THE DANIEL McALPINE MEMORIAL LECTURE 1985

Professor Bob McIntosh and Dr Colin Wellings, The University of Sydney, and The Department of Agriculture, NSW 

Wheat Rust Resistance -- The Continuing Challenge

Introduction

Mr. Chairman, members of APPS. It is an honour to be invited to deliver the fifth Daniel McAlpine Memorial Lecture at this first meeting of APPS to be held in New Zealand.
We fail to be unmoved when we read of the many and varied achievements of "the Father of Plant Pathology in Australia" and certain of his contemporaries, including the wheat breeder, W.J. Farrer, with whom McAlpine had close personal contact. Probably few are aware that McAlpine had a very distinguished scientific (in Scotland) and teaching (Ormand College, Melbourne) career before his appointment as Vegetable Pathologist with the Victorian Department of Agriculture in 1890 "to attend to any disease that might form the subject of enquiry". Fish (6) further stated "there is little doubt that his appointment . . . was a result of the 1889 rust epiphytotic". McAlpine participated in four of the five historic "Rust in Wheat" Conferences held during the 1890's. Needless to say, the rusts of Australia were an early challenge for him. By 1906, he had described 75 new species of rust pathogens.
Today, we propose to discuss the development of current knowledge of host-pathogen genetics in relation to the rust diseases of wheat. In the latter part of the lecture, we will review a rust disease that is relatively new to our geographic area, namely stripe rust of wheat.
Cereal rust problems are not new. The threat of rust was so important to the Romans that they held a feast and made sacrifices on April 25 in order to placate Rubigus, the Corn God. While we presume that similar events are no longer commemorated, the threat of rust epidemics continues.
There are three rust diseases of wheat, viz., stem or black rust, leaf or brown rust and stripe or yellow rust. Current methods of breeding rust resistant wheats are largely dependent on a theoretical basis that has developed since the mid-1800's. We propose to briefly discuss the various developments which culminated in the gene-for-gene theory of host-pathogen genetics, and will attempt to assess where this theory has taken us.

Landmarks of Wheat Rust Genetics

Our current knowledge of the biology of wheat rusts began with the formal description of the heteroecious nature of cereal rust fungi by de Bary and colleagues. However, French farmers had long recognised the benefits of barberry eradication in reducing epidemics and yield losses due to stem rust in areas where today rust incidence is low (12).
In the 1890's, Eriksson and colleagues - contemporaries of McAlpine and Farrer- showed that the stem rust and stripe rust pathogens were specialized in ability to attack different graminaceous species. About 25 years later, Stakman and colleagues demonstrated a further dimension of pathogenic specialization within formae speciales. Puccinia graminis triciti was divided into races, (or strains or pathotypes) on the basis of ability to attack individual wheat genotypes (17). In the 1920's, Craigie (4) discovered the role of pycnia in the sexual cycle on barberry, the alternate host for P. graminis.
Thus the scene was set for the most significant development in host-pathogen genetics when, in the 1940's and 1950's, Flor made concurrent studies of the inheritance of reaction in the flax host and of the inheritance of pathogenicity in the flax rust pathogen (5, 7s 13). He firstly showed that resistance in a selected flax host was determined by a single dominant allele, and that avirulence in Melampsora lini with respect to the particular flax resistance gene was also inherited as a single dominant allele. Thus, incomparability between host and pathogen occurred only in the presence of the host gene for resistance and the pathogen gene for avirulence (Fig. 1). It should be noted that this also established that resistance and susceptibility, and avirulence and virulence, were allelic states. When two genes were studied jointly, it was possible to demonstrate that each interacted only with its corresponding counterpart (see Fig. 2). Moreover, the presence of only one corresponding host resistance gene-pathogen avirulence gene pair was sufficient to produce an incompatible interaction, irrespective of the genotypes at other corresponding gene sites.
It should be remembered that the phenotypes (and often, genotypes) for both host and pathogen are deduced from the disease phenotype. The interaction can be compared to a lichen, a complex organism consisting of a fungus and an algae. Hence the genetics of disease might be considered analogous to the genetics of a lichen.
For genetics we must have words describing the character of interest and we must have words describing the contrasting phenotypes we wish to examine (Fig. 3). The gene-for-gene relationship has been demonstrated for various plant diseases and pests (for examples, see (5)). Moreover, the types of data available allow us to assume its application to other systems; for example, stripe rust of wheat where there is no known sexual stage in the pathogen.
Allow us to restate the genetic rules that are comparable with Mendel's laws: 1 . Low infection type or incomparability is the consequence of interaction of a host gene for low reaction (resistance) and the corresponding pathogen gene for low pathogenicity (avirulence) i.e., LIT = LR:LP (Fig. 1). 2. When more than one corresponding pair of low reaction and low pathogenicity genes are involved, the low infection type that is expressed is as low as, or lower than, the lowest of the single corresponding gene pair interactions, i.e., LIT 1 +2 @LIT 1 where LIT 1 <LIT 2 (Fig. 2). One example of a two-corresponding-gene-pair interaction in wheat stem rust where increased incompatability occurs is Srl5Sr22:P15P22.

