This project for improvement of waterlogging tolerance of wheat in India and Australia, officially commenced January 1, 2001, and it was initiated with a Work Plan Meeting held at the Central Soil Salinity Research Institute (CSSRI), Karnal, India, June 6-8, 2001. Experimental work on the project commenced after June, 2001, with the first field season in Australia over June to December, and the first field season in India over November to March, 2002. To date, the project is on schedule with approximately 6 months of experimental work and good progress in physiology and breeding.

The Work Plan Meeting, which initiated project work, was attended by representatives of all collaborating institutions:
Department of Agriculture of Western Australia (WADA),
The University of Western Australia (UWA),
The Victorian Institute of Dryland Agriculture (VIDA),
Directorate of Wheat Research (DWR, India),
Narendra Deva University of Agriculture and Technology (NDUAT, India), and
Central Soil Salinity Research Institute (CSSRI, India).
Key outputs of the Work Plan Meeting were an interaction of staff; the development of experimental Milestone Reports for all partners, Experiment Information Sheets, and a Minimum Data Set for environmental characterisation; and agreement on varieties for germplasm exchange, crossing programs and multidisciplinary interactions (Section 3.3(ii)).

Scientific exchange visits have resulted in key periods of intensive project work during 2001. The first research exchange visit of staff commenced on June 18, with Dr. K.N. Singh (CSSRI) to WADA; and the second research exchange visit was of Dr. J. Rane on October 15 to UWA. Both visits were successful. Dr. K.N. Singh learned new techniques for producing doubled haploid (DH) wheat lines, and he produced over 600 DH lines from 3 crosses (Section 3.2(ii) Breeding and Section 4.3, Dr. K. N. Singh). Dr. Rane learned new techniques for assessment of physiological traits associated with waterlogging and anoxia tolerance. He concluded that for specific wheat lines grown in deoxygenated nutrient solution simulating waterlogged soil, the potential of plants to recover after waterlogging, rather than withstand O2 deficiency during waterlogging, may be the reason for differential grain yield response of wheat under field conditions (Section 4.3, Dr. J. Rane).

Research was initiated at all collaborating institutions and this is described in general in Section 3.2, and in detail in the 14 Experiment Information Sheets (EIS) for 2001 (Appendix, Section 4.2) and from statements submitted to the Project Coordination Committee (PCC) in India (Section 3.3.(i)). This first PCC meeting for the project was held on December 3, 2001, in New Delhi (Section 4.3, Dr. T. Setter to India (December 2 to 17, 2001)) to oversee the direction and progress of project research in India. Significant additional supporting achievements occurred through links of this project with GRDC projects supervised by Dr. Colmer and Dr. Setter (Section 3.2). These linkages are expected to increase with new projects currently under development (Section 3.2). Research progress has exceeded the project Work Plan for 2001, including development of 3 DH populations, development of 25 crosses of Australian and Indian germplasm, and confirmation of genetic diversity for waterlogging tolerance in the set of wheat genotypes targeted for this project (Section 3.2).

Future directions of project research include (i) initiation of work focusing on environmental characterisation, (ii) continued work on physiology and breeding components as described in the Project Outputs (ACIAR Project Document, Section 1.15, Output Table), (iii) organisation of exchange visits in 2002 for project scientific staff, two collaborating institution Directors, and several ICAR staff (Section 3.3.(iv)).

The aim of this project is to produce waterlogging tolerant breeding lines of wheat for Australian and Indian target environments by identifying and evaluating the genetic basis of physiological traits conferring waterlogging tolerance in wheat , so that traits can be combined in improved breeding lines. The objectives and outputs to achieve this aim are:

1. Environmental Characterisation Outputs
Characterise the nature of waterlogging in wheat fields in India and Australia
Prioritise waterlogging environments and regimes
2. Physiology Outputs
Develop an improved physiological understanding of waterlogging tolerance in wheat
Identify physiological traits useable by plant breeders
3. Germplasm Evaluation Output
Identify waterlogging tolerant wheat germplasm
4. Germpalsm Development Output
Develop new germplasm based on physiological traits
5. Research Capacity Output
Exchange visits

Good progress has been made in all objectives during 2002, with key information coming from environmental characterisation of waterlogging prone areas, germplasm evaluation and development, and exchange visits (Outputs 1, 3, 4 and 5).

