小麦品种贵农22号抗条锈基因遗传分析

贵农22号是利用簇毛麦(Haynaldia villosa)、硬粒小麦(Triticum durum)及普通小麦(Triticum aestuvum)杂交而育成的普通小麦品种,其抗中国目前流行和出现的条锈菌小种,已成为目前重要的抗小麦条锈病抗源。为了明确该品种抗锈遗传规律并进行应用前景评价,用一个流行的强毒性小种条中31号和一个突变弱毒性小种CY29-mut3,分别接种贵农22与国际已知抗锈基因品种Moro及感病品种辉县红双列杂交F2、F2代各株系幼苗,对贵农22号进行了抗锈性遗传分析,以便于在抗病育种中进一步应用。研究结果表明,贵农22号有三对独立遗传的抗条锈基因,暂定名为YrGui 1、YrGui 2和YrGui 3,它们表达稳定,不受亲本正反交影响,而并不具有Yr 10。Yr10基因载体品种Moto中有二或四对基因抗中国不同的条锈菌小种,不同小种及正反交对基因的表达有影响,为父本时其对CY29-mut3小种有两对完全显性基因、一对中度抗病基因及一对隐性抗病基因,而为母本时有一对完全显性基因和一对中度抗病基因起抗病作用;对条中31号,其为父本时有一对显性基因和一对隐性基因,为母本时可能存在两对累加作用基因或两对隐性抗病基因控制抗痫作用。     

Number of genes controlling slow rusting resistance to leaf rust in five spring wheat cultivars

The number of genes controlling slow rusting resistance to leaf rust (Puccinia triticina) was estimated in five spring wheat (Triticum aestivum) cultivars using quantitative formulae. Parents and F₆ families were evaluated in replicated field trials under epidemics initiated by artificial inoculation. The F₆ families resulted from a diallel cross involving the fast-rusting cultivar Yecora 70 and five slow-rusting wheat cultivars: Sonoita 81, Tanager ‘S’, Galvez 87, Ures 81, and Moncho ‘S’. The area under the disease progress curve (AUDPC) was used to measure leaf rust severity over time. Results indicate that cultivar Sonoita 81 has three or four genes, Tanager ‘S’ has two or three genes, Galvez 87 has three genes, and both Ures 81 and Moncho ‘S’ have two genes for slow rusting resistance to leaf rust. Based on this result and previously reported moderate to high narrow-sense heritability estimates for slow rusting resistance in these materials, early-generation selection for slow leaf rusting would be effective.     

河南省16个主栽小麦品种抗叶锈基因分析

为了明确河南省小麦品种的抗叶锈性及抗叶锈基因的分布,为小麦品种推广与合理布局、叶锈病防治及抗病育种提供依据,本研究利用2015年采自河南省的5个小麦叶锈菌流行小种混合菌株,对近几年河南省16个主栽小麦品种进行了苗期抗性鉴定,然后选用12个小麦叶锈菌生理小种对这些品种进行苗期基因推导,同时利用与24个小麦抗叶锈基因紧密连锁(或共分离)的30个分子标记对该16个品种进行了抗叶锈基因分子检测。结果显示,供试品种苗期对小麦叶锈菌混合流行小种均表现高度感病;基因推导与分子检测结果表明,供试品种可能含有Lr1、Lr16、Lr26和Lr30这4个抗叶锈基因,其中先麦8号含有Lr1和Lr26;郑麦366和郑麦9023含有Lr1;西农979和怀川916含有Lr16;中麦895、偃展4110、郑麦7698、平安8号、众麦1号、周麦16、衡观35和矮抗58含有Lr26;周麦22中含有Lr26,还可能含有Lr1和Lr30;豫麦49-198和洛麦23可能含有本研究中检测以外的其他抗叶锈基因。因此,河南省主栽小麦品种的抗叶锈基因丰富度较低,今后育种工作应注重引入其他抗叶锈性基因,提高抗叶锈性,有效控制小麦叶锈病。     

Intercellular chitinase and peroxidase activities associated with resistance conferred by geneLr35to leaf rust of wheat

