Conformity and genetic relatedness estimation in crop species having a narrow genetic base: the case of cucumber (Cucumis sativus L.)*

A set of 155 SSR (107) and SCAR (48) markers were used to evaluate 53 cucumber (Cucumis sativus L.) accessions of diverse origin to characterize genetic relationships and to define a standard marker array that was most effective in detecting genetic differences in this germplasm array. A multivariate marker‐based analysis of diverse germplasm using this standard marker array (17 SSR and 5 SCAR markers) was compared with results from a set of 70 previously reported RAPD markers, and then used to explore the potential value of these genetic markers for plant variety protection (PVP) and the establishment of essential derivation (ED) threshold values in this species using elite lines and hybrids and backcross progeny. Diversity analysis allowed identification of distinctly different lines that were used for the construction of three sets of backcross families (BC₁‐BC₃). While general genetic relationships among accessions were similar in SSR/SCAR analyses (r_s= 0.65) using two genetic distance (GD) estimators, differences in accession relationships were detected between RAPD and SSR/SCAR marker evaluations regardless of the estimator used. The GDs among elite germplasm with known pedigrees were relatively small (0.06‐0.23 for any pairwise comparison). GD values decreased and degree of fixation (at three to seven loci depending on the mating) increased with increased backcrossing such that recurrent parent allelic fixation occurred in least one family of each of the BC₃ families. In many instances the degree of fixation of loci was not uniformly achieved in the BC₃. Although the level of genetic polymorphisms will likely restrict the use of molecular markers for PVP and the establishment of ED values, the use of single nucleotide differences will likely provide opportunities to define specific functional distances that have potential for PVP in cucumber. Nevertheless, without an expanded, genetically robust standard marker array (e.g. 50 codominant markers), ED threshold values will be difficult to define in this species, and perhaps will require the appraisal of single nucleotide polymorphisms as discriminators of difference in this species.     

Direct plant regeneration from leaf explants in cucumber (Cucumis sativus L.) is free of stable genetic variation

A new procedure is described for rapid and efficient plant regeneration from leaf explants of Cucumis sativus and C. anguria. The following factors were most important: young leaves, the NO₃: NH₄ ratio in the plant induction medium, small explants and the growth regulator combination. After about 4 weeks of culture under optimal growth conditions, the frequency of regeneration was 10–100% of explants. Six to 7 weeks were required to obtain well-rooted plants, which were mostly able to survive after transfer into soil. From a single young leaf of C. sativus cv. ‘Borszezagowski’, 135 plants could be regenerated. All plants transferred to a greenhouse were free of morphological or physiological abnormalities, flowered normally and bore fruits. The analysis of R₀ plants showed no genetic variation, whereas in the R₁ two new phenotypes, which were not transmittable to the R₂, were observed. This procedure is recommended for its production of homogeneous cucumber plants.     

New C-banding pattern for chromosome identification in cucumber (Cucumis sativus L.)

A chromosome study of cucumber, C. sativus L., was performed using orcein and C-banding techniques. The diploid and tetraploid plants investigated here showed the somatic chromosome numbers 2n=14 and 28, respectively. The haploid chromosome complement was composed of five metacentric and two submetacentric chromosomes. All C. sativus chromosomes had clearly visible C-bands, and each chromosome could be identified unequivocally after C-banding staining, with 13 C-bands appearing in the haploid complement. The haploid complement had a 44.9% ratio of total C-band length to total chromosome length. Chromosomes 1, 2, 4, 5 and 7 had stable C-bands. Three large, dark C-bands appeared at the proximal regions of chromosomes 1 and 2. Chromosome I had quite a large C-band and with a 68.4% ratio of C-band length to short arm length. Chromosome 2 also had quite a large C-band in the pericentromeric region with a 57.6% ratio of C-band length to the full length of this chromosome and possessed an elongated primary constriction in early metaphase. In prometaphase, chromosome 2 showed that the long arm was completely separated from the short arm. The number of secondary constrictions could not be clearly observed because these chromosomes are small and they could not be counted in every metaphase cell. However, six chromosomes seemed to have secondary constrictions in the diploid plants. Two silver-stained bands were observed at primary constrictions of two of the large chromosomes.     

Analysis of generation means and components of variance for parthenocarpy in cucumber (Cucumis sativus L.)

