Chunji Liu

Pathogen population structure and epidemiology are keys to wheat crown rot and Fusarium head blight management

This paper summarises the key findings from recent research on the population genetics and epidemiology of Fusarium pathogens causing head blight and crown rot of wheat in Australia and how this information has enabled the screening and selection of wheat germplasm with improved resistance to Fusarium. By relating new findings to the current state of knowledge, the paper serves as a timely and critical review of the international literature. In Australia, both Fusarium pseudograminearum and F. graminearum can cause both crown rot and Fusarium head blight under artificial inoculation. However, the former species is more widespread and is predominantly associated with crown rot whereas F. graminearum is mainly associated with Fusarium head blight, with limited geographical distribution in and around the Liverpool Plains in northern New South Wales. Studies of population structure and genetics have revealed that both species are genotypically diverse with similar levels of genetic recombination despite Gibberella zeae, the teleomorph of F. graminearum, being homothallic and G. coronicola, the teleomorph of F. pseudograminearum, being heterothallic. A high-throughput and reliable crown rot bioassay has been developed and used to screen over 1500 wheat germplasms to select 17 lines with putative crown rot resistance. Key differences in pathogen biology and epidemiology between Australia and the USA have emerged from other recent collaborative studies, which show that macroconidia constitute the bulk of aerial Fusarium head blight inoculum in Australia, whereas ascospores are the dominant primary inoculum for Fusarium head blight worldwide. The limited spread of splash-dispersed macroconidia of F. graminearum probably explains the restricted geographical distribution of this species in Australia. Other research collaboration has compared the aggressiveness, mycotoxin production and genotypic polymorphisms of the pathogen population from Australia and the USA. These and other differences in pathogen adaptation emphasise that research outcomes from elsewhere must be tested for relevance before applying them to Australian farming systems.

Identification and validation of a major QTL conferring crown rot resistance in hexaploid wheat

Crown rot (CR), caused by various Fusarium species, is a chronic wheat disease in Australia. As part of our objective of improving the efficiency of breeding CR resistant wheat varieties, we have been searching for novel sources of resistance. This paper reports on the genetic control of one of these newly identified resistant genotypes, ‘CSCR6’. A population derived from a cross between CSCR6 and an Australian variety ‘Lang’ was analyzed using two Fusarium isolates belonging to two different species, one Fusarium pseudograminearum and the other Fusarium graminearum. The two isolates detected QTL with the same chromosomal locations and comparable magnitudes, indicating that CR resistance is not species-specific. The resistant allele of one of the QTL was derived from ‘CSCR6’. This QTL, designated as Qcrs.cpi-3B, was located on the long arm of chromosome 3B and explains up to 48.8% of the phenotypic variance based on interval mapping analysis. Another QTL, with resistant allele from the variety ‘Lang’, was located on chromosome 4B. This QTL explained up to 22.8% of the phenotypic variance. A strong interaction between Qcsr.cpi-3B and Qcsr.cpi-4B was detected, reducing the maximum effect of Qcrs.cpi-3B to 43.1%. The effects of Qcrs.cpi-3B were further validated in four additional populations and the presence of this single QTL reduced CR severity by up to 42.1%. The fact that significant effects of Qcrs.cpi-3B were detected across all trials with different genetic backgrounds and with the use of isolates belonging to two different Fusarium species make it an ideal target for breeding programs as well as for further characterization of the gene(s) involved in its resistance.

Genetic relationships between resistances to Fusarium head blight and crown rot in bread wheat (Triticum aestivum L.)

Fusarium head blight (FHB) and crown rot (CR) are two wheat diseases caused by the same Fusarium pathogens. Progress towards CR resistance could benefit from FHB-resistant germplasm if the same genes are involved in resistance to these two different diseases. Two independent studies were conducted to investigate the relationship between host resistances to these two diseases. In the first study 32 genotypes were assessed and no significant correlation between their reactions to FHB and CR was detected. The second study was based on a QTL analysis of a doubled haploid population derived from a variety with resistance to both diseases. Results from this study showed that loci conferring resistance to FHB and CR are located on different chromosomes. Together, these results suggest that, despite a common aetiology, different host genes are involved in the resistance against FHB and CR in wheat. Thus, although it is possible that genes affecting both diseases may exist in other germplasm or under different conditions, separate screening seems to be needed in identifying sources of CR resistance.

