Griffith University

The project inception meeting was held in Jakarta on 14 and 15 April 2010, with 5 delegates from Australia and 12 from Indonesia. The meeting started with a discussion of the lessons learnt from the previous ACIAR project on fruit flies in Indonesia. The prospective treatment areas containing mangoes in Indramayu (West Java) were discussed with 40 ha in Lobener (Jatibarang) and 60 ha in neighbouring Sliyeg considered suitable, together with a 20-30 ha control (untreated) area in Cikedung about 25 km away. The monitoring activities for 2010 were then negotiated. Funding limitations in 2010 restricted activities to host mapping, limited male trapping (starting July 2010) and host fruit collection (October 2010) to provide pre-treatment data. Treatment requirements and supplies were discussed including the approximate volumes of protein bait and numbers of male annihilation blocks required. The meeting also discussed the communication and extension requirements, particularly community and farmer engagement, and a risk management strategy. A work plan was developed for 2010 and 2011.
The pre-treatment trapping with both methyl eugenol and cuelure traps commenced in July 2010 in the areas designated above. Trap catches ranged from 20 flies/trap/day to >100 between July and October (the mango production period). The 1000 ripe and near ripe mango fruit sampled from the Cikedung area in October had an average infestation level of 4.5%. Maps of mango tree distribution in the treatment and control areas have been prepared and progressively refined over the past 10 months.
A pre-treatment coordination meeting with key staff was held in Indramayu in February 2011. Male annihilation treatments commenced in April 2011 when approximately 500 blocks containing methyl eugenol and fipronil were deployed through the Lobener/Krasak and Sliyeg areas. The blocks will be replaced every 2 months. An increased number of male traps have also been deployed through the designated areas, partly to counteract the influence of the blocks. All traps have GPS coordinates.
This season's protein bait shipment has been received and is in storage in Indramayu. Baiting will commence in June 2011 to coincide with the young fruit stage in mangoes and after some staff training in application techniques.
Recent changes in positions within the Indonesian Ministry of Agriculture have seen Ir. Soekirno replaced in the project by Ir. Soesilo, Director of Horticulture Protection, and Ir. Cahyaniati, Head of the Sub-Directorate of Fruit Protection and entomologist overseeing the project in Indonesia, replaced by Dr. Ir. Dwi Iswari.
In Australia, Gail Lowe was appointed as the project technician during 2010. The 2010/2011 mango season was poor through a number of production areas including north Queensland. This was due to a mild wet winter and its impact on flowering. This affected some of the project research in Australia.
No data were obtained on the kill efficacy of three different protein baits, but data were collected on fruit blemishing with one bait producing limited effect compared to the other two.
The first season's work on the relationship between fruit maturity and fruit fly infestation examined early season Kensington Pride (KP) types and later maturing Keitts and other varieties. There were no fruit fly control treatments in place. Fruit maturity was measured with a hand-held spectrophotometer to measure skin and flesh colour, a refractometer to estimate % Brix and through flesh sub sampling to calculate dry weight. Fruit were examined for fruit fly stings and suspect fruit were held to rear any flies through. No fruit fly stings or infestation could be ascertained in the KP type fruit. However, in the Keitts and later varieties there were fruit fly stings on and flies reared from both maturing and ripe fruit. Most were in ripe fruit with a reasonable correlation between the numbers stings and increasing Brix. The later varieties are subject to greater fruit fly pressure than the KP types.
Further data were collected on the efficacy of ammonium bicarbonate, torula yeast and fruit volatilises as female attractants. These were tested in both early and late mango varieties. During KP mango maturation, 96.7% of flies caught in traps with attractants were females and 98.4% of these were pest species. The results for the fruit volatilises were varied and confirmed that they perform best when they do not have to compete with the odours produced by a large amount of ripe fruit. Ways are currently being considered to extend attractant life in the field without impeding attractancy.

The protein bait commercialization project has been ongoing for just over 1 year, commencing with the inauguration of the Ento-pro production facility in Bac Giang province. The anticipated completion date of the project has been extended by six months in order to allow completion of a promising field demonstration and for extended media promotion of Ento-pro.
The majority of project activities have been completed, with the remaining activities proposed to be (i) field demonstration of Ento-pro in Hai Phong province (ii) training of distribution staff in three provinces and (iii) ongoing media promotion.
The commercialization of Ento-pro is proceeding relatively well, with production and sales of more than 14,000 kilograms (litres) of Ento-pro in 2007-2008, protecting an estimated more than 1000 hectares of fruit in the Northern and Central provinces from fruit fly infestation. This is delivering substantial economic, social and environmental benefits to smallholder fruit farmers.
The successful demonstrations, distribution staff training and media promotion have set a good base for the further expansion of Ento-pro production and use in fruit protection in the North and Centre of Vietnam.

Vietnam is one of many countries in South East Asia which experience serious preharvest fruit and vegetable losses to fruit flies ranging from 70 - 100%. As a result, fruit flies are seen as a major contributor to the ongoing problems of hunger, poor nutrition and poverty, especially in the rural communities, which comprise over 75% of Vietnam's population. The purpose of the project is to develop and introduce simple yet practical, in-field solutions to the fruit fly problem that will result in a direct and positive influence on household and food security for Vietnam.

The key collaborating institutions in Vietnam are the Ministry of Agriculture and Rural Development (MARD) through the National Institute of Plant Protection (NIPP) in Hanoi, the Southern Fruits Research Institute (SOFRI) at Long Dinh, the Provincial Plant Protection Departments (PPPD), Foster's Asia (Tien Giang Brewery) and AVENTIS Crop Science (Vietnam office). Collaboration within Australia is with Foster's Brewery, Melbourne, AVENTIS Crop Sciences and the Crawford Fund.

The various components of the project are progressing well and have yielded good results as follows:
Surveys of fruit flies in Vietnam are being conducted through trapping and host fruit collecting. These surveys have revealed nine species of fruit flies as being of major economic importance, along with another 20 non-pest species in Vietnam.
Studies to define the pest species of fruit flies, their geographic and host ranges, and levels of damage to major food crops have shown that crops such as gourds and water apple experience losses in excess of 95% in unprotected situations.
The construction of a protein manufacturing plant at Foster's Brewery, Tien Giang, although initially delayed, has now been completed and the necessary equipment (yeast waste evaporator, digestor and drum dryer) has been installed. Production of protein from the brewery yeast waste is expected to commence in the final quarter of 2002.
As an interim measure, Vietnamese collaborators were taught the procedures and have begun processing Foster's Brewery waste into protein bait in the laboratory at SOFRI. This laboratory prepared bait is being used for the scheduled laboratory and field tests to determine the correct formulations and application rates for the new protein product for efficient use in fruit fly control programs.

