基于SSR标记分析大豆疫霉根腐病抗源的遗传多样性

丰富的遗传多样性可为大豆育种提供宽阔的遗传基础,本研究基于35对SSR标记,对60份东北地区大豆疫霉根腐病抗性品种进行了遗传多样性分析,共检测到189个等位基因,平均每个位点等位变异数5.4个,多态性信息含量指数(PIC)为0.1550~0.8195,平均为0.6636;遗传相似系数的变异范围为0.31~0.74。利用5对高多态性SSR引物构建了60份抗性材料的指纹图谱,这5对SSR引物构建的指纹图谱可以将60份疫霉根腐病抗性材料逐一区分开。采用NTSYS2.10基于遗传距离的聚类分析,将60份抗性材料分为7个类群,其中78.33%的抗性品种(系)的遗传相似系数在0.45~0.74间,表明遗传差异相对较窄,品种间遗传多样性水平较低。聚类分析与群体遗传结构分析结果有部分重合,均反映出不同地区的抗性材料间存在一定的渗透和交流。     

Phytophthora cryptogea root rot of tomato in rock woo] nutrient culture. II. Effect of root zone temperature on infection, sporulation and symptom development

The effect of root-zone temperature on Phytophthora cryptogea root rot was studied in tomato cv. Counter grown under winter and summer conditions in rockwool culture. A nutrient temperature of 25°C resulted in increased root initiation and growth, higher in winter-grown than in summer-grown plants. Rhizosphere zoospore populations were greatly reduced at 25°C and above. Growth of P. cryptogea in vitro was optimal between 20°C and 25°C and completely suppressed at 30°C. Encystment was enhanced by increased temperatures above 20°C. Zoospore release in vitro occurred in cultures maintained at constant temperatures in the absence of the normal chilling stimulus. Optimal release was at 10°C; no zoospores were released at 30°C. Inoculated, winter-grown tomato plants maintained at 15°C developed acute aerial symptoms and died after 21 days. Comparable plants grown at a root-zone temperature of 25°C remained symptomless for the 3-months duration of the experiment. Summer-grown infected plants at the higher root temperature wilted but did not die. Enhanced temperature was ineffective as a curative treatment in summer-grown plants with established infection. Aerial symptoms of Phytophthora infection are seen as a function of the net amount of available healthy root. With high root zone temperatures this is determined by new root production and decreased inoculum and infection.     

Phytophthora cryptogea root rot of tomato in rockwool nutrient culture:

The effect of root temperature on growth and yield of rockwool-grown tomato plants infected with Phytophthora cryptogea was investigated. Measurements of shoot and root growth were taken at high (25^oC) and low (15^oC) root temperatures during the generative phase of growth. The growth of roots of healthy and P. cryptogea-infected tomato plants in rockwool blocks was higher in plants grown with roots at 25^oC than at 15^oC after 60 days and a similar effect was found in slabs after 98 days. Under sub-optimal conditions for growth the disease became severe when root temperatures were low. Growth of roots was greatest when roots were maintained at a high temperature in combination with an ambient air temperature of c. 15^oC and the response was greater in cv. Counter than cvs Calypso and Marathon. Water-soluble carbohydrates of roots were higher in those produced in blocks than slabs and were reduced by infection compared to healthy plants with roots at 15^oC and 25^oC. Reduced transpiration rates were found 17 days after inoculation in symptomless plants grown at a root temperature of 25^oC. Infection, regardless of the temperature of the roots or cultivar, led to reduced stem growth. The plants grown at 25^oC were taller than those with a root temperature of 15^oC. After 9 wk of harvest, the cumulative fruit yields in infected cvs Counter and Calypso grown at 25^oC were comparable to that in healthy plants grown at either temperature and cumulative fruit numbers followed a similar pattern. High root temperatures led to delayed fruit ripening between weeks 3–10 and a larger number of unripe fruit. The weight of unripe fruit from infected plants grown at 25^oC at the terminal harvest was higher than from healthy plants with roots maintained at 15^oC.     

