PI09004 New Generation Pineapple Breeding Project

To work with a major industry production/marketing unit to develop new pineapple varieties with good plant vigour, high yields and post-harvest attributes and eating quality equal to or better than the standard industry varieties 73-50 and MD2 under commercial production systems. The project uses previously developed germplasm as parental material.

The application of DArTseq technology to pineapple

DArTseq technology is potentially the most appropriate system to discover hundreds of polymorphic genomic loci, scoring thousands of unique genomic-wide DNA fragments in one single experiment, without requiring existing DNA sequence information. The DArT complexity reduction approach in combination with Illumina short read sequencing (Hiseq2000) was applied. To test the application of DArTseq technology in pineapple, a reference population of 13 Ananas genotypes from primitive wild accessions to modern cultivars was used. In a comparison of 3 systems, the combination of restriction enzymes PstI and MseI performed the best producing 18,900 DArT markers and close to 20,000 SNPs. Based on these markers genetic relationships between the samples were identified and a dendrogram was generated. The topography of the tree corresponds with our understanding of the genetic relationships between the genotypes. Importantly, the replicated samples of all genotypes have a dissimilarity of close to 0.0 and occupy the same positions on the tree, confirming high reproducibility of the markers detected. Eventually it is planned that molecular markers will be identified that are associated with resistance to Phytophthora cinnamomi (Pc), the most economically important pathogen of pineapple in Australia, as genetic resistance is known to exist within the Ananas. Marker assisted selection can then be utilized in a pineapple breeding program to develop cultivars resistant to Pc.

Review of genetic improvement of pineapple.

World consumption of fresh pineapple has quadrupled in less than 15 years (Loeillet and Pacqui, 2009). This phenomenal event started around 1996 when the first dedicated fresh market pineapple, '73-114', was released by Del Monte Inc. This was the culmination of somewhere in the vicinity of 34 years of breeding and selection and comprised 24 individual parent combinations (Anon., PRI breeding records). This demonstrates the difficulty of breeding new pineapple cultivars but also the value of a successful program. The success of '73-114' and the competitive nature of world pineapple markets have provided impetus for pineapple breeding programs. However, the highly heterozygous nature and self-incompatibility of pineapple limit breeding strategy options. This review looks at the collective experience in pineapple genetic improvement both conventional and using biotechnology tools, with an emphasis on fresh market pineapple. It focus on relevant pineapple reproductive biology, breeding strategies, parent cultivars and the relevance of biotechnology.

Measuring nitrous oxide emissions from conventional and controlled release fertilisers in south-east Queensland pineapple production

Nitrous oxide (N2O) is a potent greenhouse gas with a global warming potential 298 times higher than carbon dioxide. Soils are a natural source of N2O, contributing 65% of global emissions. This paper is the first in Australia to measure and compare N2O emissions from pre-plant controlled release (CR) and conventional granular (CV) fertilisers in pineapple production using static PVC chambers to capture N2O emissions. Farm 1 cumulative emissions from the CR fertiliser were 3.22 kg ha-1 compared to 6.09 kg ha-1 produced by the CV. At farm 2 the CV blend emitted 2.36 kg ha-1 in comparison to the CR blend of 2.92 kg ha-1. Daily N2O flux rates showed a relationship of direct response to rainfall and soil moisture availability. High emissions were observed for wheel tracks where increased N2O emissions may be linked to soil compaction and waterlogging that creates anaerobic conditions after rain events. Emission measurements over three months highlighted the inconsistencies found in other studies relative to reducing emissions through controlled release nitrogen. More investigations are required to verify the benefits associated with controlled release fertiliser use in pineapples, placement and seasonal timing to address N2O emissions in pineapples.