The effect of leaf rust (Puccinia triticina) infection on intercellular chitinase (EC 3.2.1.14) and peroxidase (EC 1.11.1.7) activities was studied in resistant [RL 6082 (Thatcher/Lr35)] and susceptible (Thatcher) near isogenic wheat (Triticum aestivum L.) lines at seedling, stem elongation and flag leaf stages of plant growth. The levels of activity of these enzymes were low during the seedling and stem elongation stages. Resistant plants at the flag leaf stage, during which the Lr35 resistance gene was maximally expressed, exhibited high constitutive levels of chitinase and peroxidase activities, in contrast to the lower constitutive levels of susceptible plants. The results suggest that chitinase and peroxidase, constitutively present in the intercellular spaces of Thatcher/Lr35 wheat leaves, may play a role in Lr35 mediated resistance to leaf rust.     

山东省12个主栽小麦品种(系)抗叶锈性分析

本研究旨在明确山东省12个小麦主栽品种(系)抗叶锈性及抗叶锈基因,为小麦品种推广与合理布局、叶锈病防治及抗病育种提供依据。利用2015年采自山东省的5个小麦叶锈菌流行小种的混合小种对这些材料进行苗期抗性鉴定,然后选用15个小麦叶锈菌生理小种对这些品种(系)进行苗期基因推导,并利用与24个小麦抗叶锈基因紧密连锁(或共分离)的30个分子标记对其进行抗叶锈基因分子检测。结果显示,山东省12个主栽小麦品种(系)苗期对该省2015年的5个小麦叶锈菌混合流行小种均表现高度感病。通过基因推导与分子检测发现,济南17含有Lr16,矮抗58和山农20含有Lr26,其余济麦系列、烟农系列、良星系列等9个品种(系)均未检测到所供试标记片段。此外,本研究还对山东省3个非主栽品种进行了检测,结果发现,中麦175含有抗叶锈基因Lr1和Lr37,含有成株抗性基因;皖麦38只检测到Lr26,济麦20未检测到所供试标记片段。综合以上结果,山东省主栽小麦品种(系)所含抗叶锈基因丰富度较低,尤其不含有对我国小麦叶锈菌流行小种有效的抗锈基因,应该引起高度重视,今后育种工作应注重引入其他抗叶锈基因,提高抗叶锈性。     

Genetics of adult plant stripe rust resistance in CSP44, a selection from Australian wheat

Wheat line CSP44, a selection from an Australian bread wheat cultivar Condor, has shown resistance to stripe rust in India since the last twenty years. Seedlings and adult plants of CSP44 showed susceptible infection types against stripe rust race 46S119 but displayed average terminal disease severity of 2.67 on adult plants against this race as compared to 70.33 of susceptible Indian cultivar, WL711. This suggests the presence of nonhypersensitive adult plant stripe rust resistance in the line CSP44. The evaluation of F₁, F₂ and F₃ generations and F₆ SSD families from the cross of CSP44 with susceptible wheat cultivar WL711 for stripe rust severity indicated that the resistance in CSP44 is based on two genes showing additive effect. One of these two genes isYr18 and the second gene is not yet described.     

Genetic research as the valid base of strategies for breeding rust resistant wheats

It is known that few wheat cultivars maintain their resistance to rust diseases for a long period of time, particularly when crop populations become genetically more uniform. A number of genetically diverse, so far unexploited, sources of rust resistance in the natural as well as mutagenized population of wheat cultivars were identified. Several of these genes were placed in agronomically superior well-adapted backgrounds so that they could be used as pre-breeding stocks for introducing genetic diversity for resistance in a crop population. Some of these stocks when employed as parents in several cross combinations in a breeding programme have generated a number of promising cultivars with diversity for resistance.Many presently grown wheats in India, near-isogenic lines each with Lr14b, Lr14ab, Lr30 and certain international cultivars were identified as possessing diverse sources of adult plant resistance (APR) to leaf rust. Prolonged leaf rust resistance in some of the Indian cultivars was attributed to the likely presence of Lr34 either alone or in combination with other APR components. Tests of allelism carried out in certain cultivars that continue to show adequate levels of field resistance confirm the presence of Lr34, which explains the role that this gene has played in imparting durability for resistance to leaf rust. Also, Lr34 in combination with other APR components increases the levels of resistance, which suggests that combination of certain APR components should be another important strategy for breeding cultivars conferring durable and adequate levels of resistance. A new adult plant leaf rust resistance source that seems to be associated with durability in Arjun has been postulated. Likewise, cultivars possessing Sr2 in combination with certain other specific genes have maintained resistance to stem rust.Further, non-specific resistances that were transferred across widely different genotypes into two of the popular Indian wheats provided easily usable materials to the national breeding programmes for imparting durable resistance to stripe rust.     

