Differential gene expression in loblolly pine (Pinus taeda L.) challenged with the fusiform rust fungus,Cronartium quercuum f.sp. fusiforme

Cronartium quercuum f.sp. fusiforme is the pathogen that incites fusiform rust disease of southern pine species. To date, a number of host resistance genes have been mapped. Although genomic mapping studies have provided valuable information on the genetic basis of disease interactions in this pine-rust pathosystem, the interaction at the molecular level is poorly understood. To further our understanding of this interaction, we implemented a microarray study to examine the differential expression of genes in pathogen-challenged progeny of a full-sib loblolly pine family known to be segregating at a single dominant resistance gene (Fr1). Statistical analyses revealed shifts in gene expression that may reflect discrete stages of gall development.     

Variation in the pine stem rust fungus Peridermium pini in the United Kingdom

Peridermium pini aeciospores from lesions on Scots pine ( Pinus sylvestris ) in Scotland and in Thetford Forest, East Anglia, differed in terms of germ tube morphology and cytology. The former produced long aseptate germ tubes containing a pair of nuclei. The latter produced short septate germ tubes, each cell containing a single nucleus. One third of the 'Scottish' germ tubes ended in a vesicle. Nuclei could be counted in only a small proportion of ungerminated aeciospores. On average 83% of Scottish spores had two nuclei and 4% had one. The equivalent figures for Thetford spores were 66 and 20%. A small number of aeciospore samples from other parts of England resembled Scottish spores rather than Thetford spores in germ tube morphology and cytology.
Both Scottish and Thetford spores proved capable of causing infection of wounded Scots pine shoots. In addition, some Thetford spores caused infection on leaf discs taken from certain Paeonia cultivars. From these results it is concluded that two distinct forms of Peridermium pini exist in the UK. The implications of this finding are discussed in relation to the history of pine stem rust in the country and the genetics and taxonomy of P. pini.     

Influence of host resistance on the genetic structure of the white pine blister rust fungus in the western United States

Cronartium ribicola, the causal agent of white pine blister rust, has been devastating to five-needled white pines in North America since its introduction nearly a century ago. However, dynamic and complex interactions occur among C. ribicola, five-needled white pines, and the environment. To examine potential evolutionary influences on genetic structure and diversity of C. ribicola in western United States, population genetic analyses of C. ribicola were conducted using amplified fragment length polymorphism (AFLP) molecular markers. The fungus was sampled at six sites. Collections for two of the six sites were from separate plantings of resistant-selected western white pine and sugar pine. Heterozygosity based on polymorphic loci among populations ranged from 0.28 to 0.40, with resistant-selected plantations at the extremes. Genetic differentiation was also highest between these two populations. Principal coordinates analysis and Bayesian assignment placed most isolates that are putative carriers of virulence to major-gene resistance into a discernable cluster, while other isolates showed no clustering by site or host species. These results indicate that C. ribicola in western North America is not genetically uniform, despite its presumed single site of introduction and relatively brief residence. Moreover, major-gene resistance appears to have imposed strong selection on the rust, resulting in reduced genetic diversity. In contrast, no evidence of selection was observed in C. ribicola from hosts that exhibit only multigenic resistance.     

A major gene for resistance to white pine blister rust in Western white pine from the Western cascade range

ABSTRACT A dominant gene for resistance to white pine blister was indicated by Mendelian segregation in full-sib families of western white pine parent trees selected for phenotypic resistance in six heavily infected stands in the Western Cascades of Oregon and Washington. Seedlings were artificially inoculated three times between 1959 and 1964 and observed for development of stem infection. Segregation at this locus (Cr2) occurred in only two of the six parent populations sampled: one a natural stand, Champion Mine (CM), and the other a plantation of unknown seed origin. At CM, reduced penetrance of this gene was expressed by altered Mendelian ratios (mostly less-than-expected resistant phenotypes) in families of specific combinations of certain parents, indicating the presence of modifier genes with effects that ranged from mild to almost complete suppression of Cr2. Between 1968 and 1994, an apparent shift in virulence at CM caused all of the resistant selections to become infected and die. Recent inoculations of many of the same or related families from these parents, made from grafted ramets in a seed orchard, showed that Cr2 conditions a classical hypersensitive reaction (HR) in needle tissues, the primary infection courts. In the latter tests, seedlings were challenged with wild-type and four other sources of inoculum at and near CM that were also suspected of having wider virulence than wild type. No seedlings segregating for HR that were inoculated with wild type subsequently developed stem symptoms, but the other inocula induced both susceptible and HR needle spots on Cr2- genotypes, and many of these seedlings did develop stem infections. This implied that spore genotypes with specific virulence to Cr2 are carried in these inocula.     