Leadership training and regular communication identified as key strategies for the development of the Australian pineapple industry

Changes in the circumstances of the Australian pineapple industry left growers with a leadership vacuum, limited technical support and no funds for conducting research and marketing. Inspirational leadership training together with regular district farm meetings were used to assist the Australian pineapple industry to successfully adapt to these challenges. All growers were assigned to one of a number of regional grower study groups and regular on-farm meetings commenced to facilitate communication between growers, transfer of technology, awareness of industry affairs and an opportunity to become involved in industry business. A leader was appointed within each study group and these leaders attended a leadership course consisting of three, three-day modules. These original course graduates formed the nucleus of a new grower representative group which subsequently instigated levies to fund research and marketing. Two more courses have since been conducted to provide the depth of leadership to satisfy the growers' desire to rotate industry leadership on a regular basis.

In Vitro Conditions for a Successful Biolistics transformation System of Pineapples (ananas comosus merr.), in 'Contributing to a Sustainable Future'

In order to develop an efficient and reliable biolistics transformation system for pineapples parameters need to be optimised for growth, survival and development of explants pre- and post transformation. We have optimised in vitro conditions for culture media for the various stages of plant and callus initiation and development, and for effective selection of putative transgenic material. Shoot multiplication and proliferation is best on medium containing MS basic nutrients and vitamins with the addition of 0.1 mg/L myo-inositol, 20 g/L sucrose, 2.5 mg/L BAP and 3 g/L Phytagel, followed by transfer to basic MS medium for further development. Callus production on leaf base explants is best on MS nutrients and vitamins, to which 10 mg/L of BAP and NAA each was added. Optimum explant age for bombardment is 17-35 week old callus, while a pre-bombardment osmoticum treatment in the medium is not required. By comparing several antibiotics as selective agent, it has been established that a two-step selection of 2 fortnightly sub-cultures on 50 μg/mL of geneticin in the culture medium, followed by monthly sub-cultures on 100 μg/mL geneticin is optimal for survival of transgenic callus. Shoot regeneration from callus cultures is optimal on medium containing MS nutrients and vitamins, 5% coconut water and 400 mg/L casein hydrolysate. Plants can be readily regenerated and multiplied from transgenic callus through organogenesis. Rooting of shoots does not require any additional plant hormones to the medium. A transformation efficiency of 1 – 3.5% can be achieved, depending on the gene construct applied.

The genetic diversity of ampeloviruses in Australian pineapples and their association with mealybug wilt disease

Pineapple mealybug wilt-associated virus 1 (PMWaV-1), 2 (PMWaV-2) and -3 (PMWaV-3) have been detected in Australian commercial pineapple crops, along with a previously undescribed ampelovirus, for which the name Pineapple mealybug wilt-associated virus 5 (PMWaV-5) is proposed. Partial sequences extending from open reading frame 1b through to the heat shock protein homologue were obtained for PMWaV-1, -3 and -5. Phylogenetic analyses of selected regions of these sequences indicated that PMWaV-5 is a distinct species and most closely related to PMWaV-1. The amino acid sequence variation observed in the RNA-dependent RNA polymerase region of PMWaV-1 isolates was 95.8–98.4% and of PMWaV-3 isolates was 92.2–99.5%. In surveys of mealybug wilt disease (MWD) affected crops, none of the four viruses was clearly associated with the disease at all survey sites. A statistically significant association (P < 0.001) between the presence of PMWaV-2 and symptoms was observed at one survey site (site 3), but the virus was at a low incidence at the remaining three survey sites. By contrast, although PMWaV-1 and -3 were equally distributed between symptomless and MWD-affected plants at site 3, there was a statistically significant (P < 0.001) association between each of these two viruses and MWD at sites 1 and 4. At site 2, there was a statistically significant (P < 0.001) association only between PMWaV-3 and MWD. PMWaV-1 was the most commonly found of the four viruses and conversely PMWaV-5 was only occasionally found. Australian isolates of PMWaV-1, -2 and -3 were transmitted by the mealybug species Dysmicoccus brevipes.

