Mapping of Aegilops umbellulata‐derived leaf rust and stripe rust resistance loci in wheat

Aegilops umbellulata, a non‐progenitor diploid species, is an excellent source of resistance to various wheat diseases. Leaf rust and stripe rust resistance genes from A. umbellulata were transferred to the susceptible wheat cultivar WL711 through induced homoeologous pairing. A doubly resistant introgression line IL 393‐4 was crossed with wheat cultivar PBW343 to develop a mapping population. Tests on BC₂F₇ RILs indicated monogenic inheritance of seedling leaf rust and stripe rust resistance in IL 393‐4 and the respective co‐segregating genes were tentatively named LrUmb and YrUmb. Bulked segregant analysis placed LrUmb and YrUmb in chromosome 5DS, 7.6 cM distal to gwm190. Aegilops geniculata‐derived and completely linked leaf rust and stripe rust resistance genes Lr57 and Yr40 were previously located in chromosome 5DS. STS marker Lr57/Yr40MAS‐CAPS16 (Lr57/Yr40‐CAPS16), linked with Lr57/Yr40 (T756) also co‐segregated with LrUmb/YrUmb. Seedling infection types differentiated LrUmb from Lr57. Absence of leaf rust‐susceptible segregants among F₃ families of the intercross (IL 393‐4/T756) indicated repulsion linkage between LrUmb and Lr57. YrUmb expressed a consistently low seedling response under greenhouse conditions, whereas Yr40 expressed a higher seedling response. Based on the origin of LrUmb/YrUmb from the U genome and Lr57/Yr40 from the M genome, as well as phenotypic differences, LrUmb and YrUmb were formally named Lr76 and Yr70, respectively. These genes have been transferred to Indian wheat cultivars PBW343 and PBW550, and advanced breeding lines are being tested in state and national trials.     

Screening of weed extracts for antifungal properties against Colletotrichum lagenarium,the causal agent of anthracnose in cucumber

The antifungal activity of the leaf extracts from 203 weed species was investigated by performing a bioassay using cucumber plants and Colletotrichum orbiculare. The leaf extracts from four families, namely, Urticaceae, Onagraceae, Commelinaceae, and Solanaceae, showed a relatively stronger inhibition of the anthracnose lesions in cucumber plants when compared with the other families investigated in the study. A remarkable inhibition of anthracnose infection in cucumber leaves was observed with the extracts from the following 19 weed species: Boehmeria nipononivea and Boehmeria longispica, Persicaria scabra, Ranunculus japonicus and Ranunculus sceleratus, Cardamine flexuosa, Oenothera biennis, Aeschynomene indica, Indigofera pseudo‐tinctoria, Torilis scabra, Calystegia japonica, Solanum americanum, Bidens pilosa, Gnaphalium japonicum, Kalimeris yomena, Bromus catharticus, Cynodon dactylon, Alopecurus aequalis, and Scirpus tabernaemontani. In particular, it is noteworthy that the extracts from C. dactylon, K. yomena, and S. americanum completely inhibited anthracnose infection in cucumber.     

Functional analysis of the 3′ end of avrBs3/pthA genes from two Xanthomonas species

The phytopathogens Xanthomonas oryzae pathovar (pv.) oryzae and Xanthomonas axonopodis pv. citri each contain several avrBs3/pthA family genes. Structural features of these genes important for avirulence and/or virulence functions include a central region of multiple direct repeats and three nuclear localization signals (NLSs) and an acidic activation domain (AAD) at the 3′ end. To identify other regions critical to function in the 3′ ends of these genes, we constructed several chimeras using apl1 and apl2 from X. axonopodis pv. citri and avrXa10 and avrXa7 from X. oryzae pv. oryzae and evaluated their functions by inoculation to citrus and rice. The apl1 and avrXa7 genes are major virulence determinants in citrus and rice, respectively, while the contributions of apl2 and avrXa10 to virulence are negligible or not measurable. Constructs that contained a 417 bp HincII-SphI fragment from the 3′ end of apl1 in combination with the repeats from avrXa7, avrXa10, and apl1 caused a canker phenotype on citrus. Interchange of the HincII-SphI fragment between avrXa7 and avrXa10 abolishes avrXa7 avirulence function and reduces its virulence but it does not affect avrXa10 avirulence function in rice. avrXa7 caused a hypersensitive response (HR) in citrus and replacement of it's 3′ end with that of apl1 resulted in loss of canker and induction of HR. Thus, the HincII-SphI fragment of the avrBs3/pthA gene family is important for avirulence and virulence functions in two different plant species, Oryza sativa and Citrus natsudaidai HAYATA.     