基于基因混池测序技术的小麦条锈菌对三唑酮抗性位点相关分析

为探究小麦条锈菌Puccinia striiformis f. sp. tritici对三唑类杀菌剂产生抗药性的相关机理,以小麦条锈菌抗药性菌株YQ324与敏感性菌株贵1-2在转主寄主堆花小檗Berberis aggregata上构建的有性遗传群体为材料,通过离体叶段法对F₁代杂交及F₂代自交后代的夏孢子群体进行三唑酮敏感性生物学测定,并在F₂代夏孢子群体中筛选20株抗药性菌株和20株敏感性菌株分别构建DNA抗、感池。在对抗、感混池进行高通量测序之后,通过质量检测和数据分析得到2个极端混池中的变异信息。根据ΔIndex为1的筛选标准在小麦条锈菌中鉴定出67个与三唑酮抗性显著相关的单核苷酸多态性（single nucleotide polymorphism,SNP）位点;利用SNPEff软件对这些SNP位点引起氨基酸非同义突变所在的基因序列进行功能注释,最终在pcontig_030上筛选到Pst104E_09436和Pst104E_09437这2个基因作为小麦条锈菌对三唑酮抗性相关的候选基因。     

Leaf rust resistance gene lr34 associated with nonsuppression of stem rust resistance in the wheat cultivar canthatch

ABSTRACT The common wheat cultivar Thatcher and the backcross derivative Canthatch are moderately or fully susceptible to several races of stem rust because of a suppressor on chromosome 7DL that inhibits the expression of the relevant resistance gene(s). The incorporation of leaf rust resistance gene Lr34 into 'Thatcher' is known to enhance stem rust resistance. The effect of this gene in a 'Canthatch' background and its relationship with the 7DL suppressor were determined by replacing chromosome 7D of 'Canthatch' with 7D of 'Chinese Spring', which possesses Lr34 on the short arm. 'Canthatch' nullisomic 7D was crossed with 'Chinese Spring', followed by a succession of backcrosses to the nullisomic recurrent parent. Homozygous resistant disomic and monosomic substitution lines were recovered that exhibited the same resistant reaction as that of 'Thatcher' possessing Lr34 and as that of 'Canthatch' nullisomic 7D, in which the 7DL suppressor is absent. The results indicate that, in 'Canthatch', Lr34 permits expression of resistance genes normally inhibited by the 7DL suppressor. Furthermore, it is likely that, in 'Thatcher' and 'Thatcher' back-cross derivatives, Lr34 inactivates the 7DL suppressor.     

Molecular mapping of the crown rust resistance gene rpc1 in barley

ABSTRACT Crown rust of barley, caused by Puccinia coronata var. hordei, occurs sporadically and sometimes may cause yield and quality reductions in the Great Plains region of the United States and Canada. The incompletely dominant resistance allele Rpc1 confers resistance to P. coronata in barley. Two generations, F(2) and F(2:3), developed from a cross between the resistant line Hor2596 (CIho 1243) and the susceptible line Bowman (PI 483237), were used in this study. Bulked segregant analysis combined with random amplified polymorphic DNA (RAPD) primers were used to identify molecular markers linked to Rpc1. DNA genotypes produced by 500 RAPD primers, 200 microsatellites (SSRs), and 71 restriction fragment length polymorphism (RFLP) probes were applied to map Rpc1. Of these, 15 RAPD primers identified polymorphisms between resistant and susceptible bulks, and 62 SSR markers and 32 RFLP markers identified polymorphisms between the resistant and susceptible parents. The polymorphic markers were applied to 97 F(2) individuals and F(2:3) families. These markers identified 112 polymorphisms and were used for primary linkage mapping to Rpc1 using Map Manager QT. Two RFLP and five SSR markers spanning the centromere on chromosome 3H and one RAPD marker (OPO08-700) were linked with Rpc1 and, thus, used to construct a 30-centimorgan (cM) linkage map containing the Rpc1 locus. The genetic distance between Rpc1 and the closest marker, RAPD OPO08-700, was 2.5 cM. The linked markers will be useful for incorporating this crown rust resistance gene into barley breeding lines.     

