A simplified PCR test for early detection of dwarf off-types in micropropagated Cavendish bananas (Musa spp. AAA)

The dwarf somaclonal variant is a major problem affecting micropropagation of the banana cultivar Williams (Musa spp. AAA; subgroup Cavendish). This problem arises from genetic changes that occur during the tissue culture process. Early identification of this problem is difficult and propagators must wait until plants are ex vitro in order to visualise the dwarfism phenotype. In this study, we have improved a SCAR-based molecular diagnostic technique, developed by Damasco et al. [Acta Hortic. 461 (1997) 157], for the early identification of dwarf off-types. We have included a positive internal control in a multiplex PCR and adapted the technique for use with small amounts of fresh in vitro leaf material as PCR template. The control product is a 500 bp fragment from 18S rRNA and is amplified in all tissues irrespective of phenotype. The use of small in vitro leaf material removing the need for genomic DNA extraction.

Characterisation and early detection of an offtype from micropropagated Lady Finger bananas

An offtype has been identified from micropropagated Lady Finger bananas (Musa spp., AAB group, Pome subgroup) that is characterised by its slow growth and poor bunch size. Bunch weights were approximately 25% those of normal Lady Finger plants and all of the fruit produced was unmarketable. This particular offtype is the most commonly encountered from micropropagated Lady Finger plants and, in 2 instances, blocks of 3000 and 1500 plants were entirely comprised of this single offtype. Detection of offtype plants was possible during establishment and growth of plants in the glasshouse by the presence of chlorotic streaks in the leaves. In more severe cases the streaks coalesced into chlorotic patches that developed thin, necrotic areas that eventually produced holes or splits in the leaves. Symptom expression was not ameliorated by the addition of fertiliser and even though symptoms were similar to severe Ca and B deficiency, both normal and offtype plants had similar levels of these elements in the leaves. The offtype plants were also slow growing in the glasshouse and produced significantly (P<0.05) smaller pseudostems and leaves than normal plants. Offtype plants could be readily detected after 4 weeks deflasking using the presence of chlorotic streaks in the leaves as the main selection criterion. Maximum discrimination was possible between weeks 5–7 and at the 6-leaf stage when all of the offtypes could be detected.

Inconsistent Transmission of Banana Bunchy Top Virus in Micropropagated Bananas and its Implication for Germplasm Screening

Banana bunchy top virus (BBTV) was readily transmitted through tissue culture in banana (Mum sp.) cv. Lady finger (AAB) and Cavendish cv. Williams (AAA). Lines derived from infected and healthy field plants had similar in vitro multiplication rates. BBTV infected in vitro cultures displayed symptoms of stunting, leaf curling, chlorotic and green flecks, and poor root growth. Symptoms became milder with time, and were often difficult to discern in older, rapidly multiplying cultures. A triple antibody sandwich ELISA using polyclonal and monoclonal antibodies was very efficient for detecting BBTV in vitro. Symptomless, ELISA-negative plants arose in 10 out of 11 lines derived from BBTV-infected field plants and first appeared after 9 months continuous in vitro culture at a constant 28OC. Meristem tip culture or heat therapy was not used. These plants remained symptomless and ELISA-negative after planting out in the glasshouse (individual plants checked for up to 16 months). The implications of this inconsistent transmission of BBTV for germplasm indexing and exchange are discussed.

ACIAR Integrated crop production of bananas

Integrated crop production of bananas to manage wilt diseases for improved livelihoods in Indonesia and Australia.

Influence of soil organic amendments on suppression of the burrowing nematode, Radopholus similis, on the growth of bananas.

Radopholus similis is a major constraint to banana production in Australia and growers have relied on nematicides to manage production losses. The use of organic amendments is one method that may reduce the need for nematicides, but there is limited knowledge of the influence of organic amendments on endo-migratory nematodes, such as R. similis. Nine different amendments, namely, mill mud, mill ash, biosolids, municipal waste compost, banana residue, grass hay, legume hay, molasses and calcium silicate were applied to the three major soil types of the wet tropics region used for banana production. The nutrient content of the amendments was also determined. Banana plants were inoculated with R. similis and grown in the soil-amendment mix for 12-weeks in a glasshouse experiment. Assessments of plant growth, plant-parasitic nematodes and soil nematode community characteristics were made at the termination of the experiment. Significant suppression of plant-parasitic nematodes occurred in soils amended with legume hay, grass hay, banana residue and mill mud relative to untreated soil. These amendments were found to have the highest N and C content. The application of banana residue and mill mud significantly increased shoot dry weight at the termination of the experiment relative to untreated soil. Furthermore, the applications of banana residue, grass hay, mill mud and municipal waste compost increased the potential for suppression of plant-parasitic nematodes through antagonistic activity. The application of amendments that are high in C and N appeared to be able to induce suppression of plant-parasitic nematodes in bananas, by developing a more favourable environment for antagonistic organisms.