Environmental information has enabled protocols to be developed for controlled waterlogging experiments in Australia and India. These protocols differ in different locations due to major differences in environmental factors including temperature, soil biological activity, organic matter and nutrition (see review by Setter and Waters, 2003; Plant and Soil, In Press).

Our approach of yearly field evaluations in Australia and India, integrated with laboratory and glasshouse based studies under controlled conditions, provides feedback for: (i) identification and development of suitable donor parents for future breeding programs in Australia and India, and (ii) further refinements of our breeding objectives.

Germplasm evaluation has been the largest single activity in 2002 across all partners. Results demonstrate that the project has provided Indian partners access to Australian germplasm which has the highest level of waterlogging tolerance of any variety tested for waterlogging conditions in neutral soils in India; these varieties are also well known for good grain quality in Australian environments. Selected Indian varieties have also performed well in controlled waterlogging experiments in Australia using duplex soils. These varieties have already been used as parental lines with Australian varieties for the production of three doubled haploid populations, and about 200 lines of the first population are due to be sent to Indian partners in February, 2003.

Plant breeding continues to be led by the Department of Agriculture, Western Australia (DAWA), with the development of new genetically fixed doubled haploid populations and segregating populations. Major breeding programs are also expected to be initiated by at least two Indian partners in 2003/04.

During 2002, controlled waterlogging facilities were constructed at Katanning, Western Australia, and these were particularly successful in identifying up to 5-fold differences in waterlogging tolerance based on shoot dry weights of plants after 6 weeks waterlogging. Doubled haploid wheat lines used in this project were the most waterlogging tolerant genotypes. These lines are now being assessed for physiological traits, and molecular markers for waterlogging tolerance are being developed in another project linked to this work.

Recent results highlight the importance of microelement toxicities as one of the key factors for some waterlogging prone soils in Australia. Microelement toxicities (Mn2+, Fe2+ and Al) may occur during long term waterlogging due to severe reduction in soils, and increased solubility and uptake of reduced microelements by plants. If confirmed, the subsequent work to increase tolerance to specific microelement toxicities could be the first step in pyramiding genes for waterlogging tolerance of varieties targeted to these soils. This work could also build on the large amount of published information for tolerance of cereals to specific microelement toxicities.

With at least 24 mechanisms of waterlogging tolerance identified for cereals by Setter and Waters (2003), the process of identifying key mechanisms in specific parental lines or populations is expected to take time. Information currently available indicates good prospects for pinpointing key physiological mechanisms during 2003/04.

Our research resulted in four publications in 2002, due to synchronising the ACIAR Annual Project Meeting, with the 12th Australasian Plant Breeding Conference in Perth, WA. A review on waterlogging tolerance in cereals is now "in press" in Plant and Soil; ACIAR supported research is specifically highlighted in the review.

At the end of this project, it is expected that the mechanistic approach for pyramiding genes for waterlogging tolerance based on physiological mechanisms will result in the utilisation of new selection criteria and new tools in cereal breeding programs; and the development of new germplasm (breeding lines) will provide a substantial, sustainable improvement in waterlogging tolerance of wheat for Australian and Indian farming systems.