7种林木植物对华北大黑鳃金龟取食和繁殖的影响

为明确华北大黑鳃金龟Holotrichia oblita(Faldermann)的嗜食植物,优化其室内人工饲养技术及为诱虫植物提供候选材料,以花生叶片为对照,研究了7种林木植物叶片对华北大黑鳃金龟取食和繁殖的影响。结果表明,华北大黑鳃金龟对8种植物日均取食量由高到低依次为金叶女贞、刺槐、核桃、榆树、花生、柳树、香椿、毛白杨,其中金叶女贞、香椿和毛白杨与花生间有极显著差异,金叶女贞与刺槐、核桃、榆树间无显著差异;单雌产卵量由高到低依次为金叶女贞、柳树、核桃、香椿、花生、榆树、毛白杨、刺槐,其中金叶女贞、柳树、核桃、刺槐与花生间有极显著差异,而金叶女贞与柳树间无显著差异。研究表明,饲养华北大黑鳃金龟成虫适宜饲料植物为金叶女贞,日均取食量最高为4.70 g,产卵期最长为56.00 d,单雌产卵量最多为58.88个,其次为核桃和柳树。     

Transient expression of pepper Bs2 gene in Citrus limon as an approach to evaluate its utility for management of citrus canker disease

The pepper Bs2 gene confers resistance to Xanthomonas campestris pv. vesicatoria (Xcv) pathogenic strains containing the avrBs2 avirulence gene in susceptible pepper and tomato. The avrBs2 gene is highly conserved in the Xanthomonas genus and when bacteria lack this gene their growth in a susceptible host is diminished, indicating that the avrBs2 gene product could confer an adaptive advantage to the pathogen. The avrBs2 of Xanthomonas citri subsp. citri (Xcc), cause of citrus canker, shares 96% homology with avrBs2 of Xcv. To evaluate if Bs2 could recognize avrBs2 of Xcc in citrus plants and thereby activate plant defence mechanisms to increase resistance to canker, transient expression experiments were conducted using Agrobacterium tumefaciens in lemon plants subsequently challenged with wildtype Xcc. The results showed that transient expression of Bs2 reduced canker formation in lemon and induced plant defence mechanisms, as shown by callose deposition and PR‐1 expression. Moreover, when an avrBs2 mutant of Xcc was used, no decrease in disease symptoms was observed. This work shows that the Bs2 gene from Solanaceae is functional in lemon, a member of the Rutaceae family. Therefore, Bs2 is a potential candidate gene for stable expression in transgenic citrus plants in order to improve resistance to canker disease.     

Quick assessment of the invasiveness of non‐native plant species by using eco‐physiological parameters in Tram Chim National Park,Vietnam

Using eco‐physiological parameters, a quick assessment of the invasiveness of non‐native plant species was conducted in Tram Chim National Park, a Ramsar site that is located in the Mekong River Delta region of Vietnam. An investigation of non‐native species and a vegetation analysis were carried out along 25 line transects and in 50 quadrats by using the Braun–Blanquet method. The researchers identified 84 non‐native plant species but only 31 species were naturalized in the wetland ecosystems. Twenty of those 31 species with a high importance value index were screened by using a parameter that was obtained from chlorophyll a fluorescence measurement, the performance index. Five species were identified as invasive and five others were predicted to be potentially invasive. The first group of five species were: M imosa pigra, P anicum repens, E ichhornia crassipes, S alvinia cucullata and L eersia hexandra, which already had been confirmed as important weeds in the national park by previous studies. From the second group, two species ( L udwigia hyssopifolia and S accharum spontaneum) already are becoming prominent species in some locations. The three remaining species ( M onochoria hastata, I sachne globosa and M arsilea quadrifolia) are likely to become invasive in the future.     