The first of three training workshops planned for the project was conducted in SOFRI from 3 - 7 June 2002. The purpose of the workshop was to train a core group of Vietnamese trainers who will then conduct further training for other Vietnamese staff, especially Plant Protection Sub-Department staff. A comprehensive training manual covering morphology, taxonomy and management of fruit flies in Vietnam was prepared and distributed to all participants. A total of 27 participants from 12 different agencies in Vietnam attended the training workshop. A major outcome of the workshop was the design of a brochure in the Vietnamese language by workshop participants for local farmers on fruit fly identification, biology and management. This brochure is to be produced by NIPP and SOFRI and widely distributed to Vietnamese farmers. The circulation of this farmer brochure will be made in conjunction with the release of the new protein bait from Foster's Brewery at Tien Giang.

Extensive collections of adult fruit flies collected from male lure traps and reared from host fruits (cultivated and wild) over 23 provinces covering north, central and south Vietnam have revealed that there are nine species of fruit flies are of economic importance to horticultural production and export trade in Vietnam. These are Bactrocera dorsalis, B. carambolae, B. correcta, B. cucurbitae, B. diversa, B. latifrons, B. pyrifoliae, B. zonata and B. tau. The species causing the greatest damage in north Vietnam are B. dorsalis, B. pyrifoliae and B. cucurbitae, whereas in south Vietnam the species causing greatest damage are B. dorsalis, B. correcta and B. cucurbitae. Crop losses ranging from 40 to 100% are being recorded in a wide range of fruits and vegetables when no control measures are applied.

To assist with field control studies, laboratory colonies (rearing on artificial diet) of B. cucurbitae have been established at the National Institute of Plant Protection (NIPP). Colonies of two other major pest species-B. dorsalis reared from litchi and B. pyrifoliae reared from peach are also being set up at NIPP. At the Southern Fruits Research Institute (SOFRI) colonies of pest species B. correcta and B. dorsalis have been successfully established. These laboratory colonies currently provide adult flies for laboratory and field testing of the new protein bait produced at Foster's Tien Giang brewery.

The production plant at Foster's Brewery at Tien Giang to process brewery waste into a fruit fly bait has been fully commissioned. Batches of protein bait produced at the plant are currently being evaluated by NIPP for field control of fruit flies infesting peach and bitter luffa, and by SOFRI on water apple and guava.

An extensive training program for on the biology and control of fruit flies for Provincial Plant Protection Department (PPPD) staff, as well as for farmers has been successfully implemented in various provinces around Vietnam. Project staff from Brisbane initially ran a week-long workshop in June 2002 to train a core group of trainers from NIPP, SOFRI, PPD and selected universities in Vietnam. These staff members from NIPP and SOFRI have subsequently completed training a total of 177 PPPD staff as well as 1600 farmers from 16 provinces across Vietnam. The training was accompanied by the distribution of over 3000 illustrated brochures in the Vietnamese language on the biology and management of fruit flies, incorporating the new bait spray technology.

Continued collections of adult fruit flies collected from male lure traps and reared from host fruits (cultivated and wild) over various provinces covering north, central and south Vietnam have not revealed any new pest species beyond the nine that have so far in the project been identified as being of economic importance to horticultural production and export trade in Vietnam. These are Bactrocera dorsalis, B. carambolae, B. correcta, B. cucurbitae, B. diversa, B. latifrons, B. pyrifoliae, B. zonata and B. tau. In the current sampling period, however, B. correcta has also been recorded in North Vietnam where it was not recorded before. Crop losses ranging from 40 - 100% are still being recorded in a wide range of fruits and vegetables when no control measures are applied.

Laboratory colonies (rearing on artificial diet) of B. cucurbitae at the National Institute of Plant Protection (NIPP) and B. correcta and B. dorsalis at the Southern Fruits Research Institute (SOFRI) continue to be maintained. These laboratory colonies currently provide adult flies for laboratory and field testing of the new protein bait produced at Foster's Tien Giang brewery.

The protein bait production plant at Foster's Tien Giang has been fully commissioned and was officially launched together with the bait called SOFRI PROTEIN on 16 April 2004 by the Australian Ambassador to Vietnam, the Hon Joe Thwaites. The high profile ceremony was attended by senior officials from the Tien Giang provincial government, Ministry of Agriculture and Rural Development (MARD) Vietnam, Fosters Brewing International, Griffith University, Queensland Government, AusAID and ACIAR.

The one-day training program for farmers on the biology and control of fruit flies for Provincial Plant Protection Department (PPPD) staff, as well as for farmers has been successfully continued. In South Vietnam, training in this reporting period has focussed on Barbados Cherry farmers with whom an large area fruit fly control program is being organised. Over 4,000 copies of farmer extension leaflets have been printed and distributed by NIPP and SOFRI.

To ensure more effective control of fruit flies, two large-area fruit fly control programs have been implemented, one on peach grown by minority hill tribe people in Moc Chau, North Vietnam, and the second in Barbados cherry grown largely in Tien Giang province in South Vietnam. These trials combine the use of male lures with SOFRI PROTEIN bait sprays for more effective fruit fly population suppression. Farmers are organised in large groups of 25 or more and apply these treatments in a coordinated manner. Initial results show excellent suppression of pest fruit fly populations and very low levels of fruit damage to the benefit of a large number of farmers in an area.

SUMMARY FROM FINAL REPORT:
Extensive collections of adult fruit flies obtained from male lure traps and reared from host fruits (cultivated and wild) over 23 provinces covering north, central and south Vietnam have revealed that there are eight species of fruit flies are of economic importance to horticultural production and export trade in Vietnam. These are Bactrocera dorsalis, B. carambolae, B. correcta, B. cucurbitae, B. latifrons, B. pyrifoliae, B. zonata and B. tau. The species causing the greatest damage in north Vietnam are B. dorsalis, B. pyrifoliae and B. cucurbitae, whereas in south Vietnam the species causing greatest damage are B. dorsalis, B. correcta and B. cucurbitae. Crop losses ranging from 40 - 100% are being recorded in a wide range of fruits and vegetables when no control measures are applied.

To assist with field control studies and other ACIAR funded projects to improve postharvest fruit quality, laboratory colonies (rearing on artifical diet) of B. cucurbitae B. dorsalis and B. pyrifoliae were established at the National Institute of Plant Protection (NIPP). At the Southern Fruits Research Institute (SOFRI) colonies of pest species B. correcta and B. dorsalis were successfully established. These laboratory colonies provide adult flies for laboratory and field attractancy testing for on-going quality control of the new protein bait produced at Foster's Tien Giang brewery.