Experimentation

The above attributes of the gene-for-gene relationship are used in everyday experimental designs (1). There are four main designs that vary with the extent of knowledge of the genetics of host and pathogen (Table 1).
Design 1 applies to situations where no genetic information is available with respect to either host or pathogen. A data matrix is generated from tests of an array of host lines with an array of pathogen isolates. By inspection of rows and columns, one can postulate hypothetical genotypes and can select representative host lines and pathogen isolates for further study. Design 2 is the basis for pathotype surveys. A set of genetically, or phenotypically, distinct lines is chosen as testers for pathotyping unknown collections of the pathogen. If the purpose of the survey is to relate pathogen variation to the performance of commercial cultivars, then commercial cultivars, or stocks with the genes possessed by commercial cultivars, must be used as the testers. Design 3 permits host materials to be tested with a selected group of distinctive pathotypes. This permits postulation of host genotypes and, in a breeding context, allows the selection of host lines most likely to extend the range of host variation. Design 4 forms the basis for physiological and biochemical experiments. Various workers have emphasised the significance of the quadratic check (Fig. 1) in providing the essential experimental controls for fundamental studies.
The genetics of host-pathogen interactions tell us that both the host allele for resistance and the pathogen allele for avirulence produce active products. Only a few days ago, I attended an Australian-U.S. Workshop on genetic engineering in plants. I believe there was a consensus of opinion indicating that the time is ripe for someone to clone a resistance gene (or a pathogen avirulence gene), to establish its DNA base sequence, to isolate or synthesise the gene product, and to transfer DNA possessing a resistance gene from a resistant host to a susceptible host, either within a host species or between related host species. It is most gratifying that the experimental systems being used by molecular geneticists are precisely those predicted by the gene-for-gene relationship.

Rust Resistant Cultivare in Agriculture

Our laboratory at Castle Hill is committed to the development of rust resistant wheats for Australia. For this, a multidisciplinary approach is essential in order to combine rust resistance with the many other criteria needed in a modern wheat cultivar that is to be sold into a competitive international market. In order to achieve rust resistance, we must be the wearers of several hats: as geneticists we are interested in the inheritance of phenotypic contrasts; we often make special crosses for this, as pathologists we must ensure that resistance sources provide adequate protection at critical stages of epidemics, as breeders we require simple, fast and lowcost methods for screening and selection; we must be able to combine resistance with the other attributes required in a cultivar, as agronomists we must be able to introduce and recommend the resistant cultivar to agriculture, as extension workers we must provide information to farmers enabling them to assess risk and to take management decisions. The information must be relevant to current pathotypes and should provide an adequate

Fig. 1. Australasian Plant Pathology Vol. 15 (1) 1986

description of expected cultivar responses. Our laboratory has recently adopted the principles of listing recommended cultivars in genetic diversity groups and in one of four disease response groups. In rust terms, response groups 1,2,3 and 4 equate with resistant, moderately resistant, moderately susceptible and susceptible.

