ITEMS FROM AUSTRALIA

CSIRO Division of Plant Industry

North Ryde (Sydney) 2113 and Canberra 2600, Australia.

Program leader Dr. R. Appels.

Wheat germplasm and grain quality program.

This is one of several research programs of the CSIRO Division of Plant Industry, based in Canberra, Australia. Staff of the program are located at both the Grain Quality Research Laboratory, in Sydney, and at the division's headquarters in Canberra, permitting the integrated study of wheat quality at technological, chemical, and genetic levels. The participation of this research program in the recently created Cooperative Research Centre for Quality Wheat

Products and Processes will expand opportunities for collaboration with other research groups and with the wheat-growing and processing industries.

Molecular genetics.

An important achievement of the past year has been the transformation of wheat with the regeneration of phenotypically normal transgenic plants and also contributions to the transformation of barley. We recognise the need to identify the most worthwhile genes to target for application of the technology (a major goal of our other research), as well as suitable promoter sequences. Steps towards these goals involve the application of gene probing. Such studies have involved the development of new approaches to identify sources of pathogen resistance, new information about the relatives of wheat, and methods of genotype identification.

Wheat-grain proteins and dough function.

Our very-small-scale dough testing equipment is helping breeders to test at an earlier stage and is an important research tool for identifying the proteins involved in specific aspects of dough function. Study of the contributions of specific proteins has included the role of rye proteins in 1RS lines and study of durum wheats. Beyond the level of identifying coding genes and specific polypeptides, their polymerisation into macromolecules has been identified as critical to their functional properties in dough. New approaches to analysing size distribution have been developed to further elucidate this relationship.

Direct observation of the contributions to dough properties of individual polypeptides have been made in the 2-gram Mixograph, after initially developing a procedure for breaking and reforming disulfide bonds, in the process of incorporating added glutenin polypeptides into the macromolecular structure. The study of individual polypeptides has included those purified from flour and also proteins expressed heterologously in bacteria from the isolated gluten genes. These mixing studies h also ave included elucidation of the role of the `improver', bromate.

The development of antibodies specific to particular groups of gluten proteins has provided the opportunity of identifying amino acid sequences expected to relate to dough-quality attributes. Such antibodies have been used to provide predictions of dough strength, to identify the presence of rye protein in wheat-rye translocation lines, and to identify other alien-chromosome substitutions.

Molecular studies on starch and grain hardness.

A starch-granule protein associated with the soft-grain character in wheat has been further characterised, and the genes for its synthesis have been cloned. The locus for this protein has been studied in the wheat progenitor Triticum tauschii. This species also has been used to study the mechanisms of starch synthesis.

Wheat-starch structure is particularly important for noodle-processing quality, and a predictive system of quality evaluation has been developed based on chemical analysis. For industrial use of wheat starch, a high paste viscosity is important. We have shown that loss of viscosity may be due to the presence of residual FONT SIZE=2 FACE="WP Greek Century""-amylase after washing.

Modification of grain quality by growth environment.

Starch and protein quality are affected by growing conditions. In particular, a few days of heat stress can produce an increase in the proportion of large (A-type) starch granules and a weakening of dough properties. On the other hand, modest temperature increases (15 to 35 C) during grain filling appear to cause a strengthening in dough properties. Because genotypes differ in their reactions to temperature change, we have examined a range of wheats for quality changes to identify those that are more tolerant to heat stress. Another approach to overcoming the problems of

quality variation due to environmental factors is to predict the extent of these variations. We have focused on the protein content of wheat and barley, for which winter rainfall and spring heat stress were identified as major factors.

Quality protection during storage .

Beyond harvest, the conditions of grain storage and transport are critical to providing the market with grain of suitable quality. Moisture content and temperature are the critical factors taken into account in modeling the rates of quality change for a range of grains. These models have been used to develop predictive software to better manage grain storage, even for rain-damaged grain.

Pesticide treatment is still an important means of protecting stored grain from insect attack, but there is a need to monitor treatment to ensure that minimum residue levels are not exceeded. We have developed antibody-based test kits to determine the levels of specific pesticides in grain - fenitrothion, chlorpyrifos-methyl, bioresmethrin, and several others. The development of these test systems requires the application of sophisticated organic chemistry, but the immunoassay principle is capable of delivering results fast (within 15 minutes) under field conditions. The tests have been used to determine the fate of pesticides during milling and baking. Immunoassay systems also have been developed for agrochemicals used in irrigated agriculture, particularly for the pesticide endosulfan. The speed and portability of test kits are already improving the management of irrigation waters.

Rapid determination of grain quality attributes.