Construction of bacterial artificial chromosome libraries and their application in developing PCR-based markers closely linked to a major locus conditioning bruchid resistance in mungbean (Vigna radiata L. Wilczek)

Bacterial artificial chromosome (BAC) libraries have been widely used in different aspects of genome research. In this paper we report the construction of the first mungbean (Vigna radiata L. Wilczek) BAC libraries. These BAC clones were obtained from two ligations and represent an estimated 3.5 genome equivalents. This correlated well with the screening of nine random single-copy restriction fragment length polymorphism probes, which detected on average three BACs each. These mungbean clones were successfully used in the development of two PCR-based markers linked closely with a major locus conditioning bruchid (Callosobruchus chinesis) resistance. These markers will be invaluable in facilitating the introgression of bruchid resistance into breeding programmes as well as the further characterisation of the resistance locus.

A major QTL conferring crown rot resistance in barley and its association with plant height.

Crown rot (CR) is one of the most destructive diseases of barley and wheat. Fusarium species causing CR survive in crop residue and a growing acceptance of stubble retention practices has exacerbated disease severity and yield loss. Growing resistant cultivars has long been recognised as the most effective way to reduce CR damage but these are not available in barley. In a routine screening of germplasm, a barley landrace from China gave the best CR resistance among the genotypes tested. Using a doubled haploid population derived from this landrace crossed to Franklin, we demonstrate that the CR resistance of TX9425 was conditioned by a major QTL. The QTL, designated as Qcrs.cpi-3H, was mapped near the centromere on the long arm of chromosome 3H. Its effect is highly significant, accounting for up to 63.3% of the phenotypic variation with a LOD value of 14.8. The location of Qcrs.cpi-3H was coincident with a major QTL conferring plant height (PH) and the effect of PH on CR reaction was also highly significant. When the effect of PH was accounted for by covariance analysis, the Qcrs.cpi-3H QTL remained highly significant, accounting for over 40% of the phenotypic variation. The existence of such a major QTL implies that breeding barley cultivars with enhanced CR resistance should be feasible.

Geographical distribution of genetic variation in Stylosanthes scabra revealed by RAPD analysis

A large number of S. scabra accessions have been accumulated worldwide. The majority of them were collected from Brazil and most of the others came from either Colombia or Venezuela. One hundred of these accessions, selected to represent the geographical distribution of the S. scabra collection held at the Australian Tropical Forages Genetic Resource Centre, were analysed using RAPD as markers. Seven of these accessions were found not to be S. scabra. Of the S. scabra accessions, the average dissimilarity value among Brazilian accessions (0.053) was much lower than that among Colombian (0.074) or Venezuelan (0.088) accessions, with an overall dissimilarity value of 0.059 among all the S. scabra accessions. Based on their dissimilarity values, most of these accessions could be separated into five groups. Geographical distributions for most accessions in each of these groups were well defined. Limited long distance introductions/dispersions of S. scabra between these regions were detected and they were mainly confined to Brazilian genotypes. The clustering results based on RAPD were compared with those based on morphological-agronomical characters, and the groups produced by the two different methods did not always match. Possible reasons for this discrepancy are discussed.

Sequence-Based Analysis of Translocations and Inversions in Bread Wheat (Triticum aestivum L.)

Structural changes of chromosomes are a primary mechanism of genome rearrangement over the course of evolution and detailed knowledge of such changes in a given species and its close relatives should increase the efficiency and precision of chromosome engineering in crop improvement. We have identified sequences bordering each of the main translocation and inversion breakpoints on chromosomes 4A, 5A and 7B of the modern bread wheat genome. The locations of these breakpoints allow, for the first time, a detailed description of the evolutionary origins of these chromosomes at the gene level. Results from this study also demonstrate that, although the strategy of exploiting sorted chromosome arms has dramatically simplified the efforts of wheat genome sequencing, simultaneous analysis of sequences from homoeologous and non-homoeologous chromosomes is essential in understanding the origins of DNA sequences in polyploid species.