Parthenocarpy (seedless fruit) has potential for increasing yield in cucumber (Cucumis sativus var. sativus L.). To determine the inheritance of parthenocarpy in gynoecious cucumber, P₁, P₂, F₁, F₂, BC₁P₁, and BC₁P₂ generations derived from crossing two non‐parthenocarpic gynoecious inbred lines [Gy8 (P₂; processing type) and ‘Marketmore 80’ (P₂; MM, fresh market type)] with a highly parthenocarpic inbred line [2A (P₁; processing type)] were evaluated for fruit number in a greenhouse at Arlington, Wisc. in 1999 (designated 2A × Gy8 1999) and in the open‐field at Hancock, Wisc. in 2000 (designated 2A × Gy8 2000 and 2A × MM 2000). There were significant location and location × generation interaction effects, and therefore generation means analyses were conducted separately for each location. The minimum numbers of effective factors controlling parthenocarpy were estimated to be at least one (2A × Gy8 1999), two (2A × Gy8 2000) and four (2A × MM 2000). Results suggest that selection for parthenocarpy for multiple hand harvest operations will likely be more effective than that for once‐over machine harvest operations. However, the selection efficiency will likely vary across different populations and environments.     

Identification of quantitative trait loci for powdery mildew resistance in highly resistant cucumber (Cucumis sativus L.) using ddRAD-seq analysis

Powdery mildew, caused by Podosphaera xanthii (syn. Sphaerotheca fuliginea ex Fr. Poll.), is one of the most economically important foliar diseases in cucumber (Cucumis sativus L.). Cucumber parental line ‘Kyuri Chukanbohon Nou 5 Go’, developed from weedy cucumber line CS-PMR1, is highly resistant to powdery mildew and is promising breeding material. We performed quantitative trait locus (QTL) analysis using double-digest restriction-site-associated DNA sequencing (ddRAD-Seq) in a population from a cross between ‘Kyuri Chukanbohon Nou 5 Go’ and the Japanese native cultivar ‘Kaga-aonaga-fushinari’, which is susceptible to powdery mildew. The resistance of the population and its parents was evaluated using leaf disc assays and image analysis. We detected one major QTL on Chr. 5 that was effective at both 20°C and 25°C and one minor QTL on Chr. 1 effective at 20°C. We detected two additional QTLs in subpopulation: one on Chr. 3 effective at 20°C and one on Chr. 5 effective at both 20°C and 25°C in a position different from the major QTL. The resistance alleles at all four QTLs were contributed by ‘Kyuri Chukanbohon Nou 5 Go’. The results of this study can be used to develop practical DNA markers tightly linked to genes for powdery mildew resistance.     

Biochemical affinities between Cucumis hystrix Chakr. and two cultivated Cucumis species (C. sativus L. and C. melo L.) based on isozyme analysis

Isozymes of eight enzymes in Cucumis hystrix Chakr. and two cultivated Cucumis species (C. sativus L. and C. melo L.) were analysed electrophoretically to investigate the biosystematics of these three species. Cluster analysis using data from six enzymes indicated that considerable genetic distance existed between both C. hystrix and melon and between C. hystrix and cucumber. C. hystrix might be a key species for studying the evolution and taxonomy of genus Cucumis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of QTLs controlling low‐temperature tolerance during the germination stage in cucumber (Cucumis sativus L.)

Poor seed germination of cucumber at suboptimal temperatures is a great concern for growers wishing to take advantage of the early market. The development of low‐temperature‐tolerant varieties would be aided by understanding the inheritance of the trait and mapping its locus. In this study, a set of 140 recombinant inbred lines (RILs) derived from a cross between 65G (low‐temperature‐tolerant) and 02245 (low‐temperature‐sensitive) was used to identify the QTLs linked with low‐temperature tolerance. A linkage map was developed using 135 simple sequence repeat (SSR) markers and five insertion–deletion (Indel) markers, and three QTLs were identified, qLTG1.1, qLTG2.1 and qLTG4.1. qLTG1.1, the major one for germination rate, germination energy and germination index, explained more than 50% of the observed phenotypic variability. The major QTL for radicle length, qLTG4.1, explained 13.8% of the phenotypic variability. The results showed that qLTG1.1 and qLTG4.1 play an important role in low‐temperature tolerance during seed germination of cucumber and provide a basis for further fine mapping to determine the molecular mechanism for this trait.     