Effects of three esca-associated fungi on Vitis vinifera L.: III. Enzymes produced by the pathogens and their role in fungus-to-plant or in fungus-to-fungus interactions

When the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme) were grown in liquid stationary cultures, it was seen that they were able to live in media containing resveratrol (RES) or tannic acid (TA) as the sole carbon source and that the fungi were able to convert both compounds. Particular attention is paid here to detecting RES and TA conversion. Pch, Tmi and Fme were partially inhibited by RES or TA. Pch, Tmi and Fme produced extracellular tannase, laccase and peroxidase enzymes in liquid or agarized cultures, whether glucose was present or not. When colonies of Pch, Tmi and Fme were confronted, they showed spatially and temporally heterogeneous patterns of laccase and peroxidase activity. The results indicate the non-synergistic, competitive association of Pch and Tmi and the inhibition of Fme growth. Muconic acid, a well-known intermediate in a large number of lignin and phenol oxidative processes, can partly or completely inhibit the lignolytic agent Fme, but is tolerated by Pch and Tmi. An explanation for wood pigmentation patterns by Pch, Tmi and Fme is given.     

PCR-based differentiation of Fusarium oxysporum ff. sp. lycopersici and radicis-lycopersici and races of F. oxysporum f. sp. lycopersici

The pathogenic type (form and race) of Fusarium oxysporum, which generates wilt symptoms on tomato, was rapidly identified with a polymerase chain reaction (PCR)-based technique. We compared the partial nucleotide sequences of endo polygalacturonase (pg1) and exo polygalacturonase (pgx4) genes from isolates of F. oxysporum ff. sp. lycopersici (FOL) and radicis-lycopersici (FORL) from Japan and designed specific primer sets (uni, sp13, sp23, and sprl) based on the nucleotide differences that appeared among the pathogenic types. PCR with the uni primer set amplified a 670∼672-bp fragment from all isolates of FOL and FORL. With the sp13 primer set, an amplicon of 445 bp was obtained only from isolates of FOL race 1 and 3. With the sp23 primer set, a 518-bp fragment was obtained from isolates of FOL race 2 and 3. The sprl primer set yielded a 947-bp fragment from isolates of FORL, but not from FOL. A combination of amplifications with these primer sets effectively differentiated the pathogenic types of F. oxysporum in tomato.     

Gene Flow Analysis of Phytophthora porri Reveals a New Species: Phytophthora brassicae Sp. Nov.

Isozyme analysis and sequence analysis of the internal transcribed spacer regions (ITS-1 and ITS-2) and the 5.8S subunit of the ribosomal DNA gene repeat were used to examine whether isolates of Phytophthora porri from Allium and Brassica represent a single homogeneous species. Twenty-six strains of P. porri, 16 strains isolated from the genus Allium, and 10 strains isolated from the genus Brassica, were analyzed using malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH) and lactate dehydrogenase (LDH), represented altogether by four putative loci (Mdh-2, Idh-1, Idh-2, and Ldh-2). Isozyme analysis revealed that strains isolated from Allium contained five private alleles at three isozyme loci (Ldh-2 ⁸³, Ldh-2 ¹⁰⁴, Idh-1 ¹⁰⁸, Idh-1 ¹¹², and Idh-2 ⁹⁸), whereas six different alleles were observed at four isozyme loci (Ldh-2 ⁸⁵, Ldh-2 ¹⁰⁰, Ldh-2 ¹¹⁴, Idh-1 ¹⁰⁰, Idh-2 ¹⁰⁰, and Mdh-2 ¹¹¹) in strains obtained from Brassica. The heterozygosity at the Ldh-2 locus, differing in allele composition, however, between strains from Allium and Brassica, was present in all strains, indicating that it is probably fixed. Sequence analysis of the ITS regions and the 5.8S subunit showed consistent differences between isolates from Allium and isolates from Brassica. Based on isozyme data, ITS sequence analysis and formerly published differences in restriction enzyme patterns of mitochondrial DNA, morphology and pathogenicity, it was concluded that the isolates of P. porri Foister did not represent a homogeneous species. Isolates from Brassica constitute a distinct species which is described here as P. brassicae sp. nov. It was inferred from isozyme patterns, which were in no case intermediate between the two species, that P. porri and P. brassicae do not hybridize and are reproductively isolated by barriers to gene flow.     