Good progress has been made in all objectives during 2003, with key information coming from environmental characterisation of waterlogging (WL) prone areas, physiology, germplasm evaluation and development. Further environmental characterisation has:
provided a better understanding of the nature of constraints that wheat crops experience during WL and also the extent and severity of the problem
led to identification of two distinct WL environments based on soil parameters and their interaction with genotypes in Australia and India.
Knowledge on the WL environments will be useful in improving the already developed protocols for controlled WL screening experiments in Australia and India. These protocols differ in separate locations due to major discrepancies in environmental factors including temperature, soil constraints and its biological activity, organic matter and nutrition.
Field evaluations in Australia and India integrated with laboratory and glasshouse based studies under controlled conditions revealed:
identification of genotypes with tolerance to WL to be used either as donor parents in breeding programs or production of further DH populations. These genotypes are from a doubled haploid DH population derived from Ducula-4/2* Brookton, varieties and breeding lines sourced from participating project partners
further refinements of breeding objectives based on continued environmental characterisation and the demonstration of its interaction with genotypes during WL.
Germplasm evaluation was the major activity in 2003 for all partners. The project has provided Indian partners with access to germplasm produced in Australia that has moderate WL tolerance in reclaimed soils in India; these varieties are also well known for good grain quality in Australian environments. The identification of further tolerant genotypes to WL and sodicity continued with13 breeding lines reputed to have good tolerance to WL in sodic soil introduced from India for use as parental lines in production of new germplasm.
Plant breeding continues to be led by the Department of Agriculture, Western Australia (DAWA), with the development of both new, genetically fixed DH and segregating populations. Their distribution was coupled with statistical design to maintain uniformity and accuracy of field trials across partners. Good progress was also made with major breeding activities by two Indian partners (NDUAT and CSSRI) in 2003.
More than 200 genotypes were sown in over 4,000 pots to evaluate waterlogging tolerance and recovery ability under controlled conditions. Results demonstrate that WL accentuates the accumulation of high to toxic concentrations of some micronutrients in wheat seedlings in some WL prone soils in Australia and India. This occurs during long term WL due to severe reduction in soils, and increased solubility and uptake of reduced microelements by plants. The results so far have revealed that tolerance to aluminium (Al) might determine performance of wheat in some WL soils in
Significant evidence of differential tolerance to either manganese (Mn) or iron (Fe) and WL tolerance was not observed, although all tested genotypes had a moderate level of tolerance to Mn in Australia. The current hypothesis is that enhanced tolerance to Al confers an improved tolerance to WL in Australian soils, possibly by exclusion of Al which is more available during WL. Note that these results seem to be unique for Australian soils; preliminary results indicate an important role for other microelement toxicities (sodium and boron) in waterlogged soils of India.
Good progress has been made in evaluations of aerenchyma, and biochemical traits for recovery from anoxia and protection against oxidative damage. Some of the recommendations for waterlogging tolerant varieties generated through this project have already been communicated and extended to farmers in India.

The aim of this project is to produce waterlogging (WL) tolerant breeding lines of wheat for Australian and Indian target environments by identifying and evaluating the genetic basis of physiological traits conferring WL tolerance in wheat, so that traits can be combined in improved breeding lines.

Good progress has been made in all objectives during 2004, with key information coming from environmental characterisation, physiology, germplasm evaluation, breeding, and exchange visits (Outputs 1-5).

Environmental characterisation has:
(i) Confirmed that waterlogging is a "hidden constraint" to wheat production in Australia and India;
(ii) Led to identification of multiple WL environments based on the predisposition of soils to microelement toxicities.
(iii) Identified a major new constraint of B (Mn and Fe) toxicity likely to affect large areas of wheat production in sodic soils exposed to waterlogging in India.

Germplasm evaluation was the major activity in 2004 for all partners. Results demonstrate that the project has provided Indian partners with access to germplasm produced in Australia that has moderate WL tolerance in reclaimed soils in India; these varieties are also well known for good grain quality in Australia. In 2004, partners were provided with DH lines derived from HD2329/2*Wyalkatchem and WAWHT2531/NW1014. The key germplasm screening activity this year was to screen the Ducula-4/2*Brookton population (190 lines) in replicated trials with waterlogged and drained treatments. Largest trials were at CSSRI (2 sites) and DWR; a reduced trial was screened at NDUAT. UWA completed physiological analyses, and DAWA completed a large screening under controlled waterlogging conditions in the field. Data for 2004 will be used to (i) validate previous years data, (ii) contrast with other sites, and (iii) form the basis for developing QTLs for waterlogging tolerance through new linkages proposed with the Molecular Plant Breeding Cooperative Research Centre.