The protein bait production plant at Foster's Tien Giang was officially launched on 16 April 2004 by the Australian Ambassador to Vietnam, the Hon Joe Thwaites. The ceremony was attended by senior officials from the Tien Giang provincial government, Ministry of Agriculture and Rural Development (MARD) Vietnam, Fosters Brewing International, Griffith University, Queensland Government, AusAID and ACIAR, as well as over 200 local farmers from the surrounding provinces. The protein product has been officially registered in Vietnam under the trade name SOFRI Protein 10DDD (yeast protein + fipronil insecticide) and is being made available for sale to farmers through a local pesticide distribution company that has numerous outlets in the Mekong delta. The price has been kept very low and is controlled by a steering committee comprising Griffith University, ACIAR Vietnam, SOFRI, Foster's Tien Giang and Cantho Pesticide Company. The protein production plant at Foster's Tien Giang has an annual production capacity of about 50,000 litres of protein.

The use of SOFRI Protein 10DD both in small farms and over large areas has provided excellent control of pest fruit flies across a range of fruit and fruiting vegetable crops in Vietnam, resulting in major increase in incomes for farmers. For example, H'Mong minority hill tribe people in the northern province of Son La, Moc Chau district cultivate peach as their main crop but have always had to harvest the fruits hard green because ripe fruits are 100 per cent infested by fruit flies. Using the spot spray protein bait technology, fruit fly damage was reduced to less than 5 per cent, resulting in farmers harvesting ripe fruits from which they obtained higher yields and better prices. Overall this resulted in a 4-fold increase in incomes in one season for the peach farmers. Barbados cherry farmers in Go Cong province in the Mekong Delta have also experienced a similar benefit with a 2-fold increase in incomes resulting from higher yields in orchards using SOFRI Protein 10DD. Control trials in other crops like guava, jujube, luffa and bitter gourd have reduced fruit fly damage from over 70 per cent to less than 5 per cent. The protein bait spot spray technique also provides major health and environmental benefits by utilizing extremely low amounts of pesticide and spray volumes in comparison with cover sprays of insecticides. The Ministry of Agriculture and Rural Development now plans to set up more protein production plants in other breweries across the country and extend the benefits of the protein spot spray bait technology to the wider farming community in Vietnam.

An extensive training program on the biology and control of fruit flies for Provincial Plant Protection Department (PPPD) staff, as well as for farmers has been successfully implemented in various provinces around Vietnam. Project staff from Brisbane initially ran a week long workshop in June 2002 at SOFRI to train a core group of trainers from NIPP, SOFRI, PPD and selected Universities in Vietnam. These staff from NIPP and SOFRI have subsequently completed training a total of 290 PPPD and sub-PPPD staff as well as 4445 farmers from 16 provinces across Vietnam. The training was accompanied by the distribution of over 5000 illustrated brochures in the Vietnamese language on the biology and management of fruit flies, incorporating the new protein bait spray technology introduced by the project.

Year 5 report not sent due to a gap between final report and new variation.

Year 6 progress summary is for the period 1 May 2006 - 30 April 2007.
1. Pairs of male lure traps (methyl eugenol and cue-lure) were set up in provinces that were not surveyed before. These are Kien Giang, Thua Thien Hue, Ca Mau, Tra Vinh, An Giang and Binh Phouc in South Vietnam, and Son La, Thai Nguyen, Tuyen Quang and Ha Giang in north Vietnam. No new species of flies have been recovered so far and the species composition remains the same as for the provinces surveyed in the earlier project. Additional samples of host fruit have been collected to verify uncertain host records.

2. Trials on peach and plum integrating fruit fly control using spot sprays of protein baits with improved crop management techniques and post-harvest handing introduced through ACIAR project CP/2002/086 - Improving postharvest quality of temperate fruits in Vietnam and Australia were planned for various locations in Moc Chau province. However, these trails could not be implemented this year because of high variability in fruiting and fruit load within some of the experimental sites. It is thus proposed to conduct these trials during the next fruiting season in March - June 2008.

In the Mekong Delta, trials were planned to integrate fruit fly control using spot sprays of protein baits with improved control of pests other than fruit flies using white mineral oils introduced through ACIAR Project CP/2000/043: Huanglongbing management for Indonesia, Vietnam and Australia. One trial on Barbados cherry has been established to evaluate the effectiveness of integrating SOFRI Protein and methyl eugenol trapping for fruit fly control, and the mineral oil SK Enspray 99 for other such as aphids, mites, scales and mealy bugs. The trial is in progress.

A new protein bait plant has been constructed at An Thinh Brewery in Hanoi, and the facility was officially launched on 9 May 2007.

3. The farmer training programme is continuing with another 2950 farmers having received training in fruitfly biology and field pest management in the provinces of Hue, Quang Nai, Quang Nam, Lam Dong, Contum, An Giang, Kien Giang, Ca Mau, Soc Trang and Bac Lieu.

The list of fruit fly species occurring in Vietnam has been completed and this will be of great value to Vietnam in meeting with the SPS requirements, and to quarantine authorities and in trade negotiations.

A major development in the project has been the construction of a protein bait plant at An Thinh Brewery near Hanoi that has a capacity to produce 100,000 litres of bait a year. This plant was constructed by the Vietnamese using their own funds. The bait is being marketed under the trade name Ento Pro 10DD and is available to farmers in north Vietnam.

Farmer training continues to extend the protein bait spot spray technology to a number of new crops like guava, sapota, dragon fruit, mango and pomelo, all of which are in the national priority list for export development.

The tsunami in December 2004 that devastated Aceh and other provinces in Sumatra resulted in a delay in project implementation as planned because the resources of both collaborating agencies in Indonesia were directed towards relief and reconstruction in the tsunami affected areas for the next few months. Nevertheless, two senior Indonesian project staff from the Department of Horticulture protection (DHP) and the Agency for Agricultural Quarantine (AAQ) were brought to Brisbane from 31 January to 7 February 7 2006 to plan the fruit fly survey work in Indonesia and also to undergo training with Griffith University staff on trapping and surveillance procedures. On their return to Indonesia, the project staff then conducted a training workshop from 26 - 30 June 2005 in Yogyakarta, on fruit fly surveillance techniques, identification to species, field pest management and techniques for training farmers. The training course was attended by 47 participants representing 18 provinces in Sumatera, Java and Kalimantan. Following the training workshop, field staff from DHP and AAQ established a trapping network in the provinces of Banten, West Java, DKI Jakarta, Central Java, D.I. Yogyakarta, East Java, West Kalimantan, Central Kalimantan, East Kalimantan and South Kalimantan. Major fruits and vegetables of economic importance such as mango, citrus, guava, mangosteen, chilli, tomato, gourds, and many others have also been collected to determine the infesting fruit fly species. Trapped and host reared specimens have been sent to Griffith University where they are being identified, pinned and labelled. A full set of accurately identified specimens are to be returned to Indonesia. The first year of implementation of the project has greatly strengthened the capacity of Indonesian quarantine to meet WTO-SPS requirements in relation to fruit flies.

All the samples of fruit flies collected in Java and Kalimantan during Year 1 of the fruit fly survey and that were sent to ICMPFF at Griffith University have been identified and a list of fruit flies occurring in Java and Kalimantan has been compiled. This list together with two sets of authoritatively identified fruit fly specimens have been returned to DHP and AAQ in Indonesia respectively for their reference collections.