Nitrogen leaching from the root zone of sugarcane and bananas in the humid tropics of Australia

Loss of nitrogen in deep drainage from agriculture is an important issue for environmental and economic reasons, but limited field data is available for tropical crops. In this study, nitrogen (N) loads leaving the root zone of two major humid tropical crops in Australia, sugarcane and bananas, were measured. The two field sites, 57 km apart, had a similar soil type (a well drained Dermosol) and rainfall (∼2700 mm year -1) but contrasting crops and management. A sugarcane crop in a commercial field received 136-148 kg N ha -1 year -1 applied in one application each year and was monitored for 3 years (first to third ratoon crops). N treatments of 0-600 kg ha -1 year -1 were applied to a plant and following ratoon crop of bananas. N was applied as urea throughout the growing season in irrigation water through mini-sprinklers. Low-suction lysimeters were installed at a depth of 1 m under both crops to monitor loads of N in deep drainage. Drainage at 1 m depth in the sugarcane crops was 22-37% of rainfall. Under bananas, drainage in the row was 65% of rainfall plus irrigation for the plant crop, and 37% for the ratoon. Nitrogen leaching loads were low under sugarcane (<1-9 kg ha -1 year -1) possibly reflecting the N fertiliser applications being reasonably matched to crop requirements and at least 26 days between fertiliser application and deep drainage. Under bananas, there were large loads of N in deep drainage when N application rates were in excess of plant demand, even when applied fortnightly. The deep drainage loss of N attributable to N fertiliser, calculated by subtracting the loss from unfertilised plots, was 246 and 641 kg ha -1 over 2 crop cycles, which was equivalent to 37 and 63% of the fertiliser application for treatments receiving 710 and 1065 kg ha -1, respectively. Those rates of fertiliser application resulted in soil acidification to a depth of 0.6 m by as much as 0.6 of a unit at 0.1-0.2 m depth. The higher leaching losses from bananas indicated that they should be a priority for improved N management. Crown Copyright © 2012.

Ground cover management alters development of Fusarium wilt symptoms in Ducasse bananas

Many banana producing regions around the world experience climate variability as a result of seasonal rainfall and temperature conditions, which result in sub-optimal conditions for banana production. This can create periods of plant stress which impact on plant growth, development and yields. Furthermore, diseases such as Fusarium wilt caused by Fusarium oxysporum f. sp. cubense, can become more predominant following periods of environmental stress, particularly for many culturally significant cultivars such as Ducasse (synonym Pisang Awak) (Musa ABB). The aim of this experiment was to determine if expression of symptoms of Fusarium wilt of bananas in a susceptible cultivar could be explained by environmental conditions, and if soil management could reduce the impact of the disease and increase production. An experiment was established in an abandoned commercial field of Ducasse bananas with a high incidence of Fusarium wilt. Vegetated ground cover was maintained around the base of banana plants and compared with plants grown in bare soil for changes in growth, production and disease symptoms. Expression of Fusarium wilt was found to be a function of water stress potential and the heat unit requirement for bananas. The inclusion of vegetative ground cover around the base of the banana plants significantly reduced the severity and incidence of Fusarium wilt by 20 % and altered the periods of symptom development. The growth of bananas and development of the bunch followed the accumulated heat units, with a greater number of bunched plants evident during warmer periods of the year. The weight of bunches harvested in a second crop cycle was increased when banana plants were grown in areas with vegetative ground cover, with fewer losses of plants due to Fusarium wilt.

Growth, yield and Fusarium wilt resistance of six FHIA tetraploid bananas (Musa spp.) grown in the Australian subtropics

Six tetraploid hybrids from Fundación Hondureña de Investigación Agrícola (FHIA) were evaluated in Australia over a five year period. They included three AAAA hybrids (FHIA-02, FHIA-17 and FHIA-23) and three AAAB hybrids (FHIA-01, FHIA-18 and SH-3640.10) and they were compared with industry standards, ‘Williams’ (AAA, Cavendish subgroup) and ‘Lady Finger’ (AAB, Pome subgroup). They were screened for their resistance to Fusarium wilt race 1 and subtropical race 4 caused by the pathogen Fusarium oxysporum f.sp. cubense and they were also grown for several cycles on farms not infested with Fusarium wilt to record their agronomic characteristics. The AAAB hybrids, all derived from female parent ‘Prata Anã’ (AAB, Pome subgroup) were the most resistant to both races of Fusarium wilt and were very productive in the subtropics. They were significantly more productive than ‘Lady Finger’, which was susceptible to both races of Fusarium wilt. The AAAA hybrids, with the exception of FHIA-02 which was very susceptible to Fusarium wilt and displayed the poorest agronomic traits of the six hybrids, produced bunch weights as good as Cavendish but were significantly slower to cycle. FHIA-17 and FHIA-23, both derived from the female parent ‘Highgate’ (AAA, Gros Michel subgroup), were also significantly more resistant to Fusarium wilt than ‘Gros Michel’, while FHIA-17 demonstrated a level of resistance similar to ‘Williams’ and FHIA-23 was intermediate between ‘Lady Finger’ and ‘Williams’

