How Specific is Non-Hypersensitive Host and Nonhost Resistance of Barley to Rust and Mildew Fungi？

Full nonhost resistance can be defined as immunity, displayed by an entire plant species against all genotypes of a plant pathogen. Interesting biological questions are, whether the genes responsible for the nonhost status of a plant species have a general or a specific effectiveness to heterologous （＂nonhost＂） pathogens？ Is the nonhost resistance to pathogens of plant species that are related to the nonhost based on R-genes or on other types of genes？ We study this question in barley （Hordeum vulgare L.）, which is a near-nonhost to several rusts （Puccinia） of cereals and grasses. By crosses and selection we accumulated susceptibility and developed an experimental line, SusPtrit, with high susceptibility to at least nine different heterologous rust taxa such as the wheat and Agropyron leaf rusts （P. triticina and P. persistens, respectively）. At the microscopic level there is also some variation among barley accessions in the degree that the heterologous wheat powdery mildew （Blumeria graminis f.sp. tritici） is able to form haustoria in epidermal cells. So, also the genetics of the variation in level of nonhost resistance to heterologous mildew fungi can be studied in barley. Our data obtained on mapping populations involving three regular nonhost-immune accessions （Vada, Cebada Capa and Golden Promise） suggest that nonhost resistance is the joined effect of multiple, quantitative genes （QTLs） and very occasionally a major gene （R-gene？） is involved. Most QTLs have effect to only one or two heterologous rusts, but some have a wider spectrum. This was confirmed in a set of QTL-NILs. Those QTL-NILs are used to fine-map the effective genes. In some cases, a QTL region with effectiveness to several heterologous rusts might be a cluster of genes with a more narrow spectrum of effectiveness. Our evidence suggests that nonhost resistance in barley to rust and powdery mildew fungi of related Gramineae is not due to R-genes, but to pathogen species-specific quantitative resistance genes.     

NBS-LRR Proteins and Their Partners:Molecular Switches of Plant Defense

Specificity of the plant innate immune system is often conferred by resistance（R）proteins.Most plant disease resistance （R）proteins contain a series of leucine-rich repeats（LRRs）,a nucleotide-binding site（NBS）,and a putative amino-terminal signaling domain.They are termed NBS-LRR proteins.The LRRs are mainly involved in recognition,and the amino-terminal domain determines signaling specificity,whereas the NBS domain presumably functions as a molecular switch.During the past years,the most important discoveries are the role of partners in NBS-LRR gene mediated defenses,mounting support for the so-called＂guard hypothesis＂of R gene function,and providing evidence for intramolecular interactions and intermolecular interactions within NBS- LRR proteins as a mode of signaling regulation.The outcome of these interactions determines whether a plant activates its defense responses.     

Progress Towards Genetics and Breeding for Minor Genes Based Resistance to Ug99 and Other Rusts in CIMMYT High-Yielding Spring Wheat

Wheat rusts continue to cause significant losses worldwide despite major efforts given to their genetic control. This is due to frequent evolution and selection of virulence in pathogen overcoming the deployed race-specific resistance genes. Although the life of effective race-specific resistance genes can be prolonged by using gene combinations, an alternative approach being implemented at CIMMYT is to deploy varieties that posses adult plant resistance （APR） based on combinations of minor, slow rusting genes. When present alone, the APR genes do not confer adequate resistance especially under high disease pressure; however, combinations of 4 or 5 minor genes usually result in ＂near-immunity＂ or a high level of resistance. Although only a few APR genes are catalogued, various APR QTLs are now known and could lead to further characterization of additional genes. Four characterized genes have pleiotropic effects in conferring partial APR to all 3 rusts and powdery mildew, thus simplifying the task of breeding wheat varieties that are resistant to multiple diseases. Significant progress was made recently in developing high-yielding wheat germplasm that possesses high levels of APR to all three rusts by implementing a Mexico- Kenya shuttle breeding scheme. Parents with APR to Ug99 were hybridized with high-yielding parents that had adequate to high levels of APR to leaf rust and yellow rust. Segregating populations and advanced lines from these crosses were selected under high rust pressures in Mexico （leaf rust and yellow rust） and Kenya （Ug99 stem rust and yellow rust） to identify high- yielding progenies that possess high to adequate APR to all three rusts. International distribution of these high-yielding wheats is underway through CIMMYT intemational yield trials and screening nurseries. It is expected that several wheat varieties with APR to three rusts will be released and grown in various countries in the near-future that will allow determining the durability of resistance.     