Development of near-isogenic lines for a major QTL on 3BL conferring Fusarium crown rot resistance in hexaploid wheat

By essentially fixing the genetic background, near-isogenic lines (NILs) are ideal for studies of the function of specific loci. We report in this paper the development of NILs for a major QTL located on the long arm of chromosome 3B conferring Fusarium crown rot (FCR) resistance in hexaploid wheat. These NILs were generated based on the method of the heterogeneous inbred family analysis. 13 heterozygous lines were initially selected from three segregating populations using a single SSR marker linked with the major FCR QTL. The two isolines for each of the putative NILs obtained showed no obvious morphological differences, but differences among the NIL pairs were large. Significant differences in FCR resistance between the isolines were detected for nine of the 13 putative NIL pairs. The presence of the FCR allele from the resistant parent reduced FCR severity by 29.3–63.9% with an average of 45.2% across these NILs. These NILs will be invaluable in further characterising this major FCR locus, in studying the mechanism of FCR resistance and in investigating possible interactions between FCR resistance and other traits of agronomic importance.

Quantitative trait loci for root lesion nematode (Pratylenchus thornei) resistance in Middle-Eastern landraces and their potential for introgression into Australian bread wheat

Plant parasitic nematodes are a major biotic cause of wheat yield loss in temperate wheat-growing regions. Previous studies using Australian germplasm and/or synthetic hexaploid lines have identified quantitative trait loci (QTLs) for root lesion nematode resistance on chromosomes 2B, 6D, and 7A. This study examines Pratylenchus thornei resistance in 2 Middle-Eastern landraces (AUS13124 and AUS4926), using doubled haploid populations generated by crossing with the susceptible Australian cultivar Janz. Single marker regression and QTL analysis identified resistance loci on chromosomes 2B, 3B, 6D, and 7A, and a susceptibility locus on chromosome 1B. The 2B and 6D loci, which have been reported to explain up to 19% and 24% of variation, respectively, in previous studies, made smaller contributions in the Middle-Eastern varieties, explaining 2–13% (2B) and 1–6% (6D) of phenotypic variation in these populations. The previously reported 7A locus (P. neglectus resistance) was detected through single marker regression only (AUS13124 × Janz – LRS = 4.1, P = 0.04292; AUS4926 × Janz – LRS = 9.6, P = 0.00195), with genotype at the microsatellite marker Xgwm350.3 accounting for 3–23% of phenotypic variation. The previously unreported resistance QTL, located on chromosome 3B, explained up to 24% of phenotypic variation, and the susceptibility locus on chromosome 1B explained up to 21%. The 3B locus was detected in both the AUS13124 × Janz (max. LRS = 20.13) and AUS4926 × Janz (max. LRS = 11.19) populations, and the 1B locus was detected in the AUS4926 × Janz population (max. LRS = 18.82) only.

Transcriptome Sequencing of Mung Bean (Vigna radiate L.) Genes and the Identification of EST-SSR Markers

Mung bean (Vigna radiate (L.) Wilczek) is an important traditional food legume crop, with high economic and nutritional value. It is widely grown in China and other Asian countries. Despite its importance, genomic information is currently unavailable for this crop plant species or some of its close relatives in the Vigna genus. In this study, more than 103 million high quality cDNA sequence reads were obtained from mung bean using Illumina paired-end sequencing technology. The processed reads were assembled into 48,693 unigenes with an average length of 874 bp. Of these unigenes, 25,820 (53.0%) and 23,235 (47.7%) showed significant similarity to proteins in the NCBI non-redundant protein and nucleotide sequence databases, respectively. Furthermore, 19,242 (39.5%) could be classified into gene ontology categories, 18,316 (37.6%) into Swiss-Prot categories and 10,918 (22.4%) into KOG database categories (E-value < 1.0E-5). A total of 6,585 (8.3%) were mapped onto 244 pathways using the Kyoto Encyclopedia of Genes and Genome (KEGG) pathway database. Among the unigenes, 10,053 sequences contained a unique simple sequence repeat (SSR), and 2,303 sequences contained more than one SSR together in the same expressed sequence tag (EST). A total of 13,134 EST-SSRs were identified as potential molecular markers, with mono-nucleotide A/T repeats being the most abundant motif class and G/C repeats being rare. In this SSR analysis, we found five main repeat motifs: AG/CT (30.8%), GAA/TTC (12.6%), AAAT/ATTT (6.8%), AAAAT/ATTTT (6.2%) and AAAAAT/ATTTTT (1.9%). A total of 200 SSR loci were randomly selected for validation by PCR amplification as EST-SSR markers. Of these, 66 marker primer pairs produced reproducible amplicons that were polymorphic among 31 mung bean accessions selected from diverse geographical locations. The large number of SSR-containing sequences found in this study will be valuable for the construction of a high-resolution genetic linkage maps, association or comparative mapping and genetic analyses of various Vigna species.