Susceptible Isolines Research Articles

Proteomic analysis of near-isogenic lines reveals key biomarkers on wheat chromosome 4B conferring drought tolerance.

Drought is a major constraint for wheat production that is receiving increased attention due to global climate change. This study conducted isobaric tags for relative and absolute quantitation proteomic analysis on near-isogenic lines to shed light on the underlying mechanism of qDSI.4B.1 quantitative trait loci (QTL) on the short arm of chromosome 4B conferring drought tolerance in wheat. Comparing tolerant with susceptible isolines, 41 differentially expressed proteins were identified to be responsible for drought tolerance with a p-value of < 0.05 and fold change >1.3 or <0.7. These proteins were mainly enriched in hydrogen peroxide metabolic activity, reactive oxygen species metabolic activity, photosynthetic activity, intracellular protein transport, cellular macromolecule localization, and response to oxidative stress. Prediction of protein interactions and pathways analysis revealed the interaction between transcription, translation, protein export, photosynthesis, and carbohydrate metabolism as the most important pathways responsible for drought tolerance. The five proteins, including 30S ribosomal protein S15, SRP54 domain-containing protein, auxin-repressed protein, serine hydroxymethyltransferase, and an uncharacterized protein with encoding genes on 4BS, were suggested as candidate proteins responsible for drought tolerance in qDSI.4B.1 QTL. The gene coding SRP54 protein was also one of the differentially expressed genes in our previous transcriptomic study.

Identification of KASP markers and putative genes for pre-harvest sprouting resistance in common wheat (Triticum aestivum L.)

Common wheat (Triticum aestivum L.) is the most important crop in the world and a typical allopolyploid with a large and complex genome. Pre-harvest sprouting (PHS) leads to a significant reduction in grain quality worldwide. PHS is a complex trait with related QTL located on different chromosomes. However, the study of markers and genes related to PHS resistance is limited especially for white-grained wheat. Four pairs of near isogenic lines (NILs) from a white-grained wheat cross of Chara × DM5637B*8 targeting a major QTL for PHS resistance (Qphs.ccsu-3A.1) on wheat chromosme 3AL were genotyped using the 90K SNP Illumina iSelect array. Ten SNPs were identified, with a 75%–100% consistency between genotype and phenotype in the resistant or susceptible isolines. The 10 SNPs were converted to cost-effective kompetitive allele-specific PCR (KASP) markers. Screening of 48 wheat cultivars with different phenotypes of PHS identified four KASP markers with 81.3%–85.4% conformity between genotype and phenotype. Further investigation revealed that the four SNPs (BS00022245_51, Kukri_c49927_151, BS00022884_51 and BS00110550_51) corresponding to the four validated KASP markers are residing in three independent genes (TraesCS3A03G1072800, TraesCS3A03G1072400, TraesCS3A03G1071800) close to each other with a distance of 4.28–4.48 Mb to the targeted QTL. These three annotated genes have potential functions related to PHS resistance. Our study revealed that combined use of NILs and the 90K SNP chip is a powerful approach for developing KASP markers and mining functional genes in wheat. The KASP markers for PHS resistance on chromosome 3AL are useful for high-throughput evaluation and marker-assisted selection, and the three identified genes could lead to a better understanding of the genetic pathways controlling PHS.

Transcriptome Analyses of Near Isogenic Lines Reveal Putative Drought Tolerance Controlling Genes in Wheat.

