This article appeared in the May 2009 edition of the Kondinin Groups monthly magazine Farming Ahead.

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cropping
Disease control

Gene for wheat rust resistance discovered

Evans Lagudah Jill Griffiths
For CSIRO pLANT INdUSTRY
photos: carl Davies, cSiro

Molecular geneticist: Dr Evans Lagudah, CSIRO Plant Industry, examines wheat breeding lines with the Lr34 gene in the greenhouse. Team shot: Dr Evans Lagudah and his team, Wolfgang Spielmeyer, Libby Viccars, Sutha Chandramahan and Terese Richardson. (Absent: Helen McFadden.)

A gene that gives wheat resistance to rust has been isolated by an international team of scientists, including researchers from cSiro plant industry. The wheat gene known as Lr34 encodes a transporter protein that confers protection against leaf rust, stripe rust and powdery mildew, which in epidemic years cost wheat growers worldwide in excess of $7.8 billion.
In recent years, when wheat production in Australia has been affected by drought, it is estimated that $100 million has been spent on controlling stripe rust. Stripe rust is an aggressive disease; by the time grain is beginning to form, the plants leaves are dead.
evolving to overcome the genetic resistance. The rust pathogen needs living tissue to survive so must evolve to overcome resistance for its survival. From an agricultural perspective, growing crop varieties with genetic resistance is more environmentally friendly and less costly than strategies such as spraying pesticides. But, when crops are produced on a large scale, there is increased pressure on the pathogens to evolve to overcame plant resistance. Plants have two types of genetic resistance seedling resistance and adult plant resistance. Seedling resistance is where the seedling is resistant to the pathogen but the adult plant is not. Seedling resistance provides strong resistance against a limited number of strains of disease. In a commercial crop, seedling resistance generally only lasts about 35 years before new pathogen strains appear. Adult plant resistance, such as Lr34, lasts longer. Adult resistance operates past the seedling stage and is more evident in the plants reproductive phase when grain is forming. Lr34 confers this type of resistance and provides partial resistance against all strains of leaf and stripe rust.

At a glance
CSIRO researchers, in collaboration with the University of Zurich and CIMMYT, have discovered a wheat gene sequence that confers protection against leaf rust, stripe rust and powdery mildew. Controlling rust pathogens costs dearly. It is estimated that, in Australia, $100 million is spent annually controlling stripe rust alone. The gene, known as Lr34, occurs in many wheat varieties grown in Victoria, New South Wales and Queensland. Farmers can expect to see more wheat varieties carrying Lr34 rust resistance in the future.

Leaf rust, stripe (or yellow) rust and powdery mildew threaten the productivity and profitability of wheat crops around the world. All three diseases are caused by fungal pathogens.
The fungi that cause rust diseases are very adaptable and can rapidly evolve to overcome resistant cereal varieties. Commonly, scientists and farmers can only respond to a rust outbreak after it has passed. New resistant cereal varieties and new sources of rust resistant genes are continually sought to combat the latest rust fungi threat.

In-built defence mechanisms Plants are unable to move away from threats and need built-in mechanisms called genetic resistance to fight diseases. Fungal pathogens and plants co-evolve, in an on-going arms race. As the plants build resistance, the fungal pathogens continue

Just what is Lr34? For some time, scientists have been aware that some wheat varieties have genetic resistance to leaf rust, stripe rust and powdery mildew. Some wheat varieties were 43

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the research increased, happily resulting in recent success.

When cloning Lr34, it was discovered that there were not three separate genes but a single gene, providing resistance against three different pathogens.
This enabled them to obtain the exact sequence of this gene. Lr34 codes for a protein called an ABC transporter. This type of protein, as the name suggests, transports molecules across cell membranes. The transporter produced by Lr34 may transport anti-microbial compounds that provide a defence against leaf rust, stripe rust and powdery mildew.

photo: rp Singh, ciMMYT, Mexico

Field site in Mexico: The four rows on the left are wheat plants with the Lr34 gene, which have clearly been protected from the severe effects of leaf rust infection in contrast to the four rows on the right which lack the Lr34 gene.

known to show adult plant resistance to leaf rust. The gene conferring this resistance was designated Lr34 (leaf rust gene 34). It was noticed that these plants also carried adult resistance to stripe or yellow rust. This gene was called Yr18 (yellow rust 18). Later, researchers observed that the wheat that carried these genes was also resistant to powdery mildew. That gene was called Pm38. The location and sequences of these genes was unknown and scientists in various parts of the world set out to find, clone and use them. It was assumed that three genes were involved, each giving resistance to a single pathogen.

Although the approximate location of the gene was previously known, Dr Lagudah and his collaborators have now been able to refine the location of Lr34 on the wheat genome.
Worldwide, researchers set out to find and sequence this valuable gene. The field of scientists working on it dwindled over the years as the search was difficult and seemed fruitless. Three teams one at the University of Zurich, one at CSIRO Plant Industry and one at CIMMYT, Mexico continued the search and decided to collaborate in sequencing the Lr34 gene region. With funding by the Grains Research and Development Corporation, the collaboration led to the recent discovery of the location and sequence of Lr34.

