马铃薯帚顶病毒研究进展

综述了近年来国内外马铃薯帚顶病毒(PMTV)所取得的研究进展;简述了PMTV的分布与传播、分子特征、病理特征、致病机理及其检测方法,认为PMTV主要出现在气温冷凉的高纬度、高海拔地区的马铃薯上,能通过种薯和粉痂菌传播,诱导马铃薯块茎发生环状坏死;分析了该病毒的特殊性及防治的难点;认为今后要深入研究粉痂菌、马铃薯、PMTV三者之间的相互作用,挖掘PMTV抗性基因,开展马铃薯抗PMTV育种,建立快速准确的PMTV检测方法,探讨同时防治粉痂病和PMTV的途径。     

Investigation of long-term field experiments on response of breeding lines to common scab in a potato breeding program

Common scab, caused by Streptomyces scabies, is a disease that produces scab-like surface lesions on potato tubers. Testing for susceptibility/resistance of breeding lines at the Potato Research Centre of Agriculture & Agri-Food Canada (AFC) is carried out in a scab nursery maintained at AFC for the annual scab evaluation field trials. A replicated field trial routinely consists of breeding lines from previous testing season(s) plus newly selected lines. Data of scab scores generated from long-term experiments thus formulate an incomplete 2-way table over combinations of breeding lines (genotypes) and trials (years). This requires an advanced statistical method to estimate genetic parameters for evaluation purposes. A data base with 1,435 scab index scores from 344 breeding lines were extracted from 5 years (1995–1999) of field experiments in the scab nursery maintained at AFC. The statistical method Residual Maximum Likelihood (REML) was employed to estimate variance components of the breeding population and Best Linear Unbiased Predictor (BLUP) was used to predict genetic merit of breeding lines. High heritability was obtained from variance components estimated by REML. The BLUP scores of breeding lines provided reliable evaluation of their responses to common scab. Two data base sub-sets were separately formulated from the original data base for those parents and grandparents of the breeding lines having pedigree records available. They were again subjected to REML and BLUP analyses to compare the responses to common scab and identify sources of resistance at the parental and grandparental levels. Two random data sets with equal sized samples of breeding lines were also generated from the over-all data base. The two sets of BLUP scores between corresponding breeding lines and parents showed high association which provides an assessment of the validity of the evaluation process.     

Potato genetics,genomics, and applications

Potato has a variety of reproductive uniquenesses besides its clonal propagation by tubers. These traits are controlled by a different kind of genetic control. The reproductive information has been applied to enable interspecific hybridization to enhance valuable traits, such as disease and pest resistances, from the tuber-bearing Solanum gene pool. While progress has been made in potato breeding, many resources have been invested due to the requirements of large populations and long time frame. This is not only due to the general pitfalls in plant breeding, but also due to the complexity of polyploid genetics. Tetraploid genetics is the most prominent aspect associated with potato breeding. Genetic maps and markers have contributed to potato breeding, and genome information further elucidates questions in potato evolution and supports comprehensive potato breeding. Challenges yet remain on recognizing intellectual property rights to breeding and germplasm, and also on regulatory aspects to incorporate modern biotechnology for increasing genetic variation in potato breeding.     

Inheritance of resistance to Fusarium head blight in the wheat lines 'CJ 9306' and 'CJ 9403'

Fusarium head blight (FHB or scab) caused by Fusarium graminearum is a worldwide serious disease in wheat. Exploitation and genetic studies of elite resistance sources can speed up the development of resistant cultivars. To characterize the inheritance of host plant resistance in two new lines, ‘CJ 9306’ and ‘CJ 9403’, developed from a recurrent selection programme in China, six generations P₁, P₂, F₁, F₂, B₁ and B₂ of four crosses and 137 F_{6 : 7} recombinant inbred lines (RILs) from one cross were evaluated in the greenhouse for scab resistance using single‐floret inoculation. The data of area under disease progress curve (AUDPC) in F₂, backcross (BC) and RIL populations exhibited mono‐modal distributions without clear‐cut demarcations and skewing towards resistance. An additive–dominance model was well‐fitted, additive effects played a predominating role, and dominance effects were also significant. Continuous distributions with two major peaks and one minor peak for the number or percentage of scabby spikelets (NSS or PSS) in segregating populations implied the existence of major genes or quantitative trait loci (QTL) for resistance. The estimates of broad‐sense and narrow‐sense heritabilities based on the six‐generation experiment were 56–76% and 26–67% respectively. The estimates of broad‐sense heritabilities based on anova with RILs were 89–90%. These two improved lines with excellent scab resistance and good agronomic traits are of interest for wheat breeding and production.     