Types of resistance in winter wheat cultivars against Puccinia recondita Rob. ex Desm. in the North Caucasus region

Abstract

The results of studying the types of resistance in 44 recognized and promising winter wheat cultivars for the North Caucasus region are presented. Relatively high levels of race-specific resistance at the germination stage were found in the cultivars Batko, Bystritsa, Krasota, Knyazhna, Polovchanka, Fisht and Yara; low race-specific resistance levels were reported for the most of cultivars studied. Under simulated infection conditions of wheat plants in fields, most cultivars had resistance similar to that one in adult plants. The cultivar Rufa demonstrated slow leaf rust progress, while the cultivar Yuna was highly susceptible to the North Caucasus leaf rust population. The analysis of leaf rust resistance in the cultivars tested makes it possible for agricultural producers to continuously improve their zoning structure according to preferable types of genetic resistance control.     

Identification of molecular markers for the detection of the yellow rust resistance gene Yr17 in wheat

The Yr17 gene, which is present in many European wheat cultivars, displays yellow rust resistance at the seedling stage. The gene introduced into chromosome 2A from Aegilops ventricosa was previously found to be closely linked (0.5 cM) to leaf and stem rust resistance genes Lr37 and Sr38, respectively. The objective of this study was to identify molecular markers linked to the Yr17 gene. We screened with RAPD primers, for polymorphism, the DNAs of cv. Thatcher and the leaf rust-resistant near-isogenic line (NIL) RL 6081 of cv. Thatcher carrying the Lr37 gene. Using a F2 progeny of the cross between VPM1 (resistant) and Thésée (susceptible), the RAPD marker OP-Y15580 was found to be closely linked to the Yr17 gene. We converted the OP- Y15580 RAPD marker into a sequence characterized amplified region (SCAR). This SCAR marker (SC-Y15) was linked at 0.8 ± 0.7 cM to the Yr17 resistance gene. We tested the SC-Y15 marker over a survey of 37 wheat cultivars in order to verify its consistency in different genetic backgrounds and to explain the resistance of some cultivars against yellow rust. Moreover, we showed that the Xpsr150-2Mv locus marker of Lr gene described by Bonhomme et al. [6] which possesses A. ventricosa introgression on the 2A chromosome was also closely linked to the Yr17 gene. Both the SCAR SC-Y15 and Xpsr150-2Mv markers should be used in breeding programmes in order to detect the cluster of the three genes Yr17, Lr37 and Sr38 in cross progenies. This revised version was published online in June 2006 with corrections to the Cover Date.     

The potential of hydroxamic acids in tetraploid and hexaploid wheat varieties as resistance factors against the bird‐cherry oat aphid,Rhopalosiphum padi

The potential for exploiting natural wheat resistance to control the cereal aphid Rhopalosiphum padi, the most important aphid pest of small grain cereals in the UK, was investigated as an alternative approach to the use of insecticides. The investigation focussed on a group of secondary metabolites, the hydroxamic acids or benzoxazinones, present naturally as glucosides, but which hydrolyse on tissue damage to give biologically active aglycones, e.g. 2,4‐dihydroxy‐7‐methoxy‐1,4‐benzoxazin‐3‐one (DIMBOA) which are associated with natural plant defence. These can be important for resistance against insects, fungi, bacteria and nematodes for a range of cultivated monocotyledonous plants and could ultimately be combined with other defence mechanisms to provide a general approach to cereal aphid control. Levels of hydroxamic acids, particularly DIMBOA‐glucoside, were determined in hexaploid (Triticum aestivum) and tetraploid (Triticum durum) wheat varieties and differences were found between species and varieties. The effect of feeding by R. padi on the level of hydroxamic acids in the leaf tissue was also investigated. Thus, after 24 h of aphid feeding, as an apparently localised hydrolytic defence reaction in the leaf, levels of DIMBOA‐glucoside decreased noticeably. When aphids were fed on sucrose solution containing low doses of DIMBOA there was a significant mortality compared to the sucrose control. However, the levels of and variation in hydroxamic acids in the wheat varieties investigated were insufficient for significant differences in aphid behaviour and development.     

