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Global warming is a serious threat to food security and severely affects plant growth, developmental processes, and, eventually, crop productivity. Respiratory metabolism plays a critical role in the adaptation of diverse stress in plants. Aconitase (ACO) is the main enzyme, which catalyzes the revocable isomerization of citrate to isocitrate in the Krebs cycle. The function of ACO gene family members has been extensively studied in model plants, for instance Arabidopsis. However, their role in plant developmental processes and various stress conditions largely remained unknown in other plant species. Thus, we identified 15 ACO genes in wheat to elucidate their function in plant developmental processes and different stress environments. The phylogenetic tree revealed that

Current nomenclature for the taxonomic classification of melon cultivars (Cucumis melo L.) at the horticultural group level relies on morphological variation in certain key characters. However, the reliability of current infraspecific classification scheme in considering horticultural groups as botanical taxa was not fully understood. In the present study, the information of horticultural group classification in melon was assessed at the molecular level using genome-wide single nucleotide polymorphisms (SNPs). A total of 143 melon accessions of 15 horticultural groups in two subspecies, subsp. melo and subsp. agresti were collected and genotyped by using Genotyping-By-Sequencing (GBS). From the filtering of resultant sequence variants, 10,949 SNPs were selected and used for downstream genetic analysis including population structure, principle component analysis, and hierarchical clustering of 143 melon accessions. Our genetic analyses indicated that the distribution of accessions at the molecular level generally matched the subspecies classification and no substantial contradictions existed between the division of accessions based on horticultural group information and genetic relatedness revealed by the GBS. However, the distinction between horticultural groups was not clear-cut, implying the limitation of considering horticultural groups as botanical taxa. To improve the resolution of horticultural group classification in melon, our SNP data may be useful as supporting information in conjunction with morphological characters.

Proline-rich extensin-like receptor kinases (PERKs) are a class of receptor kinases implicated in multiple cellular processes in plants. However, there is a lack of information on the PERK gene family in wheat. Therefore, we identified 37 PERK genes in wheat to understand their role in various developmental processes and stress conditions. Phylogenetic analysis of PERK genes from Arabidopsis thaliana, Oryza sativa, Glycine max, and T. aestivum grouped them into eight well-defined classes. Furthermore, synteny analysis revealed 275 orthologous gene pairs in B. distachyon, Ae. tauschii, T. dicoccoides, O. sativa and A. thaliana. Ka/Ks values showed that most TaPERK genes, except TaPERK1, TaPERK2, TaPERK17, and TaPERK26, underwent strong purifying selection during evolutionary processes. Several cis-acting regulatory elements, essential for plant growth and development and the response to light, phytohormones, and diverse biotic and abiotic stresses, were predicted in the promoter regions of TaPERK genes. In addition, the expression profile of the TaPERK gene family revealed differential expression of TaPERK genes in various tissues and developmental stages. Furthermore, TaPERK gene expression was induced by various biotic and abiotic stresses. The RT-qPCR analysis also revealed similar results with slight variation. Therefore, this study’s outcome provides valuable information for elucidating the precise functions of TaPERK in developmental processes and diverse stress conditions in wheat.

The world has faced the challenges of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) for the last two years, first diagnosed at the end of 2019 in Wuhan and widely distributed worldwide. As a result, the WHO has proclaimed the illness brought on by this virus to be a global pandemic. To combat COVID-19, researcher communities continuously develop and implement rapid diagnoses, safe and effective vaccinations and other alternative therapeutic procedures. However, synthetic drug-related side effects and high costs have piqued scientists’ interest in natural product-based therapies and medicines. In this regard, antiviral substances derived from natural resources and some medicines have seen a boom in popularity. For instance, algae are a rich source of compounds such as lectins and sulfated polysaccharides, which have potent antiviral and immunity-boosting properties. Moreover, Algae-derived compounds or metabolites can be used as antibodies and vaccine raw materials against COVID-19. Furthermore, some algal species can boost immunity, reduce viral activity in humans and be recommended for usage as a COVID-19 preventative measure. However, this field of study is still in its early stages of development. Therefore, this review addresses critical characteristics of algal metabolites, their antioxidant potential and therapeutic potential in COVID-19.

Lettuce is one of the economically important leaf vegetables and is cultivated mainly in temperate climate areas. Cultivar identification based on the distinctness, uniformity, and stability (DUS) test is a prerequisite for new cultivar registration. However, DUS testing based on morphological features is time-consuming, labor-intensive, and costly, and can also be influenced by environmental factors. Thus, molecular markers have also been used for the identification of genetic diversity as an effective, accurate, and stable method. Currently, genome-wide single nucleotide polymorphisms (SNPs) using next-generation sequencing technology are commonly applied in genetic research on diverse plant species. This study aimed to establish an effective and high-throughput cultivar identification system for lettuce using core sets of SNP markers developed by genotyping by sequencing (GBS). GBS identified 17 877 high-quality SNPs for 90 commercial lettuce cultivars. Genetic differentiation analyses based on the selected SNPs classified the lettuce cultivars into three main groups. Core sets of 192, 96, 48, and 24 markers were further selected and validated using the Fluidigm platform. Phylogenetic analyses based on all core sets of SNPs successfully discriminated individual cultivars that have been currently recognized. These core sets of SNP markers will support the construction of a DNA database of lettuce that can be useful for cultivar identification and purity testing, as well as DUS testing in the plant variety protection system. Additionally, this work will facilitate genetic research to improve breeding in lettuce.

Salt stress is a severe type of environmental stress. It adversely affects agricultural production worldwide. The overproduction of reactive oxygen species (ROS) is the most frequent phenomenon during salt stress. ROS are extremely reactive and, in high amounts, noxious, leading to destructive processes and causing cellular damage. However, at lower concentrations, ROS function as secondary messengers, playing a critical role as signaling molecules, ensuring regulation of growth and adjustment to multifactorial stresses. Plants contain several enzymatic and non-enzymatic antioxidants that can detoxify ROS. The production of ROS and their scavenging are important aspects of the plant’s normal response to adverse conditions. Recently, this field has attracted immense attention from plant scientists; however, ROS-induced signaling pathways during salt stress remain largely unknown. In this review, we will discuss the critical role of different antioxidants in salt stress tolerance. We also summarize the recent advances on the detrimental effects of ROS, on the antioxidant machinery scavenging ROS under salt stress, and on the crosstalk between ROS and other various signaling molecules, including nitric oxide, hydrogen sulfide, calcium, and phytohormones. Moreover, the utilization of “-omic” approaches to improve the ROS-regulating antioxidant system during the adaptation process to salt stress is also described.