Drought stress, especially at the grain-filling stage, is a major constraint for wheat production. Drought tolerance is a complex trait controlled by a large array of genes and pathways. This study conducted gene expression profiling on two pairs of near-isogenic lines (NILs) for an important qDSI.4B.1 QTL conferring drought tolerance on the short arm of chromosome 4B in wheat. Analysis showed 1,614 genome-wide differentially expressed genes (DEGs) between the tolerant and susceptible isolines in both NIL pairs. Six common DEGs were found between NIL1 and NIL2 at both 7 and 14 days after stress induction, with two of them having single nucleotide polymorphism (SNP) variants. These six genes that were confirmed by quantitative real-time PCR (qRT-PCR) expression analysis are considered candidate genes for drought tolerance mediated by qDSI.4B.1 QTL with their main contributions to gene regulation, cell elongation, protein quality control, secondary metabolism, and hormone signaling. These six candidate genes and the highest number of DEGs and variants (SNPs/indels) were located between 49 and 137 Mbp of 4BS, making this interval the most probable location for the qDSI.4B.1 locus. Additionally, 765 and 84 DEGs were detected as responsive genes to drought stress in tolerant and susceptible isolines, respectively. According to gene ontology (GO), protein phosphorylation, oxidation reduction, and regulation of transcription were top biological processes involved in the drought response and tolerance. These results provide insights into stress responses regulated by the 4BS locus and have identified candidate genes and genetic markers that can be used for fine mapping of the qDSI.4B.1 locus and, ultimately, in wheat breeding programs for drought tolerance.

Comparing transcriptional responses to Fusarium crown rot in wheat and barley identified an important relationship between disease resistance and drought tolerance

BackgroundFusarium crown rot (FCR) is a chronic disease in cereal production worldwide. The impact of this disease is highly environmentally dependant and significant yield losses occur mainly in drought-affected crops.ResultsIn the study reported here, we evaluated possible relationships between genes conferring FCR resistance and drought tolerance using two approaches. The first approach studied FCR induced differentially expressed genes (DEGs) targeting two barley and one wheat loci against a panel of genes curated from the literature based on known functions in drought tolerance. Of the 149 curated genes, 61.0% were responsive to FCR infection across the three loci. The second approach was a comparison of the global DEGs induced by FCR infection with the global transcriptomic responses under drought in wheat. This analysis found that approximately 48.0% of the DEGs detected one week following drought treatment and 74.4% of the DEGs detected three weeks following drought treatment were also differentially expressed between the susceptible and resistant isolines under FCR infection at one or more timepoints. As for the results from the first approach, the vast majority of common DEGs were downregulated under drought and expressed more highly in the resistant isoline than the sensitive isoline under FCR infection.ConclusionsResults from this study suggest that the resistant isoline in wheat was experiencing less drought stress, which could contribute to the stronger defence response than the sensitive isoline. However, most of the genes induced by drought stress in barley were more highly expressed in the susceptible isolines than the resistant isolines under infection, indicating that genes conferring drought tolerance and FCR resistance may interact differently between these two crop species. Nevertheless, the strong relationship between FCR resistance and drought responsiveness provides further evidence indicating the possibility to enhance FCR resistance by manipulating genes conferring drought tolerance.

The auxin response factor gene family in wheat (Triticum aestivum L.): Genome-wide identification, characterization and expression analyses in response to leaf rust

Auxin response factors (ARFs) are plant-specific transcription factors that bind explicitly to Auxin Response Elements and mediate downstream expression of auxin responsive genes involved at various stages of plant growth and development. This study identified 52 ARF genes located within the wheat genome. Based on phylogenetic analysis, the identified ARF proteins of wheat were classified into four classes (class I, II, III and IV). Synteny analysis revealed the orthologous relation with other monocots. The maximum numbers of predicted ARF genes were mapped on wheat chromosome 3B Ten conserved motifs of ARF was determined on all the identified sequences. Based on amino acid composition of the middle region of the ARF proteins, nine ARF proteins were transcription activators and the remaining were transcription repressors. The ARF genes of wheat contained 1–17 introns and deduced proteins of the genes having similar pattern of intron-exon distribution clustered within the same clade in the phylogenetic tree. TaARF1, TaARF4, TaARF7, TaARF34, TaARF39 were predicted to be targeted by the stress responsive miRNA Tae-miRNA160. The identified proteins were mostly localized within the nucleus and had sequence specific DNA binding activity. They were annotated in regulation of expression of early auxin responsive genes in auxin signaling pathway. Thirty-one TaARF proteins interacted and exhibited co-expressed regulation with other TaARF and Aux/IAA proteins. Increased expression of TaARF1 and TaARF23 at 120 h post inoculation (hpi) and 72 hpi in resistant and susceptible isolines respectively, specifies their roles in providing better resistance to the plant during leaf-rust infection.