Ultraviolet radiation effects on fruit surface respiration and chlorophyll fluorescence

High-value fruit crops are exposed to a range of environmental conditions that can reduce fruit quality. Solar injury (SI) or sunburn is a common disorder in tropical, sub-tropical, and temperate climates and is related to: 1) high fruit surface temperature; 2) high visible light intensity; and, 3) ultraviolet radiation (UV). Positional changes in fruit that are caused by increased weight or abrupt changes that result from summer pruning, limb breakage, or other damage to the canopy can expose fruit to high solar radiation levels, increased fruit surface temperatures, and increased UV exposure that are higher than the conditions to which they are adapted. In our studies, we examined the effects of high fruit surface temperature, saturating photosynthetically-active radiation (PAR), and short-term UV exposure on chlorophyll fluorescence, respiration, and photosynthesis of fruit peel tissues from tropical and temperate fruit in a simulation of these acute environmental changes. All tropical fruits (citrus, macadamia, avocado, pineapple, and custard apple) and the apple cultivars 'Gala', 'Gold Rush', and 'Granny Smith' increased dark respiration (A0) when exposed to UV, suggesting that UV repair mechanisms were induced. The maximum quantum efficiency of photosystem II (Fv/Fm) and the quantum efficiency of photosystem II (ΦII) were unaffected, indicating no adverse effects on photosystem II (PSII). In contrast, 'Braeburn' apple had a reduced Fv/Fm with no increase in A0 on all sampling dates. There was a consistent pattern in all studies. When Fv/Fm was unaffected by UV treatment, A0 increased significantly. Conversely, when Fv/Fm was reduced by UV treatment, then A0 was unaffected. The pattern suggests that when UV repair mechanisms are effective, PSII is adequately protected, and that this protection occurs at the cost of higher respiration. However, when the UV repair mechanisms are ineffective, not only is PSII damaged, but there is additional short-term damage to the repair mechanisms, indicated by a lack of respiration to provide energy.

Cucumber mosaic virus infection of kava (Piper methysticum) and implications for cultural control of kava dieback disease

Cucumber mosaic virus (CMV) was found by reverse transcription polymerase chain reaction (RT-PCR) to be not fully systemic in naturally infected kava (Piper methysticum) plants in Fiji. Twenty-six of 48 samples (54%) from various tissues of three recently infected plants were CMV-positive compared with 7/51 samples (14%) from three long-term infections (plants affected by dieback for more than 1 year). The virus was also found to have a limited ability to move into newly formed stems. CMV was detected in only 2/23 samples taken from re-growth stems arising from known CMV infected/dieback affected plants. Mechanical inoculation experiments conducted in Fiji indicate that the known kava intercrop plants banana (Musa spp.), pineapple (Ananas comosus), peanut (Arachis hypogaea) and the common weed Mikania micrantha are potential hosts for a dieback-causing strain of CMV It was not possible to transmit the virus mechanically to the common kava intercrop plants taro (Colocasia esculenta), Xanthosoma sp., sweet potato (Ipomoea batatas), yam (Dioscorea alata), papaya (Carica papaya) or the weed Momordica charantia. Implications of the results of this research on a possible integrated disease management strategy are discussed.

Micropropagation of vegetatively propagated crops: accelerating release of new cultivars and providing an important source of clean planting material

Micropropagation is unequalled for the rapid clonal propagation of improved cultivars from several Australian breeding programmes. This has been particularly true of the pineapple breeding programme, but it has also found an important role in the strawberry breeding programme where high-health mother stock is of paramount concern. In the banana and ginger industries, while access to new cultivars has been of importance, micropropagation has been adopted by the industry to ensure that planting materials are free from serious pests and diseases. Bananas can be used as planting material as early as the first generation ex vitro and is responsible for the establishment of laboratories and nurseries specializing in the production of pathogen-tested plants. The ginger industry, on the other hand, has used micropropagated plants as a source of disease and pest-free stock to establish a clean 'seed' scheme based on the production of conventional planting material.