Global warming is a serious threat to food security and severely affects plant growth, developmental processes, and, eventually, crop productivity. Respiratory metabolism plays a critical role in the adaptation of diverse stress in plants. Aconitase (ACO) is the main enzyme, which catalyzes the revocable isomerization of citrate to isocitrate in the Krebs cycle. The function of ACO gene family members has been extensively studied in model plants, for instance Arabidopsis. However, their role in plant developmental processes and various stress conditions largely remained unknown in other plant species. Thus, we identified 15 ACO genes in wheat to elucidate their function in plant developmental processes and different stress environments. The phylogenetic tree revealed that TaACO genes were classified into six groups. Further, gene structure analysis of TaACOs has shown a distinctive evolutionary path. Synteny analysis showed the 84 orthologous gene pairs in Brachypodium distachyon, Aegilops tauschii, Triticum dicoccoides, Oryza sativa, and Arabidopsis thaliana. Furthermore, Ka/Ks ratio revealed that most TaACO genes experienced strong purifying selection during evolution. Numerous cis-acting regulatory elements were detected in the TaACO promoters, which play a crucial role in plant development processes, phytohormone signaling, and are related to defense and stress. To understand the function of TaACO genes, the expression profiling of TaACO genes were investigated in different tissues, developmental stages, and stress conditions. The transcript per million values of TaACOs genes were retrieved from the Wheat Expression Browser Database. We noticed the differential expression of the TaACO genes in different tissues and various stress conditions. Moreover, gene ontology analysis has shown enrichment in the tricarboxylic acid metabolic process (GO:0072350), citrate metabolic process (GO:0006101), isocitrate metabolic process GO:0006102, carbohydrate metabolic (GO:0005975), and glyoxylate metabolic process (GO:0046487). Therefore, this study provided valuable insight into the ACO gene family in wheat and contributed to the further functional characterization of TaACO during different plant development processes and various stress conditions.

In the current scenario of changing climatic conditions and the rising global population, there is an urgent need to explore novel, efficient, and economical natural products for the benefit of humankind. Biosurfactants are one of the latest explored microbial synthesized biomolecules that have been used in numerous fields, including agriculture, pharmaceuticals, cosmetics, food processing, and environment-cleaning industries, as a source of raw materials, for the lubrication, wetting, foaming, emulsions formulations, and as stabilizing dispersions. The amphiphilic nature of biosurfactants have shown to be a great advantage, distributing themselves into two immiscible surfaces by reducing the interfacial surface tension and increasing the solubility of hydrophobic compounds. Furthermore, their eco-friendly nature, low or even no toxic nature, durability at higher temperatures, and ability to withstand a wide range of pH fluctuations make microbial surfactants preferable compared to their chemical counterparts. Additionally, biosurfactants can obviate the oxidation flow by eliciting antioxidant properties, antimicrobial and anticancer activities, and drug delivery systems, further broadening their applicability in the food and pharmaceutical industries. Nowadays, biosurfactants have been broadly utilized to improve the soil quality by improving the concentration of trace elements and have either been mixed with pesticides or applied singly on the plant surfaces for plant disease management. In the present review, we summarize the latest research on microbial synthesized biosurfactant compounds, the limiting factors of biosurfactant production, their application in improving soil quality and plant disease management, and their use as antioxidant or antimicrobial compounds in the pharmaceutical industries.

The objective of this study was to use ‘Danta PR’, NGS (Next Generation Sequencing) technology for genome resequencing to develop polymorphic makers between Chinese oriental melon, ‘Hyangseo 1’ and Korean oriental melon. From the resequencing data that covered about 81 times of the genome size, 104,357 of SSR motifs and Indel, and 1,092,436 of SNPs were identified. 299 SSR and 307 Indel markers were chosen to cover each chromosome with 25 markers. These markers were subsequently used to identify genotypes of ‘Danta PR’ BC1 (F1 x ‘Danta PR’) population and a genetic linkage map was constructed. SSR, Indel, and SNPs identified in this study would be useful as a breeding tool to develop new oriental melon varieties.

PIN-FORMED (PIN) genes play a crucial role in regulating polar auxin distribution in diverse developmental processes, including tropic responses, embryogenesis, tissue differentiation, and organogenesis. However, the role of PIN-mediated auxin transport in various plant species is poorly understood. Currently, no information is available about this gene family in wheat (Triticum aestivum L.). In the present investigation, we identified the PIN gene family in wheat to understand the evolution of PIN-mediated auxin transport and its role in various developmental processes and under different biotic and abiotic stress conditions. In this study, we performed genome-wide analysis of the PIN gene family in common wheat and identified 44 TaPIN genes through a homology search, further characterizing them to understand their structure, function, and distribution across various tissues. Phylogenetic analyses led to the classification of TaPIN genes into seven different groups, providing evidence of an evolutionary relationship with Arabidopsis thaliana and Oryza sativa. A gene exon/intron structure analysis showed a distinct evolutionary path and predicted the possible gene duplication events. Further, the physical and biochemical properties, conserved motifs, chromosomal, subcellular localization, transmembrane domains, and three-dimensional (3D) structure were also examined using various computational approaches. Cis-elements analysis of TaPIN genes showed that TaPIN promoters consist of phytohormone, plant growth and development, and stress-related cis-elements. In addition, expression profile analysis also revealed that the expression patterns of the TaPIN genes were different in different tissues and developmental stages. Several members of the TaPIN family were induced during biotic and abiotic stress. Moreover, the expression patterns of TaPIN genes were verified by qRT-PCR. The qRT-PCR results also show a similar expression with slight variation. Therefore, the outcome of this study provides basic genomic information on the expression of the TaPIN gene family and will pave the way for dissecting the precise role of TaPINs in plant developmental processes and different stress conditions.

Lettuce is one of the economically important leaf vegetables and is cultivated mainly in temperate climate areas. Cultivar identification based on the distinctness, uniformity, and stability (DUS) test is a prerequisite for new cultivar registration. However, DUS testing based on morphological features is time-consuming, labor-intensive, and costly, and can also be influenced by environmental factors. Thus, molecular markers have also been used for the identification of genetic diversity as an effective, accurate, and stable method. Currently, genome-wide single nucleotide polymorphisms (SNPs) using next-generation sequencing technology are commonly applied in genetic research on diverse plant species. This study aimed to establish an effective and high-throughput cultivar identification system for lettuce using core sets of SNP markers developed by genotyping by sequencing (GBS). GBS identified 17 877 high-quality SNPs for 90 commercial lettuce cultivars. Genetic differentiation analyses based on the selected SNPs classified the lettuce cultivars into three main groups. Core sets of 192, 96, 48, and 24 markers were further selected and validated using the Fluidigm platform. Phylogenetic analyses based on all core sets of SNPs successfully discriminated individual cultivars that have been currently recognized. These core sets of SNP markers will support the construction of a DNA database of lettuce that can be useful for cultivar identification and purity testing, as well as DUS testing in the plant variety protection system. Additionally, this work will facilitate genetic research to improve breeding in lettuce.