Insights of Lr28 mediated wheat leaf rust resistance: Transcriptomic approach

Leaf rust is a fungal disease that causes severe yield losses in wheat. Resistant varieties with major and quantitative resistance genes are the most effective method to control the disease. However, the main problem is inadequate information for understanding resistance mechanism and its usefulness. This paper presents Lr28 mediated genome-wide response of known and unknown genes during wheat–Puccinia triticina interaction. In this study, we prepared Serial Analysis of Gene Expression (SAGE) libraries using seedling wheat mRNA for infected and mock conditions. The libraries were sequenced on Sequencing by Oligonucleotide Ligation and Detection (SOLiD) system generating 37–48 million reads. After mapping and gene expression analysis of ~6–12 million trimmed reads/library, we revealed five major categories comprised of Lr28 controlled transcripts in resistant (+Lr28) isoline (39), transcripts specific to susceptible (−Lr28) isoline (785), transcripts specific to hypersensitive-response (HR) (375), transcripts specific for basal-defense (153) and transcripts for establishment of pathogen (1616). We estimated the impact of specific genes and pathways through mapping on plant resistant gene database (PRGdb), reactive oxygen species (ROS) and phytohormone database. Functional annotation results revealed, receptor binding, homeostatic processes and cytoskeletal components as the major discriminating factors between susceptibility and resistance. We validated 28 key genes using qRT-PCR and found positive results. These findings were projected on hypothetical interaction model to demonstrate interaction mechanism. The study might have significant impact on future rust-resistance breeding through knowledge based smart genetic selection of quantitative resistance genes besides major effect R-gene.

Identification and molecular characterization of a trans-acting small interfering RNA producing locus regulating leaf rust responsive gene expression in wheat (Triticum aestivum L.).

A novel leaf rust responsive ta-siRNA-producing locus was identified in wheat showing similarity to 28S rRNA and generated four differentially expressing ta-siRNAs by phasing which targeted stress responsive genes. Trans-acting-small interfering RNAs (Ta-siRNAs) are plant specific molecules generally involved in development and are also stress responsive. Ta-siRNAs identified in wheat till date are all responsive to abiotic stress only. Wheat cultivation is severely affected by rusts and leaf rust particularly affects grain filling. This study reports a novel ta-siRNA producing locus (TAS) in wheat which is a segment of 28S ribosomal RNA but shows differential expression during leaf rust infestation. Four small RNA libraries prepared from wheat Near Isogenic Lines were treated with leaf rust pathogen and compared with untreated controls. A TAS with the ability to generate four ta-siRNAs by phasing events was identified along with the microRNA TamiR16 as the phase initiator. The targets of the ta-siRNAs included α-gliadin, leucine rich repeat, trans-membrane proteins, glutathione-S-transferase, and fatty acid desaturase among others, which are either stress responsive genes or are essential for normal growth and development of plants. Expression of the TAS, its generated ta-siRNAs, and their target genes were profiled at five different time points after pathogen inoculation of susceptible and resistant wheat isolines and compared with mock-inoculated controls. Comparative analysis of expression unveiled differential and reciprocal relationship as well as discrete patterns between susceptible and resistant isolines. The expression profiles of the target genes of the identified ta-siRNAs advocate more towards effector triggered susceptibility favouring pathogenesis. The study helps in discerning the functions of wheat genes regulated by ta-siRNAs in response to leaf rust.

Uncovering leaf rust responsive miRNAs in wheat (Triticum aestivum L.) using high-throughput sequencing and prediction of their targets through degradome analysis.