Table 1. Experimental designs for genotypes used in host-pathogen genetics.

Design Host Pathogen
1. Unknown Unknown
2. Known Unknown
3. Unknown Known
4. Known Known

It is not difficult to breed for resistance. The main problem in respect of rust, and certain other diseases, is that pathogens acquire the ability to neutralize the resistances with which they are challenged.
Australasia forms a geographic region with respect to variation in cereal rust pathogens. New pathotypes arise (20) as a consequence of introduction of inoculum to a particular area: Australasia forms a unique geographical area for plant pathogens and insect pests. Nevertheless, distinctive new rust pathotypes - indeed, new pathogens among which P. striiformis tritici must be included - may be introduced from time to time (23). Within the geographical area, air-borne re-distributions of pathotypes usually reflect the directions of predominant weather patterns. For example, pathotypes occurring in Western Australia are soon found in the east and in New Zealand. On the other hand, those occurring in eastern Australia only slowly find their way to W.A., and the occurrence in eastern Australia of pathotypes previously unique to New Zealand, is rare.
Sexual hybridization: This is not important in Australasia.
Asexual hybridization: While this appears to be an infrequent event, it is believed to have contributed to at least one important group of strains attacking wheat (3v 21) and to putative hybrids between certain formae speciales (2).
Mutation: Mutations are the most obvious events contributing to agriculturally important variation in rust fungi in Australasia. Examples of recent mutational changes in two groups of P. graminis tritici pathotypes are shown in Fig. 4. In 1984, there were two limited, but different, stem rust epidemics in Queensland. The first involved Oxiey wheat and the high inoculum levels that developed on that cultivar probably contributed to the occurrence of a new pathotype on cuitivar Cook; the second epidemic involved Satu triticale which had been recommended as a replacement for the previously susceptible cultivar Coorong (15).

Progressive increases in virulence: Longterm studies at Castle Hill have shown that differences between avirulent and virulent pathogen isolates with respect to a particular host line carrying a single gene for resistance are not always bimorphic and clear-cut (22). As many as five distinctive phenotypes have been distinguished. Neither the genetic basis for this variation, nor its significance in pathogen evolution is understood.

Disease Control

Plant disease control strategies are directed at: - reducing the probability of epidemics - reducing the magnitude of losses There is no doubt that the probability of epidemics can be reduced with resistant cultivars; this has been clearly demonstrated in the northern wheat areas of Australia where there has been no major stern rust epidemic for 30 years, an area where, previously, the probability of a stem rust epidemic was about one year in four. The probability of epidemics can also be reduced by adopting procedures that reduce carryover inoculum, and this may be a community problem since both farms and public areas such as roads and railway lines can be involved.
The magnitude of losses is reduced by resistant cultivars, but especially by the use of genetically diverse resistances so that failure of one resistance source does not have excessively large affects. Extreme levels of susceptibility "rust suckers" - should be avoided, either as

Fig. 2. Australasian Plant Pathology Vol. 15 (1) 1986

cultivars or as genetic backgrounds for a breeding program based on backcrossing. Specified minimum disease standards before cultivar recommendation in some countries takes account of knowledge that some susceptible cultivars are far more susceptible than others.

Resistance

In broadest terms, two types of resistance have been contrasted, but there are no genetic criteria that clearly distinguish them. Specific host resistance is dependent on the concurrent presence of the corresponding gene(s) for aviruience in the pathogen population. Several features are said to warn that currently effective specific resistance genes will prove transient; these include:
- Simple inheritance
- Seedling-stage effectiveness
- Hypersensitivity
On the other hand, non-specific resistance has the contrasting features. Most of all, non-specific resistance will be durable, but to demonstrate durability, the resistance source must be widely tested over area and time (g).
Hence our recipe for the prevention of widespread losses as a consequence of rust epidemics is -
- Avoidance of ultra-susceptibility
- Genetic diversity of resistance, either between or within crops
- Use of geneticaily complex resistances
- Use of durable resistance