Brassinosteroids (BRs) play crucial roles in various biological processes, including plant developmental processes and response to diverse biotic and abiotic stresses. However, no information is currently available about this gene family in wheat (Triticum aestivum L.). In the present investigation, we identified the BZR gene family in wheat to understand the evolution and their role in diverse developmental processes and under different stress conditions. In this study, we performed the genome-wide analysis of the BZR gene family in the bread wheat and identified 20 TaBZR genes through a homology search and further characterized them to understand their structure, function, and distribution across various tissues. Phylogenetic analyses lead to the classification of TaBZR genes into five different groups or subfamilies, providing evidence of evolutionary relationship with Arabidopsis thaliana, Zea mays, Glycine max, and Oryza sativa. A gene exon/intron structure analysis showed a distinct evolutionary path and predicted the possible gene duplication events. Further, the physical and biochemical properties, conserved motifs, chromosomal, subcellular localization, and cis-acting regulatory elements were also examined using various computational approaches. In addition, an analysis of public RNA-seq data also shows that TaBZR genes may be involved in diverse developmental processes and stress tolerance mechanisms. Moreover, qRT-PCR results also showed similar expression with slight variation. Collectively, these results suggest that TaBZR genes might play an important role in plant developmental processes and various stress conditions. Therefore, this work provides valuable information for further elucidate the precise role of BZR family members in wheat.

Many pathogenic viral pandemics have caused threats to global health; the COVID-19 pandemic is the latest. Its transmission is growing exponentially all around the globe, putting constraints on the health system worldwide. A novel coronavirus, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), causes this pandemic. Many candidate vaccines are available at this time for COVID-19, and there is a massive international race underway to procure as many vaccines as possible for each country. However, due to heavy global demand, there are strains in global vaccine production. The use of a plant biotechnology-based expression system for vaccine production also represents one part of this international effort, which is to develop plant-based heterologous expression systems, virus-like particles (VLPs)-vaccines, antiviral drugs, and a rapid supply of antigen-antibodies for detecting kits and plant origin bioactive compounds that boost the immunity and provide tolerance to fight against the virus infection. This review will look at the plant biotechnology platform that can provide the best fight against this global pandemic.

PIN-FORMED (PIN) genes play a crucial role in regulating polar auxin distribution in diverse developmental processes, including tropic responses, embryogenesis, tissue differentiation, and organogenesis. However, the role of PIN-mediated auxin transport in various plant species is poorly understood. Currently, no information is available about this gene family in wheat (Triticum aestivum L.). In the present investigation, we identified the PIN gene family in wheat to understand the evolution of PIN-mediated auxin transport and its role in various developmental processes and under different biotic and abiotic stress conditions. In this study, we performed genome-wide analysis of the PIN gene family in common wheat and identified 44 TaPIN genes through a homology search, further characterizing them to understand their structure, function, and distribution across various tissues. Phylogenetic analyses led to the classification of TaPIN genes into seven different groups, providing evidence of an evolutionary relationship with Arabidopsis thaliana and Oryza sativa. A gene exon/intron structure analysis showed a distinct evolutionary path and predicted the possible gene duplication events. Further, the physical and biochemical properties, conserved motifs, chromosomal, subcellular localization, transmembrane domains, and three-dimensional (3D) structure were also examined using various computational approaches. Cis-elements analysis of TaPIN genes showed that TaPIN promoters consist of phytohormone, plant growth and development, and stress-related cis-elements. In addition, expression profile analysis also revealed that the expression patterns of the TaPIN genes were different in different tissues and developmental stages. Several members of the TaPIN family were induced during biotic and abiotic stress. Moreover, the expression patterns of TaPIN genes were verified by qRT-PCR. The qRT-PCR results also show a similar expression with slight variation. Therefore, the outcome of this study provides basic genomic information on the expression of the TaPIN gene family and will pave the way for dissecting the precise role of TaPINs in plant developmental processes and different stress conditions.

In this study, we have introduced newly synthesized substituted benzothiazole based berberine derivatives that have been analyzed for their in vitro and in silico biological properties. The activity towards various kinds of influenza virus strains by employing the cytopathic effect (CPE) and sulforhodamine B (SRB) assay. Several berberine–benzothiazole derivatives (BBDs), such as BBD1, BBD3, BBD4, BBD5, BBD7, and BBD11, demonstrated interesting anti-influenza virus activity on influenza A viruses (A/PR/8/34, A/Vic/3/75) and influenza B viral (B/Lee/40, and B/Maryland/1/59) strain, respectively. Furthermore, by testing neuraminidase activity (NA) with the neuraminidase assay kit, it was identified that BBD7 has potent neuraminidase activity. The molecular docking analysis further suggests that the BBD1–BBD14 compounds’ antiviral activity may be because of interaction with residues of NA, and the same as in oseltamivir.

Three pumpkin species Cucurbita maxima, C. moschata, and C. pepo are commonly cultivated worldwide. To identify genome-wide SNPs in these cultivated pumpkin species, we collected 48 F1 cultivars consisting of 40 intraspecific hybrids (15 C. maxima, 18 C. moschata, and 7 C. pepo) and 8 interspecific hybrids (C. maxima x C. moschata). Genotyping by sequencing identified a total of 37,869 confident SNPs in this collection. These SNPs were filtered to generate a subset of 400 SNPs based on polymorphism and genome distribution. Of the 400 SNPs, 288 were used to genotype an additional 188 accessions (94 F1 cultivars, 50 breeding lines, and 44 landraces) with a SNP array-based platform. Reliable polymorphisms were observed in 224 SNPs (78.0%) and were used to assess genetic variations between and within the four predefined populations in 223 cultivated pumpkin accessions. Both principal component analysis and UPGMA clustering found four major clusters representing three pumpkin species and interspecific hybrids. This genetic differentiation was supported by pairwise Fst and Nei’s genetic distance. The interspecific hybrids showed a higher level of genetic diversity relative to the other three populations. Of the 224 SNPs, five subsets of 192, 96, 48, 24, and 12 markers were evaluated for variety identification. The 192, 96, and 48 marker sets identified 204 (91.5%), 190 (85.2%), and 141 (63.2%) of the 223 accessions, respectively, while other subsets showed

Potato is an important source of food in South Korea, and viruses represent a significant threat to sustainable and profitable potato production. However, information about viruses affecting the potato crop in South Korea is limited. In 2017, potato plants of five cultivars exhibiting foliar mosaic, crinkling, and mottle were collected in two seed potato production areas, in Gangwon-do and Jeollabuk-do Provinces, and subjected to virus testing and characterization. Potato virus Y (PVY) was found associated with mosaic symptoms, and samples were characterized using reverse transcription polymerase chain reaction (RT-PCR) and whole genome sequencing. All analyzed PVY-positive samples were found to represent the same recombinant PVY strain: PVYNTN. Three PVY isolates were subjected to whole genome sequencing using overlapping RT-PCR fragments and Sanger methodology, and all three were confirmed to represent strain PVYNTNa after a recombination analysis of the complete genomes. In phylogenetic analysis, the three South Korean isolates were placed most closely to several PVYNTNa isolates reported from Japan and Vietnam, suggesting a common source of infection. This is the first report and complete molecular characterization of a PVYNTN strain present in the country, and because this strain induces tuber necrotic ringspot disease in susceptible cultivars of potato, appropriate management tools need to be implemented to mitigate potential tuber quality losses.