Genetic differentiation of the wheat leaf rust fungus Puccinia triticina in Europe

The objective of this study was to determine whether genetically differentiated groups of Puccinia triticina are present in Europe. In total, 133 isolates of P. triticina collected from western Europe, central Europe and Turkey were tested for virulence on 20 lines of wheat with single leaf rust resistance genes, and for molecular genotypes with 23 simple sequence repeat (SSR) markers. After removal of isolates with identical virulence and SSR genotype within countries, 121 isolates were retained for further analysis. Isolates were grouped based on SSR genotypes using a Bayesian approach and a genetic distance method. Both methods optimally placed the isolates into eight European (EU) groups of P. triticina SSR genotypes. Seven of the groups had virulence characteristics of isolates collected from common hexaploid wheat, and one of the groups had virulence characteristics of isolates from tetraploid durum wheat. There was a significant correlation between the SSR genotypes and virulence phenotypes of the isolates. All EU groups had observed values of heterozygosity greater than expected and significant fixation values, which indicated the clonal reproduction of urediniospores in the overall population. Linkage disequilibria for SSR genotypes were high across the entire population and within countries. The overall values of R_ST and F_ST were lower when isolates were grouped by country, which indicated the migration of isolates within Europe. The European population of P. triticina had higher levels of genetic differentiation compared to other continental populations.     

中粱93447抗条锈病基因遗传分析和分子作图

 用小麦条锈菌生理小种对中梁93447与感病品种铭贤169的杂交后代F₁、F₂和F₃代进行苗期温室抗条锈性遗传分析,结果表明中梁93447对CYR30的抗病性由1对显性基因控制。用中梁93447×铭贤169 F₂代分离群体建立抗、感DNA池,在F₂代群体中通过SSR标记技术寻找与抗病基因连锁的分子标记,发现7个位于5BS上的标记Xwmc813、Xwmc740、Xgwm159、Xbarc4、Xwmc616、Xwmc363和Xbarc89与目的基因连锁,遗传距离分别为17、12.9、8.3、4.5、3.7、9.1和20.8cM。系谱分析及分子标记分析表明,该基因可能来自中间偃麦草,暂命名为YrZL93447。与已定位于5B染色体上的抗条锈病基因的比较研究表明,YrZL93447可能是1个新的抗条锈病基因。用SSR引物BARC4和WMC616检测43个黄淮麦区主栽品种,其中7%的黄淮麦区主栽品种具有与YrZL93447基因相同的标记位点。     

Molecular marker mapping of leaf rust resistance gene lr46 and its association with stripe rust resistance gene yr29 in wheat

ABSTRACT Leaf and stripe rusts, caused by Puccinia triticina and P. striiformis, respectively, are globally important fungal diseases of wheat that cause significant annual yield losses. A gene that confers slow rusting resistance to leaf rust, designated as Lr46, has recently been located on wheat chromosome 1B. The objectives of our study were to establish the precise genomic location of gene Lr46 using molecular approaches and to determine if there was an association of this locus with adult plant resistance to stripe rust. A population of 146 F(5) and F(6) lines produced from the cross of susceptible 'Avocet S' with resistant 'Pavon 76' was developed and classified for leaf rust and stripe rust severity for three seasons. Using patterns of segregation for the two diseases, we estimated that at least two genes with additive effects conferred resistance to leaf rust and three to four genes conferred resistance to stripe rust. Bulked segregant analysis and linkage mapping using amplified fragment length polymorphisms with the 'Avocet' x 'Pavon 76' population, F(3) progeny lines of a single chromosome recombinant line population from the cross 'Lalbahadur' x 'Lalbahadur (Pavon 1B)', and the International Triticeae Mapping Initiative population established the genomic location of Lr46 at the distal end of the long arm of wheat chromosome 1B. A gene that is closely linked to Lr46 and confers moderate levels of adult plant resistance to stripe rust is identified and designated as Yr29.     