Postulation of Seedling Resistance Genes in 20 Wheat Cultivars to Yellow Rust （ Puccinia striiformis f. sp. triticl）

The gene postulation, cluster analysis and pedigree analysis of 20 wheat cultivars to 28 physiological races of yellow rust （Puccinia striiformis） were conducted under greenhouse conditions, The spectrum of their resistance were compared each other. None of the 20 cultivars were resistant to all the test pathogens. The cultivars containing resistance genes Yr5 and Yr24 were not find and genes Yr8, Yrl9 and Yr27 could not be postulated from cultivars tested due to the susceptibility to all isolates used. A total of 6 probale seedling yellow rust resistance genes or gene combinations （Yrl, Yr2, Yr2 ＋ YrHVII, Yr3 ＋ unknown, Yr3 ＋ Yr4, YrAlba） were postulated in the wheat cultivars （Atou, Flanders, Maris Huntsman, Bouquet, Holdfast, Elite Lepeuple, and Vilmorin 27）. The gene combination Yr2＋YrHVII with the highest frequency （35%） was present in 7 cultivars. The Yr genes present in some wheat cultivars could not be postulated because of non-matching virulence combinations with any of known genes. Cluster result showed that Yr2 and Yr3 are the most important genes in the cultivars. The 13 cultivars are believed to have the pedigree of Noe, which was selected from South Russian wheat. These results will be useful for wheat breeding and provide information about genetic control of wheat yellow rust.     

Blast-Resistance Inheritance of Space-Induced Rice Lines and Their Genomic Polymorphism by Microsatellite Markers

To understand the resistance inheritance basis of space-induced rice lines to blast, and to probe mutants＇ genomic DNA polymorphism compared with ground control by microsatellite markers, three space-induced lines were crossed with a highly susceptible variety LTH, and their F1 and F2 populations were inoculated by two representative blast isolates with broad pathogenicity to analyze their resistance inheritance basis. Meanwhile three mutant lines and the ground control were analyzed by 225 rice SSR （simple sequence repeat） primer pairs selected throughout the 12 chromosomes of whole rice genome, to scan the mutagenesis in genome of the mutant lines. The results indicated the blast-resistant genes harbored in these mutant lines were dominant. It was demonstrated that the resistance of mutant H1 to isolate GD0193 and GD3286 was controlled by a single gene, respectively; while mutants H2 and H3 were controlled by two pairs of major genes against isolate GD3286 and H2 showed complicated genetic mechanism to isolate GD0193. H3＇s resistance to isolate GD0193 was verified to be controlled by a single gene. According to the results of SSR analysis, three mutant lines showed different mutant rates as compared with the ground control, and the mutant rates also varied. Resistance genes can be induced from rice by space mutation, and different genomic variations were detected in blast-resistant lines.     

Genome Array on Differentially Expressed Genes of Skin Tissue in Cashmere Goat at Early Anagen of Cashmere Growth Cycle Using DNA Microarray

In order to study the molecular mechanism involved in cashmere regeneration, this study investigated the gene expression profile of skin tissue at various stages of the cashmere growth cycle and screen differentially expressed genes at proangen in 10 cashmere goats at 2 years of age using agilent sheep oligo microarray. Significance analysis of microarray （SAM） methods was used to identify the differentially expressed genes, Hierarchical clustering was performed to clarify these genes in association with different cashmere growth stages, and GO （Gene ontology） and the pathway analyses were con-ducted by a free web-based Molecular Annotation System3.0 （MAS 3.0）. Approximately 10200 probe sets were detected in skin tissue of 2-yr-old cashmere goat. After SAM analysis of the microarray data, totally 417 genes were shown to be differentially expressed at different cashmere growth stages, and 24 genes are significantly up-regulated （21） or down-regulated （3） at proangen concurrently compared to angen and telogen. Hierarchical clustering analysis clearly distinguished the differentially expressed genes of each stage. GO analysis indicated that these altered genes at proangen were predominantly involved in collagen fibril organization, integrin-mediated signaling pathway, cell-matrix adhesion, cell adhesion, transforming growth factor-β （TGF-β） receptor signaling pathway, regulation of cell growth. Kyoto encyclopedia of genes and genomes （KEGG） analysis showed that the significant pathways involved mainly included focal adhesion and extracellular matrixc （ECM）-receptor interaction. Some important genes involved in these biological processes, such as COL1A1, COL1A2, COL3A1, SPARC, CYR61 and CTGF, were related to tissue remolding and repairing and detected by more than one probe with similar expression trends at different stages of cashmere growth cycle. The different expression of these genes may contribute to understanding the molecular mechanism of cashmere regeneration.     