Function and evolution of allelic variations of Sr13 conferring resistance to stem rust in tetraploid wheat (Triticum turgidum L.)

The resistance gene Sr13 is one of the most important genes in durum wheat for controlling stem rust caused by Puccinia graminis f. sp. tritici (Pgt). The Sr13 functional gene CNL13 has haplotypes R1, R2 and R3. The R1/R3 and R2 haplotypes were originally designated as alleles Sr13a and Sr13b, respectively. To detect additional Sr13 alleles, we developed Kompetitive allele specific PCR (KASP™) marker KASPSr13 and four semi-thermal asymmetric reverse PCR markers, rwgsnp37–rwgsnp40, based on the CNL13 sequence. These markers were shown to detect R1, R2 and R3 haplotypes in a panel of diverse tetraploid wheat accessions. We also observed the presence of Sr13 in durum line CAT-A1, although it lacked any of the known haplotypes. Sequence analysis revealed that CNL13 of CAT-A1 differed from the susceptible haplotype S1 by a single nucleotide (C2200T) in the leucine-rich repeat region and differed from the other three R haplotypes by one or two additional nucleotides, confirming that CAT-A1 carries a new (R4) haplotype. Stem rust tests on the monogenic, transgenic and mutant lines showed that R1 differed from R3 in its susceptibility to races TCMJC and THTSC, whereas R4 differed from all other haplotypes for susceptibility to TTKSK, TPPKC and TCCJC. Based on these differences, we designate the R1, R3 and R4 haplotypes as alleles Sr13a, Sr13c and Sr13d, respectively. This study indicates that Sr13d may be the primitive functional allele originating from the S1 haplotype via a point mutation, with the other three R alleles probably being derived from Sr13d through one or two additional point mutations.     

Molecular markers for the detection of the wheat leaf rust resistance gene Lr10 in diverse genetic backgrounds

We recently showed that the Lr10 wheat leaf rust resistance gene cosegregated with the candidate resistance gene Lrk10 which encodes a putative receptor-like kinase. The aim of this study was to develop Lrk10-derived molecular markers for the detection of the Lr10 gene in breeding material. Different subfragments of Lrk10 were tested as RFLP markers for the Lr10 resistance gene. The most specific fragment (Lrk10-6) was converted into the PCR-based STS marker STSLrk10-6. Both the RFLP and the STS marker did not give a signal with near isogenic lines containing a different Lr gene. The applicability of these markers for the detection of Lr10 in genetically diverse material was tested with 62 wheat and spelt breeding lines, mostly from European breeding programmes. Twelve varieties known to have Lr10 showed the same alleles as the originally characterized line ThatcherLr10. Most of the lines with unknown composition at the Lr10 locus had a null allele with both the RFLP marker Lrk10-6 and the marker STSLrk10-6 whereas 20% of the lines had a different allele. For six lines, including a traditional spelt variety derived from a landrace, both markers showed the same allele as Thatcher Lr10. Artificial infections of these lines with an isolate avirulent on Lr10 resulted in a hypersensitive reaction of all these lines, indicating also the presence of the Lr10 resistance gene. These data demonstrate that the markers derived from sequences of Lrk10 are highly specific for the Lr10 gene in breeding material of very diverse genetic origin. The markers will allow the defined deployment of Lr10 in wheat breeding programmes and will contribute to the elucidation of the role of Lr10 in polygenic resistances against leaf rust.     