Tuberculosis (TB) is a recurrent and progressive disease, with high mortality rates worldwide. The drug-resistance phenomenon of Mycobacterium tuberculosis is a major obstruction of allelopathy treatment. An adverse side effect of allelopathic treatment is that it causes serious health complications. The search for suitable alternatives of conventional regimens is needed, i.e., by considering medicinal plant secondary metabolites to explore anti-TB drugs, targeting the action site of M. tuberculosis. Nowadays, plant-derived secondary metabolites are widely known for their beneficial uses, i.e., as antioxidants, antimicrobial agents, and in the treatment of a wide range of chronic human diseases (e.g., tuberculosis), and are known to “thwart” disease virulence. In this regard, in silico studies can reveal the inhibitory potential of plant-derived secondary metabolites against Mycobacterium at the very early stage of infection. Computational approaches based on different algorithms could play a significant role in screening plant metabolites against disease virulence of tuberculosis for drug designing.

Brassinosteroids (BRs) play crucial roles in various biological processes, including plant developmental processes and response to diverse biotic and abiotic stresses. However, no information is currently available about this gene family in wheat (Triticum aestivum L.). In the present investigation, we identified the BZR gene family in wheat to understand the evolution and their role in diverse developmental processes and under different stress conditions. In this study, we performed the genome-wide analysis of the BZR gene family in the bread wheat and identified 20 TaBZR genes through a homology search and further characterized them to understand their structure, function, and distribution across various tissues. Phylogenetic analyses lead to the classification of TaBZR genes into five different groups or subfamilies, providing evidence of evolutionary relationship with Arabidopsis thaliana, Zea mays, Glycine max, and Oryza sativa. A gene exon/intron structure analysis showed a distinct evolutionary path and predicted the possible gene duplication events. Further, the physical and biochemical properties, conserved motifs, chromosomal, subcellular localization, and cis-acting regulatory elements were also examined using various computational approaches. In addition, an analysis of public RNA-seq data also shows that TaBZR genes may be involved in diverse developmental processes and stress tolerance mechanisms. Moreover, qRT-PCR results also showed similar expression with slight variation. Collectively, these results suggest that TaBZR genes might play an important role in plant developmental processes and various stress conditions. Therefore, this work provides valuable information for further elucidate the precise role of BZR family members in wheat.

Genetic diversity analysis and cultivar identification were performed using a core set of single nucleotide polymorphisms (SNPs) in cucumber (Cucumis sativus L.). For the genetic diversity study, 280 cucumber accessions collected from four continents (Asia, Europe, America, and Africa) by the National Agrobiodiversity Center of the Rural Development Administration in South Korea and 20 Korean commercial F1 hybrids were genotyped using 151 Fluidigm SNP assay sets. The heterozygosity of the SNP loci per accession ranged from 4.76 to 82.76%, with an average of 32.1%. Population genetics analysis was performed using population structure analysis and hierarchical clustering (HC), which indicated that these accessions were classified mainly into four subpopulations or clusters according to their geographical origins. The subpopulations for Asian and European accessions were clearly distinguished from each other (FST value = 0.47), while the subpopulations for Korean F1 hybrids and Asian accessions were closely related (FST = 0.34). The highest differentiation was observed between American and European accessions (FST = 0.41). Nei’s genetic distance among the 280 accessions was 0.414 on average. In addition, 95 commercial F1 hybrids of three cultivar groups (Baekdadagi-, Gasi-, and Nakhap-types) were genotyped using 82 Fluidigm SNP assay sets for cultivar identification. These 82 SNPs differentiated all cultivars, except seven. The heterozygosity of the SNP loci per cultivar ranged from 12.20 to 69.14%, with an average of 34.2%. Principal component analysis and HC demonstrated that most cultivars were clustered based on their cultivar groups. The Baekdadagi- and Gasi-types were clearly distinguished, while the Nakhap-type was closely related to the Baekdadagi-type. Our results obtained using core Fluidigm SNP assay sets provide useful information for germplasm assessment and cultivar identification, which are essential for breeding and intellectual right protection in cucumber.

Single nucleotide polymorphisms (SNPs) have been widely used as a molecular marker in crop species with advances in next-generation sequencing technology. The use of SNP markers for variety identification is a cost-effective strategy to protect breeder’s intellectual property rights. This study was conducted to identify genome-wide SNPs and develop core marker sets for assessing genetic variations in commercial tomato cultivars. A total of 10,615 confident SNPs was generated from genotyping by sequencing for 48 F1 cultivars representing four market classes (large-fruited fresh market, cherry, grape, and rootstock). Of these, 288 SNPs across 12 chromosomes were selected to genotype additional 94 F1 cultivars using the Fluidigm assay and 224 SNPs showed reliable polymorphisms in 91 F1 cultivars. Both model-based and hierarchical clustering analyses with these markers found that the large-fruited fresh market cultivars were significantly distinct from the cherry and grape cultivars. In addition, the cherry and grape cultivars showed higher levels of genetic diversity relative to the large-fruited fresh market cultivars. The 224 SNP markers were also effective in differentiating all 139 F1 cultivars and Heinz 1706 (an inbred for the tomato reference genome). Of the five subsets, the 192 and 96 markers identified all of these tomato cultivars, respectively. Furthermore, the other three subsets of 48, 24, and 12 markers showed 80.0–93.6 % of identification rates. These results demonstrate that all five subsets of SNP markers will be useful in developing a high-throughput DNA barcoding system for variety identification in commercial tomato cultivars.

The Oriental melon is an important delicious fruit crop across East Asia. Single-nucleotide polymorphisms (SNPs) are considered to be a useful genotyping tool both to detect genetic diversity and to protect breeder rights. This study used genotyping-by-sequencing (GBS) to detect genome-wide SNPs by analyzing 48 commercial Oriental melon varieties to generate 5640 filtered SNPs. Based on the high polymorphism information content (PIC), genetic and physical distances, a subset of 192 SNPs was selected as putative SNPs for validation via the Fluidigm JunoTM system. Of these, 164 SNPs were successfully validated in 87 Oriental melon varieties with a validation ratio of 85.41 %. Furthermore, these 87 Oriental melon varieties were classified into ten distinct groups, based on 164 SNP markers. Therefore, these large set of SNP markers detected here has several application, such as genetic diversity studies, varietal identification and marker-assisted selection (MAS) in the Oriental melon. Genome-wide associations (GWAS) analyses detected significant 18 SNPs associated with various morphological traits, including two novel SNPs for sex expression on chromosomes 1 and 8 that were not colocalized with previous studies. The four potential candidate genes such as MELO3C015898, MELO3C015904, MELO3C024563, and MELO3C024565 were predicted within the GWAS-SNPs regions for sex expression in the Oriental melon. Hence, identification of these candidate genes has provided the foundation to study the molecular genetic mechanism of sex expression trait in the Oriental melon.