Plant breeding
Benefits of project work on germplasm improvement are already evident. DAWA has created 11 doubled haploid (DH) populations using waterlogging and sodicity/salinity tolerant Indian wheats crossed with Australian varieties. In 2004, four DH lines from the Ducula-4/2*Brookton population produced at DAWA were given KRL names at CSSRI, and they were advanced for All-of-India Coordinated Trials for 2005. This Ducula-4/2*Brookton cross was made by Mr. Robin Wilson, and developed by Sue Broughton's team at DAWA.

Breeding trial recommendations on advanced breeding lines grown in India were used in support of a case to release the newest wheat variety, KRL35, by Dr. K.N. Singh at CSSRI in 2004. This variety has been evaluated as part of ACIAR project work. This variety has recently been introduced to DAWA, and it is currently being used to develop six new DH populations in crosses with Australian varieties to increase waterlogging and salinity tolerance and grain quality. These new populations will be returned to India in 2005/06.

Physiology work has focused on critical evaluation of mechanisms of tolerance to waterlogging. This year the project has reviewed major physiological traits including Na and B toxicities (CSSRI), Al, Mn and Fe toxicities (DAWA), aerenchyma development and recovery ability (DWR), alcoholic fermentation, carbohydrate accumulation, anoxia tolerance and antioxidant metabolism (UWA), and agronomic traits, including tillering and plant height (CSSRI, DWR, NDUAT, UWA and DAWA). Mechanisms of tolerance which relate most to waterlogging tolerance are those associated with tolerance to a wide range of microelements. The current hypothesis is that waterlogging tolerance is a product of (1) tolerance/avoidance of anaerobiosis and (2) tolerance to microelements (Setter et al., 2004). Interactions with different microelements in different environments, explain why WL tolerance and ranking of germplasm often varies between environments.

A major contribution of physiology work in 2004 has been to clarify simple, non-destructive, selection criteria as screening protocols for waterlogging tolerance. Research in India and Australia confirms that selection for maintenance of high tiller number during waterlogging gives one of the best correlations to waterlogging tolerance; maintenance of plant height is also a suitable criterion for waterlogging tolerance.

At the end of this project, it is expected that the mechanistic approach for pyramiding genes for WL tolerance based on physiological mechanisms will result in utilisation of new selection criteria and new tools in cereal breeding programs. Our research resulted in 8 publications in 2004, including 1 PhD thesis and 6 papers under submission/preparation. Some of the recommendations for waterlogging tolerant varieties generated through this project have already been extended to farmers in India through work at NDUAT.

At the Final Review of this project proposed for June, 2005, a case will be presented for project extension to capture and extend the benefits of the existing achievements.

Research on waterlogging tolerance of wheat for 2005 was completed successfully, resulting in the compilation of data for the Final Review held at the Central Soil Salinity Research Institute (CSSRI), Karnal, India, over March 6-8, 2006. Research covered in this period consists of 1) concluding research from the final six months (January 1 to June 30, 2005) on Project CS1/1996/025, and 2) the first six months research (July 1 to December 31, 2005) on the "12 Month Project Extension to ACIAR Project CS1/1996/025 on Waterlogging Tolerance of Wheat" (Project CMS/1996/025).