Breeding Ananas for the cut-flower and garden markets

Only small quantities of Ananas have been marketed as cut flowers or as potted plants for garden use in Australia. Worldwide there have, until very recent times, been no breeding programs to develop ornamental characteristics and hence the choice of cultivars has been limited mainly to semi-domesticated selections or those developed by amateur enthusiasts. Interest in developing Ananas selections specifically for the ornamental market is now increasing. A small program has operated in Australia since 1995. In this program, a total of 4,700 seedlings were generated over three generations using various parental combinations of Ananas comosus var. comosus, A. comosus var. bracteatus, A. comosus var. ananassoides 'FRF223', A. comosus var. erectifolious 'Selvagem 6' and Ananas macrodontes 'I.26-803'. Several selections have been developed for the garden and or cut-flower market. Characteristics represented include a pink or red syncarp, dark red-brown foliage and a dwarf, clumping growth habit. While a surprising display of ornamental diversity exists within Ananas, the genus is limited in comparison to the other bromeliad genera. Opportunity might exist however to introgress characteristics such as additional foliage colours, plant morphology and syncarp colours from other genera into Ananas.

Management Guidelines for New Warm-Season Grasses in Australia

After more than 30 years in which ‘Tifgreen’ and ‘Tifdwarf’ were the only greens-quality varieties available, the choice for golf courses and bowls clubs in northern Australia has been expanded to include six new Cynodon hybrids [Cynodon dactylon (L.) Pers x Cynodon transvaalensis Burtt-Davy]. Five of these – ‘Champion Dwarf’ (Texas), ‘MS-Supreme’ (Mississippi), FloraDwarf™ (Florida), ‘TifEagle’ (Georgia), MiniVerde™ (Arizona) - are from US breeding programs, while the sixth, ‘TL2’ (marketed as Novotek™) was selected in north Queensland. The finer, denser and lower growing habit of the “ultradwarf” cultivars allows very low mowing heights (e.g. 2.5 mm) to be imposed, resulting in denser and smoother putting and bowls surfaces. In addition to the Cynodon hybrids, four new greens quality seashore paspalum (Paspalum vaginatum O. Swartz) cultivars including ‘Sea Isle 2000’, Sea Isle Supreme™, Velvetene™ and Sea Dwarf™ (where tolerance of salty water is required) expands the range of choices for greens in difficult environments. The project was developed to determine (a) the appropriate choice of cultivar for different environments and budgets, and (b) best management practices for the new cultivars which differ from the Cynodon hybrid industry standards ‘Tifgreen’ and ‘Tifdwarf’. Management practices, particularly fertilising, mowing heights and frequency, and thatch control were investigated to determine optimum management inputs and provide high quality playing surfaces with the new grasses. To enable effective trialling of these new and old cultivars it was essential to have a number of regional sites participating in the study. Drought and financial hardship of many clubs presented an initial setback with numerous clubs wanting to be involved in the study but were unable to commit due to their financial position at the time. The study was fortunate to have seven regional sites from Queensland, New South Wales, Victoria and South Australia volunteer to be involved in the study which would add to the results being collected at the centralised test facility being constructed at DEEDI’s Redlands Research Station. The major research findings acquired from the eight trial sites included: • All of the new second generation “ultradwarf” couchgrasses