In addition to their use in agrochemical analysis, antibodies have proved valuable for determining dough strength, the presence of rye proteins, and varietal identity. Near-infrared spectroscopy also has been used for grain-quality testing, originally for moisture and protein analysis. More recently, we have improved on calibration procedures for these analyses and extended the capability of this technology towards dough-property analysis and distinguishing red-grained wheats from white wheat.

The need to make this distinction arises from the recent registration of red wheats for growing in Australia where wheat has exclusively been white grained. Therefore, we have examined a range of methods for visually distinguishing red from white wheat with a reflection colour meter or by image analysis. Image analysis also has been used to identify yellowed grains in samples of milled rice.

Because genotype is a major indicator of grain quality for all species, variety is used widely as a basis for quality segregation, and there is a pressing need for better methods and strategies for variety identification. Visual methods are limited. Analysis of protein composition has proved adequate for many grain varieties, using immunoassay, column chromatography, and gel electrophoresis. DNA probing methods now are being developed for the direct identification of genotype for grains.

Annual report available.

Full details of the program's research results are provided in our publications. The details of 150 papers, for the year July, 1993, to June, 1994, are given in our Annual Report, available on request from Colin Wrigley (Fax: 612 887 8419 or E-mail: c.wrigley@pi.csiro.au).

QUEENSLAND

Queensland Wheat Research Institute

PO Box 2282, Toowoomba Q 4350, Australia.

Wheat breeding.

P.S. Brennan, P.M. Banks, J.A. Sheppard, and M. Cooper.

The drought that commenced in the summer of 1990-91 continued throughout 1994, when it was at its most severe. Queensland produced 180,000 t of wheat in 1994. The long-term average is 1.4 m t. The most recent occasions when Queensland has produced less wheat than this were in 1970 and 1946. In those years, the area that could be sown to wheat was less than half of what it is today.

One set of yield trials was established and maintained with irrigation, and all others were either not planted or failed to develop secondary roots. It is clear from previous experience with such trials that yield prediction is poor.

For a number of years, we have advocated the use of BC1F2 populations as the starting point for a breeding program. The recurrent parent would have the greater number of favourable alleles. This approach has been emphasised in the QWRI breeding program for sometime, and late generation material derived from this type of population has been evaluated over the last few years.

This material is exhibiting very large advances for yield, while combining the better quality and disease resistance attributes of both parents. These results have been obtained consistently over 2 or more years. This type of outcome is consistent with the predictions made from theoretical considerations. We are now using BC1F2-derived populations, almost exclusively, as the starting point for our wheat breeding activities in our pedigree program.

The much maligned backcross procedure has been used to transfer genes for a number of economically important diseases into adapted cultivars. Two varieties were released last year from this program. Both varieties exhibited nontarget improvements of economic importance. This probably was due to the evaluation of fairly large populations at the BC3 level and to using a number of resistant BC1 lines as donor parents.

In 1995, we anticipate the release of two more backcross derived lines. QT5793 is based on Hartog and has yellow (tan) spot resistance from CNT2. The second is based on Ford and has mildew and rust resistance from 1BL-1RS. This will provide a stem, leaf, and stripe rust resistant hay-wheat for Queensland.

Staff changes. Mr. Ma Wujun was awarded a PhD scholarship through the University of Southern Queensland to work on the location of molecular markers for genes for high protein.

Ms. Tanya Pearce will continue the development and assessment of wheat/barley translocations for the transfer of the barley FONT SIZE=2 FACE="WP Greek Century""-amylase inhibitor from barley to wheat. This work was commenced by Mr. Ian Haak who has completed writing up the work for a master's degree. Ian is now involved in a project to locate molecular markers for a number of disease resistance genes. The disease resistances selected for this work are effective, but selection on a single plant basis is difficult because of low heritability. Double haploids will be used.

Russian wheat aphid. For some time, we have been backcrossing resistance to the Russian wheat aphid into varieties adapted to Australia. This has been accomplished by resistance testing at Colorado State University by Professor Jim

Quick. Agronomically acceptable lines have been identified in a number of populations and will be yield and quality tested in the near future.

Mr. Steve Kammholz has developed a small double-haploid population from a `Turcikum 57/Hartog' cross. This population segregates in a 1:1 manner, suggesting a single gene for RWA resistance in Turcikum 57.

Double haploids.

Steve Kammholz.

A considerable effort has been committed to developing double haploids in wheat using pollination with maize and chromosome doubling with colchicine. The technical problems associated with the development of haploids were overcome, and large populations developed. The initial doubling was unsuccessful because of high glasshouse temperatures. The process was repeated in 1994 using much cooler conditions and was successful.

This procedure is now routine and is being applied to a number of research projects.

Funds are being sought to determine the utility of double haploids in a breeding program.