Detection and identification of the crucifer pathogen, <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> pv. <Emphasis Type="Italic">campestris</Emphasis>, by PCR amplification of the conserved Hrp/type III secretion system gene <Emphasis Type="Italic">hrcC</Emphasis>

The development of a rapid detection method for Xanthomonas campestris pv. campestris (Xcc) in crucifer seeds and plants is essential for high-throughput certification purposes. Here we describe a diagnostic protocol for the identification/detection of Xcc by PCR amplification of fragments from the pathogenicity-associated gene hrcC. Under stringent conditions of amplification, a PCR product of 519 bp from hrcC was obtained from a collection of 46 isolates of Xcc, with the exception of two isolates from radish. No amplicons were obtained from 39 pure cultures of the phytopathogenic bacteria Xanthomonas campestris pv. cerealicola, X. campestris pv. juglandis, X. campestris pv. pelargonii, X. campestris pv. vitians, X. arboricola pv. pruni, X. axonopodis pv. phaseoli, X. axonopodis pv. vesicatoria, X. vesicatoria, Pseudomonas syringae pv. phaseolicola, P. syringae pv. syringae, P. syringae pv. tomato, P. fluorescens, P. marginalis, Pectobacterium atrosepticum, P. carotovorum subsp. carotovorum. In addition, PCR reactions were negative for fifty unidentified environmental isolates purified from the surface of crucifers. The PCR fragment was obtained from four strains previously classified as X. campestris pv. aberrans, X. campestris pv. armorociae, X. campestris pv. barbarae and X. campestris pv. incanae using pathogenicity assays. Our PCR protocol specifically detected Xcc in inoculated leaves, seeds and naturally infected leaves of crucifers.     

Antinematodal activity of some Malaysian plant extracts against the pine wood nematode,Bursaphelenchus xylophilus

Methanolic extracts of 79 Malaysian plants representing 42 families were assessed for antinematodal activity against Bursaphelenchus xylophilus using a fungal-feeding assay. Extracts of 27 plants from 19 families showed antinematodal activity, while 52 species were inactive. Five extracts (Sauropus androgynus, Eugenia polyantha, Areca catechu, Piper betle and Piper nigrum) exhibited very strong activity against Bursaphelenchus xylophilus at a minimum effective dose (MED) of 0·625 mg per ball. Strong antinematodal activity (MED: 1·25–2·5 mg per ball) was shown by the extracts of Spondias cyntherea, Codiageum variegatum, Euodia glabra and Cicca acida. Eleven extracts (Carica papaya, Ipomoea aquatica, Ocimum basilicum, Leea gigantea, Pithecellobium jiringa, Crypteronia paniculata, Myristica fragrans, Murraya koenigii, Leucaena leucocephala, Melastoma malabathricum and Morinda citrifolia) demonstrated moderate activity between MED of 5 and 10 mg per ball, and weak activity was observed in seven extracts (Ipomoea batatas, Cymbopogon citratus, Garcinia atroviridis, Psophocarpus tetragonolobus, Tamarindus indica, Allium odorum and Stenochalaena palustris). © 1997 SCI     

Tagging and mapping a second resistance gene for <Emphasis Type="Italic">Fusarium</Emphasis> wilt race 0 in chickpea