A training workshop on "Fruit Flies of Indonesia: Their Identification and Pest Status" was held in Darwin at the AQIS Training Facility from 27 - 31 March 2006. The aim of the workshop was to provide training on the identification and pest status of flies collected during Year 1 of the survey in Kalimantan and Java. The workshop was attended by 17 Indonesian field staff who had participated in the Year 1 fruit fly survey in Kalimantan and Java. The training workshop was also of benefit to another 5 Australians, 4 of whom were from the Northern Australian Quarantine Strategy (NAQS), AQIS and 1 from the Northern Territory Department of Primary Industries, Fisheries and Mines.

Project staff in Indonesia organised the second training workshop on fruit fly surveillance techniques for project field staff from who were to be involved in the Year 2 fruit fly survey in the islands of Bali, Sulawesi and Sumatera. The training workshop was conducted in Bali from 9 - 14 July 2006 and was attended by 32 project field staff from Bali, Sulawesi and Sumatera. Following this training workshop, the field staff have established a fruit fly trapping network using methyl eugenol and cue-lure traps in 17 provinces in Bali, Sulawesi and Sumatera.

Crop damage assessments have been initiated in West, Central and East Java as well as in East Kalimantan. A controlled environment fruit fly rearing facility has been established at the Pest Forecasting Centre in Jatisari, West Java with flies reared from mango and citrus.

All samples of fruit flies collected in 16 provinces from the islands of Bali, Sulawesi and Sumatera during Year 2 of the fruit fly survey, and that were sent to ICMPFF at Griffith University have been identified and a list of fruit flies occurring in these regions has been compiled.

Project staff in Indonesia organised the third and final training workshop on fruit fly surveillance techniques for staff involved in Year 3 of the fruit fly survey. The workshop was conducted from 11 - 16 May, 2007 in Mataram, West Nusa Tenggara and was attended by 13 staff from the Food Crops and Horticulture Protection Centre. Following this training workshop, the field staff have established a fruit fly trapping network using methyl eugenol and cue-lure traps in 6 provinces, i.e. West Nusa Tenggara, East Nusa Tenggara, North Maulku, Maluku, Papua and West Irian Jaya. A separate training workshop on Indentification of Fruit Flies in Indonesia, funded by the Indonesian Ministry of Agriculture, was also conducted at Jatisari, Java, for 10 technical staff from the Food Crops and Horticulture Protection Centre.

In conjunction with the ACIAR project, a workshop on Fruit Fly Pest Free Areas was conducted in Jakarta from 18-22 June 2007. Invited speakers from Griffith University, Biosecurity Australia, NSW and Victoria Departments of Primary Industries and AQIS, provided in-depth presentations on fruit fly area freedom as practised in Australia. The workshop was attended by 44 senior scientists and quarantine staff from various agencies in Indonesia.

Crop damage assessments have also been initiated in West Nusa Tenggara, East Nusa Tenggara, North Maulku, Maluku, Papua and West Irian Jaya.. The controlled environment fruit fly rearing facility established earlier at the Pest Forecasting Centre in Jatisari is now able to supply laboratory reared flies for attractancy testing of protein baits.

Fabrication and installation of the protein bait plant at PT MultiBintang Brewery in Tangerang, Java began in July 2007 and is expected to be completed by early 2008.

All samples of fruit flies collected during Year 3 of the fruit fly trapping survey conducted in the provinces of Nusa Tenggara, Maluku, Papua and West Irian Jaya and sent to Griffith University have been identified, and a list of species occurring in these provinces has been compiled. Fruit fly damage assessments on fruits of economic importance is still in progress in these provinces.

The second training workshop on "Fruit Flies of Indonesia: Their Identification, Pest Status and Pest Management" was held in Darwin from,10 - 14 March 2008. The workshop provided training to 21 staff members from the Department of Horticulture Protection and the Agency for Agricultural Quarantine in Indonesia, particularly on the identification of fruit flies collected in the nationwide survey of fruit flies. With the aid of an illustrated key that was prepared and provided, trained staff now has the independent capacity to carry out accurate identifications of fruit flies in Indonesia. In addition to the Indonesian staff, the training workshop was also of great benefit to 3 participants from the Northern Australia Quarantine Strategy, Darwin, Broome and Mareeba, and 3 participants from the Northern Territory Department of Primary Industry and Mines, all of whom were able to obtain the most up to date knowledge on fruit flies of quarantine importance to Australia.

In conjunction with the ACIAR project, a workshop on Fruit Fly Pest Free Areas was conducted in Jakarta from 18-22 June 2007. Invited speakers from Griffith University, Biosecurity Australia, NSW and Victoria Departments of Primary Industries and AQIS, provided in-depth presentations on fruit fly area freedom as practised in Australia. The workshop was attended by 44 senior scientists and quarantine staff from various agencies in Indonesia.

Construction of the protein bait plant at PT MultiBintang Brewery in Tangerang, Java was completed in February 2008 and the plant was commissioned in March 2008. Once registration has been obtained, production of the protein bait will begin in Indonesia.

A list of fruit fly species occurring in West Nusa Tenggara, East Nusa Tenggara, North Maluku, Maluku, Papua and West Papua has been completed after extensive sorting and study of thousands of fruit fly specimens sent to ICMPFF at Griffith University during the third year of the project. This survey exercise now provides a complete picture of the fruit flies of the Indonesian archipelago. The 7 species of economic importance are Bactrocera albistrigata, B. carambolae, B. cucurbitae, B. latifrons, B. occipitalis (only in Kalimantan), B. papayae, and B. umbrosa.

The third and final training workshop on "Fruit Flies of Indonesia: Their Identification, Pest Status and Pest Management" was held in Brisbane from,16 - 20 February 2009. The workshop provided training to another 14 staff members from the Directorate of Horticulture Protection and the Agency for Agricultural Quarantine in Indonesia, particularly on the identification of fruit fly collected in the nationwide survey of fruit flies. With the aid of an illustrated key that was prepared and provided, Indonesian project staffs now have the independent capacity to carry out accurate identifications of fruit flies in Indonesia.

The protein bait plant at PT Multi Bintang Brewery in Tangerang, West Java which was commissioned in March 2008 is now producing fruit fly bait for the field control trials in Indonesia. The bait has been registered under the trade name Indo Prima and is selling at a price of 40,000 Rupiah per litre. Imported protein baits cost more than 3 times this amount, making Indo Prima very affordable to Indonesian farmers.