Stripe Rust

Wheat stripe rust was first found in southeastern Australia in October, 1979. When first reported, it was already widespread, but crop surveys indicated that the outbreak was centred around Chariton in north-central Victoria. The initial outbreak revealed certain significant features. Firstly, crop losses in susceptible cultivars such as Zenith - then the leading cultivar in Victoria - were as high as 50-60%. Losses in less susceptible wheats such as Egret, Halberd and Condor were considerably less. Secondly, uredospores moved over relatively long distances. This was evident not only in 1979, if we assume that as the year of introduction, but again in 1980, when stripe rust appeared in New Zealand, apparently having been windborne from Australia.

When it first appeared, experience and knowledge of this disease were superficial. No suitable temperature-controlled facilities for infection and incubation were available. Moreover, the possibility that the disease might not survive the dry summers put a necessary constraint upon study, especially in laboratories located in wheatgrowing areas. The first preparations for research came with a recommendation by the National Wheat Rust Control Committee that the University of Sydney make special application to Wheat Industry Research Council for funds. Support for a greenhouse facility was provided from reserve funds, and C.R. Wellings was seconded to the program from the N.S.W. Department of Agriculture.
A further recommendation by the Committee was that it would be consistent with the other wheat rust diseases to adopt the American name of stripe rust, rather than the European name, yellow rust. Stripe rust was the adopted name throughout Australia and New Zealand.

Pathotype Analysis

Using the International and European sets of host differentials (10) obtained from R. Johnson, P.B.I., Cambridge, the initial P.striiformis tritici pathotype in Australia was identified as 104 E137. During the late 1970's, pathotype 104 E137 was present in southern and central Europe. We therefore assume that the pathogen was transported to Australia by man, either on his person or his goods.
During its short history in this region, four single-step mutational variants have been identified in Australia, and another has been found in New Zealand (Fig. 5). Clearly, the pathogen has maintained its reputation for variability, despite the absence of a sexual reproductive phase. Although temperatures during the Australian wheat-growing season, i.e., from May until November, are ideal for infection and spread of stripe rust, its greatest barrier to survival is the dry summers. However, experience has shown that the disease does survive at low levels and apparently at more-or-less random locations. Unfortunately, it has not been possible to find infections during summer. While several grasses, as well as barley and triticale, may become infected, survival almost certainly occurs on self-sown wheat.

Stripe Rust Control

John S. Brown and colleagues at Crops Research Institute, Horsham, have studied crop loss and chemical control. They have produced a model suggesting that crop losses are directly related to the amount of disease 2-3 weeks after flowering.

Table 2. Responses of Australian wheat cultivars to P. striiformis tritici pathotype 104 E137A+i based on results from Disease Progress Nurseries and field reports.

Response
Group Cultivar

1 Banks, Bass, Bindawarral, Cook, Corelial, Cranbrook3, Hartog 1 2@ Jacupl, Milingl, Miliewal, Oxiey', Quarrion, Sunkota 2 , Takaril,
Vascol.
2. Blade, Cocamba, Condor, Festiguay, Gatcher 2 4 , Gutha, Kite, Matong,
Meering, Osprey, Sundor, Suneca, Suneig, Sunstar, Torres, Vufcan.
3. Egret, Flinders, Gamenya, Dagger, Halberd, Harrier, Hyden, King, Katyil, Kewell, Machete, Madden, Mokoan, Olympic, Songlen, Warimba, Warigal, Wialki, Wyuna.
4. Aroona, Avocet, Bayonet, Bodailin, Eradu, lsis, Lance, Teal, Tincurrin, Zenith.