Phytophthora cryptogea root rot of tomato in rockwool nutrient culture. I. Analysis of root infection

The spatial and temporal colonisation of tomato plants by Phytophthora cryptogea was studied in rockwool nutrient culture. Root growth and the distribution and progress of infection were measured on dissected root fragments obtained from a detailed destructive sampling of the substrate. Dissected fragments of root or roots and fibres were plated on BNPRA agar, a selective Phytophthora medium. Roots colonised all parts of the rockwool substrate 60 days after planting with the exception of surface marginal and central areas of the slab which had a lower solution content. Most root biomass occurred in and immediately beneath the original growing block. The distribution of P. cryptogea closely followed the pattern of root colonisation. An alternative, novel method for root analysis involved the dissolution of the mineral fibre and its formate bonding resin by digestion in 1 m H₃PO₄ for 45 min. Comparative recoveries of P. cryptogea from plated fragments of dissected root and fibre or comminuted samples of acid-released or dissected root showed that the acid treatment initially reduced the number of Phytophthora colonies in block and slab roots by 67% and 61% respectively. After 28 days, colony recovery from acid-released roots in the rockwool slab increased and was between 4% and 13% lower than from other plating methods. Since 1 m H₃PO₄ was lethal to zoospores and surface sporangia, the colonies recovered were interpreted as originating exclusively from root lesions. Root fresh weight of healthy and inoculated plants declined during the initial period of fruit formation. P. cryptogea infection led to a progressive reduction in root weight in the growing block and main slab and 28 days after inoculation, was approximately 50% of the controls. The acid digestion of rockwool fibre is proposed as a new approach to the problem of root and pathogen analysis in this substrate.     

Peroxidase isoenzymes in the defense response of Capsicum annuum to Phytophthora capsici

Quantitative and qualitative changes in isoperoxidase patterns from stems of three cultivars of pepper ( Capsicum annuum L.). one susceptible, one intermediate and one resistant, were found upon inoculation with Phytophthora capsici using a decapitation method. The peroxidase activity was determined in the intercellular fluid as well as in the cytosolic fraction of the necrotic, healthy and intermediate zones of stems of the three cultivars, 6 days after inoculation. In the intercellular fluid, peroxidase activity of the susceptible cv. Yolo Wonder increased somewhat from 4.7 (healthy zone) to 12.9 (intermediate zone) μmol mg⁻¹ protein min⁻¹, whereas in the intermediate cv. Americano, the peroxidase activity decreased from 123 (healthy zone) to 78 (intermediate zone) μmol mg⁻¹ protein min⁻¹. The most dramatic increase (5.7 to 662 μmol mg⁻¹ protein min⁻¹) in intercellular peroxidase activity was found in the resistant cv. Smith-5. This, in conjunction with the appearance of an additional acidic isoperoxidase (pI 4.4) specific for the cv. Smith-5, could be the reason for the resistance of this cultivar against the fungus attack. The release of peroxidase into the intercellular space as a defense reaction was confirmed by histochemical analysis, showing that peroxidase activity occurred in the intercellular spaces of those stems of the resistant cultivar that had not yet been invaded by the fungus, but was detected neither in the other cultivars nor in the intercellular spaces of such stems of the intermediate and susceptible cultivars that contained growing mycelium of P. capsici. The lack of staining in the intercellular spaces of the susceptible cultivars could be attributed to their low content in peroxidase.     

The relationship between colonisation and crown rot symptoms in strawberry plants infected with Phytophthora cactorum

Strawberry tissues infected with Phytophthora cactorum were comminuted and plated in a selective antibiotic agar medium to determine levels of tissue colonisation as indicated by the number of colony forming units (CFU) recovered per gramme of infected tissue. The number of CFU recovered per gramme of tissue increased logarithmically with the amount of necrosis in infected crown, leaf and petiole tissues. Under the conditions of enhanced susceptibility to infection and colonisation caused by cold storage treatments, this relationship between colonisation and necrosis was not significantly altered in the susceptible cv. Tamella. A recovery index was used to determine the effect of infected tissues on the recovery of CFU. This indicated that increasing levels of host colonisation stimulated CFU recovery and may partly explain the large increase in CFU g^-1 with larger amounts of necrosis. The amount of tissue colonisation was greater in inoculated plants of the susceptible cv. Tamella than in less susceptible cv. Cambridge Favourite, although the necrotic tissues of the latter contained more CFU g^-1, indicating a greater level of tolerance to colonisation. In cv. Tamella small amounts of colonisation were capable of causing wilt symptoms, although no wilted plants contained less than 200 CFU g^-1. Conversely, plants containing more than 1000 CFU g^-1 always wilted. In the early stages of infection, low levels of colonisation could be detected in strawberry crowns in the absence of symptoms. Dormant strawberry plants of cv. Tamella were readily infected by P. cactorum zoospore inoculations but, unlike actively growing plants, the majority of infections remained latent. These latent infections exhibited little or no symptoms and CFU recoveries from infected tissues were always below 100 CFU g^-1.     