Activities of enzymes involved in lignification during the postharvest storage of etiolated asparagus spears

The content of lignin and the activities of 5 enzymes involved in lignification were monitored along the length of etiolated spears of asparagus ( Asparagus officinalis L., INRA Fl male hybrid n°156) stored for 22 h with their base in air (control), water or water containing the ethylene antagonist, silver thiosulfate (STS). At the time of harvest the lignin content increased basipetally, as did the activity of all the enzymes studied, viz., phenylalanine ammonia lyase (PAL; EC 4.3.1.5), hydroxycinnamate: CoA ligase (HCoAL; EC 6.2.1.12), cinnamoyl-CoA reductase (CCR: EC 1.2.1.44), cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) and syringaldazine oxidase (SyrOx. a peroxidase [POD; EC 1.11.1.7] with syringaldazine as substrate). Neither the lignin content nor the activity of any enzyme changed in the spear apex during storage, regardless of treatment. In the spear base. all enzyme activities decrased during the first 2 to 4 h in every storage treatment. Subsequently. PAL and HCoAL activities remained constant. whereas the activities of CAD and SyrOx gradually increased. Lignification in the spear base was not affected by storage in air. However, storage in water increased lignin formation and SyrOx activity, whereas treatment with STS prevented both of these increases. The results indicate that postharvest lignification in etiolated asparagus spears is caused primarily by enhanced SyrOx activity, and that ethylene is involved in the control of this activity.     

The involvement of reactive oxygen species in defense of wheat lines with the genes introgressed from <Emphasis Type="Italic">Agropyron</Emphasis> species contributing the resistance against brown rust

The role of reactive oxygen species (ROS) in the defense of nearly isogenic lines of common wheat (Triticum aestivum L., cv. Thatcher) with the genes of resistance to brown rust introgressed from Agropyron species was studied using light microscopy. This disease is induced by the fungus Puccinia triticina Erikss. The presence of superoxide anion in the sites of infection was detected with the dye nitro blue tetrazolium. In addition, we studied fungus development on plants treated with the inhibitor of Ca²⁺-channels, verapamil, disturbing penetration into the cells of Ca²⁺ required for ROS generation. During fungus development in the immune line with the Lr38 resistance gene (from A. intermedium (Host) Beuv.), oxidative burst developed at the sites of contacts of appressoria with stomata and exerted a fungicidic effect. When ROS generation was suppressed, the fungus developed haustoria in the mesophyll cells. In plants with the Lr19 gene (from A. elongatum (Host) Beuv.), only moderate amount of superoxide anion accumulated on the cell walls of stomatal guard cells and in the infection structures when the fungus penetrated into the substomatal cavity and in mesophyll cells. In plants with the Lr24 gene (from A. elongatum), superoxide anion was detected only around haustoria. Suppression of ROS generation in plants harboring the Lr19 and Lr24 genes did not affect fungus entrance into the substomatal cavity but facilitated penetration of haustoria into the mesophyll cells. At the same time, in the lines with the Lr1 gene (from T. aestivum), cytological examination did not detect O ₂ ^? accumulation in plant cells, whereas treatment with verapamil enhanced mycelium development. In all lines, the suppression of oxidative burst slowed the development of hypersensitive response.     

QTL identification and microphenotype characterisation of the developmentally regulated yellow rust resistance in the UK wheat cultivar Guardian

Yellow rust (causal agent: Puccinia striiformis f.sp. tritici) resistance in the UK wheat cultivar Guardian is developmentally regulated, resistance increasing as the plant matures. Yellow rust resistance was assessed under field conditions on plants after ear emergence to ensure maximum expression of resistance. Three quantitative trait loci (QTL) for yellow rust resistance were identified, being located on chromosomes 1B (QPst.jic-1B), 2D (QPst.jic-2D) and 4B (QPst.jic-4B). The largest resistance effect, QPst.jic-1B located to the same position on the long arm of chromosome 1B as the known durable source of yellow rust resistance, Yr29. Microscopic studies were carried out to determine what effect the resistance in Guardian had on the development of P. striiformis f.sp. tritici. While the adult plant resistance in Guardian did not prevent germinated urediniospores from establishing an effective infection site, the growth of hyphae within flag leaf tissue was significantly inhibited, slowing the development of microcolonies. 3,3-diaminabenzadine (DAB) and trypan blue staining indicated that this inhibition of hyphal growth was not associated with hydrogen peroxide accumulation or extensive plant cell death.     