Origin and distribution of cr2, a gene for resistance to white pine blister rust in natural populations of Western white pine

ABSTRACT The distribution and frequency of the Cr2 gene for resistance to white pine blister rust (Cronartium ribicola) in western white pine (Pinus monticola) was surveyed in natural populations of the host by inoculation of open-pollinated seedlings from 687 individual seed parents from throughout most of the species' range. Because Cr2 is dominant and results in a conspicuous hypersensitive reaction (HR) in pine needles, the phenotype can readily be detected in offspring of susceptible seed parents fertilized by unknown Cr2 donors in the ambient pollen cloud. Gametic frequencies of Cr2 were thus determined as the proportion of total challenged seedlings that were pollen receptors exhibiting the Cr2 phenotype. Zygotic frequencies, the proportion of seed parents with progeny that segregated in Mendelian ratios for the Cr2 phenotype to the total number of parents, were a complementary, though less precise, measure. Cr2 frequency was rare overall, ranging from 0.004 to 0.008 in the Sierra Nevada to about 0.001 in the central Cascade Range; it was undetectable further north in the Cascades, as well as in the Rocky Mountains and Coast Mountains of the United States and Canada. The diminishing frequency of Cr2 from the southern and central Sierra Nevada northward mirrors that of Cr1 in sugar pine (P. lambertiana) and points to this region as the origin of both genes. We rationalize that this coincidence may have resulted from protection that these genes may have conferred on both species to an endemic pine stem rust congeneric with C. ribicola (C. occidentale) in recent geologic epochs.     

小偃9366抗条锈病基因的遗传分析及分子作图

为了明确小偃9366抗条锈病遗传特点,对小偃9366与铭贤169及其杂交F₁、F₂、F₃和BC₁F₁代进行温室苗期抗条锈性遗传分析,选取400余对SSR引物对接种CYR31的群体进行分子标记,并利用目标基因的侧翼引物分析99个黄淮麦区主栽小麦品种。小偃9366对CYR25的抗病性由1显、1隐2对基因独立控制,对CYR27的抗病性由3对显性基因控制,其中2对基因表现累加作用,对CYR30 和CYR31的抗病性均由1对显性基因独立控制,对Su11-4的抗性由2对隐性基因独立控制。位于2AL上的6个标记Xwmc794、Xwmc455、Xwmc261、Xgwm47、Xgwm294和Xcfd168与抗CYR31基因(暂命名Yrxy9366)连锁,与目的基因的遗传距离分别为10.8、6.5、3.2、4.4、16.0和32.8 cM,将Yrxy9366定位在2AL上。利用Xwmc261、Xgwm47两个引物分析99个黄淮麦区主栽小麦品种,仅5%检测到同源片段。研究表明Yrxy9366是一个新的抗病基因。     

欧洲小麦品种Mega抗条锈病基因的遗传分析及分子标记

 本研究表明欧洲小麦品种Mega对我国小麦条锈病重要流行小种CYR30、CYR31、CYR32、Su-4和Su-14在苗期都具有良好的抗病性。采用小麦条锈菌小种CYR30对Mega与感病小麦品种铭贤169杂交的F₁、F₂和BC₁代及双亲进行苗期抗病性遗传分析,结果表明,Mega对CYR30的抗性由1对显性基因独立控制。采用SSR标记技术对其携带的抗性基因进行分子标记,在237对SSR引物中,发现位于5BL上的2个SSR引物位点Barc232、Wmc640在双亲和抗、感池间能扩增出稳定的特异性片段,与抗病基因连锁的遗传距离分别是3.7cM和8.6cM,暂命名为YrMe。本研究结果为科学利用Mega抗条锈基因培育抗病品种提供了依据。     