利用抑制差减杂交技术分离马铃薯晚疫病抗性相关基因

以晚疫病病原菌混合小种接种处理48h的马铃薯水平抗性材料(R-gene-free)叶片为目的材料,以未处理材料作为对照,用抑制差减杂交技术构建了一个富集晚疫病抗性相关基因的差减文库。应用反向Northern技术对840个克隆进行斑点杂交筛选,筛选出150个病原诱导后信号明显增强的克隆。26个片段测序结果表明：部分片段基因功能与抗病性明显相关。7个差异表达片段与GenBank EST数据库中已有晚疫病原诱导马铃薯叶片得到的EST有很高同源性(达95％～100％);部分片段核苷酸或氨基酸序列分别与番茄、烟草、拟南芥等的EST序列或氨基酸序列有较高同源性;另有4个基因片段在GenBank EST数据库中未找到明显的同源序列,可能为新发现的基因片段。     

Interaction Between Root and Leaf Disease Development in Barley Cultivars after Inoculation with Different Isolates of Bipolaris sorokiniana

The relationship between root and leaf infection in 11 cultivars of barley ( Hordeum vulgare ) by different isolates of Bipolaris sorokiniana was investigated in young plants. Roots of 10-day-old seedlings, grown in filterpaper rolls, and the third leaf of 17-day-old seedlings were inoculated with the different isolates and a Disease Development Index (DDI) was calculated.
The rate of lesion development in leaves was higher than in roots, indicated by generally higher DDI after leaf inoculation than after root inoculation. Significant differences in resistance were found among the barley cultivars. Inoculation with different isolates of B. sorokiniana caused significant differences in DDI for both roots and leaves. In the leaves, but not in the roots, a significant cultivar–isolate interaction was found. No significant correlations, neither in isolate aggressiveness nor in cultivar reaction between root and leaf, were observed.     

Estimates of Disease Effects on Soybean Yields in the United States 2003 to 2005

Research must focus on management of diseases that cause extensive losses, especially when funds for research are limited. Knowledge of the losses caused by various soybean diseases is essential when prioritizing research budgets. The objective of this project was to compile estimates of soybean yields suppressed due to diseases for each soybean-producing state in the US from 2003 to 2005. The goal was to provide this information to help funding agencies and scientists prioritize research objectives and budgets. Yield suppression due to individual diseases varied among regions in the US, and the total of soybean yields suppressed due to diseases in the US varied among years. Soybean cyst nematode suppressed US soybean yield more during 2003 to 2005 than any other disease. Phytophthora root and stem rot, sudden death syndrome, and seedling diseases ranked in the top four on the list of diseases that suppressed soybean yield during these years. This is the first report of soybean yield suppression due to Asian soybean rust in the United States.     

Peroxidase, catalase, amine oxidase and acid phosphatase activities in Pisum sativum during infection with Fusarium oxysporum and F. solani

Two genotypes (cv. Smaragd and line DP1059) of Pisum sativum with different susceptibility to Fusarium oxysporum and F. solani and influence of pathogenesis on enzyme activities were studied. The increase of activity of studied enzymes was mostly observed in both roots and shoots during pathogenesis. Only activity of acid phosphatase decreased in the root and increased in shoots. The correlation between enzyme activity change and susceptibility of pea cultivars to F. oxysporum or F. solani was observed.     

High-resolution Mapping and Analysis of the Resistance Locus <Emphasis Type="Italic">Rpi-abpt</Emphasis> Against <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in Potato

Introduction of more durable resistance against Phytophthora infestans causing late blight into the cultivated potato is of importance for sustainable agriculture. We identified a new monogenically inherited resistance locus that is localized on chromosome 4. The resistance is derived from an ABPT clone, which is originally a complex quadruple hybrid in which Solanum acaule, S. bulbocastanum, S. phureja and S. tuberosum were involved. Resistance data of the original resistant accessions of the wild species and analysis of mobility of AFLP markers linked to the resistance locus suggest that the resistance locus is originating from S. bulbocastanum. A population of 1383 genotypes was screened with two AFLP markers flanking the Rpi-abpt locus and 98 recombinants were identified. An accurate high-resolution map was constructed and the Rpi-abpt locus was localized in a 0.5 cM interval. One AFLP marker was found to co-segregate with the Rpi-abpt locus. Its DNA sequence was highly similar with sequences found on a tomato BAC containing several resistance gene analogues on chromosome 4 and its translated protein sequence appeared to be homologous to several disease resistance related proteins. The results indicated that the Rpi-abpt gene is a member of an R gene cluster.