Development of Sequence Characterized Amplified Region （SCAR） Primers for the Detection of Resistance to Sporisorium reiliana in Maize

Head smut of maize （Zea mays L.）, which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of resistance genes and MAS. Two sets of BC3 progenies, one （BC3Q） derived from the cross Qi319 （resistance）×Huangzao 4 （susceptible）, the other （BC3M） from Mol7 （resistance）× Huangzao 4 （susceptible）, were generated. Huangzao 4 was the recurrent parent in both progenies. A combination of BSA （bulked segregant analysis） with AFLP （amplified fragment length polymorphism） method was applied to map the genes involving the resistance to S. reiliana, and corresponding resistant and susceptible bulks and their parental lines were used for screening polymorphic AFLP primer pairs. One fragment of PI3M61-152 was converted into SCAR （sequence charactered amplified fragment） marker S130. The marker was mapped at chromosome bin 2.09, the interval of a major QTL region previously reported to contribute to S. reiliana resistance. Furthermore, S130 was highly and facilitate map-based cloni associated with resistance to S. reiliana, and could be useful for marker-assisted selection ng of resistance genes.     

Fine Mapping and Candidate Gene Analysis of Resistance Gene RSC3Q to Soybean mosaic virus in Qihuang 1

Soybean mosaic virus （SMV） disease is one of the most destructive viral diseases in soybean （Glycine max （L.） Merr.）. SMV strain SC3 is the major prevalent strain in huang-huai and Yangtze valleys, China. The soybean cultivar Qihuang 1 is of a rich resistance spectrum and has a wide range of application in breeding programs in China. In this study, F1, F2 and F2：3 from Qihuang 1×nannong 1138-2 were used to study inheritance and linkage mapping of the SC3 resistance gene in Qihuang 1. The secondary F2 population and near isogenic lines （nILs） derived from residual heterozygous lines （RhLs） of Qihuang 1×nannong 1138-2 were separatively used in the ifne mapping and candidate gene analysis of the resistance gene. Results indicated that a single dominant gene （designated RSC3Q） controls resistance, which was located on chromosome 13. Two genomic-simple sequence repeat （SSR） markers BARCSOYSSR_13_1114 and BARCSOYSSR_13_1136 were found lfanking the two sides of the RSC3Q. The interval between the two markers was 651 kb. Quantitative real-time PCR analysis of the candidate genes showed that ifve genes （Glyma13g25730, 25750, 25950, 25970 and 26000） were likely involved in soybean SMV resistance. These results would have utility in cloning of RSC3Q resistance candidate gene and marker-assisted selection （MaS） in resistance breeding to SMV.     

Transgenic Expression of a Functional Fragment of Harpin Protein Hpa1 in Wheat Represses English Grain Aphid Infestation

The harpin protein Hpa1 produced by the rice bacterial blight pathogen promotes plant growth and induces plant resistance to pathogens and insect pests. The region of 10-42 residues （Hpa110-42） in the Hpa1 sequence is critical as the isolated Hpa110-42 fragment is 1.3-7.5-fold more effective than the full length in inducing plant growth and resistance. Here we report that transgenic expression of Hpa110-42 in wheat induces resistance to English grain aphid, a dominant species of wheat aphids. Hpa110-42-induced resistance is effective to inhibit the aphid behavior in plant preference at the initial colonization stage and repress aphid performances in the reproduction, nymph growth, and instar development on transgenic plants. The resistance characters are correlated with enhanced expression of defense-regulatory genes （EIN2, PP2-A, and GSL10） and consistent with induced expression of defense response genes （Hel, PDF1.2, PR-1b, and PR-2b）. As a result, aphid infestations are alleviated in transgenic plants. The level of Hpa110-42-induced resistance in regard to repression of aphid infestations is equivalent to the effect of chemical control provided by an insecticide. These results suggested that the defensive role of Hpa110-42 can be integrated into breeding germplasm of the agriculturally signiifcant crop with a great potential of the agricultural application.     

Genetic Analysis and Molecular Mapping of an All-Stage Stripe Rust Resistance Gene in Triticum aestivum-Haynaldia villosa Translocation Line V3