The project has been operational since April 2008. The initial phase of the project has been successfully completed with demonstration trials established at three Rural Training Centres (RTCs): Tabaka RTC, Western Province; Airahu RTC, Malaita Province and; St Dominic's RTC, Western Province. These initial demonstration plots will be further expanded in the first half of 2009.
Work has begun on developing curricular materials for use by the RTCs at the curriculum development unit of the Solomon Islands College of Higher Education (SICHE). This work is being partially funded through the European Union and is a close collaboration with a similar project introducing smallholder timber plantings into the State School system.
Whilst the plan to establish fully-replicated scientific trials at the Solomon Islands College of Higher Education land on Guadalcanal has not gone ahead, an alternative site has been identified and SICHE students and the students from St Josephs School at Tenaru, east Honiara, are working to prepare the site which will be planted in June 2009. Further scientific trials have been established at Poitete and Ringgi Cove on Kolombangara Island in Western Province. The trial at Poitete will be managed by the trainees of the SICHE Forestry Training Institute when the programme re-commences following refurbishment in late 2009. The trial at Ringgi will be under the management of Kolombangara Forest Products Ltd and will receive normal, operational maintenance.
Thinning trials have also been established on community teak plantations in Western Province and Guadalcanal with further trials to be established on Malaita and Kolombangara. These trials have included students from SICHE, trainee extension officers from the Ministry of Forestry and local landholders as part of a larger effort to introduce thinning into the established community plantings throughout the Solomon Islands.
The timber testing programme which will provide information on the timber properties of different age classes of Solomon Islands provenance teak and compare this with Australian-grown teak from north Queensland has commenced with the delivery of the N. Qld teak to the DEEDI (formerly DPI) timber testing facility at Salisbury in Brisbane.
An addition to the original proposal has seen the involvement of the Hopevale Aboriginal Community in Queensland with the identification and clearing of a mixed species trial area which will feature Teak and African Mahogany grown as the high value species with Eucalyptus pellita (red mahogany) grown as the inter planted species. The project is working in conjunction with Mareeba Campus of the Australian Agricultural College to provide training opportunities in forestry and silviculture related activities suitable for the trainees to find employment in the local plantation forestry industry.

A fertile PRSV-P resistant plant was produced in a previous project when a C. papaya x C. quercifolia hybrid was backcrossed to C. papaya clone 2.001. This plant is numbered clone 54 for ease of identification. In the previous 12 months clone 54 has been used to backcross to C. papaya clone 2.001 (2nd backcross) and to C. papaya lines 16 (a SE Qld variety), Solo (developed in Hawaii and used extensively in the tropics) and GD 319-2 (the parent of a hybrid seed line from North Queensland). Clone 54 and seed of these crosses has been sent to the Philippines. Problems have been experienced with seed germination of these crosses in both countries (particularly the 2nd backcross to 2.001), however seedlings populations of each cross have been established in a glasshouse at Griffith University.

Sub-Project 2. Selection of backcross generations for PRSV-P resistance and good agronomic traits.
Two hundred plants of a backcross generation between clone 54 and papaya line 16 have been grown in a glasshouse at Griffith University. These plants have been manually inoculated 5 times with PRSV-P over the 12-month period. Twenty-six plants have remained free of symptoms after 12 months, and no virus has been detected in them by ELISA. Virus infected all control plants after 2 inoculations. They will be planted in the field in 2003 where they will be subjected to aphid transmission of the virus. Other populations representing backcrosses between 54 and other C. papaya lines will be inoculated with PRSV-P in 2003.

Sub-Project 3. Backcross fertile resistant C. papaya x C. quercifolia F1 hybrids to C. papaya.

Australia
Of the large number of F1 hybrids between C. papaya x C. quercifolia, generated in a previous ACIAR project (CS1/1991/007), 4 have been identified as being PRSV-P resistant and having some pollen fertility. These have been given the clone numbers 410, 468 [parent of 54], 469 and 507. Pollen from these hybrids has been used to pollinate female flowers of C. papaya clone 2.001 and other C. papaya varieties in SE Qld. Embryo rescue will be attempted on seeds from resultants fruit between Jan and April in 2003.

Philippines
Seven inbred lines developed by the Institute of Plant Breeding and 5 F1 interspecific hybrids (401, 410, 468, 469 and 507) from Australia were planted in the field last Oct. 18, 2002. Table 1 shows the fruit characteristics and reaction to PRSV of the different inbred lines. Flowering has commenced and backcrossing activities have been started. Tissue culture activities to mass propagate the F1 interspecific hybrids through nodal cuttings is also in progress. These materials will be used to set-up duplicate hybridisation blocks in the islands of Davao in Mindanao, and Guimaras Island in the Visayas.

Table 1. Reaction to papaya ringspot virus (PRSV) and fruit characteristics of selected inbred lines developed at the Institute of Plant Breeding used in the experiment.

Inbred Line Reaction to PRSV Fruit flesh color Height to flower (cm) TSS (B)
Py 2 (4108) moderately tolerant red orange 67.30 10.8
Py 3 (4172) moderately tolerant red orange 121. 35 11.6
Py 4 (4308) severe red orange 74.54 11.1
Py 5 (5648) moderately tolerant yellow orange 71.46 11.8
Py 6 (5893) moderately tolerant red orange 73.70 11.0
Py 7 (3878) severe yellow orange 67.54 11.4
Py 10 (F2- derived) moderately tolerant red orange 105.80 11.7

Sub-Project 4. Development of molecular markers for PRSV-P resistance.
We decided to work initially on the C. pubescens x C. parviflora population as we have produced a large F2 population (300 plants in the glasshouse) that is segregating for resistance to PRSV-P. DNA has been extracted from F1 and F2 plants, and from plants of C. pubescens and C. parviflora, and is being compared using RAF markers. RAF is a new technique that produces more polymorphisms than AFLPs. A genetic map of the two species is being developed. Multiple primers will be screened across two F2 populations (one resistant and one susceptible to PRSV-P) using the RAF marker technique during the first 6 months of 2003. We hope to develop a marker for PRSV-P resistance using this approach. If a marker linked to PRSV-P resistance is developed, it will be used to screen the C. papaya x C. quercifolia and backcross populations.

Sub-Project 5. Micropropagation of elite papaya genotypes.
Clones of all C. papaya genotypes, fertile resistant C. papaya x C. quercifolia hybrids, and some backcross plants (including 54) are being maintained in vitro. Clones have been transferred to the Philippines and are being multiplied in the tissue culture laboratory at IPB.

Sub-Project 1. Incorporation of PRSV-P resistance genes into elite papaya genotypes, using fertile resistant backcross plants.