A second gene conferring resistance to the chickpea wilt pathogen, Fusarium oxysporum f. sp ciceris race 0, has been mapped to linkage group 2 (LG2) of the chickpea genetic map. Resistance to race 0 is controlled by two genes which segregate independently; one present in accession JG62 (Foc0 ₁ /foc0 ₁ ) and mapping to LG5 and the second present in accession CA2139 (Foc0 ₂ /foc0 ₂ ) but remaining unmapped. Both genes separately confer complete resistance to race 0 of the wilt pathogen. Using a Recombinant Inbred Line (RIL) population that segregated for both genes (CA2139 × JG62) and the genotypic information provided by two markers flanking Foc0 ₁ /foc0 ₁ ten resistant lines containing the resistant allele Foc0 ₂ /foc0 ₂ were selected. Genotypic analysis using these ten resistant lines paired with ten susceptible RILs, selected in the same population, revealed that sequence tagged microsatellite sites (STMS) markers sited on LG2 were strongly associated with Foc0 ₂ /foc0 ₂ . Linkage analysis, using data from two mapping populations (CA2139/JG62 and CA2156/JG62), located Foc0 ₂ /foc0 ₂ in a region where genes for resistance to wilt races 1, 2, 3, 4 and 5 have previously been reported and which is highly saturated with tightly-linked STMS markers that could be used in marker-assisted selection (MAS).     

Interaction of benzimidazole-N-sulfonamides with the cytochrome b/c1 complex of Pythium aphanidermatum

Experiments with intact cells and submitochondrial fractions of Pythium aphanidermatum (Edson) Fitz. indicated an interference of benzimidazole-N-sulfonamides with the NADH- or succinate-driven electron transport system between cytochromes b and c. Comparison with Ustilago maydis (DC) Corda and Botrytis cinerea Pers. ex Fr. revealed that this effect is Oomycetes specific. The molecular interaction between benzimidazole-N-sulfonamides and the mitochondrial cytochrome b/c₁ complex from P. aphanidermatum has been investigated. Binding assays with [¹⁴C]52232 RP (dimefluazole) indicated a time- and dose-dependent labelling of two proteins. The molecular mass of one labelled protein and the competition of the binding with antimycin A suggest that benzimidazole-N-sulfonamides interact with the Q₁-centre of cytochrome b. Furthermore, experiments with doubly labelled [³H][¹⁴C]CGA 323103 revealed a possible irreversible inactivation of the b/c₁ complex leading to covalent linkage of the dimethylsulfonamoyl moiety to the target site.     

A contribution to the knowledge of the Turkish aphid (Hemiptera: Aphidoidea) fauna

As a result of the study carried out between 2007 and 2009 in the eastern part of the Black Sea Region of Turkey, 4 genera and 17 species were determined as new records for the Turkish aphid fauna. New recorded species are Aphis arbuti, Aphis cytisorum, Aphis kachkoulii, Cavariella digitata, Chaitophorus kapuri, Chaitophorus longisetosus, Cinara pinivora, Illinoia lambersi, Myzus beybienkoi, Neobetulaphis pusilla, Phyllaphis fagifoliae, Pseudessigella brachychaeta, Schizaphis rotundiventris, Symdobious oblongus, Thecabius lysimachiae, Thelaxes californica and Tinocallis takachihoensis. With these new records, the number of genera increased to 134 and the number of the species in the Turkish aphid fauna amounted to about 480. Since Turkey has a huge range of agricultural crops and a great diversity in its natural flora, research conducted on the Turkish aphid fauna has current and future indications for plant protection.     