小麦品系中梁93444的抗条锈性遗传分析与分子作图

为明确抗锈品种中梁93444抗条锈基因及遗传特点,用CYR30、CYR31、CYR32对该品种、铭贤169及杂交组合进行遗传分析,用SSR技术对分离家系F_3-3进行PCR扩增和电泳分析。结果显示,中梁93444对CYR30、CYR31的抗病性均由1对显性和1对隐性基因控制,对CYR32由2对显性互补基因控制;F_3-3分离家系对CYR32的抗病性由1对显性基因控制,该基因暂命名为Yr93444。对F_3-3分离群体进行SSR标记,建立了与该基因连锁的8个标记Xgwm122、Xwmc702、Xwmc644、Xwmc794、Xgwm328、Xwmc455、Xgwm372、Xwmc819,遗传距离分别为38.1、30.7、22.9、15.6、10.0、6.9、3.5和2.8 cM。 应用SSR标记、中国春及缺四体将Yr93444定位于2AL上。系谱分析和SSR分子标记检测表明,该基因是来自中间偃麦草的新抗条锈基因。用与该基因紧密连锁的SSR标记Xgwm372和Xwmc819检测中梁品种和黄淮麦区主栽品种,发现89%中梁品种含该抗病基因,而86%黄淮麦区主栽品种不含该抗病基因,表明该基因应用潜力很大。     

普通小麦-柔软滨麦草易位系M852-1抗条锈基因的遗传分析和分子作图

 M852-1是经杂交和回交培育的普通小麦-柔软滨麦草易位系,苗期对我国小麦条锈菌流行小种均表现良好抗性。为明确其抗条锈性遗传规律,本研究选用条锈菌流行小种（类型）CYR29、CYR32、CYR33和Su11-7的单孢菌系对其与铭贤169杂交F₁、F₂、F₃及BC₁代群体进行遗传分析, 同时应用420对SSR引物对接种CYR32的M852-1/铭贤169 F₂代144个单株作图群体进行抗病基因定位。结果表明,M852-1对供试小种均表现免疫或近免疫,对CYR29的抗锈性由1对显性基因控制,对CYR32、CYR33和Su11-7的抗锈性均由1对隐性基因控制。筛选到3个与抗CYR32基因连锁的SSR标记Xbarc124、Xbarc200和Xgwm429,遗传距离分别为6.3、5.6 和 9.7 cM。根据SSR标记锚定性将该基因定位于小麦2BS染色体,暂命名为YrM852。基因来源、分子标记检测及染色体位点分析表明,YrM852很可能是1个不同于目前已知抗条锈病基因的新基因。     

Genetic relationship between the adult plant resistance gene Lr12 and the complementary gene Lr31 for seedling resistance to leaf rust in common wheat

A resistant phenotype similar to that conferred in wheat by the complementary genes Lr27  +  Lr31 was produced in the progeny of intercrosses of cultivars carrying Lr27 and a line possessing Lr12 . This confirms that Lr12 is either completely linked with Lr31 or is the same gene. On the basis of these findings and that Lr31 is located on chromosome 4BS, it is concluded that Lr12 must also be located on 4BS. Adult-plant genetic tests confirm that the Australian wheat cultivar Timgalen carries Lr12 , and stocks with Lr12 alone were established from this cultivar.     