黄瓜育种中"血缘"遗传关系分析研究

采用RAPD技术研究了华北型与欧洲温室型品种的杂交后代的遗传漂移情况，同时根据供试亲本材料差异位点谱带在F2的分离情况，进行了初步的遗传分析以及F2个体的基因型分析，目的是以遗传距离和说带基因型为依据，分析在育种过程中判断血缘成分的可能性及基本方法，以便最终能够进行遗传关系控制。     

Development and Characterization of Microsatellite Markers from an Enriched Genomic Library of Cucumber (Cucumis sativus)

The development, characterization and application of cucumber ( Cucumis sativus L.) microsatellite markers was accomplished using a library-enrichment procedure. Fifty-seven primer pairs flanking the microsatellite repeats were used for DNA amplification. Sixteen C. sativus accessions were assessed for polymorphisms using 45 primer pairs. The average number of alleles per locus was 3.6, and up to seven alleles were found at one locus. The maximum polymorphism information content value was 0.78 with an average of 0.47. The cucumber microsatellite makers could be useful for seed purity control in hybridity testing. Some of these cucumber markers were transferable to other cucurbit species (i.e. melon, watermelon, pumpkin and bitter gourd).     

Inheritance of firmness in raw cucumber (Cucumis sativus L.) fruit

Summary Inheritance of raw cucumber fruit texture (Magness-Taylor Fruit Pressure Tester firmness) was investigated over a 4-year period from 1971–1974. Results from 2 separate but related experiments suggested that firmness was quantitatively inherited with sufficient additive effects to permit gain from selection. In a selection study within 4 F₂ populations derived from crosses between firm (Chipper and Gy3) and soft (Mincu and Green F) fruit type cultivars, variation among and within F₃ and F₄ families was significant but overall family means were not significantly higher than the high parent in any of the 4 crosses. Narrow sense heritability estimates for fruit texture were 0.80 in the Mincu × Chipper population and 0.77 in the Green F × Chipper, Mincu × Gy3, and Gy3 × Green F crosses. In a separate experiment, generation means analysis was used to assess the mode of gene action in 2 crosses: Green F × Chipper, and Gy3 × Green F. Additive genetic effects accounted for 98.8% and 99.3% of the total genetic variation within each cross, respectively.Scientific Journal Series Paper No. 9794.     

Gynoecious sex expression and stability in cucumber (Cucumis sativus L.)

Summary Results indicated that gynoecious stability in the F₁ is governed by partial dominance. The degree of gynoecious stability is dependent upon genotypes, stages of plant for AgNO₃ application and number of sprays. Depending upon the genotypes, the one-true-leaf stage and one spray of 250 ppm AgNO₃ showed more gynoecious stability in the F₁. Twice application of AgNO₃ at the two-true-leaf stage produced the maximum number of male flowers. Plants exposed to light for longer time (10.00 h) after AgNO₃ application produced less flowers than when exposed for shorter time (15.00 and 20.00 h).Present address: Division of Vegetable Crops, Indian Agricultural Research Institute, New Delhi-110012, India.     

The relationship between the regeneration system and genetic variability in the cucumber (Cucumis sativus L.)

Durum wheat has been subjected to intense breeding in Italy due to its local economic importance. Four groups of Italian cultivars representative of different breeding eras were compared in northern Syria for yield potential and morphophysiological features at a moderately favourable site, and drought tolerance at a stressful site. Group 1 included indigenous landraces; Group 2 comprised genotypes selected from exotic landraces (released in 1920's–1930's); Group 3 included genotypes selected from crosses or mutagenesis involving Group 2 materials (1950's–1960's); Group 4 comprised genotypes selected from crosses between CIMMYT and Group 2 materials (from 1970's). Under moderately favourable conditions, a yield increase of 1.03 t ha–¹ was observed from Group 1 through Group 4, corresponding to a genetic gain of about 0.017 t ha–¹ per year. Such increase was only partly accounted for by a parallel increase in spike fertility and seed weight. Plant stature decreased dramatically from Group 1 to Group 4; a remarkable reduction of height was already attained in Group 3, before the introduction of dwarfing genes from bread wheat. Earliness of heading and grain filling duration increased consistently across breeding phases, the length of crop cycle remaining almost unaltered. No significant improvement of drought tolerance resulted from the breeding activity, suggesting the need to put more emphasis on selection for real stress tolerance rather than escape. Overall variation for morphophysiological traits, assessed by a principal components analysis, highlighted the great diversity among the Group 1 cultivars, while variation within Groups 3 and 4 was lower. Indigenous landraces, little used in the breeding history, as well as novel, unexploited exotic germplasm sources could contribute to broaden the crop genetic base in the region.     

