Genetic diversity revealed in the apomictic fruit species Garcinia mangostana L. (mangosteen)

The novel molecular marker technique Randomly Amplified DNA Fingerprinting (RAF) was used to survey genetic relationships between 37 accessions of the tropical fruit G. mangostana (mangosteen) and among 11 accessions from eight other Garcinia species. Although mangosteen is believed to reproduce exclusively through apomixis, our results show that considerable genetic diversity exists within G. mangostana and between other Garcinia species. Among the 37 G. mangostana accessions examined, nine different genotypes were identified which clustered into three distinct groups based on correspondence analysis (reciprocal averaging). For 26 (70%) of the accessions no marker variation was detected over 530 loci screened. A further eight (22%) accessions exhibited very low levels of variation (0.2-1%) suggesting at least one well conservedm angosteen genotype. The remaining three accessions (8%) showed extensive variation (22-31%) compared with the majority of accessions. The three mangosteen groups were 63-70% dissimilar to the other Garcinia species investigated. The genetic diversity identified in this research will assist in the conservation of Garcinia germplasm and provides a valuable framework for the genetic improvement of mangosteen.

Manipulating avocado fruit ripening with 1-methylcyclopropene

Previous investigations with 1-methylcyclopropene (1-MCP) on avocado (Persea americana Mill.) fruit have focussed mainly on improving storage life by reducing the severity of disorders causing discolouration of the flesh. Development of 1-MCP and ethylene treatments, which also help control the time to reach the eating ripe stage, may confer additional practical benefits. In this context, the current study investigated the potential of 1-MCP to accurately manipulate ripening of non-stored 'Hass' avocado fruit by treatment before or after ethylene and at different times during ripening. To investigate this, 500 nL L-1 1-MCP was applied within 1 day after harvest, followed by ethylene 0-14 days after 1-MCP. In addition, fruit were treated with ethylene, then 1-MCP 0-8 days after ethylene. Treatment of fruit with 500 nL L-1 1-MCP for 18 h at 20 degreesC provided the maximum effect by increasing the days from harvest to ripe (DTR) from 8 (with no 1-MCP) to 20. Fruit treated with 500 nL L-1 1-MCP for 18 h at 20 degreesC remained insensitive to 100 muL L-1 ethylene applied between 0 and 14 days after 1-MCP for 24 h at 20 degreesC. Ripening of fruit exposed to 100 muL L-1 ethylene for 24 h at 20 degreesC could be delayed by up to 3.3 days by applying 500 nL L-1 1-MCP for 18 h at 20 degreesC up to 2 days after ethylene treatment. However, once the fruit started to soften (sprung) there was little effect of 1-MCP on DTR, compared with no 1-MCP. 1-MCP treatment was associated with increased severity of body rots (caused mainly by Colletotrichum spp.) and stem-end rots (caused mainly by Dothiorella spp.), which was likely due to the increased DTR in these treatments. Significant differences in disease severity were found between orchards (replications), with replicates with low disease severity being less affected by 1-MCP treatment. These results indicate that 1-MCP can delay ripening, but careful sourcing of fruit is required to reduce the risk of diseases in ripe fruit. There is some capacity to delay ripening using 1-MCP after ethylene. There is little potential to control ripening using ethylene after treatment with 500 nL L-1 1-1-MCP, but lower concentrations may be more effective. (C) 2004 Elsevier B.V. All rights reserved.

Effect of modified atmosphere packaging on the quality of minimally processed pineapple cv.'Smooth Cayenne' fruit

The effects of modified atmosphere (MA) conditions on the quality of minimally processed pineapple slices were determined. Commercial pineapple slice packs sealed with 40 pm thick polyester film were kept at 4.5 degrees C for 14 d. The oxygen transmission rate of the film was 23 ml m(-2) day(-1) atm(-1) (at 25 degrees C, 75% RH). In-built atmospheres and the quality of the products were determined. O-2 concentrations within the packs stabilised at 2%, while CO2 concentrations increased to 70% by day 14. The high CO2 level suggested an inappropriate lidding film permeability for the product, and hence affected its quality. Three batches of pineapple slices were packed in the laboratory using lidding films with oxygen transmission rate of 75, 2790 or 5000 ml m(-2) day(-1) atm(-1) (at 23 degrees C, 0% RH). Headspace atmospheres from laboratory-packed pineapple slices suggested an optimum equilibrium modified atmosphere of ca. 2% O-2 and 15% CO2. Respiration data from the laboratory-prepared packs were pooled together and used to develop a correlation model relating respiration rates to O-2 and CO2 concentrations. The model showed a decrease in respiration rate with decreasing O-2 and increasing CO2 concentrations. Respiration rate stabilised at 2% 02 and 10% CO2. The high concentrations of CO2 observed in the commercial packs did not fit the range in the respiration model. The model could aid in selection of MA conditions for minimally processed pineapple fruit.