西科麦2028抗条锈病的遗传特点

<正>小麦条锈病是由小麦条锈菌Puccinia striiformis f.sp.tritici引起的小麦叶部病害,是全世界范围内小麦上最重要的病害之一[1,2]。在我国,西北冬春麦区、西南冬麦区、黄淮海华北、淮北等地冬麦区,都发生过小麦条锈病的暴发流行,给小麦生产带来了巨大损失。2002年,生理小种CYR32在我国引起小麦条锈病暴发流行,造成小麦产量损失1.3亿多吨,有的省份甚至有10%~20%以上的小麦     

Allele sequencing of the barley stem rust resistance gene Rpg1 identifies regions relevant to disease resistance

The stem rust resistance gene Rpg1 has protected North American barley cultivars from significant yield losses for over 65 years. The remarkable durability of this gene warrants further study as to its possible origin and allelic variation. Eight Swiss barley (Hordeum vulgare) landraces and eight wild barley (H. vulgare subsp. spontaneum) accessions from diverse geographic regions were analyzed to uncover new alleles of Rpg1 and learn about its possible origin. The two germplasm groups included accessions that were resistant and susceptible to Puccinia graminis f. sp. tritici pathotype MCCF. Allele-specific primers were utilized to amplify 1 kbp overlapping fragments spanning the Rpg1 gene and sequenced if a polymerase chain reaction (PCR) fragment was generated. Variation among the PCR products revealed significant polymorphisms among these Hordeum accessions. Landraces and wild barley accessions susceptible to pathotype MCCF exhibited the highest degree of Rpg1 polymorphism. One resistant landrace (Hv672) and one resistant wild barley accession (WBDC040) yielded all seven Rpg1-specific PCR fragments, but only landrace Hv672 coded for an apparently functional Rpg1 as determined by comparison to previously characterized resistant and susceptible alleles and also resistance to HKHJ, a stem rust pathotype that can specifically detect Rpg1 in the presence of other resistance genes. Accessions resistant to stem rust pathotype MCCF, but completely lacking Rpg1-specific PCR amplification and hybridization with an Rpg1-specific probe, suggested the presence of stem rust resistant gene(s) different from Rpg1 in the Hordeum germplasm pool. Some Rpg1 alleles that retained the ability to autophosphorylate did not confer resistance to Puccinia graminis f. sp. tritici pathotype MCCF, confirming our previous observations that autophosphorylation is essential, but not sufficient for disease resistance. Thus, the RPG1 protein plays a complex role in the stem rust disease resistance-signaling pathway.     

Genetic variability and the inheritance of resistance to cedar-apple rust in apple

Thirteen apple cultivars and selections were evaluated in a greenhouse for their reaction to cedar-apple rust inoculation Five of the cultivars selections developed aecial lesions on their leaves whereas the sixth cultivar Jonathan, showed only pycnial lesions. The responses of the remaining seven resistant cultivars selections ranged from a lack of any visible symptoms to the presence of discrete flecks or continuous chlorotic mottles Quantitative parameters were used for statistical analysis; these included the highest number of pycnial lesions on a leaf/plant ( N ). the largest diameter of pycnial lesions on any leaf/plant (D)), and the calculated infection rating ( IR = ND ). Significant differences were observed among cultivars selections for variables N. D. and IR . Segregation ratios among progenies derived from 16 controlled crosses indicated that resistance to cedar-apple rust was conditioned by two major genes, and both were required in dominant form to confer resistance. However, in three crosses the dual-gene hypothesis was inadequate to explain the reaction of seedlings.     

小麦持久抗病品种N. Strampelli的抗条锈病基因分析和微卫星标记

 N. Strampelli是由意大利引入我国的小麦持久抗病性品种,对我国目前多数的条锈菌流行小种均有良好的抗性。为了明确其抗条锈病基因的遗传机制,利用小麦条锈病小种CYR30、CYR31、Su-4和Su-14对N. Strampelli与中国春杂交后代进行遗传分析,结果表明N. Strampelli对CYR30、CYR31的抗病性均由1对显性基因和1对隐性基因互补控制,对Su-14、Su-4的抗病性各由1对隐性基因控制,将其中控制Su-14抗病性的隐性基因暂时命名为YrNS-1。利用分离群体分析法(BSA)对接种Su-14的正交F₂代群体进行SSR分子标记,在1BL上找到4个与YrNS-1紧密连锁的微卫星标记Xwmc719、Xgwm124、Xwmc44和Xcfa2147,遗传距离分别为3.2、4.6、5.7和10.3cM。与已知位于1BL染色体上的抗条锈基因比较分析表明,YrNS-1可能是1个新的抗条锈病基因。