1 Yr6.
2 Yr7.
3 Unknown seedling resistance.
4 Genetically heterogeneous for indicated gene.

While chemical control of stripe rust is very effective, the relatively low wheat yields in Australia will usually limit applications to a single foliar spray. In addition, seed dressings give several weeks of post-emergent protection. The farmer must, therefore, have access to reliable information on expected cultivar responses and anticipated losses. Furthermore, he must be alerted as soon as possible in the event of pathotype changes, even though these are not always easily confirmed.
The good news for breeders is the wealth of genetic resistance to stripe rust. The first impression one gains from a stripe rust disease nursery is the continuous range of host responses covering a wide array of germplasm. In addition, many Australian wheats show some resistance. Part of this undoubtedly reflects the widespread use of CIMMYT germplasm, especially in eastern Australia. Interestingly, stripe rust has not become a problem in South Australia and Western Australia, despite the significantly more susceptible cultivars.
For convenience, the first subdivision of resistance types are those occurring at the seedling stage and effective for the entire growth cycle. The second subdivision includes the postseedling types which become effective at various growth stages and may be influenced by temperature. However, under Australian conditions, resistance may not occur early enough to prevent some damage. Because of variable adult plant responses, we have directed considerable effort at placing cultivars into response groups (Table 2).

Implications for Resistance Breeding

In New South Wales in particular, there are increasing demands for wheats with stronger winter growth habit, permitting earlier and more flexible sowing, and possibly higher yields as a consequence. The implications of this in relation to rust are that the oversummer survival period for the pathogen will be shortened and there will be a longer growing season for development of epidemics. To compensate, it may be essential to have more effective resistance in cultivars designated for early sowing.
How do we obtain that resistance? We can try seedling resistance either as single genes, or as gene combinations, but be warned, these genes may have short-term effectiveness.
we can use post-seedling resistances. Again, various sources of this kind of resistance are highly specific whereas other sources have proved durable. The problem is how to distinguish between them, especially in the short term.
In addition to the pragmatic approach of using whatever is currently available, we at Castle Hill, are exploring two breeding approaches based on overseas experience. Studies in the U.S. (8@l 1916) and U.K. (19) suggest that improved levels of resistance can be obtained from intercrosses among moderately resistant and moderately susceptible wheats. Since such transgressive segregation to stripe rust occurs fairly frequently,
Australian wheats, established sources of durable resistance. Unfortunately, most sources are poorly adapted to Australian wheat-growing conditions, and are red seeded and stem rust susceptible.
For the future, there are three immediate areas requiring more information. Firstly, we need to establish how and where oversummering occurs. Secondly, we need to assess seed treatment as a

Fig. 4. Australasian Plant Pathology Vol. 15 (1) 1986

Fig. 5. Australasian Plant Pathology Vol. 15 (1) 1986

it seems reasonable to expect it to occur in intercrosses of adapted Australian cultivars. If it occurs, such resistance should be determined by additive gene effects, and therefore unlikely to be neutralized as a consequence of a single mutational event in the pathogen. The second approach is to transfer., by backcrossing to means of controlling early infection and to develop firmer guidelines for foliar sprays. Thirdly, we need to refine our pathotype surveys in order to more closely monitor adult resistances; currently, only seedling resistances are being monitored.

Conclusion

Throughout this presentation, we have attempted to emphasise the multi-disciplinary problems involved in the study of rust resistance, and in the exploitation of resistance in agriculture. Whereas pathologists such as de Bary, McAlpine, Cobb, Stakman and Flor laid the foundations for host-pathogen genetics, many of the agriculturally significant contributions came from breeder/pathologists including Farrer, Waterhouse, Watson, Caldwell and Borlaug. Today, there is a more-than-ever need for a multi-disciplinary approach involving genetics, pathology and breeding. We believe we are now at the beginning of the most exciting stage in the history of plant pathology. The next great advance will come with the DNA-base sequencing of resistance (and avirulence) genes, with a consequent understanding of what they do and how they do it. Most of all, we should have increased knowledge of how resistance genes might be manipulated or engineered in our constant efforts to keep ahead of rapidly evolving pathogens.
Finally, Mr. Chairman, allow us to express our gratitude to all members of the Castle Hill group who have contributed in various ways to wheat rust research ' Our thanks go to John S. Brown for supplying data and reports from studies in Victoria. In addition, we acknowledge the long-term support that has been provided by Wheat Industry Research Council.

References

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