampled feeding area appears to support multiple breeding populations, with two aggregations dominated by a local breeding population. Geographic distance between breeding and feeding habitat strongly influenced whether a breeding population contributed to a feeding ground (wi = 0.654); however, neither distance nor size of a breeding population was a good predictor of the extent of their contribution. The differential proportional contributions suggest the impact of anthropogenic mortality at feeding grounds should be assessed on a case-by-case basis.

The Relationship Between the Stay-Green Trait and Grain Yield in Elite Sorghum Hybrids Grown in a Range of Environments

The stay-green drought adaptation mechanism has been widely promoted as a way of improving grain yield and lodging resistance in sorghum [Sorghum bicolor (L.) Moench] and as a result has been the subject of many physiological and genetic studies. The relevance of these studies to elite sorghum hybrids is not clear given that they sample a limited number of environments and were conducted using inbred lines or relatively small numbers of experimental F-1 hybrids. In this study we investigated the relationship between stay-green and yield using data from breeding trials that sampled 1668 unique hybrid combinations and 23 environments whose mean yields varied from 2.3 to 10.5 t ha(-1). The strength and direction of the association between stay-green and grain yield varied with both environment and genetic background (male tester). The majority of associations were positive, particularly in environments with yields below 6 t ha(-1). As trial mean yield increased above 6 t ha(-1) there was a trend toward an increased number of negative associations; however, the number and magnitude of the positive associations were larger. Given that post-flowering drought is very commonly experienced by sorghum crops world wide and average yields are 1.2 and 2.5 t ha(-1) for the world and Australia, respectively, our results indicate that selection for stay-green in elite sorghum hybrids may be broadly beneficial for increasing yield in a wide range of environments.