Melon is an important horticultural crop across the globe, which consisted of several subspecies. Single nucleotide polymorphisms (SNPs) are the most preferred molecular markers in genetic diversity and genomic studies. Recently, Genotyping-by-sequencing (GBS) has produced sequences that detect genome-wide distribution of these SNP markers. In this study, we identified genome-wide highly efficient SNP markers to differentiate the cultivars of the melon using GBS. Here we detected 9018 SNPs by the analysis of 48 commercialized Korean melon cultivars, which released by various seed companies in South Korea. Further, precise filtering resulted in 5644 high-quality SNPs. Of these, 5293 SNPs and 351 were identified in the genic region and intergenic region, respectively. A total of 192 SNPs were identified as putative SNPs, on the basis of high polymorphism information content (PIC) and physical distance. These SNPs were further analyzed to distinguish 95 commercial melon cultivars via high-throughput Fluidigm system, thus revealed 95.83 % (184 SNPs) of validation ratio. The summary of SNP genotyping revealed estimates of PIC ranged from 0.02 to 0.53 and 179 SNPs (97.28 %) showed more than 0.3 of PIC. This is the first study in melon to develop and validate genome-wide SNP markers using GBS and Fluidigm platform, respectively. The SNP markers reported in this study will be a valuable tool for cultivar identification, genetic diversity, and marker-trait association studies in melon.

Tomato (Solanum lycopersicum L.) is an economically important vegetable crop worldwide. Recently, a high-density single nucleotide polymorphism (SNP) array was developed based on genome-wide SNPs in tomato. In this study, we genotyped a collection of 48 Korean elite tomato varieties (26 fresh market and 22 cultivated cherry) using 7,720 SNPs of this array. Out of 6,652 polymorphic SNPs (86.1%) in the entire collection, there were 6,589 SNPs with < 10% missing data. The number of polymorphic SNPs in the fresh market and cultivated cherry subpopulations were 4,733 (61.3%) and 6,087 (78.8%), respectively. To examine the genetic variation between sub-populations, the SNP genotypes of the Korean tomato germplasm were analyzed along with the previously reported data on SNPs of the 277 Solanaceae Agricultural Coordinated Project (SolCAP) varieties (109 fresh market, 27 cultivated cherry, and 141 processing). Principal component analysis, pairwise F st, and Nei’s standard genetic distance revealed genetic differentiation between these five sub-populations. Moreover, we validated another division within the Korean cherry varieties using the unweighted pair group mean algorithm (UPGMA). The genetic diversity of each sub-population was estimated based on allelic richness and expected heterozygosity. The fresh market and cultivated cherry sub-populations in the Korean tomato germplasm showed similar levels of genetic diversity as the corresponding SolCAP sub-populations. Visualization of the polymorphic information revealed genomic regions that differed between the two sub-populations in the Korean tomato germplasm. These results suggest that diversifying selection for market niches and environmental adaptation has led to allelic variation in cultivated tomatoes in Korea.

Most melon (Cucumis melo L.) breeding lines in South Korea display andromonoecious sex expression, which necessitates laborious hand emasculation during F1 hybrid seed production. Thus, there is a need to develop monoecious sex types in elite germplasm to obviate self-pollination. Sex expression is associated with floral ethylene production, which, in monecious melon plants, is associated with the A locus. Our study was conducted to develop molecular markers for selection of monoecious plants based on sequence variation inherent in the CmACS-7 gene [encoding 1-aminocyclopropane-1-carboxylic acid synthase (ACS)] that is associated with ethylene production. Full-length CmACS-7 sequences were cloned from a monoecious (MO23) and an andromonoecious (AM24) line. The alignment of those CmACS-7 sequences revealed a single nucleotide polymorphism (SNP; C170T) in exon 1 and an 18 bp indel in the 3′-untranslated region (UTR) of between MO23 and AM24, which was then used to develop a cleaved amplified polymorphic sequence (CAPS) (EX1-C170T) and a sequence characterized amplified region (SCAR) marker (T1ex), respectively. The sex expression and the T1ex SCAR-based genotype of 442 F2 plants derived from a MO23 × AM24 cross was determined. Monoecy and andromonoecy segregated in a 3:1 ratio in F2 progeny, where the sex type of 429 plants (13 plants not classified) co-segregated with the SCAR marker, demonstrating that sex expression regulated by CmACS-7 is controlled by a single dominant gene and that it confers monoecy in line MO23. Allelic variation in 112 geographically diverse melon lines for CmACS-7 as accessed by CAPS EX1-C170T and SCAR T1ex markers indicated that the: 1) exon 1 of CmACS-7 is highly conserved and the SNP/sex expression association detected is highly predictable making it potentially useful for marker-based selection of monoecious plants, and; 2) 18 bp indel mutation in the 3′-UTR was present in various lengths depending on different monoecious melon germplasm.

Genetic transformation using foreign genes and the subsequent development of transgenic plants has been employed to develop enhanced elite germplasm. Although some skepticism exits regarding pollen tube-mediated gene transfer (PTT), reports demonstrating improved transformation efficiency with PTT systems are increasing and encouraging and the adoption of increasingly refined pollen-mediated methodologies may lead to species-dependent improvements in breeding. Here, we highlight PTT technology as an alternative to genetic transformation.

The responses to chilling temperature of 12 Korean cucumber varieties was compared to those of two U.S.A. (previously determined cold tolerant NC76 and ‘Chipper’), and Chinese and Japanese germplasms. Seedlings of each entry were exposed to 4°C (Experiment 1) and 1°C (Experiments 2 and 3) at the first-true leaf stage for eight and nine hours, respectively, under 80% relative humidity (RH) and 149 μmoles?m -2 ?s -1 photosynthetic photon flux (PPF). The chilling response [damage rating (DR)] of each accession was based on visual ratings (1 to 5) after treatment, where 1 = no damage, 2 = slight, 3 = moderate, 4 = advanced, and 5 = severe damage. Predictably the cumulative average DR of chilling tolerant line NC76 and ‘Chipper’ after chilling w as 1 a nd 1 .1, respectively . Korean ‘Nacdongchungjang’ w as m ost sensitive to c hilling temperatures [DR = 2 .3] when compared to the other entries examined. The sensitivity to chilling of ‘Nacdongchungjang’ was followed by Chinese ‘Dongguan’ [DR = 1.7]. In contrast, ‘Saeronchungjang’ (DR = 1) and ‘Janghyungnachap’ (DR = 1) were the most chilling tolerant of the Korean accessions examined and equivalent to the response of line NC76 and ‘Chipper’. Nevertheless, chloroplast type genotyping of these accessions with known chilling-linked sdCAPS genomic markers revealed genotypic differences between chilling tolerant lines (NC76 and ‘Chipper’) and all Korean lines examined.