黄瓜(Cucumis sativus L.)种子含油量性状的QTL定位与分析

应用SRAP和SSR技术,对黄瓜种子高含油量品系Ma7与低含油量品系M6杂交组合的F2群体进行检测,获得102个分子标记,构建了7个连锁群组成的分子标记遗传图谱;图谱总长764 cM,标记间平均长度7.49 cM.应用Windows QTL Cartographer 2.5对种子含油量性状进行QTL扫描,在2009年秋...     

黄瓜的分子标记和连锁图谱研究进展

黄瓜分子标记和遗传连锁图谱的研究工作相对番茄、小麦等作物比较落后，至今开展的分子标记研究多围绕黄瓜遗传关系分析进行，现有的几张遗传连锁图谱基本是由美国Staub研究小组完成，已被定位在图谱上的同工酶、RAPD、RFLP、SSR、AFLP等标记总数达到300多个，各标记间平均距离达到2.1cM，被整合在一起的遗传基因不足10个(F、B、de、ll、dm)。     

Cloning and Sequence Analysis of Lipoxygenase Gene cDNA from Cucumber Fruit （Cucumis sativus L.）

Lipoxygenases are nonheme-iron-containing dioxygenases that catalyze the hydroperoxidation of unsatrated fatty acids containing a cis, cis-1,4-pentadiene structure producing hydroperoxy acids with conjugated dienes. LOX activity has been found in a wide range of plants. Typical substrates for LOX in plants are linoleic acid and linilenic acid fatty acids. The function of various LOXs in plants is unknown, but their participation in all stages of plant growth and development has been suggested （Hildebrand, 1989; Siedow, 1991）. Some of the physiological processes in whicn lipoxygenses have been implicated include wounding （Saravitz and Siedow, 1996）, pathogen attack （Melan et al., 1993）, seed germination （Kato et al., 1992）, fruit ripening （Ferrie et al., 1994）, plant senescence （Paliyath and Droillard, 1992）. The study on the role of lipoxygenase in ripening and senescence fruit focused on tomato and strawberry. Cloning LOX gene of cucumber fruit will make us further understand the molecularaction mode of this enzyme during fxuit ripening and senescence. In this paper we isolated the partial nucleotide sequences of cucumber fiuit lipoxygenase gene and discuss the characterization of it.     

根结线虫侵染对黄瓜生长和内源激素含量的影响

通过酶联免疫吸附法测定根结线虫侵染后48h内的黄瓜体内的各种内源植物激素含量的变化,探索黄瓜根结线虫病害产生的生理应答机制。通过外部形态结果观察看到根结线虫侵染后,黄瓜生长缓慢,植株矮小,并且提前衰老。同时,黄瓜叶片中的内源植物激素含量在48h内也发生了较大变化:黄瓜叶片中茉莉酸的含量在线虫侵染16h内急剧上升,上升幅度高达对照的8.2倍,而水杨酸、脱落酸和异戊烯基腺苷在侵染后含量有显著上升的趋势,并在24h达到各自的峰值;但是生长素和赤霉素含量仅短暂升高,其余时间显著低于对照。通过测定浸染初期各内源植物激素之间的动态平衡,可以为研究植物诱导抗病性提供理论参考。     

Search for resistance to gummy stem blight (Didymella bryoniae) in cucumber (Cucumis sativus L.)

Summary Resistance to gummy stem blight (Didymella bryoniae) was not detected among 1208 cucumber lines of diverse origin when cotyledons of four-day-old seedlings were inoculated with an aqueous suspension of 5000 spores of D. bryoniae delivered to freshly crushed tissue, followed by 48 h incubation at 20°C and 100% RH in the dark. Among 49 lots evaluated for gummy stem blight in the field, cv. Homegreen #2 and Pl 200818 from Burma were resistant. Earliness in fruit maturity was positively correlated with susceptibility to gummy stem blight (r=0.424). Cucumber accessions differed in the incidence of stem and hypocotyl cankers. In greenhouse grown plants at the 2nd–3rd leaf stage, differentiation of resistant lines was obtained by spraying plants with an aqueous suspension of 10⁶/ml D. bryoniae spores followed by 48 h incubation at 20°C and 100% RH in the dark. One cycle of mass selection in Homegreen #2 resulted in improved gummy stem blight resistance.     

