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  • Author or Editor: Chen Wang x
  • Journal of the American Society for Horticultural Science x
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The morphological characteristics of chrysanthemum (Chrysanthemum ×morifolium) are rich in variation. However, as a result of the aneuploid polyploidy of the chrysanthemum genome and the lack of proper tools, the genomic information of this crop is limited. Development of microsatellite markers has been an increasing trend in crop genetic studies because of the applicability of these markers in breeding programs. In this study, we reported the development of a simple sequence repeat in chrysanthemums using a magnetic beads enrichment method. An enriched genomic library with AC and GT microsatellite motifs was constructed, and 53 positive clones were detected by a colony polymerase chain reaction (PCR) technique. Of these clones, 35 showed high-quality sequences, and 35 primer pairs were designed accordingly. Twenty-six (74.29%) of the 35 primer pairs revealed polymorphisms on a set of 40 chrysanthemum cultivars. There were 172 alleles amplified over 26 loci with an average of 6.615 alleles per locus. The mean values of gene diversity corrected for the sample size and the inbreeding coefficient were 0.609 and 0.119 over 26 loci, respectively, which indicated that the majority of the microsatellite loci is highly informative. Cluster analysis based on 26 polymorphic loci demonstrated that the selected cultivars were clustered according to geographical origin. This study shows the isolation efficiency of the magnetic beads technique; the abundance of microsatellites in chrysanthemum; and the potential application for the cultivar classification, the studies on genetic diversity, and molecular breeding of chrysanthemums, which is beneficial to promoting the conservation and sustainable use of this crop.

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Volatile chemicals emitted from the flowers of chinese wisteria (Wisteria sinenesis) and japanese wisteria (W. floribunda) were collected using a dynamic headspace technique and identified using gas chromatography–mass spectrometry; 28 and 22 compounds were detected from chinese wisteria and japanese wisteria flowers, respectively. These chemicals can be classified into four major classes, including fatty acid derivatives, benzenoids/phenylpropanoids, terpenoids, and nitrogen-containing compounds. Two monoterpenes, (E)-β-ocimene and linalool, belonging to the class of terpenoids, were the most abundant compounds emitted from both species. Despite strong similarity, the floral volatile profiles of the two species displayed variations in both quality and quantity. Chinese wisteria was selected as a model for further study of volatile emission from different parts of flowers, emission dynamics, and regulation of floral scent production. Although floral volatiles were detected from all flower parts, petals emitted the most. The emission of floral volatiles displayed a diurnal pattern with the maximal emissions occurring during the daytime. This rhythmic pattern was determined to be light-dependent. Regulation of floral volatile emission by exogenous chemicals, including silver thiosulphate (an ethylene inhibitor), salicylic acid, and jasmonic acid, also was analyzed. Generally, jasmonic acid promoted the emission of floral volatiles. In contrast, neither silver thiosulphate nor salicylic acid showed a significant effect on floral volatile emission. The results presented in this article suggest that wisteria can serve as a useful system for exploring novel biochemistry of floral scent biosynthesis. They also build a foundation for the study of the biological/ecological significance of floral volatiles on the reproductive biology of wisteria species.

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Watermelon (Citrullus lanatus) is an economically important cucurbit (Cucurbitaceae) crop cultivated globally for its nutritional benefits. Fruit development in watermelon is characterized by fast fruit expansion attributed to unique biological processes. Myeloblastosis (MYB) family genes play important roles in plant growth and development, especially R2R3-MYB-type genes. However, the evolution of R2R3-MYB family genes in the watermelon genome and whether they participate in the regulation of watermelon fruit development remain unknown. To address these questions, duplication modes of R2R3-MYB family genes were identified and their expression profiles were investigated during watermelon fruit development. A total of 48 duplicated gene pairs were identified among the 89 R2R3-MYBs in watermelon. Segmental and transposed duplication events play major roles in the R2R3-MYB family gene expansion process. The ratio of nonsynonymous mutation and synonymous mutation analysis indicated that all the duplicated R2R3-MYBs experienced negative selection. Gene structures and cis-element compositions in promoter sequences exhibited abundant divergences between the R2R3-MYB duplicated genes. Transcriptome analyses of seed, rind, and flesh during fruit development showed that only two duplicated gene pairs had significantly similar expression patterns, whereas divergent expression profiles were found between the remaining duplicated gene pairs. Tissue-specific and development stage-specific divergent expression patterns demonstrated that neo-functionalization occurred between watermelon R2R3-MYB duplicated genes. The current study provides valuable information for further functional analyses of R2R3-MYBs in watermelon.