Viccars, Sutha Chandramohan, Helen McFadden, Terese Richardson, in collaboration with the University of Zurich and the International Maize and Wheat Improvement Centre (CIMMYT), have identified the wheat gene, which provides resistance against leaf rust, stripe rust and powdery mildew. The gene, called Lr34, is the first of its kind to be identified in a commercial crop rather than a model plant. Model plants are plants with simple genomes that scientists study to gain broad understandings of the way plant genetics work, in much the way medical researchers study rats and mice to gain broad understandings of the way drugs may affect people. Model plants have given scientists knowledge on transporter proteins that may help with the work on wheat. Usually one gene protects against only one disease, but tests conducted after identifying the Lr34 gene sequence showed that it has provided partial but constant protection against leaf rust for more than 80 years.

Lr34 is durable Lr34 owes its longevity to the fact that it confers partial resistance, which means less evolutionary pressure comes into play. In effect, the arms race is scaled down. Lr34 allows the rust to develop slowly; by the time the wheat has matured, rust levels are building up, but do not pose a significant threat to production. At the end of the season, the wheat leaves are dying off; rust only grows on living tissue, so it too is at the end of its life-cycle. Lr34 does not kill off the pathogen completely; rather it slows its growth creating an unfavourable environment for it. Durable disease resistance that defends a plant against multiple threats from within is highly desirable in commercial crops like wheat. Plant breeders strive to combine several adult plant resistance genes together to give a plant full protection to a disease. Improved control of fungal rust diseases in cereal crops is critical not only to the financial survival of Australian farmers, but also to food security worldwide. Identifying varieties with Lr34 Under certain conditions the tip of the leaves of plants with Lr34 die back, or become necrotic, in a process called leaf tip necrosis. But this is not always a reliable indicator of the presence of Lr34. The discovery of the Lr34 sequence has now given researchers a tool to use to confirm the genes presence. It is a rapid diagnostic test and can be performed on seed, giving plant breeders a huge boost in establishing which plants will have Lr34 genetic resistance to rust. Understanding the molecular nature of this type of resistance has important implications for long-term control of rust diseases. Origins and future of Lr34 in wheat The identification of Lr34 is intriguing because it has established itself as a durable and long-lasting defence mechanism found in wheat crops from around the world.

One gene, multiple defences CSIRO Plant Industrys Dr Evans Lagudah and his team, Wolfgang Spielmeyer, Libby

Finding the new gene The wheat genome is large and complicated making specific gene identification difficult. The team successfully identified the location of Lr34 using the latest genetic techniques. Researchers took large stretches of DNA, which they cloned and sequenced. Comparisons were made between stocks known to carry Lr34 and those known not to have it. Mutated stock was also used, with researchers looking for the changes in the sequence on mutations that had lost resistance. It is a long and painstaking process. The idea of cloning Lr34 was first conceived during 1997. During the past five years, with funding from GRDC the pace of

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After identifying the Lr34 gene sequence, the research team searched wheat lines from around the world and found Lr34 in the earliest cross-bred varieties first released in Italy during the beginning of last century.

in Australia, Lr34 occurs in south-east Australian wheats, but not in commercial cultivars released in South Australia or Western Australia.
This discrepancy in the occurrence of Lr34 is caused by the variety of sources cereal breeders use to obtain germ plasm for breeding programmes. A large proportion of the varieties that come from CIMMYT origins carry Lr34. These varieties have traditionally been used to breed wheat varieties for Queensland and NSW. Similarly, Condor-derived varieties, which have been popular in Victoria, contain Lr34. But in Western Australia and South Australia, other germ plasm has been used and Lr34 is not widely found. Plant breeders will now be able to produce wheat varieties adapted for WA and SA that carry Lr34 by using the DNA sequence to select plants that retain the Lr34 gene sequence from crosses between Lr34 germplasm with WA and SA varieties.

Future research will focus on how Lr34 works to confer resistance to disease. In the meantime, current research findings have been shared with WA and SA scientists and are being used towards incorporating Lr34 resistance into wheat varieties in those states. Some varieties in eastern Australia have Lr34. Now, wheats will be bred specifically to contain Lr34. In WA the newly released wheat varieties GBA Shenton and GBA Sapphire are predicted to carry Lr34. In the future, farmers can expect to see an increase in varieties with Lr34.
AcknowledgeMenTs This work was supported in Australia by the grains Research and development corporation. The lr34 research was published in the journal science on 19 February 2009. Reference: krattinger sg, lagudah es, spielmeyer w, singh RP, Huerta-espino J, McFadden H, Bossolini e, selter ll, keller B 2009. A Putative ABc Transporter confers durable Resistance to Multiple Fungal Pathogens in wheat.

CSIRO research
conTAcT dr evans lagudah (02) 6246 5392 evans.lagudah@csiro.au www.csiro.au/pi

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