Developing pineapple fruit has a small transcriptome dominated by metallothionein

In a first step toward understanding the molecular basis of pineapple fruit development, a sequencing project was initiated to survey a range of expressed sequences from green unripe and yellow ripe fruit tissue. A highly abundant metallothionein transcript was identified during library construction, and was estimated to account for up to 50% of all EST library clones. Library clones with metallothionein subtracted were sequenced, and 408 unripe green and 1140 ripe yellow edited EST clone sequences were retrieved. Clone redundancy was high, with the combined 1548 clone sequences clustering into just 634 contigs comprising 191 consensus sequences and 443 singletons. Half of the EST clone sequences clustered within 13.5% and 9.3% of contigs from green unripe and yellow ripe libraries, respectively, indicating that a small subset of genes dominate the majority of the transcriptome. Furthermore, sequence cluster analysis, northern analysis, and functional classification revealed major differences between genes expressed in the unripe green and ripe yellow fruit tissues. Abundant genes identified from the green fruit include a fruit bromelain and a bromelain inhibitor. Abundant genes identified in the yellow fruit library include a MADS box gene, and several genes normally associated with protein synthesis, including homologues of ribosomal L10 and the translation factors SUI1 and eIF5A. Both the green unripe and yellow ripe libraries contained high proportions of clones associated with oxidative stress responses and the detoxification of free radicals.

Effects of sodium on potassium nutrition in three tropical root crop species

The potassium (K) nutrition and high K requirement of tropical root crops may be affected by their sodium (Na) status, as has been observed in a number of plant species. Solution culture was used to study the effects of K and Na supplies in tannia [Xanthosoma sagittifolium (L.) Schott.], sweetpotato [Ipomoea batatas (L.) Lam.] and taro [Colocasia esculenta (L.) Schott]. At low K supply, Na ameliorated symptoms of K deficiency and increased growth in tannia, and to a lesser extent in sweetpotato, but not in taro. None of the species responded to Na at adequate K supply. Differences in response to Na were attributed to differences in Na translocation to plant tops. At maximum Na supply, the Na concentration in index leaves averaged 1.82% in tannia, 0.205% in sweetpotato, and 0.0067% in taro. An increase in the supply of Na resulted in a shift in the critical K concentration for deficiency (i.e., 90% of maximum yield) in index leaves from 2.9% to 1.2% in tannia, and from 4.8% to 2.5% in sweetpotato. The critical K concentration in taro was 3.3%, irrespective of Na supply. To overcome the problem in tannia and sweetpotato of determining the critical concentration relevant to a leaf sample of unknown K status, a relationship was established for each species relating the critical K concentration to the concentration of Na in the index leaves.

Present and future potential of pineapple biotechnology

Pineapple is an important crop for many countries in Central and South America as well as the Asia-Pacific region. Even though the history of the crop dates to pre-Colombian times there is a remarkable lack of commercial varieties with a single cultivar ‘Smooth Cayenne’ dominating the whole industry. Variety improvement is a very difficult task for pineapple breeders and very little progress has been made in this respect when compared to other crops more suitable to classical breeding approaches. This special characteristic makes pineapple specially suited for genetic engineering approaches that can transfer specific traits from other species into pineapple. In this presentation past and present efforts to use biotechnological methods for the improvement of pineapple will be reviewed. On-going biotechnology projects include control of flowering and control of ‘blackheart’ disease. The development of pineapple biotechnology, as with any other crop, is dependent on the availability of a number of molecular tools, which will also be discussed. For pineapple, these tools can be roughly classified into three different categories: (1) availability of useful genes (2) availability of suitable promoters and (3) availability of an efficient transformation method.

The isolation, characterization, and application of WRKY genes as useful molecular markers in tropical trees

The improvement of tropical tree crops using conventional breeding methods faces challenges due to the length of time involved. Thus, like most crops, there is an effort to utilize molecular genetic markers in breeding programs to select for desirable agronomic traits. Known as marker assisted breeding or marker assisted selection, genetic markers associated with a phenotype of interest are used to screen and select material reducing the time necessary to evaluate candidates. As the focus of this research was improving disease resistance in tropical trees, the usefulness of the WRKY gene superfamily was investigated as candidates for generating useful molecular genetic markers. WRKY genes encode plant-specific transcriptional factors associated with regulating plants' responses to both biotic and abiotic stress. ^ One pair of degenerate primers amplified 48 WRKY gene fragments from three taxonomically distinct, economically important, tropical tree crop species: 18 from Theobroma cacao L., 21 from Cocos nucifera L. and 9 from Persea americana Mill. Several loci from each species were polymorphic because of single nucleotide substitutions present within a putative non-coding region of the loci. Capillary array electrophoresis-single strand conformational polymorphism (CAE-SSCP) mapped four WRKY loci onto a genetic linkage map of a T. cacao F2 population segregating for resistance to witches' broom disease. Additionally, PCR primers specific for four T. cacao loci successfully amplified WRKY loci from 15 members of the Byttneriae tribe. A method was devised to allow the reliable discrimination of alleles by CAE-SSCP using only the mobility assigned to the sample peaks. Once this method was validated, the diversity of three WRKY loci was evaluated in a germplasm collection of T. cacao . One locus displayed high diversity in the collection, with at least 18 alleles detected from mobility differences of the product peaks. The number of WRKY loci available within the genome, ease of isolation by degenerate PCR, codominant segregation demonstrated in the F2 population, and usefulness for screening germplasm collections and closely related wild species demonstrates that the WRKY superfamily of genes are excellent candidates for developing a number of genetic molecular markers for breeding purposes in tropical trees. ^