Open Access

Volatile chemicals emitted from the flowers of globe amaranth (Gomphrena globosa) were collected using a dynamic headspace technique and analyzed using gas chromatography–mass spectrometry. Among the four globe amaranth cultivars analyzed, Fireworks was the highest producer of floral volatiles. The flowers of the other three cultivars, Las Vegas White, Las Vegas Pink, and Las Vegas Purple, emit less volatiles, both qualitatively and quantitatively, than ‘Fireworks’. ‘Fireworks’ was chosen for detailed characterization of regulation of floral volatile emission. A diurnal pattern of emission of floral volatiles was observed from the flowers of ‘Fireworks’. In addition, the emission pattern was not significantly affected by light, suggesting that the circadian clock plays a major role in the regulation of volatile emission. The emission of floral volatiles from ‘Fireworks’ flowers that were treated with several chemicals was also analyzed. The treatment with silver thiosulphate, an ethylene inhibitor, led to enhanced emission of total volatiles. In contrast, the treatments with salicylic acid and jasmonic acid led to enhanced emission of total floral volatiles at 4 h but reduced emission at 24 h after the treatment. Biochemical pathways leading to the production of the major floral volatiles of globe amaranth are proposed and partly validated by cluster analysis of floral volatiles emitted from ‘Fireworks’ flowers under various conditions. The implications of the results of this study to the understanding of the reproductive biology of globe amaranth and the breeding of novel globe amaranth cultivars are discussed.

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The objective of this study was to investigate ascorbic acid (AsA) accumulation, mRNA expression of genes involved in AsA biosynthesis as well as recycling, activity of key enzymes, and the relationship of them to AsA levels during the development of apple fruit (Malus ×domestica cv. Gala). AsA concentration, which mainly depends on biosynthesis, was the highest in young fruit post-anthesis and then decreased steadily toward maturation. However, AsA continued to accumulate over time because of the increase in fruit mass. Transcript levels of guanosine diphosphate (GDP)-L-galactose phosphorylase, GDP-mannose pyrophosphorylase, D-galacturonate reductase, and the post-transcriptionally regulated L-galactono-1,4-lactone dehydrogenase were not correlated with AsA accumulation in apple. In contrast, patterns of expression for L-galactose dehydrogenase, L-galactose-1-phosphate phosphatase, and GDP-mannose-3′,5′-epimerase showed a pattern of change similar to that of AsA accumulation. Although activities and expression levels of monodehydroascorbate reductase and dehydroascorbate reductase in fruit, which had less capacity for AsA recycling, were much lower than in leaves, they were not clearly correlated with AsA level during fruit development.

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Lagerstroemia indica (crape myrtle) is a popular Chinese landscape plant with a long flowering period that contributes to its gorgeous flowers and high ornamental value, which motivate L. indica breeding. We found a wild acarpous individual of L. indica that did not bear seeds after flowering and had a significantly longer flowering period than fructiferous L. indica. This study identified differences in floral organ morphology, and stamen and pistil structure between fructiferous and acarpous L. indica through observation, paraffin sectioning, and scanning electron microscopy (SEM). The flowering time of each acarpous L. indica inflorescence lasts as long as 18 to 25 days. When a single flower withers, it falls from the pedicel without any fruit. The abortion in the floral organ of acarpous L. indica is characterized by sterile and undehisced anthers, pollen abortion, and deformed and irregularly arranged filament cells. Acarpous L. indica features short and loosely arranged papilla cells in the stigma, a flat style and narrow stylar canal, loosely arranged epidermal cells, and no obvious nuclei. No embryo sac cavity is found in acarpous L. indica ovules. In some nucelli, the egg apparatus structure can be observed indistinctly but without cell contour. In others, the egg apparatus structure is completely absent, and only flocculent tissue is observed. This study may provide a theoretical foundation for future studies on the molecular mechanisms of the mutations in acarpous L. indica.

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Anthocyanins are important molecules that are responsible for fruit color formation and are also beneficial to human health. To date, numerous structural and regulatory genes associated with anthocyanin biosynthesis in peach (Prunus persica) have been reported based on linkage analysis. In this study, we sought to identify further genes associated with anthocyanin content in peach by conducting a genome-wide association analysis of 129 peach accessions to detect markers associated with the trait. Significant association signals were detected when anthocyanin content was considered a qualitative character but not when it was considered a quantitative trait. We detected an association region located between 11.7 and 13.1 Mb in chromosome 1, a region in which only 133 of 146 genes have previously been functionally annotated. Gene ontology annotation of the genes in this region showed that membrane-associated genes (including one gene encoding a chloride channel protein and 17 sugar transport/carrier-associated genes) were significantly enriched, and we focused on these in subsequent analyses. Based on in vitro induction of anthocyanins in fruit flesh using different exogenously applied sugars and subsequent culture, we found that the expression level of 3 of the 18 membrane-associated genes, Prupe.1G156300, Prupe.1G156900, and Prupe.1G157000, increased during induction treatment. Furthermore, during the fruit development period of a white-fleshed and a red-fleshed peach cultivar, the expression of one gene encoding a transmembrane sugar transport protein was observed to be positively correlated with anthocyanin biosynthesis. These results will facilitate understanding of the molecular mechanism of anthocyanin biosynthesis in peach.