Year 2 (01/01/2003-31/12/2003)
Subproject 1: Incorporation of PRSV-P resistance genes into elite papaya genotypes, using fertile resistant backcross plants
Australia: A fertile PRSV-P resistant plant was produced in a previous project when a C. papaya x C. quercifolia hybrid was backcrossed to C. papaya clone 2.001. This plant is numbered clone 54 for ease of identification. Second generation backcross plants (BC2) from V. quercifolia have been produced when 54 was crossed with 4 papaya lines. These second-generation backcross plants are being screened for virus resistance both in Australia and the Philippines
Philippines: Clone 54 and seed of these crosses were sent to the Philippines. Both 54 and all the backcross plants were susceptible to PRSV-P in Los Banos however there was variation in symptom expression. The fertile PRSV-P resistant plant clone 54 succumbed to PRSV-P two months after transplanting in the field, with symptoms ranging from chlorotic leaves to oily streaks on the trunk.
Subproject 2: Selection of backcross generations for PRSV-P resistance and good agronomic trait.
Australia: Of the first 200 backcross (BC2) plants screened in Queensland (a backcross generation between clone 54 and papaya line 16), 26 remained symptomless after 5 manual inoculations of virus in a glasshouse. All control plants produced PRSV-P symptoms after 2 inoculations. These 26 plants were planted in the field in the presence of heavy PRSV-P infection. After 8 months, one male and one female plant remain symptomless. The other 26 vary in symptom expression from severe to mild symptoms of PRSV-P. More recently, more second-generation backcross plants have been planted in the glasshouse at GU and inoculated with PRSV-P. They represent 120 plants of 54 backcrossed to Solo, 60 of 54 backcrossed to GD319-2 (from North Queensland) and 70 plants of 54 backcrossed to papaya clone 2.001. It is too early for symptom development at this stage. To determine agronomic traits of backcross plants, 1 row (34 plants) of 54 crossed to 2.001 and 1 row of 54 crossed to Solo have been planted in the field in Southeast Queensland.
Philippines: Three BC2 populations (2.001 x 54, Sunrise Solo x 54, and GD 319-2 x 54) were planted in Mainit experimental station to evaluate the reaction of the segregating populations against PRSV-P. Symptom expression was observed two weeks after artificial inoculation. Symptom expression of ringspot disease varies in terms of onset of disease and severity of infection in each plant. Another batch of the same BC2 generations were evaluated in the glasshouse. One hundred ninety five BC2 generation between clone 54 and papaya line 2.001, 36 BC2 generation between line 54 and line GD 319-2 and 26 BC2 generation between line 54 and Sunrise Solo papaya were grown in the glasshouse at IPB. The plants were manually inoculated with PRSV-P. From the first inoculation, 74 of BC2 2.001 x 54 plants, 10 of BC2 GD 319-2 x 54 plants and two of Sunrise Solo x 54 plants remained free of symptoms and no virus has been detected by ELISA . Virus infected all control and susceptible plants two weeks after inoculation.
Subproject 3: Backcross fertile resistant C. papaya x C. quercifolia F1 hybrids to C. papaya
Australia: Of the large number of F1 hybrids between C. papaya x C. quercifolia, generated in a previous ACIAR project (CS1/1991/007), 5 were identified as being PRSV-P resistant and having some pollen fertility. These have been given the clone numbers 401,410, 468 [parent of 54], 469 and 507. Pollen from these hybrids has been used to pollinate female flowers of C. papaya clone 2.001 and other C. papaya varieties in Southeast Queensland. Seeds from 1077 fruit (from 1302 crosses) of C. papaya clone 2.001 have been dissected. From these 1077 fruit, 35,626 seed have been dissected and 185 embryos cultured in vitro. Of these, 131 have produced plants and they will be tested for resistance to PRSV-P next year.
Philippines: Four susceptible IPB inbred lines (4108, 4172, 5648 and 5893) as females and resistant F1 interspecific hybrid lines 410 and 401 as males were used in the backcrossing program to transfer PRSV resistance from Carica quercifolia to elite Philippines papaya genotypes. A total of 280 fruits were harvested from crosses generated using these inbred and F1 hybrid lines. Majority of the fruits came from 5648 (154) and 5893 (76) while the least number came from 4108 (30) and 4172 (20). Ninety to 120 day -old fruits were harvested for embryo rescue and culture. A total of 41, 553 seeds were dissected from the different crosses. Inbred 5648 produced the highest seed number of 20,395 but despite these number of seeds only 99 embryos were rescued. Ten well developed embryos were recovered from 10,877 seeds involving inbred line 5893. Twenty six embryos were rescued from 160 seeds involving inbred line 4108. On the other hand, inbred line 4172 produced only 15 seeds, however despite this number eight fully developed embryos were recovered. From the 143 embryos that were rescued from the different crosses, 90 embryos developed into plantlets and are now being maintained in the laboratory.
Subproject 4: Development of molecular markers for PRSV-P resistance
We decided to work initially on the C. pubescens x C. parviflora population as we have produced a large F2 population (300 plants in the glasshouse) that is segregating for resistance to PRSV-P. Good progress has been made on mapping of V. pubescens and V. parviflora using RAF markers. A genetic map of the two species has been developed. In recent research on a F2 population that is segregating for virus resistance, 5 molecular markers have been linked to PRSV-P resistance. Two of these, one each side of the gene are being sequenced.
Subproject 5: Micropropagation of elite papaya genotypes
Clones of all C. papaya genotypes, fertile resistant C. papaya x C. quercifolia hybrids, and some backcross plants (including 54) are being maintained in vitro. Plants obtained from the 131 backcross embryos (reported under objective 3) are being multiplied in Australia. In the Philippines, clones of fertile resistant C. papaya x C. quercifolia hybrids and backross plants which were embryo rescued are being maintained in vitro.

Sub-Project 1. Incorporation of PRSV-P resistance genes into elite papaya genotypes, using fertile resistant backcross plants.
Australia
All second backcross (BC2) plants produced from line 54 eventually succumbed to PRSV-P after 9 months in the field. However, there was large variation between plants in terms of degrees of severity to the virus symptoms and time to develop symptoms (2 to 9 months). Thus line 54 had a higher but variable level of tolerance to PRSV-P .

Philippines
Line 54 and BC2 plants produced from it have been susceptible to PRSV-P in the field. As in Australia, variation in rate and severity of symptom expression was observed.

Sub-Project 2. Selection of backcross generations for PRSV-P resistance and good agronomic traits.
A large number of BC2 plants between 54 and other papaya genotypes were tested for PRSV-P resistance. In Australia, 200 plants of 54 backcrossed to line 16, 120 plants of 54 backcrossed to Solo, 60 of 54 backcrossed to GD319-2 (from North Queensland) and 70 plants of 54 backcrossed to papaya clone 2.001. Similarly, a large number of progeny were tested in the Philippines. All were susceptible to PRSV-P.

To determine agronomic traits of backcross plants, 1 row (34 plants) of 54 crossed to 2.001 and 1 row of 54 crossed to Solo were planted in the field in SE Qld. Plants of 54 x Solo and 2.001 x 54 showed good agronomic traits. Plants tended to be more like C. papaya. High yields, good fruit shape and size and high brix levels were recorded ~14 (can be linked back to Vasconcella quercifolia, which has fruit with high sugar levels). Preliminary molecular marker analyses showed these plants had approx. 2% quercifolia DNA. This suggests that only a one or two more backcrosses would be needed to produce a commercial papaya.