Evaluation of chlorothalonil and paraffinic oil alone and in combinations with registered fungicides for the control of yellow Sigatoka (Mycosphaerella musicola) of bananas in Northern Queensland, Australia

The efficacy of chlorothalonil and paraffinic oil alone and in combinations with the registered fungicides propiconazole, tebuconazole, difenoconazole, epoxiconazole and pyrimethanil was evaluated in a field experiment over two cropping cycles in 2013 and 2014 in Northern Queensland, Australia, for control of yellow Sigatoka (caused by Mycosphaerella musicola) of banana. The predominantly applied by the banana industry treatment mancozeb with paraffinic oil was included for comparison. The results from the two cropping cycles suggested that all chemicals used with paraffinic oil were as effective or more effective than when applied with chlorothalonil, and chlorothalonil alone. Difenoconazole and epoxiconazole with paraffinic oil followed by propiconazole with paraffinic oil were the most effective treatments. Pyrimethanil and tebuconazole plus chlorothalonil were the least effective treatments. None of the chemical treatments was phytotoxic or reduced yield.

Bananas adrift in time – a case study in the Solomons

Diseases, pests and environmental constraints pose a major threat to the sustainability of banana production globally. To address these challenges, the discovery and study of new sources of genetic resistance and adaptability are required, along with the general conservation of diversity. The Solomon Islands, located in the south-western Pacific region near Papua New Guinea, are a major center of banana diversity. Some collections had been made by nationals of some of the diversity present but little was known internationally of the rich genetic resource present. Two separate visits to the Solomon Islands characterized banana collections, documented and collected germplasm, recommended conservation strategies and provided training in cultivar characterization. A remarkable range of genetic diversity was found, including: many AA and AAA cooking types somewhat like those present in Papua New Guinea; nine Fei cultivars in relatively common usage, and two undescribed wild species as well as five AAB Pacific Plantains and four ABB cooking bananas belonging to the Kalapua subgroup. About six of the unique cultivars were successfully collected and established in the regional in vitro germplasm collection of SPC in Suva, Fiji. Nine Solomon Islanders were trained in the finer points of characterizing banana cultivars. Further collecting and study/evaluation of this rich diversity will promote its appreciation and potential utilization for meeting the challenges and opportunities ahead. Future studies could also determine the spread of the Awawe species and variability of morphological traits in the population. Community-based conservation could promote awareness of dietary diversity for better nutrition, via using the Fei bananas described in this paper. Establishing a virus-free regional field collection could help in comprehensively characterizing and evaluating regional Musa genetic resources. Existing sites could embrace the broader unique diversity of the Solomon Islands, and facilitate sharing this diversity in conjunction with a regional virus-tested in vitro collection.

In vitro Induction of Banana Autotetraploids by Colchicine Treatment of Micropropagated Diploids

Alternative breeding strategies, based on colchicine-induced autotetraploids, have been proposed as a means of introducing disease resistance into banana breeding programs. This paper describes techniques for the in vitro induction of banana autotetraploids by the use of colchicine on cultured explants. The technique can be readily applied and large numbers of autotetraploids produced. The optimum treatment involved immersing shoot tips in a 0.5% w/v colchicine solution for 2 h under aseptic conditions. Dimethyl sulfoxide (DMSO) was applied with the colchicine treatments to increase cell permeability and so absorption of colchicine, resulting in the optimum treatment unchanged at 0.5% colchicine, but including the addition of 2% v/v DMSO. Of the shoot tips treated over 30% were induced to the autotetraploid level. Methods for in vitro selection of induced tetraploids from treated diploid plantlets were also developed. Tetraploid plants were more robust with thicker pseudostems, roots and broader leaves than diploids and they could be selected on these morphological characteristics. Mean stornatal lengths of diploid banana plants growing in vitro were significantly smaller (16.0 pm) than the tetraploids (26.9pm) and were used as a more reliable indicator of ploidy than morphological criteria alone. A root tip squash technique using carbol fuchsin was developed for positive confirmation of ploidy change by chromosome counts. Although chimerism and reversion to the diploid form occurred, it was not considered a problem because of the large number of autotetraploids induced. Stable autotetraploids were recovered and established in the field and were characterised by their large, drooping leaves and thick pseudostems. They have retained these characteristics for more than 3 years in the field.