Heritility of field resistance to potato leafroU virus in cultivated potato

Potato progenies in a line x tester mating design and the clonal parents were screened for field resistance to potato leafroll virus (PLRV) to determine the heritability of this trait. Twelve advanced potato clones or varieties were crossed as pistillate parents to two pollen testers. The seedling progenies and clonal parents were exposed to aphid-transmitted potato leafroll virus for two growing seasons. Cumulative infection by potato leafroll virus was determined by post-season sero-logical indexing of foliage grown from sprouted tubers after 2 years of exposure. Narrow-sense heritability was estimated from regression of mid-parent on progeny as h = 0.72. This estimate indicates a high level of useabie genetic variance for PLRV resistance in advanced breeding materials. Although variation in resistance to PLRV appears to be a quantitative trait in susceptible and moderately resistant clones, per-formance of the most resistant parents suggests that genes with major effects may be present. These results are similar to the conclusions of other researchers who found one or two genes controlling the pheno-types of extreme resistance, resistance to infection, or suppression of virus titre.     

Trial heterogeneity and variance models in the genetic evaluation of potato tuber yield

Genetic evaluation aims to identify genotypes with high empirical breeding values (EBVs) for selection as parents. In this study, 2157 potato genotypes were evaluated for tuber yield using 8 years of early‐stage trial data collected from a potato breeding programme. Using linear mixed models, spatial parameters to target greater control of localised spatial heterogeneity within trials were estimated and variance models to account for across‐trial genetic heterogeneity were tested. When spatial components improved model fit, correlations of errors were mostly small and negative for marketable tuber yield (MTY) and total tuber yield (TTY), suggesting the presence of interplot competition in some years. For the analysis of multi‐environment trials, a variance model with a simple correlation structure (with heterogeneous variances) was the most favourable variance structure fitted for TTY and PTY (per cent marketable yield). There was very little difference in model fit when comparing a factor analytic structure of order 2 (FA2) with either FA1 or simple correlation structures for MTY, indicating that simple variance models may be preferable for early‐stage genetic evaluation of potato yield.     

Approaches for breeding apples with durable disease resistance

Summary In most apple breeding programmes resistance to diseases has a high priority. The main emphasis is put on scab and mildew resistance. Recent evidence about the risk of breakdown of the Vf scab resistance demonstrates the need to search for new approaches in disease resistance breeding. The following options might be considered: discover and exploit new sources of resistance to enlarge the genetic basis of resistance; accumulate different functional resistances in one genotype; develop orchard designs in order to increase diversity in the host-pathogen interactions and establish a resistance management system. To implement these strategies the genetic structure of the resistance must be examined by using modern methods of molecular biology. Resistance donors and advanced selections should be examined under high natural inoculum pressure at several locations around the world. We examined the possibility of identifying polygenic or partial scab resistance in apple. Nineteen apple varieties and selections, among them the differential hosts for scab races 1 to 5, were artificially inoculated in the glasshouse with scab conidia. The leaf symptoms were analysed macroscopically and microscopically There were significant differences which are actually exploited in our breeding programme. Conclusions are drawn towards new approaches for breeding and growing apples with durable disease resistance.     

Mapping powdery mildew resistance gene in V97‐3000 soybean

V97‐3000 is a maturity group (MG) V soybean breeding line derived from SS 516 × V90‐2592 (Vance × V81‐1325) with high stachyose, small seed and powdery mildew resistance. A total of 53 F_2:3 families were derived from a cross between V97‐3000 and a powdery mildew susceptible line V99‐5089. The 53 F_2:3 families, each with 30 plants, were grown in the greenhouse for powdery mildew evaluation, and the corresponding 53 F₂ plants were genotyped using simple sequence repeat (SSR) markers. Results showed that the 53 F_2:3 families segregated in ratio of one resistant : two segregating : one susceptible (13 : 26 : 14) and the 26 segregating F_2:3 families each exhibited a good fit to three resistant : one susceptible, indicating that resistance to powdery mildew is conditioned by a single dominant gene. The gene for powdery mildew resistance in V97‐3000 was mapped on chromosome 16 [linkage group (LG) J] flanked by Satt547 and Sat_396 on one side and Sat_393 on the other side with 3.8 cM and 3.9 cM distance, respectively. This study provides a new source of powdery mildew resistance and information of genetic location of the resistance gene and linked markers, which is useful for breeders selecting powdery mildew resistance through marker‐assisted selection (MAS) in soybean breeding programmes.     