Compact and dwarfing vining habits in melon (Cucumis melo L.; 2n = 2x = 24) may have commercial importance since they can contribute to the promotion of concentrated fruit set and can be planted in higher plant densities than standard vining types. A study was designed to determine the genetics of dwarfism associated with a diminutive (short internodes) melon mutant line PNU-D1 (C. melo ssp. cantalupensis). PNU-D1 was crossed with inbred wild-type melon line PNU-WT1 (C. melo ssp. agrestis), and resultant F1 progeny were then self-pollinated to produce an F2 population that segregated as dwarf and vining plant types. Primary stem length of F2 progeny assessed under greenhouse conditions indicated that a single recessive gene, designated mdw1, controlled dwarfism in this population. To identify the chromosomal location associated with mdw1, an simple sequence repeat (SSR)-based genetic linkage map was constructed using 94 F2 progeny. Using 76 SSR markers positioned on 15 linkage groups spanning 462.84 cM, the location of mdw1 was localized to Chromosome 7. Using the putative dwarfing-associated genes, fine genetic mapping of the mdw1 genomic region was facilitated with 1,194 F2 progeny that defined the genetic distance between mdw1 and cytokinin oxidase gene, a candidate gene for compact growth habit (cp) in cucumber, to be 1.7 cM. The candidate gene ERECTA (serin/threonine kinase) and UBI (ubiquitin) were also mapped to genomic regions flanking mdw1 at distances of 0.6 and 1.2 cM, respectively.

Unexploded explosives that include royal demolition explosive (RDX) and trinitrotoluene (TNT) cause environmental concerns for surrounding ecosystems. Baccharis halimifolia is a plant species in the sunflower family that grows naturally near munitions sites on contaminated soils, indicating that it might have tolerance to explosives. B. halimifolia plants were grown on 100, 300, and 750 mg kg(-1) of soil amended with composition B (Comp B) explosive, a mixture of royal demolition explosive and trinitrotoluene. These concentrations are environmentally relevant to such munitions sites. The purpose of the experiment was to mimic contaminated sites to assess the plant's physiological response and uptake of explosives and to identify upregulated genes in response to explosives in order to better understand how this species copes with explosives. Stomatal conductance was not significantly reduced in any treatments. However, net photosynthesis, absorbed photons, and chlorophyll were significantly reduced in all treatments relative to the control plants. The dark-adapted parameter of photosynthesis was reduced only in the 750 mg kg(-1) Comp B treatment. Thus, we observed partial physiological tolerance to Comp B in B. halimifolia plants. We identified and cloned 11 B. halimifolia gene candidates that were orthologous to explosive-responsive genes previously identified in Arabidopsis and poplar. Nine of those genes showed more than 90% similarity to Conyza canadensis (horseweed), which is the closest relative with significant available genomics resources. The expression patterns of these genes were studied using quantitative real-time PCR. Three genes were transcriptionally upregulated in Comp B treatments, and the Cytb6f gene was found to be highly active in all the tested concentrations of Comp B. These three newly identified candidate genes of this explosives-tolerant plant species can be potentially exploited for uses in phytoremediation by overexpressing these genes in transgenic plants and, similarly, by using promoters or variants of promoters from these genes fused to reporter genes in transgenic plants for making phytosensors to report the localized presence of explosives in contaminated soils.

Chilling temperatures (

Even within closely related taxa, total length variation of PCR amplicons from chloroplast SSR must be confirmed by sequencing to avoid misinterpreting genetic relationships.

DNA markers can determine the genotype of many species. Single nucleotide polymorphism (SNP) detection is difficult without sequencing but it becomes easier with sdCAPS method. Here an experiment was performed for developing molecular markers using two SNPs, CSatpB-SNP and CSycf1-SNP, of chloroplast in cucumber plants. Properly designed primers with nucleotide sequences for restriction enzymes proved success of PCR and efficacy of digestion by the restriction enzymes. Then these markers were used to study the genotyping of cucumber breeding lines and cultivars obtained from various sources in respect of their chilling stress response. We confirmed that a U.S. cucumber line, 'NC76' known to possess a nuclear factor for the chilling tolerance showed the chloroplast genotypes related to chilling tolerance. However all Korean cucumber cultivars tested in this study showed the chloroplast genotypes related to chilling susceptibility. In conclusion, to develop chilling tolerant cucumber, both maternal and a nuclear factors related to chilling tolerance should be transferred from 'NC76' when 'NC76' is used as a female source and other elite lines as recurrent parents.

DNA marker is a powerful tool for plant genetics and breeding. In this study, 995 SSR markers were employed with chilling resistant cucumber, known as ‘NC76’, and chilling susceptible cucumber, known as ‘GY14’. Using 2% agarose gel electrophoresis, 145 SSR markers were identified as length variation markers between ‘NC76’ and ‘GY14’. The SSR markers that showed no length polymorphism were then screened using high resolution melting analysis technique and additional 30 polymorphic SSR markers were identified. As a preliminary evaluation for mapping, 20 markers among these 175 markers were employed to a F₂ population of ‘NC76’ x ‘GY14’ cross. Linkage analysis revealed 13 markers that joined into six linkage groups and seven markers that remained unlinked. This result indicates that these 175 markers could be used for construction of a genetic map using a cross between ‘NC76’ and ‘GY14’ for further investigation in developing markers related to resistance to chilling in cucumbers.

Heat-shock proteins (HSPs) and heat-shock transcription factors (HSFs) are central components of the heat-shock regulatory network and are involved in cellular responses to various forms of stresses. To examine the differences in heat shock responses (HSRs) of two comparable inbred lines of Chinese cabbage (Brassica rapa), 51 genes were selected from 130,000 Brassica rapa ESTs that belong to an HSF and six HSP families and examined their expression using RT-PCR. Two Chinese cabbage inbred lines, Chiifu and Kenshin, have different geographic origins, in that Chiifu is from temperate regions, while Kenshin is from subtropical and tropical regions. Among the 51 genes, six genes were induced, eleven were stimulated, and three were reduced in both inbred lines in response to heat shock (HS) treatment. However, eleven genes were differentially expressed between the two inbred lines. Among these genes, several appear to be involved in normal growth and chloroplast development. These data suggest that the two Chinese cabbage inbred lines have similar HSRs, but the unique HSRs allow Kenshin to develop at higher temperatures.

The low and high temperature stress might affect the yield and quality of many crop species. Alternative oxidase (AOX) gene is known as factors related to stress resistance in plants. In order to develop molecular markers related to stress resistance in Chinese cabbage, fifteen ESTs sharing sequence similarity to arabidopsis AOX genes were found using Brassica rapa EST database from NCBI. The polymorphic DNA sequences using the ESTs were then screened between Chinese cabbage, `Chiifu` and `Kenshin`. We found four ESTs that have either insertion or deletion between the two cultivars. These polymorphic sites were then targeted for development of the four PCR based molecular markers. These molecular markers developed in this study could be useful for a test of their relationship with abiotic stress resistance in Chinese cabbage.

