Variation of Fungicide Resistance in Czech Populations of Pseudoperonospora cubensis

During the growing seasons between the years 2001 and 2004, 98 isolates of Pseudoperonospora cubensis from nine regions of Czech Republic were collected and screened for tolerance/resistance to the three frequently used fungicides (propamocarb, fosetyl‐Al, metalaxyl). Fungicides were tested in five different concentrations, using a floating disc bioassay. Fungicide effectiveness varied considerably. Propamocarb appeared most effective and all the isolates collected in the years 2001–2003 were found sensitive to all tested concentrations [607–9712 μg active ingredient (a.i.)/ml]. In 2004, some strains with increased resistance to propamocarb were detected. These strains were characterized by tolerance at the lowest concentrations (607 μg a.i./ml, eventually on 1214 μg a.i./ml); however, they were controlled by 2428 μg a.i./ml. Fosetyl‐Al was effective at the recommended concentration of 1600 μg a.i./ml against all isolates. However, the occurrence of isolates (collected in 2001) which sporulated at low concentrations (400 and 800 μg a.i./ml) indicated that the selection for tolerance occurs in the pathogen population. Nevertheless, this phenomenon was not confirmed with the P. cubensis isolates collected between the years 2002 and 2004. Metalaxyl was found ineffective, because 97% of the isolates showed the resistance to the recommended concentration (200 μg a.i./ml), and the other 3% of isolates expressed tolerant response. The majority of the isolates showed profuse and/or limited sporulation at higher concentrations (400 and 800 μg a.i./ml). A substantial shift to highly metalaxyl resistant strains was evident in the Czech P. cubensis populations during 2001–2004.     

Distribution,Host Range and Disease Severity of Pseudoperonospora cubensis on Cucurbits in the Czech Republic

Cucurbit downy mildew, caused by Pseudoperonospora cubensis, is a major cucumber disease in the Czech Republic. Disease prevalence, host range and disease severity were evaluated from 2001 to 2009. The geographical distribution of P. cubensis was assessed on ca 80–100 locations per year in two main regions of the Czech Republic (central and southern Moravia, and eastern, northern and central Bohemia). Infection by P. cubensis was observed primarily on cucumber (Cucumis sativus) but only on the leaves. During the study, disease prevalence ranged from 66 to 100%. The majority of C. sativus crops were heavily infected at the end of the growing season (second half of August). Generally, P. cubensis was present at high or very high disease severity. The loss of foliage results in the reduction in the quality and quantity of marketable yield of fruit. Pseudoperonospora cubensis was widespread across the whole area of the Czech Republic studied. Very rarely, infection was recorded in muskmelon (Cucumis melo) and Cucurbita moschata. Of other pathogens, the most frequently recorded was the cucurbit powdery mildew (Golovinomyces cichoracearum and Podosphaera xanthii).     

Analysis of the interacting partners eIF4F and 3′‐CITE required for Melon necrotic spot virus cap‐independent translation

We have shown previously that the translation of Melon necrotic spot virus (MNSV, family Tombusviridae, genus Carmovirus) RNAs is controlled by a 3′‐cap‐independent translation enhancer (CITE), which is genetically and functionally dependent on the eukaryotic translation initiation factor (eIF) 4E. Here, we describe structural and functional analyses of the MNSV‐Mα5 3′‐CITE and its translation initiation factor partner. We first mapped the minimal 3′‐CITE (Ma5TE) to a 45‐nucleotide sequence, which consists of a stem‐loop structure with two internal loops, similar to other I‐shaped 3′‐CITEs. UV crosslinking, followed by gel retardation assays, indicated that Ma5TE interacts in vitro with the complex formed by eIF4E + eIF4G_980–1159 (eIF4F_p20), but not with each subunit alone or with eIF4E + eIF4G_1003–1092, suggesting binding either through interaction with eIF4E following a conformational change induced by its binding to eIF4G_980–1159, or through a double interaction with eIF4E and eIF4G_980–1159. Critical residues for this interaction reside in an internal bulge of Ma5TE, so that their mutation abolished binding to eIF4E + eIF4G_1003–1092 and cap‐independent translation. We also developed an in vivo system to test the effect of mutations in eIF4E in Ma5TE‐driven cap‐independent translation, showing that conserved amino acids in a positively charged RNA‐binding motif around amino acid position 228, implicated in eIF4E–eIF4G binding or belonging to the cap‐recognition pocket, are essential for cap‐independent translation controlled by Ma5TE, and thus for the multiplication of MNSV.     

几种瓜类作物对根际高温胁迫的生理响应

以黑籽南瓜、‘春秋王2号’黄瓜、‘傲美’苦瓜、‘兴蔬’丝瓜、‘五叶香’丝瓜为试材,采用营养液栽培法,研究了这几种瓜类作物在根际35℃高温处理下的生长量、光合参数及地上部、地下部的渗透胁迫物质的变化。结果显示：处tE5d后,两种丝瓜幼苗的株高、茎粗、叶面积、根长、地上干重及地下干重,叶片相对叶绿素含量、净光合速率（Pn）,气孔导度（Gs）、胞间CO2浓度（Gi）、蒸腾速率（Tr）与对照相比变化均不大,而另外三种瓜类的以上指标均受到明显的抑制;高温使得黑籽南瓜及黄瓜叶片和根系中都积累了大量的膜脂过氧化产物丙二醛（MDA）,渗透调节物质脯氨酸和可溶性蛋白含量在叶片中增大,但在根系中却减小,而处理下‘五叶香’丝瓜的MDA含量与对照差异不显著,且两种渗透调节物质表现为叶片中减小根系中增大;处理下‘兴蔬’丝瓜根系中MDA的减少量及可溶性蛋白的增加量都大于‘傲美’苦瓜。结果表明,根际高温下两个丝瓜品种,特别是‘五叶香’丝瓜有较高的耐受力,通过提高根系中的渗透调节物质含量来降低根际高温对植株整体造成的伤害。     