Mapping QTL involved in powdery mildew resistance of the apple clone U 211

Scab and powdery mildew, caused by Venturia inaequalis (Cke.) Wint. and Podosphaera leucotricha (Ellis et Ev.) Salm. are the most important apple diseases. The apple clone U 211 is resistant to scab and is also highly resistant to powdery mildew under field conditions. The interval mapping method was applied for the identification of genomic regions conferring U 211 resistance to powdery mildew. The genetic maps of the ‘Idared’ and U 211 genome sectors were constructed using amplified fragment lenght polymorphism and simple sequence repeat markers and 98 individuals from the progeny of the cross ‘Idared’× U 211. On the basis of the phenotypic and molecular marker data 10 powdery mildew resistance quantitative trait loci (QTL) were identified in U 211 and ‘Idared’. One of the QTL in the clone U 211 explained 48‐72% of the phenotypic variation and its effect was stable over years.     

BCL、RIP细胞凋亡基因向小麦中的导入和赤霉病抗性鉴定

利用农杆菌介导法将BCL和RIP 2个与细胞凋亡有关的基因分别导入了小麦品种扬麦10号和Bobwhite,转化效率分别为1.60%和1.25%。Southern检测结果表明,细胞凋亡基因已稳定整合到小麦染色体上,多数植株为单拷贝整合。Northern检测结果表明,细胞凋亡基因能以小麦遗传背景高水平转录为RNA。转基因植株T₁代分离比例为2.11～2.33∶1,表明外源基因在后代中能够稳定遗传。BCL转基因植株和RIP转基因植株对赤霉病均表现出一定抗性,其中BCL-20、BCL-21、RIP-8、RIP-18等15个T₁代植株的小穗发病率为5.6%~16.1%,可望进一步培育抗小麦赤霉病新材料。     

利用“永久F2”群体定位抗赤霉病QTL

为了研究抗赤霉病侵染性的遗传, 利用感赤霉病品种南大2419和抗赤霉病品种望水白杂交单粒传获得的重组自交系群体132个株系间的随机配对组合, 构建了一个包含198个株系的“永久F2”群体。通过两年抗侵染田间试验和QTL作图, 定位了6个抗侵染QTL, 其中抗性等位位点源于望水白的Qfhi.nau-4B和Qfhi.nau-5A以及源于南大2419的Qfhi.nau-2B的效应较为稳定。Qfhi.nau-4B和Qfhi.nau-5A的效应较大且以加性效应为主, 前者存在部分显性基因效应。此外, 还检测到4对显著的互作位点。这些结果进一步说明赤霉病抗性遗传的复杂性, 同时也表明在利用望水白进行抗赤霉病育种时早代选择抗侵染性是可行的。     

A partial diallel study of powdery mildew resistance in six apple cultivars under three growing conditions with different disease pressures

Summary The suitability of large-fruited apple cultivars as alternative donors of powdery mildew resistance to the crabapples carrying major gene resistances was assessed. Five accessions with varying levels of mildew resistance (‘Dülmener Rosen', ‘Liberty', ‘Sciros', ‘Prima' and ‘Red Delicious') and one susceptible cultivar (‘Cox's Orange Pippin') were mated in a partial diallel design, and sub-sets of each seedling family were evaluated for three years at three New Zealand sites (Havelock North, Riwaka and Clyde) with different climatic conditions. There was a high correlation among the powdery mildew susceptibility scores of the individual seedlings over the three years, but disease levels varied among sites, with those at Clyde being the highest. All accessions with some resistance were found to confer additive resistance to their progenies to varying degrees depending on the climatic conditions of the sites, but findings suggest that progeny selection at Clyde will be sufficient for breeding purposes. The performance of the cultivar itself was not necessarily a good predictor of the level of resistance conferred to its progenies. The mostly negative genetic correlations between powdery mildew scores and fruit quality attributes suggest that selecting for powdery mildew resistance and desirable fruit quality traits may not be difficult. However, a larger number of families may be needed for more precise estimates of the genetic correlations to strengthen confidence in this conclusion.     