Triticum aestivum-Hayaldia villosa translocation line V3 has shown effective all-stage resistance to the seven dominant pathotypes of Puccinia striiforms f.sp.tritici prevalent in China.To elucidate the genetic basis of the resistance,the segregating populations were developed from the cross between V3 and susceptible genotype Mingxian 169,seedlings of the parents and F 2 progeny were tested with six prevalent pathotypes,including CYR29,CYR31,CYR32-6,CYR33,Sun11-4,and Sun11-11,F 1 plants and F 3 lines were also inoculated with Sun11-11 to confirm the result further.The genetic studied results showed that the resistance of V3 against CYR29 was conferred by two dominant genes,independently,one dominant gene and one recessive gene conferring independently or a single dominant gene to confer resistance to CYR31,two complementary dominant genes conferring resistance to both CYR32-6 and Sun11-4,two independently dominant genes or three dominant genes（two of the genes show cumulative effect） conferring resistance to CYR33,a single dominant gene for resistance to Sun11-11.Resistance gene analog polymorphism（RGAP） and simple-sequence repeat（SSR） techniques were used to identify molecular markers linked to the single dominant gene（temporarily designated as YrV3） for resistance to Sun11-11.A linkage map of 2 RGAP and 7 SSR markers was constructed for the dominant gene using data from 221 F 2 plants and their derived F 2：3 lines tested with Sun11-11 in the greenhouse.Amplification of the complete set of nulli-tetrasomic lines of Chinese Spring with a RGAP marker RG1 mapped the gene on the chromosome 1B,and then the linked 7 SSR markers located this gene on the long arm of chromosome 1B.The linkage map spanned a genetic distance of 25.0 cM,the SSR markers Xgwm124 and Xcfa2147 closely linked to YrV3 with genetic distances of 3.0 and 3.8 cM,respectively.Based on the linkage map,it concluded that the resistance gene YrV3 was located on chromosome arm 1BL.Given chromosomal location,the reaction patterns and pedigree analysis,YrV3 should be a novel gene for resistance to stripe rust in wheat.These closely linked markers should be useful in stacking genes from different sources for wheat breeding and diversification of resistance genes against stripe rust.     

Induction and Characterization of Laboratory Mutants of Phytophthora capsici Resistant to Dimethomorph and Flumorph

Laboratory studies were conducted to evaluate the risk of Phytophthora capsici developing resistance to two morphlines, dimethomorph and flumorph. Metalaxyl, the well-known high risk of resistance fungicides, was used as reference fungicide. Resistant mutants for the three fungicides were isolated by treating mycelium with ultraviolet radiation. Metalaxyl-resistant mutants were obtained with high frequency and exhibited high level of resistance with factors more than 100 folds, while mutation frequency for dimethomorph-resistance was relatively low and the resistance factors ranged from 3.0 to 13.9 folds. Most dimethomorph-resistant mutants decreased in hyphal growth rate and the spoulation ability, which have a large impact upon the epidemic development of dimethomorph-resistant populations. These results suggested that the risk of resistant pathogen population was much lower for dimethomorph than for metalaxyl. Both the frequency of developing resistance and level of resistance （resistance factors = 1.8-14.6） to dimethomorph were similar to those of its structure analogue flumorh. Moreover, the cross-resistance were found between them, which suggested the risks of developing resistance to dimethomorph and flumorph in the pathogen were very closely related. As P. capsici can potentially develop resistance to dimethomorph and flumorph, and oomycetes usually have the high risk to develop resistance to fungicides, appropriate management against resistance development should be taken.     

Progresses in the Mechanism of Resistance to Fusarium Wilt in Cucumber （Cucumis sativus L.）

Fusarium wilt caused by Fusarium oxysporum f.sp. cucumerinum （Owen） is one of the most devastating diseases in cucumber production worldwide. Recent progresses in the mechanism of resistance to Fusarium wilt in cucumber were reviewed in this paper, including pathogenic mechanism of Fusarium oxysporum, the resistance mechanism of cucumber, the heredity of resistance, and the location of resistance genes. Following works should be the location and cloning of resistance genes with molecular biologic methods.     

Studies on Antimicrobial Resistance Transfer In vitro and Existent Selectivity of Avian Antimicrobial-Resistant Enterobacteriaccae In vivo

Increasing antimicrobial resistance （AR） has become a severe problem of public health in the world, whereas control of the AR of bacteria will be based on investigation of the AR mechanism. Furthermore, understanding the existent selectivity of AR organisms from animals can prevent the emergence and diffusion of AR effectively. PCR amplifications of gyrA and parC genes have been performed for detecting fluoroquinolones-resistance （FR） genes. A conjugational transfer test has been carried out using a donor which is resistant to tetracycline （TE）, ampicillin （AMP）, sulfamethoxazole-trimethoprim （SXT）, and a recipient which is sensitive to TE, AMP, and SXT. The AR strains have been passed 20 passages. Two groups of chicken inoculated multi-AR Escherichia coli （E. coli） and multi-AR Salmonella, respectively, are mix-fed. The result shows that amino acid codons of Ser-83 and Asp-87 are mutations from gyrA and there are no mutations from parC genes in all the FR strains. Resistance to TE, AM, and SXT can transfer among E. coli and the conjugal transfer frequency of TE is 3×10^-7. AR can inherit in 20 passages at least. The multi-AR E. coli and Salmonella can be isolated from all chickens three days after inoculation but CIP-resistant strains decrease during the time run out and disappear at 23 days after inoculation. The results indicate that the mutations of gene gyrA are correlative with the FR phenotype. AR genes that are not connected to the chromosome can transfer horizontally and vertically. AR bacteria can diffuse quickly and eliminate naturally from the host if the chicken is not under the pressure of this antibiotic.