Sub-Project 3. Backcross fertile resistant C. papaya x V. quercifolia F1 hybrids to C. papaya.

Australia
We have continued to backcross from 5 F1 hybrids that are PRSV-P resistant and having some pollen fertility, (401, 410, 468 [parent of 54], 469 and 507). 131 BC1 plants are currently being screened for PRSV-P in the field. The first 50 plants have developed viral symptoms, again with differing degrees of severity. The other 81 clones are being monitored.

Philippines
Six F1 intergeneric hybrid lines (401, 404, 410, 468, 469 and 507) of C. papaya x V. quercifolia from Australia were used as paternal parents to incorporate PRSV-P resistant gene/genes to five elite papaya inbred lines (4108, 4172, 5648, 5893 and 3878) of IPB. Intergeneric hybrid lines 410, 401, 404 and 468 had been extensively used in the backcrossing work. Pollen viability ranged from 1.2 to 2.2% under Philippine conditions. The ratio of male flower in cross-pollinating female flower was increased from 10:1 to 20:1 to compensate the low pollen viability of the hybrids. To date, fifteen backcrosses were completed with varying success in terms of viable embryos produced. Nine hundred forty (940) fruits were harvested in which 114,839 seeds were dissected under laboratory conditions. Ten backcrosses (4172 x 410, 4108 x 410, 5648 x 410, 5893 x 410, 5893 x 404 , 5648 x 404, 5648 x 468, 4108 x 468, 4172 x 404 and 3878 x 410) produced a total of 1011 embryos. Seven hundred thirty three (733) embryos had germinated into normal plantlets, each one was considered as a unique genotype. BC1 5648 x 410 comprised 46.66% of the genotypes with 342 embryos germinated, the other BC1 with embryos germinated were 4108 x 410 (132), 5893 x 410 (60), 4172 x 410 (60), 5648 x 404 (76), 3878 x 410 (44), 5893 x 404 (5), 4108 x 468 (6) and 4172 x 404 (8). Most of the genotypes were subjected to clonal micropropagation. As of now 427 genotypes had been potted out and were evaluated for PRSV-P resistance in the screenhouse. One genotype, BC1 5648 x 410 showed putative resistance to PRSV after three mechanical inoculations. This was planted in the field on Oct 6, 2004. This plant's reaction to natural infection of PRSV is currently being monitored in the field. It is still symptom and virus free as of now (5 months in the field). ELISA confirmed the resistance of this promising genotype to the presence of PRSV. This putative resistant plant is a male tree and has 65% pollen fertility. Backcrossing activities using pollen from this plant is in progress. In addition, 52 different interspecific hybrid plants were established in the field this year.

Sub-Project 4. Development of molecular markers for PRSV-P resistance.
Good progress has been made on mapping of V. pubescens and V. parviflora using RAF markers. A genetic map of the two species has been developed. Five molecular markers have been linked to PRSV-P resistance. Two of these have been sequenced, and a co-dominant SCAR marker has been developed from one. The other segregates 100% with PRSV-P resistance, and will be used in an attempt to isolate the gene for resistance in 2005.

Sub-Project 5. Micropropagation of elite papaya genotypes.
Clones of all C. papaya genotypes, fertile resistant C. papaya x V. quercifolia hybrids, and some backcross plants (including 54) are being maintained in vitro both in Australia and the Philippines. Considerable effort has been given to maintaining the 131 backcross plants (reported under objective 3) in Australia until they can be evaluated for virus resistance.

Philippines

During the three years of implementation of the project, we have reported one genotype, BC1 5648 x 410 that showed putative resistance to PRSV-P after three manual inoculations and under field condition for less than two years before it succumbed to bacterial crown rot. Backcrossing activities using the pollen from this plant were intensified during the last phase of the project.

One hundred and thirty seven (137) BC2 03R lines were generated and inoculated for screening against PRSV-P. After three (3) manual inoculations in a screenhouse, 32 plants (21 female and 11 male) were symptom-free and were transplanted to the field where susceptible papaya varieties are currently being grown. Mild PRSV symptoms were observed a month later. These observed mild symptoms were confined to the older leaves and did not progress to the younger leaves. Symptoms were localized. ELISA test was conducted and negative ELISA test results showed in 21 plants despite the mild and confined symptoms in the older leaves.

Sib-pollination of selected (male and female) BC2 03R line plants are currently in progress towards the advancement of a backcross generation. Likewise, efforts to incorporate and cross the resistant BC2 03R lines to Davao Solo is also in progress. The BC2 03R lines are also being crossed to inbred line 4172 (the male parent of hybrid 'Sinta') to reconstitute 'Sinta' and test the resulting hybrid with 'Sinta' in terms of PRSV resistance. 'Sinta' is tolerant to PRSV-P and is a cross between inbred line 5648 x 4172.

This putative resistant PRSV-P BC plant is the first conventionally bred resistant line in the Philippines.

Australia

A fertile PRSV-P resistant plant (54) was produced in a previous project when a C. papaya x C. quercifolia hybrid was backcrossed to C. papaya clone 2.001. All second backcross (BC2) plants produced from line 54 eventually succumbed to PRSV-P after 9 months in the field. However, there was large variation between plants in terms of degrees of severity to the virus symptoms and time to develop symptoms (2 to 9 months). Thus line 54 had a high level of tolerance to PRSV-P but was not resistant. BC2 plants showed good agronomic traits: high yields, good fruit shape, flavour and high brix levels. These results show that commercially acceptable fruit could be produced after 2 or 4 backcrosses.

Five F1 plants (C. papaya x V. quercifolia) that were PRSV resistant and had low pollen fertility were used to produce 131 BC1 plants. All were cloned and multiples of each clone were field tested in the presence of PRSV-P. All developed viral symptoms however symptom severity and time to infection varied between clones.

A BC2 population developed from the resistant BC1 plant produced in Los Banos have been field tested against an Australian strain of PRSV-P in SE Queensland. After 6 months in the field, 100 plants are segregating for PRSV-P resistance.

V. pubescens and V. parviflora and an F1 and F2 population derived from them were used as a mapping population. The F2 segregated in a 3:1::resistant: susceptible ratio indicating a single dominant gene controlling PRSV-P resistance in Vasconcellea. A genetic map of the two species has been developed using RAF markers. Twelve molecular markers have been linked to PRSV-P resistance. Two of these have been sequenced, and a co-dominant SCAR marker has been developed from one. The other segregates 100% with PRSV-P resistance, and is being used to identify the PRSV-P resistance gene.

Clones of all C. papaya genotypes, Vasconcellea species, fertile resistant C. papaya x V. quercifolia hybrids, and some backcross plants (including 54) are being maintained in vitro. This collection is essential for future research.

Sub-Project 1. Incorporation of PRSV-P resistance genes into elite papaya genotypes, using fertile resistant backcross plants.