The concluding research from the original project (CS1/1996/025) is reported here in the attached detailed Appendices from field work conducted in India during the 2004/05 season at CSSRI (Appendix 1), Directorate of Wheat Research (DWR; Appendix 2) and Narendra Deva University of Agriculture and Technology (NDUAT; Appendix 3). During 2005 there were no funds for continued work from UWA (whose final project achievements are summarised in the ACIAR Annual Report 2004 - Project CS1/1996/025). There were also no funds available for project staff at the Department of Agriculture and Food Western Australia (DAFWA). Therefore only a limited research program was followed in Australia primarily aimed at (i) supervising collaboration in India; (ii) extending screening methods to segregating bulk DH populations; and (ii) multiplying seed (approximately 1440 genetically fixed breeding lines) for distribution to Indian partners in 2006 (Appendix 4).

Good progress has been made during 2005 on all five objectives for the first 6 months of ACIAR Project CMS/1996/025 by both Australian and Indian partners. These project extension objectives are:
1) Develop new methods to screen DH and segregating populations as bulks.
2) Continue the development of new DH lines/populations.
3) Continue evaluation of adaptive physiological traits for waterlogging tolerance.
4) Evaluate waterlogging tolerance at the germination stage (results not presented in this first 6 mo. report)
5) Multiply seed of the available 10 DH populations for waterlogging tolerance.
Due to the 6 month delay in field season for India, results from the 12 Month Extension will only be provided here from DAFWA, while results from India will be included in the Final Report.

New methods using soil bins have been used to screen bulk DH and segregating populations in different soils in WA (DAFWA; Appendix 4). Similar approaches have been successfully evaluated in the field in India by all Indian partners (not reported here). This now extends screening protocols originally developed for varieties and genetically fixed lines to populations with an increase in screening speed of up to 100 times. The impact of this method is that now germplasm evaluation can keep pace with germplasm development. Furthermore, breeders can screen non destructively and make selections at any stage of germplasm development.

New DH lines/populations have been developed for four populations in 2005: Tammarin Rock / EGA-Bonnie Rock (~300 lines; these parents are extremes in waterlogging tolerance in Katanning soil), EGA-Bonnie Rock/Tammarin Rock (~300 lines), Chara/Camm (~200 lines; these parents show extremes in waterlogging tolerance at germination stage; Setter and Waters, 2003) and KRL35/Tammarin Rock (~200 lines).

In 2005, a small nursery was also established based on varieties and near-isogenic lines differing in tolerance to microelements. The evaluation of these genotypes in target environments where waterlogging occurs will enable determination of whether microelement toxicities are exacerbated under waterlogging in specific soils. This has been confirmed in preliminary observations. The current hypothesis is therefore supported that waterlogging adversely affects plant growth by a product of both anaerobiosis and microelement toxicities (Khabaz-Saberi et al., 2006; Setter, 2006).

Bulking seed supplies was also completed in this period. Approximately 1500 DH lines and varieties were bulked as genetic resources (25-100 g/line) for distribution to Indian partners in 2006.

A summary of achievements and impacts of project work highlighting research from 2005 was prepared for the Final Review (Setter, 2006) and this is available as a pdf file from Dr. T. Setter (tsetter [at] agric [dot] wa [dot] gov [dot] au) and attached to the electronic copy of this report. Detailed research work is now under preparation for publication for a major review on germplasm improvement for waterlogging tolerance and for separate research papers on soil science, physiology, and breeding for waterlogging tolerance.

Khabaz-Saberi, H., Setter, T.L. and Waters, I. (2006). Waterlogging induces high to toxic concentrations of iron, aluminium and manganese in wheat varieties on acidic soil. J. Plant Nutr. 29: 899-912.
Setter, T.L. and Waters, I. (2003). Review of prospects for germplasm improvement for waterlogging tolerance in wheat, barley and oats. Plant and Soil 253: 1-33.
Setter , T.L. (2006). Preliminary Report on Waterlogging Tolerance of Wheat in India and Australia. Dept. of Agriculture and Food, Western Australia; and Australian Centre for International Agricultural research, Canberra, ACT. 23 pp.