Deep sequencing identified 497 conserved and 559 novel miRNAs in wheat, while degradome analysis revealed 701 targets genes. QRT-PCR demonstrated differential expression of miRNAs during stages of leaf rust progression. Bread wheat (Triticum aestivum L.) is an important cereal food crop feeding 30% of the world population. Major threat to wheat production is the rust epidemics. This study was targeted towards identification and functional characterizations of micro(mi)RNAs and their target genes in wheat in response to leaf rust ingression. High-throughput sequencing was used for transcriptome-wide identification of miRNAs and their expression profiling in retort to leaf rust using mock and pathogen-inoculated resistant and susceptible near-isogenic wheat plants. A total of 1056 mature miRNAs were identified, of which 497 miRNAs were conserved and 559 miRNAs were novel. The pathogen-inoculated resistant plants manifested more miRNAs compared with the pathogen infected susceptible plants. The miRNA counts increased in susceptible isoline due to leaf rust, conversely, the counts decreased in the resistant isoline in response to pathogenesis illustrating precise spatial tuning of miRNAs during compatible and incompatible interaction. Stem-loop quantitative real-time PCR was used to profile 10 highly differentially expressed miRNAs obtained from high-throughput sequencing data. The spatio-temporal profiling validated the differential expression of miRNAs between the isolines as well as in retort to pathogen infection. Degradome analysis provided 701 predicted target genes associated with defense response, signal transduction, development, metabolism, and transcriptional regulation. The obtained results indicate that wheat isolines employ diverse arrays of miRNAs that modulate their target genes during compatible and incompatible interaction. Our findings contribute to increase knowledge on roles of microRNA in wheat-leaf rust interactions and could help in rust resistance breeding programs.

Cell death triggering and effector recognition by Sw-5 SD-CNL proteins from resistant and susceptible tomato isolines to Tomato spotted wilt virus.

SummaryOnly a limited number of dominant resistance genes acting against plant viruses have been cloned, and further functional studies of these have been almost entirely limited to the resistance genes Rx against Potato virus X (PVX) and N against Tobacco mosaic virus (TMV). Recently, the cell‐to‐cell movement protein (NSM) of Tomato spotted wilt virus (TSWV) has been identified as the avirulence determinant (Avr) of Sw‐5b‐mediated resistance, a dominant resistance gene which belongs to the class of SD‐CC‐NB‐LRR (Solanaceae domain‐coiled coil‐nucleotide‐binding‐leucine‐rich repeat, SD‐CNL) resistance genes. On transient expression of the NSM protein in tomato and transgenic Nicotiana benthamiana harbouring the Sw‐5b gene, a hypersensitive cell death response (HR) is triggered. Here, it is shown that high accumulation of the Sw‐5b protein in N. benthamiana leaves, achieved by co‐expression of the Sw‐5b protein with RNA silencing suppressors (RSSs), leads to auto‐activity in the absence of NSM. In a similar approach, Sw‐5a, the highest conserved paralogue of Sw‐5b from Solanum peruvianum, also triggered HR by auto‐activation, whereas the highest conserved orthologue from susceptible S. lycopersicum, named Sw‐5aS, did not. However, neither of the last two homologues was able to trigger an NSM‐dependent HR. Truncated and mutated versions of these Sw‐5 proteins revealed that the NB‐ARC [nucleotide‐binding adaptor shared by Apaf‐1 (from humans), R proteins and CED‐4 (from nematodes)] domain is sufficient for the triggering of HR and seems to be suppressed by the SD‐CC domain. Furthermore, a single mutation was sufficient to restore auto‐activity within the NB‐ARC domain of Sw‐5aS. When the latter domain was fused to the Sw‐5b LRR domain, NSM‐dependent HR triggering was regained, but not in the presence of its own Sw‐5aS LRR domain. Expression analysis in planta revealed a nucleocytoplasmic localization pattern of Sw‐5b, in which the SD‐CC domain seems to be required for nuclear translocation. Although the Sw‐5 N‐terminal CC domain, in contrast with Rx, contains an additional SD, most findings from this study support a conserved role of domains within NB‐LRR (NLR) proteins against plant viruses.

Near-isogenic lines developed for a major QTL on chromosome arm 4HL conferring Fusarium crown rot resistance in barley

Ten pairs of near-isogenic lines (NILs) were developed for a major quantitative trait locus conferring Fusarium crown rot (FCR) resistance in barley. The locus locates on the long arm of chromosome 4H and the presence of the resistance allele reduced FCR severity by between 32.8 and 63.4 % with an average of 43.9 % across these NILs. Histological and quantitative PCR analyses all confirmed that the rates of Fusarium infection and disease development were much slower in the resistant isolines compared with those in the susceptible isolines. The contrasting phenotypes in FCR severity between the isolines for each of these NIL pairs would facilitate efforts in cloning and functional analyses of genes conferring resistance to this disease.