Consistent responses of yield and resistance of wheat cultivars to the root-lesion nematode,Pratylenchus thornei,in the Australian northern subtropical region,but not in the temperate southern region

To understand the yield response of cereal cultivars to Pratylenchus thornei, eight experiments were conducted within the subtropical northern, and temperate southern grain-producing regions of Australia. Wheat cultivars (Triticum aestivum) ranging from susceptible to moderately resistant to P. thornei were grown in Year 1 to establish a range of population densities. In Year 2 before sowing, P. thornei was quantified in each plot and six cereal cultivars were each grown on a similar range of population densities (average minimum to maximum of 3.4–60.6 P. thornei/g soil); P. thornei was quantified again at harvest. In the four experiments in the northern region there was a significant, negative logarithmic response of yield of the three most intolerant/susceptible cultivars as P. thornei population densities increased (yield decreased 172–479 kg/ha per unit increase in log_e-transformed P. thornei/g soil). The responsiveness of yield to increasing P. thornei population densities diminished as the tolerance and resistance of the cultivars improved. In the southern region, there was no relationship between yield and P. thornei in three experiments and minor, positive increases in one experiment (1.6 kg/ha per unit increase in P. thornei/g soil). Across both regions, the change in P. thornei population densities from sowing to harvest was logarithmic and positive, and generally greatest in the northern region. The contrast of responses of cereal cultivars between the regions, despite similar population densities of P. thornei, is indicative of the influence of the environment particularly on tolerance, therefore management with a regional focus is essential.     

Comparison of Secreted in Xylem (SIX) genes in two fusarium wilt pathogens of ornamental palms

Fusarium oxysporum ff. sp. canariensis (Foc) and palmarum (Fop) cause lethal fusarium wilt on ornamental palms. Foc infects Phoenix canariensis and has a worldwide presence, whereas Fop primarily infects Washingtonia robusta and Syagrus romanzoffiana and is currently primarily restricted to Florida (USA). A Secreted in Xylem (SIX) gene profile was obtained for Fop and Foc isolates from the USA. SIX1, SIX7, SIX10 and SIX12 were detected in most Foc isolates and, with the exception of SIX12, with minimal sequence polymorphism. SIX8 and SIX9 were detected in all Fop isolates, with no sequence polymorphism. SIX10 was also present in seven of the Fop isolates. Phylogenetic trees were constructed using EF-1α-, SIX1, SIX7 and SIX10 gene sequences from this study and previously derived sequences from Foc isolates obtained in Australia, the Canary Islands and Japan. The topology of the housekeeping EF-1α gene indicated Foc is polyphyletic, with the Australian isolates separating into a distinct group from all other Foc isolates. However, the topologies of SIX1, SIX7 and SIX10 phylogenetic trees for Foc indicate monophyly. As has been proposed previously, this suggests the core genome of Foc evolved into a separate lineage after the SIX genes were conserved within the whole genome. This study is the first to generate SIX12 sequences for Foc. Interestingly, the topology of the SIX12 phylogenetic tree suggests polyphyly. For Fop, the topologies of the EF-1α, SIX8 and SIX9 phylogenetic trees support monophyly. The distinct SIX gene profile and EF-1α sequence of Fop demonstrates an independent evolutionary origin from Foc.     

Pathogenicity of 21 newly described Phytophthora species against seven Western Australian native plant species

The pathogenicity of some Phytophthora species recently described from Western Australia, together with P. cinnamomi as a control, was tested against seven Western Australian native plant species in the glasshouse. Host species were Banksia grandis, B. littoralis, B. occidentalis, Casuarina obesa, Corymbia calophylla, Eucalyptus marginata and Lambertia inermis. Twenty‐two Phytophthora species were grown on a vermiculite, millet seed and V8 substrate and used as soil inoculum when the plant hosts were approximately 3 months old. Pathogenicity was assessed after 6 weeks and plants were scored for death, root damage, and percentage reduction of shoot growth compared with control plants. The pathogenicity of P. cinnamomi was confirmed. Phytophthora niederhauserii was shown to be similar to P. cinnamomi in pathogenicity and of concern ecologically. Other species that killed one or more hosts were P. boodjera, P. constricta, P. elongata, P. moyootj and P. rosacearum, while P. condilina, P. gibbosa, P. gregata, P. litoralis and P. ‘personii’ caused significant reduction to shoot and/or root growth, but did not kill plants. Host species susceptible to the highest number of Phytophthora species were B. grandis, B. littoralis, B. occidentalis and E. marginata. No Phytophthora species tested killed C. calophylla.