TaADF4, an actin‐depolymerizing factor from wheat,is required for resistance to the stripe rust pathogen Puccinia striiformis f. sp. tritici

Actin filament assembly in plants is a dynamic process, requiring the activity of more than 75 actin‐binding proteins. Central to the regulation of filament assembly and stability is the activity of a conserved family of actin‐depolymerizing factors (ADFs), whose primarily function is to regulate the severing and depolymerization of actin filaments. In recent years, the activity of ADF proteins has been linked to a variety of cellular processes, including those associated with response to stress. Herein, a wheat ADF gene, TaADF4, was identified and characterized. TaADF4 encodes a 139‐amino‐acid protein containing five F‐actin‐binding sites and two G‐actin‐binding sites, and interacts with wheat (Triticum aestivum) Actin1 (TaACT1), in planta. Following treatment of wheat, separately, with jasmonic acid, abscisic acid or with the avirulent race, CYR23, of the stripe rust pathogen Puccinia striiformis f. sp. tritici, we observed a rapid induction in accumulation of TaADF4 mRNA. Interestingly, accumulation of TaADF4 mRNA was diminished in response to inoculation with a virulent race, CYR31. Silencing of TaADF4 resulted in enhanced susceptibility to CYR23, demonstrating a role for TaADF4 in defense signaling. Using a pharmacological‐based approach, coupled with an analysis of host response to pathogen infection, we observed that treatment of plants with the actin‐modifying agent latrunculin B enhanced resistance to CYR23, including increased production of reactive oxygen species and enhancement of localized hypersensitive cell death. Taken together, these data support the hypothesis that TaADF4 positively modulates plant immunity in wheat via the modulation of actin cytoskeletal organization.     

Induced chitinase activity in resistant wheat leaves inoculated with an incompatible race of Puccinia striiformis f. sp. tritici,the causal agent of yellow rust disease

Chitinase specific activity was measured spectrophotometrically in wheat leaf tissues during the compatible and incompatible interactions with Puccinia striiformis f. sp. tritici, the causal agent of yellow rust disease. The wheat cultivar, Federation^* 4/Kavkaz, was inoculated with virulent (134E134A+) or avirulent (4EOA+) races of P. striiformis f. sp. tritici in the first leaf stage. The results showed that chitinase activity pattern was similar in both compatible and incompatible interactions up to 72 hrs after inoculation. However, the specific activity increased rapidly in the incompatible reaction thereafter. In susceptible reaction, chitinase activity gradually declined after 72 hrs post-inoculation reaching a level similar to that in the control plants two weeks after inoculation. Chitinase specific activity in resistance response was at least three times greater than that in the susceptible reaction two weeks following the inoculation. Electrophoresis of native polyacrylamide gel impregnated with 0.1% (w/v) glycol chitinas the substrate revealed the presence of eight chitinase isoforms with relative electrophoretic mobility (R_m) values ranging from 0.11 to 0.64 in the resolving gel. This revised version was published online in June 2006 with corrections to the Cover Date.     

Discovery,localization, and sequence characterization of molecular markers for the crown rust resistance genes <Emphasis Type="Italic">Pc38, Pc39</Emphasis>, and <Emphasis Type="Italic">Pc48</Emphasis> in cultivated oat (<Emphasis Type="Italic">Avena sativa</Emphasis> L.)

Molecular markers for the crown rust resistance genes Pc38, Pc39, and Pc48 in cultivated oat (Avena sativa L.) were identified using near-isogenic lines and bulked segregant analysis. Six markers for Pc48, the closest being 6 cM away, were found in a Pendek-39 × Pendek-48 (Pendek3948) population, but none was found in a Pendek-48 × Pendek-38 (Pendek4838) population. Three markers for Pc39 were found in the Pendek3948 population, one of which cosegregated with the gene. This same marker was found to be 6 cM away from the gene in an OT328 × Dumont (OT328Du) population. Nine markers for Pc38 were found in the Pendek4838 population, eight of which are within 2 cM of the gene. One other marker for Pc38 was found in the OT328Du population; however, comparative mapping suggests that the Pc38 region in OT328Du is in a different location than that in Pendek4838. A number of markers unlinked to the genes under study formed linkage groups in both the Pendek3948 and Pendek4838 populations. Four of these show homology or homoeology to each other and to the Pc39 region in Pendek3948. Two RFLP clones closely linked to Pc38 code for a putative leucine-rich repeat transmembrane protein kinase and a cre3 resistance gene analogue. This study provides information to support molecular breeding in oat, and contributes to ongoing research into genomic regions associated with fungal pathogen resistance.