Transcriptome and allele specificity associated with a 3BL locus for Fusarium crown rot resistance in bread wheat.

Fusarium pathogens cause two major diseases in cereals, Fusarium crown rot (FCR) and head blight (FHB). A large-effect locus conferring resistance to FCR disease was previously located to chromosome arm 3BL (designated as Qcrs-3B) and several independent sets of near isogenic lines (NILs) have been developed for this locus. In this study, five sets of the NILs were used to examine transcriptional changes associated with the Qcrs-3B locus and to identify genes linked to the resistance locus as a step towards the isolation of the causative gene(s). Of the differentially expressed genes (DEGs) detected between the NILs, 12.7% was located on the single chromosome 3B. Of the expressed genes containing SNP (SNP-EGs) detected, 23.5% was mapped to this chromosome. Several of the DEGs and SNP-EGs are known to be involved in host-pathogen interactions, and a large number of the DEGs were among those detected for FHB in previous studies. Of the DEGs detected, 22 were mapped in the Qcrs-3B interval and they included eight which were detected in the resistant isolines only. The enrichment of DEG, and not necessarily those containing SNPs between the resistant and susceptible isolines, around the Qcrs-3B locus is suggestive of local regulation of this region by the resistance allele. Functions for 13 of these DEGs are known. Of the SNP-EGs, 28 were mapped in the Qcrs-3B interval and biological functions for 16 of them are known. These results provide insights into responses regulated by the 3BL locus and identify a tractable number of target genes for fine mapping and functional testing to identify the causative gene(s) at this QTL.

Effect of Soybean Varieties on Survival and Fecundity of Western Corn Rootworm

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a major pest of corn (Zea mays L.) in North America and has evolved resistance to crop rotation by ovipositing in alternate crops such as soybeans [Glycine max (L.) Merr.]. Through experiments with plants grown in the greenhouse and the field, we tested whether soybeans with resistance to the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), affected survival, fecundity, and consumption of soybean for D. v. virgifera. Soybean varieties tested included those types resistant to A. glycines (Rag1 and rag1/rag3) and a susceptible near isoline of the Rag1 variety. Females were provided with a diet of corn tissue for 4 d after which they were fed a diet of tissue from one of three soybean varieties for 4 d, starved for 4 d, or fed corn tissue. When fed greenhouse grown plants, strains differed significantly in survival and consumption, but consumption did not differ by variety of soybean. Diet treatment only affected fecundity; individuals fed corn continuously had greater fecundity than those individuals fed soybeans. In the experiment with plants grown in the field, leaf consumption differed among strains and individuals fed corn continuously had greater fecundity than the other treatments. Soybean varieties with Rag1 and rag1/rag3 resistance to A. glycines did not appear to affect the fitness of D. v. virgifera. Thus, planting of these A. glycines-resistant soybean varieties should not directly affect the spread of rotation-resistant D. v. virgifera.

Tolerance of banana for fusarium wilt is associated with early H 2 O 2 accumulation in the roots

Banana plants derived from a tissue culture process possess a high rate of random variations that were widely used as popular cultivars due to the new desired traits. In this study, two near-isogenic lines, one susceptible (parental Williams-8818) and the other resistant (somaclonal variation progeny Williams-8818-1 from Williams-8818) to Fusarium oxysporum f. sp. Cubense (Foc4), were inoculated with race 4 of F. oxysporum (Fox). Production of O2 • , H2O2 and MDA, as well as changes in enzyme activities, and transcript levels of SOD and CAT in root extracts were monitored every 24 h over 4 days. The histochemical location of H2O2 was also detected. In the resistant iso-line, the accumulation of O2 • and H2O2, and the activation of SOD occurred in the first 24 h, but activation of CAT reached its maximum only after 48 h. All changes were generally lower in the susceptible iso-line when compared to the resistant iso-line. SOD transcripts were further up-regulated until 72 h in the resistant iso-line, but not in the susceptible iso-line. CAT expression was not affected in any of the two iso-lines. This suggests that expressions of the two key genes in the antioxidant system are less suitable indicators for Foc resistance in banana. In contrast, the first “oxidative burst” is a better indicator for different susceptibility of banana to Foc.