tend to produce a large amount of thatch with MiniVerde™ being the greatest thatch producer, particularly compared to ‘Tifdwarf’ and ‘Tifgreen’. The maintenance of the new Cynodon hybrids will require a program of regular dethatching/grooming as well as regular light dustings of sand. Thatch prevention should begin 3 to 4 weeks after planting a new “ultradwarf” couchgrass green, with an emphasis on prevention rather than control. • The “ultradwarfs” produced faster green speeds than the current industry standards ‘Tifgreen’ and ‘Tifdwarf’. However, all Cynodon hybrids were considerably faster than the seashore paspalums (e.g. comparable to the speed diference of Bentgrass and couchgrass) under trial conditions. Green speed was fastest being cut at 3.5 mm and rolled (compared to 3.5 mm cut, no roll and 2.7 mm cut, no roll). • All trial sites reported the occurrence of disease in the Cynodon hybrids with the main incidence of disease occurring during the dormancy period (autumn and winter). The main disease issue reported was “patch diseases” which includes both Gaumannomyces and Rhizoctonia species. There was differences in the severity of the disease between cultivars, however, the severity of the disease was not consistent between cultivars and is largely attributed to an environment (location) effect. In terms of managing the occurrence of disease, the incidence of disease is less severe where there is a higher fertility rate (about 3 kgN/100m2/year) or a preventitatve fungicide program is adopted. • Cynodon hybrid and seashore paspalum cultivars maintained an acceptable to ideal surface being cut between 2.7 mm and 5.0 mm. “Ultradwarf” cultivars can tolerate mowing heights as low as 2.5 mm for short periods but places the plant under high levels of stress. Greens being maintained at a continually lower cutting height (e.g. 2.7 mm) of both species is achievable, but would need to be cut daily for best results. Seashore paspalums performed best when cut at a height of between 2.7 mm and 3.0 mm. If a lower cutting height is adopted, regular and repeated mowings are required to reduce scalping and produce a smooth surface. • At this point in time the optimum rate of nitrogen (N) for the Cynodon hybrids is 3 kg/100m2/year and while the seashore paspalums is 2 to 3 kg/100m2/year. • Dormancy occurred for all Cynodon and seashore paspalum culitvars from north in Brisbane (QLD) to south in Mornington Peninsula (VIC) and west to Novar Gardens (SA). Cynodon and Paspalum growth in both Victoria and South Australia was less favourable as a result of the cooler climates. • After combining the data collected from all eight sites, the results indicated that there can be variation (e.g. turfgrass quality, colour, disease resistance, performace) depending on the site and climatic conditions. Such evidence highlights the need to undertake genotype by environment (G x E) studies on new and old cultivars prior to conversion or establishment. • For a club looking to select either a Cynodon hybrid or seashore paspalum cultivar for use at their club they need to: - Review the research data. - Look at trial plots. - Inspect greens in play that have the new grasses. - Select 2 to 3 cultivars that are considered to be the better types. - Establish them in large (large enough to putt on) plots/nursery/practice putter. Ideally the area should be subjected to wear. - Maintain them exactly as they would be on the golf course/lawn bowls green. This is a critical aspect. Regular mowing, fertilising etc. is essential. - Assess them over at least 2 to 3 years. - Make a selection and establish it in a playing green so that it is subjected to typical wear.