小麦白粉病成株抗性和抗倒伏性及穗下节长度的QTL定位

由小麦品种花培3号和豫麦57杂交获得了DH群体168个株系, 利用305个SSR标记对白粉病成株抗性、抗倒伏性和穗下节长度进行了QTL定位研究。DH群体及两亲本于2005年和2006年种植于山东泰安, 2006年种于安徽宿州。利用基于混合线性模型的QTLNetwork 2.0软件, 共检测到12个加性效应位点和10对上位效应位点。在4D染色体上控制白粉病成株抗性的qApr4D, 贡献率为20.0%, 在各环境中稳定表达, 其抗病等位基因来源于抗病亲本豫麦57; 在7D染色体上控制小麦穗下节长度的qIlbs7D, 贡献率为12.9%, 在各环境中稳定表达。加性效应和上位效应对小麦白粉病成株抗性、抗倒伏性和穗下节长度的遗传起重要作用, 并且基因与环境常常具有互作效应。以上两个QTL可分别用于小麦白粉病成株抗性和穗下节长度的分子标记辅助选择。     

Marker assisted selection (MAS) for developing powdery mildew resistant pea cultivars

In this contribution we review the state of the art for genetic resistance to powdery mildew, caused by Erysiphe pisi, in pea (Pisum sativum L.) and potential use of marker-assisted selection (MAS) for developing disease resistant cultivars. Powdery mildew is important in many production regions worldwide and reduces yield and crop quality when present in epidemic proportions. Although genetic resistance to powdery mildew is available (er1 and er2) and has been durable since its characterization in 1969, recently a new dominant gene (Er3) has been reported in Pisum fulvum, a wild relative of pea that is different from previously reported er1 and er2. The efficacy of these genes may be at risk from the point of view of new pathotypes and pathogens. Erysiphe trifolii has been reported that was not previously known as a pathogen of pea powdery mildew. A continued search for new and diverse resistant sources remains a priority in pea breeding and special emphasis should be paid to selection of resistance that will prolong durability of existing resistance genes. Marker assisted selection is a new emerging approach for target breeding that has been intensively employed especially in cereals and has recently got popularity among legume breeders. With the advancement of genomic research, especially related to quantitative traits loci, the MAS is potentially anticipated future technique for routine plant breeding that is scarce in legumes at present. In pea, various DNA markers have been reported linked to er1, er2 and Er3 at varying distances in different mapping populations that are currently being used in breeding programs. Currently MAS of single gene is the most powerful approach and successes have been witnessed. If single marker is not close enough to the gene of interest then two flanking markers are considerably utilized to improve the correct identification that is being successfully employed in MAS for powdery mildew resistance in pea.     

Venturia inaequalis resistance in local Spanish cider apple germplasm under controlled and field conditions

Host resistance is a key method for the integrated management of apple scab caused by Venturia inaequalis, which is one of the most important diseases of apple. Artificial inoculation of 92 cider apple cultivars with a mixed inoculum of V. inaequalis identified 19 weakly resistant and 19 resistant cultivars. Twelve of these resistant cultivars were previously classified as having low susceptibility to fire blight, and four of them showed complete or weak resistance to races (1), (1, 6) and (6, 7, 13) of V. inaequalis. The analysis of a selection of 72 cultivars for 6 years under field conditions identified 14 cultivars that were classified as resistant to apple scab under high disease pressure involving one to six Mills periods of severe risk of infection each year. Eight out of these 14 cultivars previously showed high levels of resistance to fire blight, which would allow the incorporation of genetic resistance in the integrated production of cider apples in Spain through their use in breeding programs.     

Improving the yield, processing quality and disease and pest resistance of potatoes by genotypic recurrent selection

A potato (Solanum tuberosum Gp Tuberosum) breeding programme is described and analysed in which resistances to late blight [Phytophthora infestans (Mont.) de Bary] and the white potato cyst nematode (Globodera pallida) have been combined with a modest increase in yield and acceptable fry colour for processing. It began in 1991 and has involved cycles of crossing, selection between from 120 to 145 progenies (full-sib families), and clonal selection within the selected progenies. We have shown that the breeding scheme can operate on a 3-year cycle with limited within progeny selection, and on a 5- or 6-year cycle with more extensive within progeny selection. Six years are required when resistance to late blight is assessed in the tubers as well as the foliage. The more extensive within progeny selection is recommended once genes have been combined from sufficient parents to achieve one’s objectives. The yield increase after three cycles of indirect selection through breeders’ visual preference was only modest because it was operating against a decrease which would occur in the absence of selection. A practical improvement in the scheme would be to increase the number of progenies assessed to over 200, given the moderate to high heritabilities of the progeny and clonal tests. But this would require a considerable effort because the success rate achieved with the potato pollinations was typical at just over 30%. In the fourth cycle we showed how new breeding objectives and germplasm could be accommodated whilst continuing to maintain progress, something that is important in any long term breeding strategy.     