Breeding of rutabaga (Brassica napus var. napobrassica L. Reichenb.) based on biomarker selection for root maggot resistance (Delia radicum L.)

Cabbage root maggot (Delia radicum) is the most devastating and persistent pest for rutabaga (Brassica napus var. napobrassica) in all production areas in Canada. With the deregistration of terbufos (Counter®) to combat maggot attack, only chlorpyrifos (Lorsban®), an organophosphorous pesticide, remains and extensive use could lead to insecticide resistance. An unprotected crop would lead to serious domestic and export losses. Root maggot resistance from canola, that originated from the weedy crucifer, Sinapis alba, was transferred to rutabaga by standard hand crossing. A population of doubled haploids was developed from the F1s and screened in a high pressure root maggot rutabaga production field. Resistant and susceptible isolines were identified from different crossing groups and these isoline pairs were used to develop a biochemical selection protocol based on HPLC profiles where glucosinolates can be present as an aid to resistance breeding. Fourteen peaks in the HPLC profile were identified as markers and predictably varied between the more resistant and more susceptible lines. The 3–4 leaf stage was identified as the ideal stage for tissue extraction for profiling which is close to the stage when gravid female maggot flies seek host plants for oviposition utilizing olfactory signals from the host. Olfactory signals for Delia commonly are isothiocyanates which are volatile break down products of glucosinolates. The peaks in the HPLC profiles identified as markers for resistance contain glucosinolates and may be partially responsible for the plant-insect interaction. A predictive model is proposed as an aid to breeders for the development of root maggot resistant rutabaga lines.

Development of a locus-specific, co-dominant SCAR marker for assisted-selection of the Sw-5 (Tospovirus resistance) gene cluster in a wide range of tomato accessions

The best levels of broad-spectrum Tospovirus resistance reported in tomatoes thus far are conferred by the Sw-5 locus. This locus contains at least five paralogues (denoted Sw-5a through Sw-5e), of which Sw-5b represents the actual resistance gene. Here we evaluated a panel of seven PCR primer pairs matching different sequences within a genomic region spanning the Sw-5a and Sw-5b gene cluster. Primer efficiency evaluation was done employing tomato isolines with and without the Sw-5 locus. One primer pair produced a single and co-dominant polymorphism between susceptible and resistant isolines. Sequence analysis of these amplicons indicated that they were specific for the Sw-5 locus and their differences were due to insertions/deletions. The polymorphic SCAR amplicon encompass a conserved sequence of the promoter region of the functional Sw-5b gene, being located in the position −31 from its open reading frame. This primer pair was also evaluated in field assays and with a collection of accessions known to be either susceptible or resistant to tospoviruses. An almost complete correlation was found between resistance under greenhouse/field conditions and the presence of the marker. Therefore, this primer pair is a very useful tool in marker-assisted selection systems in a large range of tomato accessions.

Direct effects of soybean varietal selection and Aphis glycines-resistant soybeans on natural enemies

The direct effects of three soybean parentages, each represented by an Aphis glycines-resistant and susceptible isoline, on the fitness and performance of two key predators (Orius insidiosus and Harmonia axyridis) were evaluated in the laboratory. Predators were reared from hatch through adulthood in Petri dishes with cut trifoliolate leaves of the designated soybean variety, using eggs of Ephestia kuehniella as surrogate prey to eliminate prey-mediated effects of the host plant. Preimaginal survival and development, sex ratio, adult longevity, fecundity, and size were compared among treatments and a no-plant control. An additional experiment compared life-history parameters of predators caged with soybean versus Ipomoea hederacea (ivyleaf morning glory). Aphid resistance reduced the adult longevity of H. axyridis, but O. insidiosus was unaffected by resistance traits. However, adult O. insidiosus lived longer on soybeans with Group C base genetics than the other soybean varieties. Other parameters were not affected by soybean base genetics or resistance, but both predators generally performed worse on soybean than on I. hederacea or no-plant controls. The results suggest that soybean varietal selection, particularly with respect to A. glycines-resistance, may directly affect biological control agents. Also, implications of the generally poor suitability of soybean for natural enemies are discussed within the context of current crop production practices.