High temperature stress might affect the yield and quality of Chinese cabbage. In order to develop cultivars resistant to high temperature stress, we developed polymorphic DNA markers for trehalose synthesis genes related to abiotic stress resistance. A total of 28 Brassica rapa ESTs homologous to trehalose synthesis genes of Arabidopsis were found from the NCBI database. The polymorphic DNA sequences were searched between Chinese cabbages - Chiifu, which is relatively susceptible to high temperature stress, and Kenshin, which is tolerant to high temperature stress. Among the 28 ESTs, we found 10 ESTs that have either insertion/deletion and/or single nucleotide polymorphism between the two cultivars. Those polymorphic sites were then targeted for the development of 10 PCR based markers. These molecular markers related to trehalose genes could be used not only to test their relationship with abiotic stress resistance in Chinese cabbage, but also the development of abiotic stress resistant cultivars using MAS.

Carrot is the most economically important member of the Apiaceae family and a major source of provitamin A carotenoids in the human diet. However, carrot molecular resources are relatively underdeveloped, hampering a number of genetic studies. Here, we report on the synthesis and characterization of a bacterial artificial chromosome (BAC) library of carrot. The library is 17.3-fold redundant and consists of 92,160 clones with an average insert size of 121 kb. To provide an overview of the composition and organization of the carrot nuclear genome we generated and analyzed 2,696 BAC-end sequences (BES) from nearly 2,000 BACs, totaling 1.74 Mb of BES. This analysis revealed that 14% of the BES consists of known repetitive elements, with transposable elements representing more than 80% of this fraction. Eleven novel carrot repetitive elements were identified, covering 8.5% of the BES. Analysis of microsatellites showed a comparably low frequency for these elements in the carrot BES. Comparisons of the translated BES with protein databases indicated that approximately 10% of the carrot genome represents coding sequences. Moreover, among eight dicot species used for comparison purposes, carrot BES had highest homology to protein-coding sequences from tomato. This deep-coverage library will aid carrot breeding and genetics.

RNA interference (RNAi) is a powerful tool for functional gene analysis, which has been successfully used to down-regulate the levels of specific target genes, enabling loss-of-function studies in living cells. Hairpin (hp) RNA expression cassettes are typically constructed on binary plasmids and delivered into plant cells by Agrobacterium-mediated genetic transformation. Realizing the importance of RNAi for basic plant research, various vectors have been developed for RNAi-mediated gene silencing, allowing the silencing of single target genes in plant cells. To further expand the collection of available tools for functional genomics in plant species, we constructed a set of modular vectors suitable for hpRNA expression under various constitutive promoters. Our system allows simple cloning of the target gene sequences into two distinct multicloning sites and its modular design provides a straightforward route for replacement of the expression cassette's regulatory elements. More importantly, our system was designed to facilitate the assembly of several hpRNA expression cassettes on a single plasmid, thereby enabling the simultaneous suppression of several target genes from a single vector. We tested the functionality of our new vector system by silencing overexpressed marker genes (green fluorescent protein, DsRed2, and nptII) in transgenic plants. Various combinations of hpRNA expression cassettes were assembled in binary plasmids; all showed strong down-regulation of the reporter genes in transgenic plants. Furthermore, assembly of all three hpRNA expression cassettes, combined with a fourth cassette for the expression of a selectable marker, resulted in down-regulation of all three different marker genes in transgenic plants. This vector system provides an important addition to the plant molecular biologist's toolbox, which will significantly facilitate the use of RNAi technology for analyses of multiple gene function in plant cells.

Plant functional proteomics research is increasingly dependent upon vectors that facilitate high-throughput gene cloning and expression of fusions to autofluorescent proteins. Here, we describe the pSITE family of plasmids, a new set of Agrobacterium binary vectors, suitable for the stable integration or transient expression of various autofluorescent protein fusions in plant cells. The pSITE vectors permit single-step Gateway-mediated recombination cloning for construction of binary vectors that can be used directly in transient expression studies or for the selection of transgenic plants on media containing kanamycin. These vectors can be used to express native proteins or fusions to monmeric red fluorescent protein or the enhanced green fluorescent protein and its cyan and yellow-shifted spectral variants. We have validated the vectors for use in transient expression assays and for the generation of transgenic plants. Additionally, we have generated markers for fluorescent highlighting of actin filaments, chromatin, endoplasmic reticulum, and nucleoli. Finally, we show that pSITE vectors can be used for targeted gene expression in virus-infected cells, which should facilitate high-throughput characterization of protein dynamics in host-virus interactions.

Cucumber (Cucumis sativus L. var. sativus; 2n = 2x = 14), has a narrow genetic base (3% to 8% polymorphism). Nevertheless, several genetic maps exist for this species. It is important to know the degree of colinearity among these maps. Thus, the positions of random amplified polymorphic DNAs, sequenced characterized amplified regions, simple sequence repeat, restriction fragment length polymorphisms, and fluorescent amplified fragment length polymorphism markers were compared in four maps. A previously unreported map was constructed in a narrow cross (processing line 2A × Gy8; C. s. var. sativus; ≈7% polymorphism) and compared with the three published maps [two narrow-based (processing type; C. s. var. sativus; 8% to 12% polymorphism) and a broad-based (C. s. var. sativus × C. s. var. hardwickii (R.) Alef. ≈12%)]. Common makers were identified in seven linkage groups, providing evidence for microsynteny. These common markers were used as anchor markers for map position comparisons of yield component quantitative trait loci. The relative order of anchor markers in each of six linkage groups (linkage groups 1, 2, and 4–7) that had two or more anchor markers within each group was colinear, and instances of microsynteny were detected. Commonalities in the position of some yield component quantitative trait loci exist in linkage groups 1 and 4 of the maps examined, and the general synteny among these maps indicates that identification and mapping of additional anchor markers would lead to successful map merging to increase cucumber map saturation for use in cucumber breeding.

Parthenocarpy (seedless fruit) is an economically important yield-related trait in cucumber (Cucumis sativus L.; 2n = 2x = 14). However, the genomic locations of factors controlling parthenocarpic fruit development in this species are not known. Therefore, an F 2: 3 mating design was utilized to map quantitative trait loci (QTL) for parthenocarpy using a narrow cross employing two gynoecious, indeterminate and normal leaf lines [2A (parthenocarpic) and Gy8 (non-parthenocarpic)]. QTL detection was performed employing 2A- and Gy8-coupling phase data using the parthenocarpic yield of 126 F 3 families grown at two locations at Hancock, WI in 2000. The QTLs detected in this study were compared with the map locations of QTLs conditioning first-harvest yield of seeded cucumber characterized in a previous study. There were 10 QTLs for parthenocarpy detected defining four genomic regions, in which three QTLs also mapped to the same genomic regions as QTLs detected for fruit yield at first-harvest as reported in a previous study. The eight fluorescence amplified fragment length polymorphism (AFLP) markers linked to parthenocarpy through QTL mapping defined herein (four each in linkage groups I and 4) are candidates for use in marker-assisted selection programmes where breeding for increased levels of parthenocarpy is an objective in the elite-processing cucumber populations.

