Differential proteomic analysis of proteins in wheat spikes induced by Fusarium graminearum

Scab, caused by Fusarium graminearum, is a serious spike disease in wheat. To identify proteins in resistant wheat cultivar Wangshuibai induced by F. graminearum infection, proteins extracted from spikes 6, 12 and 24 h after inoculation were separated by 2-DE. Thirty protein spots showing 3-fold change in abundance when compared with treatment without inoculation were characterized by MALDI-TOF MS and matched to proteins by querying the mass spectra in protein databases or the Triticeae EST translation database. Based on their volume profiles, these proteins were classified into four categories. The first one fell off rapidly at the initial inoculation and then rose at 12 or 24 hai, the second one decreased considerably after inoculation and remained at low level, the third one rose at the initial inoculation and then declined at 12 or 24 hai, the forth one showed steady increase after inoculation and maintained at a high level. Many of the proteins identified in the first two categories are related to carbon metabolism and photosynthesis. While most of proteins identified in the last two categories are related to stress defense of plants, indicating that proteins associated with the defense reactions were activated or translated shortly after inoculation.     

The transcriptome of Fusarium graminearum during the infection of wheat

Fusarium graminearum causes head blight disease in wheat and barley. To help understand the infection process on wheat, we studied global gene expression of F. graminearum in a time series from 24 to 196 h after inoculation, compared with a noninoculated control. The infection was rapid and, after 48 h, over 4,000 fungal genes were expressed. The number of genes expressed increased over time up to 96 h (>8,000 genes), and then declined at the 144- and 192-h post-inoculation time points. After subtraction of genes found expressed on complete medium, during carbon or nitrogen starvation, and on barley, only 355 were found exclusively expressed in wheat, mostly genes with unknown function (72.6%). These genes were mainly found in single-nucleotide polymorphism-enriched islands on the chromosomes, suggesting a higher evolutionary selection pressure. The annotated genes were enriched in functional groups predicted to be involved in allantoin and allantoate transport, detoxification, nitrogen, sulfur and selenium metabolism, secondary metabolism, carbohydrate metabolism, and degradation of polysaccharides and ester compounds. Several putative secreted virulence factors were also found expressed in wheat.     

Phases of infection and gene expression of Fusarium graminearum during crown rot disease of wheat

Fusarium graminearum causes head blight (FHB) and crown rot (CR) diseases in wheat. Compared with FHB, CR symptom development occurs slowly, usually taking 4 to 8 weeks to become visible. To characterize CR development, we used histological and real-time quantitative polymerase chain reaction analyses to assess fungal colonization during a timecourse of infection. Three distinct phases of infection were identified: i) initial spore germination with formation of a superficial hyphal mat at the inoculation point, ii) colonization of the adaxial epidermis of the outer leaf sheath and mycelial growth from the inoculation point to the crown, concomitant with a drop in fungal biomass, and iii) extensive colonization of the internal crown tissue. Fungal gene expression was examined during each phase using Affymetrix GeneChips. In total, 1,839 F. graminearum genes were significantly upregulated, including some known FHB virulence genes (e.g., TRI5 and TRI14), and 2,649 genes were significantly downregulated in planta compared with axenically cultured mycelia. Global comparisons of fungal gene expression with published data for FHB showed significant similarities between early stages of FHB and CR. These results indicate that CR disease development involves distinct phases of colonization, each of which is associated with a different fungal gene expression program.     

Overexpression of defense response genes in transgenic wheat enhances resistance to Fusarium head blight

Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum and other Fusarium species, is a major disease problem for wheat production worldwide. To combat this problem, large-scale breeding efforts have been established. Although progress has been made through standard breeding approaches, the level of resistance attained is insufficient to withstand epidemic conditions. Genetic engineering provides an alternative approach to enhance the level of resistance. Many defense response genes are induced in wheat during F. graminearum infection and may play a role in reducing FHB. The objectives of this study were (1) to develop transgenic wheat overexpressing the defense response genes α-1-purothionin, thaumatin-like protein 1 (tlp-1), and β-1,3-glucanase; and (2) to test the resultant transgenic wheat lines against F. graminearum infection under greenhouse and field conditions. Using the wheat cultivar Bobwhite, we developed one, two, and four lines carrying the α-1-purothionin, tlp-1, and β-1,3-glucanase transgenes, respectively, that had statistically significant reductions in FHB severity in greenhouse evaluations. We tested these seven transgenic lines under field conditions for percent FHB disease severity, deoxynivalenol (DON) mycotoxin accumulation, and percent visually scabby kernels (VSK). Six of the seven lines differed from the nontransgenic parental Bobwhite line for at least one of the disease traits. A β-1,3-glucanase transgenic line had enhanced resistance, showing lower FHB severity, DON concentration, and percent VSK compared to Bobwhite. Taken together, the results showed that overexpression of defense response genes in wheat could enhance the FHB resistance in both greenhouse and field conditions.     