Sweetpotato consumer research: insights to increasing consumption

This report provides an evaluation of the behaviours and purchasing drivers of key sweetpotato consumers defined by Nielsen consumer research as Established Couples (two or more adults with no children 17 and under, and head of house 35-59), Senior Couples (two or more adults with no children 17 or under, and head of house 60 or over), and Independent Singles (one person household 35 or over, no children 17 or under). Research was qualitative in nature. Methods used included focus groups, depth interviews and shop-a-longs. The report found that preferences for sweetpotato amongst these groups were varied. In general a smaller torpedo shaped vegetable was valued for ease of preparation and the convenience of being of sufficient size for a meal for two. Satisfaction with sweetpotato was high with negative comments on quality exceedingly rare within discussions. However, shop-a-longs revealed that some quality issues were apparent at retail such as withered product, pitting and occasionally damage. A display with stock resting in any amount of water was a barrier to purchase for consumers and this was apparent on two out 15 occasions. A high quality sweetpotato was of a deep orange/red colour, had a smooth skin and was extremely dense and hard. An inferior sweetpotato was wrinkly, spongy, pitted and damaged. Awareness of sweetpotato was a relatively recent phenomenon amongst the respondents of this study with most recalling eating the vegetable in the last five to 10 years. Life-time eating patterns emerged as a consequence of childhood food experiences such as growing up with a ‘meat and three’ veg philosophy and traditional Australian meals. However, this was dependent on cultural background and those with ties to diverse cultures were more likely to have always known of the vegetable. Sweetpotato trial and consumption coincided with a breaking away from these traditional patterns, or was integrated into conventional meals such as a baked vegetable to accompany roasts. Increased health consciousness also led to awareness of the vegetable. A primary catalyst for consumption within the Established and Senior Couples groups was the health benefits associated with sweetpotato. Consumers had very little knowledge of the specific health properties of the vegetable and were surprised at the number of benefits consumption provided. Sweetpotato was important for diabetics for its low Glycemic Index status. Top-of-the-mind awareness of the vegetable resulted from the onset of the disease. Increasing fibre was a key motive for this demographic and this provided a significant link between consumption and preventing bowel cancer. For those on a weight loss regime, sweetpotato was perceived as a tasty, satisfying food that was low in carbohydrates. Swapping behaviours where white potato was replaced by sweetpotato was often a response to these health concerns. Other health properties mentioned by participants through the course of the research included the precursor β-carotene and Vitamins A & C. The sweetpotato was appreciated for its hedonic and timesaving qualities. For consumers with a high involvement in food, the vegetable was valued for its versatility in meals. These consumers took pride in cooking and the flavour and texture of sweetpotato lent itself to a variety of meals such as soups, salads, roasts, curries, tagines and so on. Participants who had little time or desire to prepare and cook meals valued sweetpotato because it was an easy way to add colour and variety to the plate and because including an orange vegetable to meals is a shortcut to ensuring vitamin intake. Several recommendations are made to the sweetpotato industry. • Vigorously promote the distinct nutritional and health properties of sweetpotatoes, particularly if they can be favourably compared to other vegetables or foods • Promote the salient properties to specific targets such as diabetics, those that are at risk to bowel cancer, and those embarking on a weight-loss regime. Utilise specialist channels of communication such as diabetic magazines and websites • Promote styles of cooking of sweetpotato that would appeal to traditionalists such as roasts and BBQs • Promote the vegetable as a low maintenance vegetable, easy to store, easy to cook and particularly focusing on it as a simple way to boost the appearance and nutritional value of meals. • Promote the vegetable to high food involvement consumers through exotic recipes and linking it to feelings of accomplishment with cooking • Promote the versatility of the vegetable • Devise promotions that link images and tone of communications with enjoying life to the fullest, having time to enjoy family and grandchildren, and of partaking in social activities • Educate retailers on consumer perceptions of quality and ensuring moisture and mould is not present at displays Qualitative information while providing a wealth of detail cannot be extrapolated to the overall target population and this may be considered a limitation to the research. However, within research theory, effective quantitative design is believed to stem from the insights developed from qualitative studies. • Develop and implement a quantitative study on sweetpotato attitudes and behaviours based on the results of this study.

Perennial pastures for marginal farming country in southern Queensland. 1. Grass establishment techniques

Efficient ways to re-establish pastures are needed on land that requires a rotation between pastures and crops. We conducted trials in southern inland Queensland with a range of tropical perennial grasses sown into wheat stubble that was modified in various ways. Differing seedbed preparations involved cultivation or herbicide sprays, with or without fertilizer at sowing. Seed was broadcast and sowing time ranged from spring through to autumn on 3 different soil types. Seed quality and post-sowing rainfall were major determinants of the density of sown grass plants in the first year. Light cultivation sometimes enhanced establishment compared with herbicide spraying of standing stubble, most often on harder-setting soils. A nitrogen + phosphorus mixed fertilizer rarely produced any improvement in sown grass establishment and sometimes increased weed competition. The effects were similar for all types of grass seed from hairy fascicles to large, smooth panicoid seeds and minute Eragrostis seeds. There was a strong inverse relationship between the initial density of sown grass established and the level of weed competition.