Coronatine Enhances Chilling Tolerance in Cucumber (Cucumis sativus L.) Seedlings by Improving the Antioxidative Defence System

Coronatine (COR) is a new plant growth regulator that mimics the biological activities of methyl jasmonate. We determined whether COR enhanced chilling tolerance of cucumber ( Cucumis sativus L. cv. Jinchun 4) seedlings and if such tolerance was correlated with changes in the activity of antioxidant enzymes. COR was applied to seedlings at two-leaf stage at 0 (Control), 0.1, 1, 10, and 100 n m . Seedlings were then subjected to chilling stress at 5 ± 1 °C for 4 days. Seedlings treated with COR showed significant higher tolerance to chilling stress and the optimal concentration was 1–10 n m . Compared with control, the chilling injury index (CII) of the seedlings treated with COR at 1 and 10 n m was decreased by 44.9 % and 24.5 %, respectively, while the membrane chilling stability (MCS) expressed as the change of relative conductance was increased by 37.2 % and 17.0 %, respectively. The malondialdehyde (MDA) content in leaves treated with COR at 1 n m was decreased by 39.7 %, and the O₂⁻ production rate and H₂O₂ content reduced by 28.6 % and 8.5 %, respectively. Treatment with COR at 1 n m increased the activities of superoxide dismutase (SOD) in leaves by 34.4 %, catalase (CAT) by 58.7 % and ascorbate peroxidase (APX) by 23.0 % under low temperature. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities were also significantly improved by 28.9 % and 21.3 % following treatment with COR at 1 and 10 n m , respectively. The overall results suggest that COR enhanced chilling tolerance in cucumber seedlings by improving the antioxidative defence system.     

Linkage of random amplified polymorphic DNA markers to downy mildew resistance in cucumber (Cucumis sativus L.)

Marker assisted selection (MAS) may improve the efficiency of breeding downy mildew resistant cucumber cultivars. A study was conducted to identify random amplified polymorphic DNA (RAPD) markers linked to the downy mildew resistance gene (dm) which would be suitable for MAS. A total of 145 F₃ families from two populations (55 from the WI 1983G × Straight 8 population and 90 from the Zudm1 × Straight 8 population) were evaluated over five locations in North America and Europe. Resistant and susceptible F₃ families were identified and mean family resistance ratings were used to type individual F₂ plants. No evidence for race differences in the pathogen (Psuedoperonospora cubensis (Berk. & Curt.) Rostow) between North America and Europe was found. Phenotypic correlations between locations ranged from 0.3 to 0.7. Of the 135 polymorphic RAPD markers identified from 960 primers, five were linked to dm - G14₈₀₀, X15₁₁₀₀, AS5₈₀₀, BC519₁₁₀₀, and BC526₁₀₀₀. In the WI 1983G × Straight 8 population, G14₈₀₀ was linked to dm at 16.5 cm, AS5₈₀₀ at 32.8 cm, BC519₁₁₀₀ at 9.9 cm, and BC526₁₀₀₀ at 19.2 cm. In the Zudm1 ×Straight 8 population, G14₈₀₀ was linked at 20.9 cm, X15₁₁₀₀at 14.8 cm, AS5₈₀₀ at 24.8 cm, and BC526_{_1000} at 32.9 cm. MarkersG14₈₀₀ and BC519₁₁₀₀ were linked in repulsion to the dm allele, and X15₁₁₀₀, AS5₈₀₀, and BC526₁₀₀₀ were linked in coupling phase. These genetic markers may be exploited to develop an efficient MAS strategy for breeding resistant cucumber cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Use of molecular marker diversity to increase forage yield in timothy (Phleum pratense L.)

Molecular marker technologies may provide a tool to overcome the forage yield plateau in timothy (Phleum pratense L.). Therefore, a study was designed to investigate the relationship between marker‐based genetic distance (GD) estimates and yield. Genetic distances were estimated using Dice coefficients framed by 28 simple sequence repeat markers. In Experiment 1, parents with high general combining ability (GCA) from two contrasting first‐generation synthetic strains (exhibiting high and low yields) were compared. Average GDs of each strain were 0.74 (high) and 0.68 (low). These differences indicated that GD may be partially the basis for contrasting yields. In Experiment 2, GDs among 40 parents of a polycross were used for calculating general genetic distance (GGD), which is indicative of their allelic complementation. Analyses revealed a significant (P < 0.01) moderate correlation (r) between GGD and GCA for yield (r = 0.45) and a significant (P < 0.01) residual mean square for the regression of yield on GGD, suggesting that considerable non‐additive effects were associated with GCA. The results are indicative of the potential use of GD estimation for yield improvement in timothy.