Cultivar-to-wild population transgene flow occurs and can pose potential socio-ecological risks. In the USA, transgenic commercial squash cultivars [i.e., Cucurbita pepo subspecies ovifera var. ovifera] are cultivated in close proximity to cross-compatible wild squash taxa (i.e., vars. ozarkana and texana) that are native to the Americas. This association provides a unique opportunity for assessing cultivar-to-wild genetic introgression in this species. Thus, experiments were conducted using previously characterized random amplified polymorphic DNA (RAPD) to determine whether gene flow has occurred and whether alleles from putative introgression from cultivar-to-wild population exists in free-living populations. Free-living populations of C. pepo from the Ozark Plateau (southern Missouri, northwestern Arkansas, northeastern Oklahoma), Mississippi, Texas, Illinois, Kentucky, and Louisiana were examined using 21 RAPD primers (23 discriminatory bands). Banding morphotypes were useful for discriminat...

Agrobacterium-mediated genetic transformation is the most widely used technology for obtaining the overexpression of recombinant proteins in plants. However, complex patent issues related to the use of Agrobacterium as a tool for plant genetic engineering and the general requirement of establishing transgenic plants can create obstacles in using this technology for speedy research and development and for agricultural improvements in many plant species. Recent studies addressing these issues have shown that virus-based vectors can be efficiently used for high transient expression of foreign proteins in transfected plants and that non-Agrobacterium bacterial species can be used for the production of transgenic plants, laying the foundation for alternative tools for future plant biotechnology.

Bottle gourd /Lagenaria siceraria (Mol.) Standley] is an edible, medicinal, and otherwise utilitarian domesticated cucurbit with an ancient pantropical distribution. This African native reached Asia and the Americas 9000 years ago, probably as a wild species whose fruits had floated across the seas. Independent domestications from wild populations are believed to have occurred in both the Old and New Worlds. However, few wild populations of L. siceraria have been found during recorded history and none has been verified or studied in detail. In 1992, Mary Wilkins-Ellert discovered an unusual free-living plant of Lagenaria in a remote region of southeastern Zimbabwe. Her morphological observations during several plantings of the collected seed, as well as results from two genetic analyses (random amplified polymorphic DNA and chloroplast sequencing), indicate that the Zimbabwe collection is part of a genetically distinct and wild lineage ofL. siceraria.

Nucleotide substitutions (i.e., point mutations) are the primary driving force in generating DNA variation upon which selection can act. Substitutions called transitions, which entail exchanges between purines (A = adenine, G = guanine) or pyrimidines (C = cytosine, T = thymine), typically outnumber transversions (e.g., exchanges between a purine and a pyrimidine) in a DNA strand. With an increasing number of plant studies revealing a transversion rather than transition bias, we chose to perform a detailed substitution analysis for the plant family Cucurbitaceae using data from several short plastid DNA sequences. We generated a phylogenetic tree for 19 taxa of the tribe Benincaseae and related genera and then scored conservative substitution changes (e.g., those not exhibiting homoplasy or reversals) from the unambiguous branches of the tree. Neither the transition nor (A+T)/(G+C) biases found in previous studies were supported by our overall data. More importantly, we found a novel and symmetrical substitution bias in which Gs had been preferentially replaced by A, As by C, Cs by T, and Ts by G, resulting in the G--A--C--T--G substitution series. Understanding this pattern will lead to new hypotheses concerning plastid evolution, which in turn will affect the choices of substitution models and other tree-building algorithms for phylogenetic analyses based on nucleotide data.

To investigate genetic relationships in Benincaseae (19 accessions), Cucurbiteae (1), Joliffieae (2), Melothrieae (2), and Sicyeae (3) tribes of the family Cucurbitaceae, consensus chloroplast simple sequence repeats (ccSSR) primer pairs obtained from tobacco (Nicotiana tabacum L.) chloroplast DNA were used. Variation in the length and putative sequence substitution events of polymerase chain reaction (PCR) products were analyzed. Sequencing of four fragments (ccSSR-1, -7, -8, and -19) revealed that convergence in fragment length occurs in more distant species comparisons. In ccSSR-1 and -8, the same fragment lengths occurred as the result of different insertion and deletion events. Nevertheless, the examination of a large number of ccSSR fragments suggested that this apparent homoplasy could be overshadowed by evolutionary relationships among taxa. This hypothesis is supported by the relative degree of positive congruence of taxon groupings after cluster and principal components analyses performed on both base pair length and sequence substitution data. Moreover, these analyses support previous biochemical and morphological data indicating that distinct lineages exist within the Benincaseae. Likewise, data support the hypotheses that the genus Benincasa is descended from an ancient African ancestor and that the progenitor of the New World Sicyeae tribe shares a common ancestor with the genus Luffa of the Old World Benincaseae.Key words: Benincaseae, chloroplast, consensus, homoplasy, microsatellite, simple sequence repeats.

Chilling temperatures (≤12°C) can cause substantial economic damage to cucumber (Cucumis sativus L.) plants. Previous studies suggest chilling tolerance trait is controlled by nuclear gene(s). To investigate inheritance of chilling injury in cucumber, cucumber lines [susceptible GY14 (P1), tolerant `Chipper' (P2), and tolerant `Little John' (P3)], and their exact reciprocal F1 and F2 cross-progeny were evaluated to determine the inheritance of chilling injury at the first true-leaf stage when challenged at 4 °C for 5.5 hours. The mean chilling ratings [1(trace) to 9(dead)] of progeny comparisons were F1(P1 × P2) = 6.2 vs. F1(P2 × P1) = 1.6; F2(P1 × P2) = 6.4 vs. F2(P2 × P1) = 2.7; F1(P1 × P3) = 5.4 vs. F1(P3 × P1) = 1.7; and F2(P1 × P3) = 5.8 vs. F2(P3 × P1) = 2.2. These data suggest that chilling tolerance was maternally inherited as is the chloroplast genome in cucumber. Parents, reciprocal F1, and F2 progeny were evaluated for variation using random amplified polymorphism DNA (RAPD). Although no maternally inherited RAPD markers were detected, polymorphic and paternally inherited RAPD bands AD21249, AV8916, and AV8969 amplified by AD2 and AV8 primers were cloned and sequenced. A BLAST search of these sequences suggested that their origin is likely cucumber mitochondrial DNA. These results indicate that the mitochondria genome is not associated with the chilling tolerant trait because this genome is paternally inherited in progeny derived from this reciprocal mating. Therefore, the results of maternally inherited chilling tolerant trait and paternally transmitted mitochondria genome support that the chilling tolerant trait as identified is likely associated with the chloroplast genome which is maternally transmitted in cucumber.