Towards the development of a Cavendish banana resistant to race 4 of fusarium wilt: gamma irradiation of micropropagated Dwarf Parfitt (Musa spp., AAA group, Cavendish subgroup)

'Dwarf parfitt', an extra-dwarf Cavendish cultivar with resistance to subtropical race 4 fusarium oxysporum f. sp. cubense 9Foc), was gamma irradiated at a dose of 20 Gy and putative mutants were recovered with improved agronomic characteristics. Further screening of putative mutants for improved yield and fruit size, as well as a degree of resistence to fusarium wilt, led to the selection of a line (DPM25) with improved productivity when grown on soils infested with subtropical race 4 Foc. DPM25 was equal to the industry standard, 'Williams', in every agronomic trait measured and it consistently showed a lower incidence of fusarium wilt. Further improvement of field resistance to race 4 Foc is needed in DPM25 and further cycles of mutation induction and selction is an option discussed.

Identification and characterization of non-pathogenic Fusarium oxysporum capable of increasing and decreasing Fusarium wilt severity

Fusarium wilt of banana is a potentially devastating disease throughout the world. Options for control of the causal organism, Fusarium oxysporum f.sp. cubense (Foc) are limited. Suppressive soil sites have previously been identified where, despite the presence of Foc, Fusarium wilt does not develop. In order to understand some aspects of this disease suppression, endophytic Fusarium oxysporum isolates were obtained from banana roots. These isolates were genetically characterized and compared with an isolate of Fusarium oxysporum previously identified as being capable of suppressing Fusarium wilt of banana in glasshouse trials. Three additional isolates were selected for glasshouse trials to assess suppression of Fusarium wilt in two different cultivars of banana, Cavendish and Lady Finger. One isolate (BRIP 29089) was identified as a potential biocontrol organism, reducing the disease severity of Fusarium wilt in Lady Finger and Cavendish cultivars. Interestingly, one isolate (BRIP 45952) increased Fusarium wilt disease severity on Cavendish. The implications of an isolate of Fusarium oxysporum, non-pathogenic on banana, increasing disease severity and the potential role of non-pathogenic isolates of Fusarium oxysporum in disease complexes are discussed.

Development of key soil health indicators for the Australian banana industry

To improve the sustainability and environmental accountability of the banana industry there is a need to develop a set of soil health indicators that integrate physical, chemical and biological soil properties. These indicators would allow banana growers, extension and research workers to improve soil health management practices. To determine changes in soil properties due to the cultivation of bananas, a paired site survey was conducted comparing soil properties under conventional banana systems to less intensively managed vegetation systems, such as pastures and forest. Measurements were made on physical, chemical and biological soil properties at seven locations in tropical and sub-tropical banana producing areas. Soil nematode community composition was used as a bioindicator of the biological properties of the soil. Soils under conventional banana production tended to have a greater soil bulk density, with less soil organic carbon (C) (both total C and labile C), greater exchangeable cations, higher extractable P, greater numbers of plant-parasitic nematodes and less nematode diversity, relative to less intensively managed plant systems. The organic banana production systems at two locations had greater labile C, relative to conventional banana systems, but there was no significant change in nematode community composition. There were significant interactions between physical, chemical and nematode community measurements in the soil, particularly with soil C measurements, confirming the need for a holistic set of indicators to aid soil management. There was no single indicator of soil health for the Australian banana industry, but a set of soil health indicators, which would allow the measurement of soil improvements should include: bulk density, soil C, pH, EC, total N, extractable P, ECEC and soil nematode community structure.

The influence of planting density on the production of 'Goldfinger' (Musa spp., AAAB) in the subtropics

'Goldfinger', a tetraploid banana produced from the Fundación Hondureña de Investigación Agrícola (FHIA) breeding program, was released to the Australian industry in 1995. It was promoted as an apple-flavoured dessert banana with resistance to Fusarium wilt race 1 and subtropical race 4, as well as resistance to black and yellow Sigatoka (Mycosphaerella fijiensis and M. musicola, respectively). This study was initiated to provide agronomic information to the banana industry, which was under threat from Fusarium wilt, on a new cultivar which could replace 'Williams' (AAA, Cavendish subgroup) or 'Lady Finger' (AAB, Pome subgroup) in those areas affected by Fusarium wilt. Also few studies had reported on the production characteristics of the new tetraploid hybrids, especially from subtropical areas, and therefore two field sites, one a steep-land farm and the other a level, more productive site, were selected for planting density and spatial arrangement treatments. The optimum density in terms of commercial production, taking into account bunch weight, finger size, length of the production cycle, plant height and ease of management, was 1680 plants/ha on the steep-land site where plants were planted in single rows with 2.5 m × 2.5 m spacings. However on the level site a double-row triangular layout with inter-row distances of 4.5 m to allow vehicular access (1724 plants/ha) gave the best results. With this arrangement plants were in an alternate, triangular arrangement along a row and a spacing of 1.5 m between plants at the points of each triangle and between each block of triangles.