Extensive backcrossing of the F1 intergeneric hybrids and inbred lines has resulted in one PRSV-P resistant backcross plant (BC1). This plant is a crossed between inbred line 5648 and intergeneric hybrid 410. This male fertile BC1 was used for further backcrossing to generate BC2 and BC3 plants.

Sub-Project 2. Selection of backcross generations for PRSV-P resistance and good agronomic traits.

A total of 137 BC2 plants were manually inoculated three times at two-week intervals in the screenhouse. Fifty plants or 38% showed typical symptoms (ranging from chlorosis to shoe-stringed leaves ) of papaya ringspot virus after inoculations. Surviving healthy plants (30 plants) were transplanted in the field and were exposed to aphid vectors from susceptible papaya (inbred 4172 and Davao Solo) plants that had a high inoculum level of PRSV-P. ELISA test results revealed that only ten out of the thirty plants tested were positive to the virus. It was also observed that four plants which showed mild infection of the virus based on visual inspection were negative to the virus. The mild infection during the first few months after field transplanting did not progress in these four plants, hence the negative ELISA result. In Southeast Queensland, approx. 25% of BC2 plants are still virus free in a field planting after >12 months in the field in the presence of PRSV-P. All controls were infected after 2 months in the field.

A total of 354 BC2 sib crosses and 124 BC3 plants which survived three manual inoculations in the screenhouse were transplanted in the field. Evaluation against natural infection to PRSV-P is in progress.

Sub-Project 3. Backcross fertile resistant C. papaya x V. quercifolia F1 hybrids to C. papaya.

IPB inbred lines (4108, 4172, 5648 and 5893), resistant BC1 and intergeneric hybrid 410 were planted in the field last Nov. 2006 for further backcrossing program.

Sub-Project 4. Development of molecular markers for PRSV-P resistance.

An attempt to develop a molecular marker is currently in progress. To confirm introgression, genetic fingerprinting of several genotypes of C. papaya and some of its wild relatives was done using simple sequence repeats (SSRs).

Preliminary result indicated introgression of a putative nucleotide fragment (150bp) from V. quercifolia, hypothesized to mark for PRSV resistant attribute of both V. quercifolia and F1 intergeneric hybrids (410, 468 and 469). In the resistant backcross progenies, a putative nucleotide fragment (100 bp) was observed to be consistent. This 100 bp could be a fragment of that putative nucleotide fragment from V. quercifolia which could
explain the resistance of the backcross plants. Sequencing of said fragments is in progress to further validate the marker.

Sub-Project 5. Micropropagation of elite papaya g
Micropropagation of some BC3 (5648 x BC2 03R-73) plantlets and clonal propagation of the remaining F1 intergeneric hybrid 410 is in progress.

Sub-project 6. Evaluation on Grower's Properties

Reconstituted 'Sinta' (BC2 03 R lines crossed with 4172) is now being tested along with the original 'Sinta' in the farmer's field in Batangas.

Sub-Project 1. Selection of backcross generations for PRSV-P resistance and good agronomic traits.
Field performance of BC3 and BC2 sib-cross plants against papaya ringspot virus (PRSV-P) was evaluated. BC3 plants are the product of introgressing the PRSV-P trait from the papaya wild type Vasconcellea quercifolia to Carica papaya. BC2 sib-crosses were developed by sib-crossing selected female and male BC2 plants. Selection was based on ELISA test result and symptom development in the field.
A total of 634 plants (88 BC3, and 546 BC2 sib-crosses) were inoculated three times at two weeks interval in the screenhouse. Three hundred twenty-five (46 BC3 and 279 BC2 sib-cross) or 51.3% showed typical symptoms, which ranged from distortion of young leaves, mosaic, chlorosis to shoe-stringed on older leaves. Plants that remained healthy and symptom free together with the susceptible check, Davao Solo, were then transplanted in the field and were assessed for resistance/susceptibility to the Philippine strain of PRSV-P. A total of 81 BC2 sib-crosses and 34 BC3 plants were planted in Mainit, Bay, Laguna. Initial results showed that, there was variation in the rate of symptom development in BC3 and BC2 sib- crosses from the control. Davao Solo produced severe symptoms after 1-2 months of transplanting in the field while there were BC3 and BC2 sib-cross plants that remained symptom free for about 7-8 months. The difference between backcross papaya and Davao solo was also evident in the ability of the trees to bear good quality fruit. Davao Solo produced a few small and unmarketable fruits. Backcross plants in contrast to the Davao Solo had the ability to recover from early infection based on visual inspection and serological test (ELISA).
Fruit qualities of some backcross plants and DS were evaluated. Fruit weight of backcross plants ranged from 834.53-754.92 grams in contrast with DS's 202.67 grams. Fruits have firm yellow orange flesh, with mild papaya aroma. TSS (0B) values of BC3, BC2 sib-cross lines and DS were 10.0, 12.2, and 9.3 respectively which corresponds to sweet taste for backcross lines and not so sweet for DS.
A total of 401 BC3 sib crosses and 355 BC4 plants are now being grown and will be inoculated manually three times in the screenhouse. The surviving plants will be transplanted in the field where its resistance/susceptibility against Philippine strain of PRSV-P will be tested.
Sub-Project 2. Backcross fertile resistant C. papaya x V. quercifolia F1 hybrids to C. papaya.
IPB inbred lines (4172, 5648 and 5893) were planted in the field last April 15, 2007, to be used in the continuing effort of introducing PRSV-P resistance gene(s) in other elite papaya lines.
Sub-Project 3. Development of molecular markers for PRSV-P resistance.
An attempt to develop a molecular marker was successfully completed. To confirm introgression, genetic fingerprinting of several genotypes of C. papaya and some of its wild relatives was carried out using simple sequence repeats (SSRs).
Preliminary result indicated introgression of a putative nucleotide fragment (150bp) from V. quercifolia, hypothesized to mark for PRSV resistant attribute of both V. quercifolia and F1 intergeneric hybrids (410, 468 and 469). In the resistant backcross progenies, a putative nucleotide fragment (100 bp) was observed to be consistent. This 100 bp could be a fragment of that putative nucleotide fragment from V. quercifolia which could explain the resistance of the backcross plants.
Sub-Project 4. Micropropagation of elite papaya genotypes.
Micropropagation of some BC3 (5648 x BC2 03R-73), BC3 sib-cross plantlets, F1 intergeneric hybrids (404, 410, 468, 469, and 507) and RR papaya cultures (cross between two Vasconcellea species) from Australia is continuing. Clonal micropropagation of BC1 is also in progress.
Sub-project 5. Evaluation on Grower's Properties
Re-constituted "Sinta" (BC2 03R lines crossed with 4172) is now being tested along with the original "Sinta" in the farmer's field in Batangas. Evaluation of resistance of these Re-constituted "Sinta" is still on-going. Fruit evaluation of this line was also Completed.