P-01 Hormones and Sexuality

The infection process was compared in interactions between Erysiphe graminis f.sp. hordei isolate A6 and two near-isogenic barley lines (the susceptible line P-01 and the resistant line P-02), and the non-host wheat. A low frequency of successful infections, measured as the percentage of germinated conidia forming primary haustoria, was detected in both susceptible (7·5%), and resistant (73%) barley isolines and wheat plants (223%). In all three types of plants, papilla formation around the sites of attempted penetration seems to be a general resistance factor and thus not the determining step for distinguishing between race-specific and non-host resistance. In wheat plants, further resistance was manifested early after penetration and characterized by hypersensitively reacting (HR) single cells, no or limited growth of elongating secondary hyphae and no visible sign of infection. The race-specific resistance of barley isoline P-02 occurred later after penetration, and was characterized by plant tissue necrosis and no or weak fungal sporulation. In the susceptible barley isoline P-01, successful infections resulted in heavily sporulating powdery mildew colonies. Putative guanidine compounds, indicated by the Sakaguchi test, occurred in papillae 4–5 h after inoculation and subsequently in HR cells too, and are apparently associated with the very early resistance responses in barley and wheat. The highly basic guanidine compounds, not identified previously in plant defence responses, were found to be bound to the cell wall, and referred to as “basic staining material”. Accumulation of polyphenolic substances and callose were demonstrated in papillae and HR cells of both barley and wheat. Lignification occurred in wheat, but did not occur or did so only minimally in barley.

The Rice Bacterial Blight Resistance Gene xa5 Encodes a Novel Form of Disease Resistance

The rice xa5 gene for disease resistance to Xanthomonas oryzae pv. oryzae has been positionally cloned and encodes the gamma subunit of transcription factor IIA (TFIIAgamma). TFIIAgamma is a general eukaryotic transcription factor with no previously known role in disease resistance. xa5 is unusual in that it is recessive and does not conform to one of the typical resistance gene structural classes. Sequencing of TFIIAgamma in resistant and susceptible isolines revealed two nucleotide substitutions resulting in an amino acid change between resistant and susceptible cultivars. This association was conserved across 27 resistant and nine susceptible rice lines in the Aus-Boro group.

Tolerance to a non-host isolate of Verticillium dahliae in tomato

Tolerance to Verticillium spp. is a condition in which a host plant develops few symptoms despite substantial colonization by the pathogen. In the present paper we have shown that Craigella tomatoes are tolerant to a non-host isolate of V. dahliae, Dvd E6. Symptom expression was used to quantify disease and quantitative PCR to assess the amount of fungus in the stems. The classical incompatible and compatible interactions between Craigella resistant or Craigella susceptible near isolines and V. dahliae, race 1 were used for comparative purposes. Additional experiments using cytological assessment and quantitative PCR showed that in the tolerant interactions one plant defence response, vascular coating, was deployed as effectively as in resistant plants, limiting pathogen distribution. However, a second defence response, which causes the cyclical elimination of fungus from the stem in the classical interactions either does not occur or is substantially delayed in tolerant plants. Thus, the Verticillium population remains stable and substantial throughout the studied time course.

Carbohydrates in near-isogenic shrunken2 sweet maize kernels

Sweet maize breeding lines are harvested at physiological maturity to provide seed for the next generation. It is important to know how sugar quality and quantity in kernels change as plants obtain physiological maturity so that superior cultivars can be developed. Kernels of iso-lines of a shrunken2 sweet maize, susceptible and resistant to the causal organism of rust disease, were harvested at the fresh-market and physiological maturity stages and assayed for content of glucose, fructose, sucrose, total sugar, starch and water soluble polysaccharides (WSP). At fresh-market the susceptible iso-line had higher levels of glucose, fructose, total sugar and WSP, while the resistant iso-line had higher levels of sucrose. There was no difference in the amount of starch in kernels. At physiological maturity kernels of the susceptible iso-line had higher levels of each sugar and their totals, but lower levels of starch and WSP than those of the resistant iso-line. Since the iso-lines appear to process sugars dififerently they can likely be manipulated to better understand how to incorporate desired traits in improved cultivars.