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Marigold (Tagetes erecta) is an important commercial plant because of its ornamental, industrial, and medicinal values. Male-sterile two-type lines are important for heterosis utilization and breeding of marigold. Mining of fertility-related genes may help to elucidate the mechanisms underlying male sterility. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) is a popular and useful tool for analyzing the expression level of a specific gene. Notably, identifying a suitable reference gene is important for data normalization because it affects the accuracy of quantitative analysis. However, at present, no reference genes are available for marigold. During the current study, 10 candidate reference genes were selected and their expression levels in different samples were analyzed by qRT-PCR. The expression level of each gene was analyzed across different developmental stages of male-sterile and male-fertile flower buds by four software programs (geNorm, NormFinder, BestKeeper, and RefFinder). The results showed that different reference genes are required for male-sterile and male-fertile samples, even if they belong to the same line. For male-sterile samples, the ribosomal protein S5/18S ribosomal RNA (RPS5/18S) gene pair was the best reference for qRT-PCR normalization, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) could be used as an alternative. For male-fertile samples, elongation factor 1-alpha (EF1α) and RPS5 were the most suitable reference genes, and Ubiquitin-conjugating enzyme (UBC) could be used as an alternative. Beta-actin (ACTB), tubulin beta (TUB), and phenylalanine ammonia-lyase (PAL) should not be used as reference genes because they were the most unstable genes in flower buds of marigold. The results of the current study may facilitate the selection of reference genes for analyzing the expression patterns of genes involved in flower development related to male sterility in marigold.

Open Access

Many reports indicate that an abundance of really interesting new gene (RING) play key roles in regulating defense responses against abiotic and biotic stresses in plants. In this study, the cloning and functional characterization of a RING gene, MaRING2, in banana (Musa acuminata) fruit are reported. MaRING2 belongs to the NEP1-interacting protein (NIP) RING-H2 finger protein family. Gene expression profiles revealed that MaRING2 was cold responsive and induced by abscisic acid (ABA) treatment during cold storage. In this study, the MaRING2 under control of the Cauliflower mosaic virus 35S (CaMV 35S) promoter was transformed to tobacco (Nicotiana benthamiana) using agrobacterium (Agrobacterium tumefaciens)-mediated transformation. The resultant MaRING2-overexpressing transgenic plants (35S:MaRING2) exhibited significantly increased tolerance to low temperatures and were hypersensitive to exogenous ABA in terms of germination and early seedling growth. In addition, overexpression of MaRING2 enhanced the expression of stress-responsive genes under normal (before cold stress) or cold conditions. These results demonstrate the biological role of MaRING2 in conferring cold tolerance. Taken together, these results suggest that MaRING2, a C3H2C3-type RING protein, is a positive regulator of the ABA-dependent stress response.

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Sacred lotus (Nelumbo nucifera) is an important aquatic ornamental plant which contains several diverse flower colors, but the underlying mechanisms of its flower coloration remain unclear. In this study, seven complementary DNA (cDNA) clones of genes involved in flavonoid biosynthesis, including chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), flavonoid 3′,5′-hydroxylase (F3′5′H), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS), were isolated and characterized. Moreover, expression patterns of these seven genes and pigment profiles were investigated across four N. nucifera cultivars with different flower colors: Zhongguohongbeijing [ZGH (red)], Xinghuafen [XHF (pink)], Molingqiuse [MLQS (yellow)], and Zhufengcuiying [ZFCY (white)]. Real-time quantitative polymerase chain reaction (qRT-PCR) analysis showed that during flower development, transcripts of early biosynthetic genes (NnCHS, NnCHI, and NnF3H) were abundant at the early stage; noticeably, highest expression of NnCHI in MLQS probably induced abundant anthoxanthin synthesis and displayed yellow. Expression of late biosynthetic genes, especially NnDFR and NnANS, was generally consistent with change patterns of anthocyanins in ZGH and XHF, but NnF3′H was barely detectable in the pink cultivars. Meanwhile, negligible expression of NnDFR and NnANS was detected in MLQS and ZFCY, respectively, which blocked their colored anthocyanin biosynthesis. Spatial expression analysis revealed that most flavonoid biosynthetic genes were highly expressed in floral tissues, rather than leaves. These results suggest that in N. nucifera cultivars with different flower colors, flavonoid biosynthesis is differentially regulated by the expression of these flavonoid biosynthetic genes, among which, NnCHI, NnF3′H, NnDFR, and NnANS are supposed to be critical for pigment accumulation, and therefore, affect different flower coloration.

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