The making of giant pumpkins: how selective breeding changed the phloem of Cucurbita maxima from source to sink

Despite the success of breeding programmes focused on increasing fruit size, relatively little is known about the anatomical and physiological changes required to increase reproductive allocation. To address this gap in knowledge, we compared fruit/ovary anatomy, vascular structure and phloem transport of two varieties of giant pumpkins, and their smaller fruited progenitor under controlled environmental conditions. We also modelled carbon transport into the fruit of competitively grown plants using data collected in the field. There was no evidence that changes in leaf area or photosynthetic capacity impacted fruit size. Instead, giant varieties differed in their ovary morphology and contained more phloem on a cross‐sectional area basis in their petioles and pedicels than the ancestral variety. These results suggest that sink activity is important in determining fruit size and that giant pumpkins have an enhanced capacity to transport carbon. The strong connection observed between carbon fixation, phloem structure and fruit growth in field‐grown plants indicates that breeding for large fruit has led to changes throughout the carbon transport system that could have important implications for how we think about phloem transport velocity and carbon allocation.     

Population genomic analyses from low‐coverage RAD‐Seq data: a case study on the non‐model cucurbit bottle gourd

Restriction site‐associated DNA sequencing (RAD‐Seq), a next‐generation sequencing‐based genome ‘complexity reduction’ protocol, has been useful in population genomics in species with a reference genome. However, the application of this protocol to natural populations of genomically underinvestigated species, particularly under low‐to‐medium sequencing depth, has not been well justified. In this study, a Bayesian method was developed for calling genotypes from an F₂ population of bottle gourd [Lagenaria siceraria (Mol.) Standl.] to construct a high‐density genetic map. Low‐depth genome shotgun sequencing allowed the assembly of scaffolds/contigs comprising approximately 50% of the estimated genome, of which 922 were anchored for identifying syntenic regions between species. RAD‐Seq genotyping of a natural population comprising 80 accessions identified 3226 single nuclear polymorphisms (SNPs), based on which two sub‐gene pools were suggested for association with fruit shape. The two sub‐gene pools were moderately differentiated, as reflected by the Hudson's F_ST value of 0.14, and they represent regions on LG7 with strikingly elevated F_ST values. Seven‐fold reduction in heterozygosity and two times increase in LD (r²) were observed in the same region for the round‐fruited sub‐gene pool. Outlier test suggested the locus LX3405 on LG7 to be a candidate site under selection. Comparative genomic analysis revealed that the cucumber genome region syntenic to the high F_ST island on LG7 harbors an ortholog of the tomato fruit shape gene OVATE. Our results point to a bright future of applying RAD‐Seq to population genomic studies for non‐model species even under low‐to‐medium sequencing efforts. The genomic resources provide valuable information for cucurbit genome research.     

根际高温胁迫对5种瓜类作物生长及叶片光合和叶绿素荧光参数的影响

采用营养液栽培法,比较分析了根际高温(35℃)处理0、3和5 d以及恢复5 d后黑籽南瓜(Cucurbita ficifolia Bouché)、‘春秋王2号’黄瓜(Cucumis sativus‘Chunqiuwang No.2’)、‘兴蔬’丝瓜(Luffa cylindrica‘Xingshu’)、‘五叶香’丝瓜(Luffa cylindrica‘Wuyexiang’)和‘傲美’苦瓜(Momordica charantia‘Aomei’)幼苗的生长及叶片光合和叶绿素荧光参数的变化。结果表明:与对照(25℃)相比,在高温处理期间及恢复期间,黑籽南瓜、‘春秋王2号’黄瓜和‘傲美’苦瓜的株高、茎粗、叶面积、叶绿素相对含量(SPAD)以及叶片净光合速率(Pn)、气孔导度(Gs)、胞间CO2浓度(Ci)、蒸腾速率(Tr)、PSⅡ的光能捕获效率(Fv’/Fm’)、PSⅡ的实际光化学效率(ΦPSⅡ)、实际光化学量子产量(Yield)、表观光合电子传递速率(ETR)、光化学猝灭系数(qP)及PSⅡ用于光化学反应的能量比例(P)总体上均显著低于对照;而‘五叶香’丝瓜的上述指标大多无明显变化,但其非光化学猝灭系数(qN)及PSⅡ用于天线色素热耗散的能量比例(D)均低于对照;‘兴蔬’丝瓜的各项指标在高温处理期间均有不同程度的降低或升高,但在恢复5 d后各项指标均接近或高于对照。研究结果表明:5种瓜类作物对根际高温的耐受性有明显差异。其中,黑籽南瓜和‘春秋王2号’黄瓜对根际高温的耐性最差;2个丝瓜品种特别是‘五叶香’丝瓜的耐性较强,可作为耐高温瓜类作物的砧木。