The development and differentiation of Gibberella zeae (anamorph: Fusarium graminearum) during colonization of wheat

Worldwide, one of the most devastating pathogens of small grains is the head blight fungus, Gibberella zeae. Ascospore-laden perithecia of this fungus develop on mature cereal crops and crop debris and provide the primary inoculum of the disease. We characterize the process of colonization of wheat tissue that leads to perithecium production. Stems were colonized systemically and extensively following inoculation of the wheat head. Haploid mycelia moved down the vascular system and pith and then colonized the stem tissue radially. Dikaryotic hyphae developed at two distinct stages: in the xylem, in support of radial hyphal growth and in the chloremchyma, in support of perithecium development. Perithecium formation was initiated in association with stomatesand silica cells. Vascular occlusions prevented mycelia from colonizing the stem in 25% of inoculated plants. Implications of these findings are discussed for developing resistant cultivars and improving chemical control of the disease.     

Inhibition of Fusarium graminearum growth and development by farnesol

The isoprenoid farnesol was previously shown to induce morphological features characteristic of apoptosis in the filamentous fungus Aspergillus nidulans. This study demonstrates that under similar liquid media growth conditions, farnesol also triggers apoptosis in the plant pathogenic fungus Fusarium graminearum. However, unlike A. nidulans, F. graminearum spores treated with farnesol exhibited altered germination patterns and most (>60%) lysed upon prolonged exposure. Given the economic importance of F. graminearum as a pathogen of small grains, this study proposes that farnesol may have potential value as an antifungal compound.     

Genomic mapping of defense response genes in wheat

 Defense response (DR) genes are a broad class involved in plant defense. In this study we mapped 36 probes representing seven classes of defense response genes. This collection of probes represents genes involved in the hypersensitive response (HR), pathogenesis-related (PR) genes, genes for the flavonoid metabolic pathway, genes encoding proline/glycine-rich proteins, ion channel regulators, lipoxygenase, lectin, and others. Using nullisomic-tetrasomic lines of ‘Chinese Spring’, we were able to assign at least 167 loci to the 21 chromosomes of wheat. Homoeologous group 7 chromosomes possessed the most DR loci followed by group 2. Sixty-two loci were placed on existing genetic linkage maps of wheat. Map locations indicated that the DR gene loci are not randomly distributed throughout the wheat genome, but rather are located in clusters and/or in distal gene-rich regions of the chromosomes. Knowledge of the chromosomal locations and genome organization of DR genes will be useful for candidate gene analysis of quantitative trait loci. Received: 12 June 1998 / Accepted: 24 July 1998     

两个与盐和赤霉病菌胁迫相关的小麦糖基转移酶基因的克隆与表达

摘要: 在植物体内, 糖基转移酶通过参与多种物质的糖基化而在植物抗逆境方面起着重要作用。为了解小麦糖基转移酶基因响应病原菌和盐胁迫的分子机制, 文章分离了两个小麦糖基转移酶基因(TaUGT1, TaUGT2)。这两个基因均编码496个氨基酸, cDNA序列相似性为90％。它们均含有一个内含子, 分别为335 bp(TaUGT1)和324 bp(TaUGT2)。序列比对分析表明, TaUGT1和TaUGT2与尿苷二磷酸葡萄糖醛酸/尿苷二磷酸葡萄糖转移酶(UDP-glucoronosyl and UDP-glucosyl transferase)基因同源性最高, 且都含有PSPG(Putative secondary plant gly-cosyltransferase)保守结构域。Real-time PCR表达分析显示, TaUGT1受赤霉病菌抑制表达, 而TaUGT2受赤霉病菌诱导表达; 在高浓度NaCl胁迫下, TaUGT1和TaUGT2的相对表达量分别为对照的2.87及4.56倍, 差异达到极显著水平。以上结果表明, TaUGT2可能与小麦赤霉病抗性有关, 而TaUGT1和TaUGT2可能共同参与了小麦对盐胁迫的响应。