Screening of pea germplasm for resistance to powdery mildew

Powdery mildew caused by Erysiphe pisi DC results heavy losses in the yield and quality of pods and seeds of pea crop. Germplasm comprising 701 accessions of garden and field pea originating from 60 countries were screened for powdery mildew resistance under natural epiphytotic conditions and 64 accessions found resistant in field screening for 2?years at one location were further screened both in field at two locations and artificially in laboratory to four isolates. The information was also obtained on the amount of genetic diversity and agronomic superiority in resistant accessions. Fifty-seven accessions showed resistant reaction for 3 consecutive years in field screening but only 14 accessions originating from 10 countries showed resistant reaction in laboratory screening against the four most prevalent isolates of E. pisi collected from different places in the area of experiment. Germplasm lines showed both complete and incomplete levels of resistance and variable reactions to different isolates. There was sufficient genetic diversity and agronomic superiority in the resistant accessions e.g. EC598655, EC598878, EC598704, IC278261, and IC218988, which may serve as useful genetic material to plant breeders for breeding pea varieties for powdery mildew resistance and high yield.     

Mapping of the apple powdery mildew resistance gene Pl1 and its genetic association with an NBS-LRR candidate resistance gene

Molecular markers for the major apple powdery mildew resistance gene Pl1 were identified and are presently used in marker-assisted selection in apple breeding. However, the precise map position of the Pl1 gene in the apple genome was not known. The objectives of this investigation were the identification of the Malus linkage group (LG) carrying the Pl1 locus, mapping of the resistance gene by simple sequence repeat (SSR) markers, and the analysis of genetic associations between the Pl1 gene and the numerous NBS-LRR resistance gene candidates already mapped in the apple genome. A two-step linkage mapping was used, based on two different apple families. The identification of LG 12 carrying Pl1 was performed indirectly by mapping the SCAR marker AT20 in an apple progeny for which there was a core genetic map but no mildew data available. Then, the position of Pl1 on LG 12 was determined by SSR markers in a second population which has been scored for mildew over 6 years in a greenhouse and in the field. The SSR Hi07f01, previously mapped on LG 12 [Tree Genet. Genomes, 2 (2006), 202] cosegregated with AT20 and was closely linked (∼1 cM) to the Pl1 gene. The TIR-NBS-LRR resistance gene analogue 15G11 mapped by the SSCP technique was also closely linked to the Pl1 resistance locus and might be a candidate for Pl1 itself, a second powdery mildew major resistance gene ( Pld , [Theor. Appl. Genet., 110 (2004), 175]), or two scab resistance genes ( Vg , [IOBC/WPRS Bull., 23 (2000), 245]; Vb , [Genome, 49 (2006), 1238]) which all seem to be located in a common R gene cluster at the distal end of apple LG 12.     

利用SNP芯片和BSA分析规模化定位小麦抗白粉病基因

规模化定位小麦品种携带的抗白粉病基因对于抗病性种质创新和新品种选育具有重要的意义。本研究采用Illumina Infinium iSelect 90k SNP芯片结合集群分离分析法(bulked segregate analysis,BSA)对36个河南省小麦新品系携带的抗白粉病基因进行了定位。SNP芯片检测表明,在24个小麦品系构建的抗、感池DNA间可检测到一个明显富集的SNP峰,表明其可能携带单一主效抗白粉病基因;在其他12个小麦品系构建的抗、感池DNA间可检测到多个SNP峰,推测其可能含多个抗白粉病基因。有26个小麦品系在2AL染色体上检测到的SNP数目最多,推测其携带位于2AL染色体上的Pm4b抗白粉病基因。开发出与2AL染色体上抗白粉病基因紧密连锁的分子标记Xwggc116,可用于这些小麦品系中抗白粉病基因的分子检测。研究结果表明高通量SNP分析技术平台可以用来规模化定位小麦品种中的抗白粉病基因,明确了河南省抗白粉病小麦品系中携带Pm2、Pm4b、Pm21和新1BL/1RS易位等有限的抗白粉病基因,抗病基因资源非常狭窄,亟需引进新的多样化抗病基因资源,拓宽遗传基础,培育抗病小麦新品种。