A model of macadamia with application to pruning in orchards

A self-organising model of macadamia, expressed using L-Systems, was used to explore aspects of canopy management. A small set of parameters control the basic architecture of the model, with a high degree of self-organisation occurring to determine the fate and growth of buds. Light was sensed at the leaf level and used to represent vigour and accumulated basipetally. Buds also sensed light so as to provide demand in the subsequent redistribution of the vigour. Empirical relationships were derived from a set of 24 completely digitised trees after conversion to multiscale tree graphs (MTG) and analysis with the OpenAlea software library. The ability to write MTG files was embedded within the model so that various tree statistics could be exported for each run of the model. To explore the parameter space a series of runs was completed using a high-throughput computing platform. When combined with MTG generation and analysis with OpenAlea it provided a convenient way in which thousands of simulations could be explored. We allowed the model trees to develop using self-organisation and simulated cultural practices such as hedging, topping, removal of the leader and limb removal within a small representation of an orchard. The model provides insight into the impact of these practices on potential for growth and the light distribution within the canopy and to the orchard floor by coupling the model with a path-tracing program to simulate the light environment. The lessons learnt from this will be applied to other evergreen, tropical fruit and nut trees.

MORPHOLOGICAL DIVERSITY OF MANGO GERMPLASM FROM THE UPPER ATHI RIVER REGION OF EASTERN KENYA: AN ANALYSIS BASED ON NON- FRUIT DESCRIPTORS

Phenotypic variation in plants can be evaluated by morphological characterization using visual attributes. Fruits have been the major descriptors for identification of different varieties of fruit crops. However, even in their absence, farmers, breeders and interested stakeholders require to distinguish between different mango varieties. This study aimed at determining diversity in mango germplasm from the Upper Athi River (UAR) and providing useful alternative descriptors for the identification of different mango varieties in the absence of fruits. A total of 20 International Plant Genetic Resources Institute (IPGRI) descriptors for mango were selected for use in the visual assessment of 98 mango accessions from 15 sites of the UAR region of eastern Kenya. Purposive sampling was used to identify farmers growing diverse varieties of mangoes. Evaluation of the descriptors was performed on-site and the data collected were then subjected to multivariate analysis including Principal Component Analysis (PCA) and Cluster analysis, one- way analysis of variance (ANOVA) and Chi square tests. Results classified the accessions into two major groups corresponding to indigenous and exotic varieties. The PCA showed the first six principal components accounting for 75.12% of the total variance. A strong and highly significant correlation was observed between the color of fully grown leaves, leaf blade width, leaf blade length and petiole length and also between the leaf attitude, color of young leaf, stem circumference, tree height, leaf margin, growth habit and fragrance. Useful descriptors for morphological evaluation were 14 out of the selected 20; however, ANOVA and Chi square test revealed that diversity in the accessions was majorly as a result of variations in color of young leaves, leaf attitude, leaf texture, growth habit, leaf blade length, leaf blade width and petiole length traits. These results reveal that mango germplasm in the UAR has significant diversity and that other morphological traits apart from fruits can be useful in morphological characterization of mango.

Adapting fruit production to climate change: how to increase intrinsic fruit quality and save water through deficit irrigation scheduling.

Fruit crops are an important resource for food security, since more than being nutrient they are also a source of natural antioxidant compounds, such as polyphenols and vitamins. However, fruit crops are also among the cultivations threatened by the harmful effects of climate change This study had the objective of investigating the physiological effects of deficit irrigation on apple (2020-2021), sour cherry (2020-2021-2022) and apricot (2021-2022) trees, with a special focus on fruit nutraceutical quality. On each trial, the main physiological parameters were monitored along the growing season: i) stem and leaf water potentials; ii) leaf gas exchanges; iii) fruit and shoot growth. At harvest, fruit quality was evaluated especially in terms of fruit size, flesh firmness and soluble solids content. Moreover, it was performed: i) total phenolic content determination; ii) anthocyanidin concentration evaluation; and iii) untargeted metabolomic study. Irrigation scheduling in apricot, apple and sour cherry is surely overestimated by the decision support system available in Emilia-Romagna region. The water stress imposed on different fruit crops, each during two years of study, showed as a general conclusion that the decrease in the irrigation water did not show a straightforward decrease in plant physiological performance. This can be due to the miscalculation of the real water needs of the considered fruit crops. For this reason, there is the need to improve this important tool for an appropriate water irrigation management. Furthermore, there is also the need to study the behaviour of fruit crops under more severe deficit irrigations. In fact, it is likely that the application of lower water amounts will enhance the synthesis of specialized metabolites, with positive repercussion on human health. These hypotheses must be verified.