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MicroRNAs (miRNAs) are important regulators of gene expression, affecting many biological processes. As yet, their roles in the response of chrysanthemum to aphid feeding have not been explored. Here, the identity and abundance of miRNAs induced by aphid infestation have been obtained using high-throughput Illumina sequencing platform. Three leaf small RNA libraries were generated, one from plants infested with the aphid Macrosiphoniella sanbourni (library A), one from plants with mock puncture treatment (library M), and the third from untreated control plants (library CK). A total of 7,944,797, 7,605,251 and 9,244,002 clean unique reads, ranging from 18 to 30 nucleotides (nt) in length, were obtained from library CK, A and M, respectively. As a result, 303 conserved miRNAs belonging to 276 miRNAs families and 234 potential novel miRNAs were detected in chrysanthemum leaf, out of which 80, 100 and 79 significantly differentially expressed miRNAs were identified in the comparison of CK-VS-A, CK-VS-M and M-VS-A, respectively. Several of the differentially abundant miRNAs (in particular miR159a, miR160a, miR393a) may be associated with the plant's response to aphid infestation.

Members of the large WRKY transcription factor family are responsible for the regulation of plant growth, development and the stress response. Here, five WRKY members were isolated from chrysanthemum. They each contained a single WRKY domain and a C2H2 zinc finger motif, so were classified into group II. Transient expression experiments demonstrated that all five were expressed in the nucleus, although CmWRKY42 was also expressed in the cytoplasm. When expressed heterologously in yeast, the products of CmWRKY22 and CmWRKY48 exhibited transactivation activity, while those of CmWRKY21, CmWRKY40 and CmWRKY42 did not. The transcription of the five CmWRKY genes was profiled when the plants were challenged with a variety of abiotic and biotic stress agents, as well as being treated with various phytohormones. CmWRKY21 proved to be markedly induced by salinity stress, and suppressed by high temperature exposure; CmWRKY22 was induced by high temperature exposure; CmWRKY40 was highly induced by salinity stress, and treatment with either abscisic acid (ABA) or methyl jasmonate (MeJA); CmWRKY42 was up-regulated by salinity stress, low temperature, ABA and MeJA treatment and aphid infestation; CmWRKY48 was induced by drought stress, ABA and MeJA treatment and aphid infestation. The function of CmWRKY48 was further investigated by over-expressing it transgenically. The constitutive expression of this transcription factor inhibited the aphids' population growth capacity, suggesting that it may represent an important component of the plant's defense machinery against aphids., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

Background: Chrysanthemum is an important ornamental plant all over the world. It is easily attacked by aphid, Macrosiphoniella sanbourni. The molecular mechanisms of plant defense responses to aphid are only partially understood. Here, we investigate the gene expression changes in response to aphid feeding in chrysanthemum leaf by RNA-Seq technology., Results: Three libraries were generated from pooled leaf tissues of Chrysanthemum morifolium 'nannongxunzhang' that were collected at different time points with (Y) or without (CK) aphid infestations and mock puncture treatment (Z), and sequenced using an Illumina HiSeqTM 2000 platform. A total of 7,363,292, 7,215,860 and 7,319,841 clean reads were obtained in library CK, Y and Z, respectively. The proportion of clean reads was >97.29% in each library. Approximately 76.35% of the clean reads were mapped to a reference gene database including all known chrysanthemum unigene sequences. 1,157, 527 and 340 differentially expressed genes (DEGs) were identified in the comparison of CK-VS-Y, CK-VS-Z and Z-VS-Y, respectively. These DEGs were involved in phytohormone signaling, cell wall biosynthesis, photosynthesis, reactive oxygen species (ROS) pathway and transcription factor regulatory networks, and so on., Conclusions: Changes in gene expression induced by aphid feeding are shown to be multifaceted. There are various forms of crosstalk between different pathways those genes belonging to, which would allow plants to fine-tune its defense responses.

Artificial aphid infestation experiments on the three chrysanthemum cultivars 'Keiun', 'Han6' and 'Jinba' showed that the three cultivars vary markedly in their resistance. Of the three cultivars, the most resistant ('Keiun') produced the longest, highest and densest trichomes, the largest and fullest gland cells, and the most wax on the lower leaf epidermis. Superoxide dismutase (EC 1.15.1.1), peroxidase (EC 1.11.1.7), ascorbate peroxidase (EC 1.11.1.11), polyphenol oxidase activity (EC 1.14.18.1) and phenylalanine ammonia lyase (EC 4.3.1.5) were enhanced by aphid herbivory. In the two more resistant cultivars ('Keiun' and 'Han6'), the activity of superoxide dismutase and ascorbate peroxidase enzymes rapidly increased following infestation, and their levels remained high from seventy-two to one hundred and sixty-eight hours after inoculation. We suggest that these two antioxidant enzymes contribute to aphid resistance of these chrysanthemum cultivars. All three cultivars showed quick responses to aphid infestation by increasing polyphenol oxidase and phenylalanine ammonia lyase activities during the early period after inoculation. Activities of these two defense enzymes were higher in the two resistant cultivars after 72h after inoculation, suggesting involvement of these two enzymes in aphid resistance., (Copyright © 2010 Elsevier GmbH. All rights reserved.)

The interspecific cross between Chrysanthemum × grandiflorum (Ramat.) Tzvel. 'rm20-12' (R, 2n = 54) and C. makinoi Matsum., and Nakai (M, 2n = 18) was achieved using embryo rescue, and a single backcross progeny using C. × grandiflorum 'rm20-12' as paternal parent was obtained. The morphology of the two independent F hybrids (MR1 and MR2) differed from that of both parents. MR1 had a larger inflorescence diameter along with narrow leaves and a reduced number of ray and tubular florets. MR2 was shorter and its inflorescence developed fewer tubular florets than either M or R. The BCF hybrid was similar to its maternal plant MR2 in terms of leaf length and width, inflorescence diameter and the number of ray florets, while it produced fewer tubular florets than either MR2 or R. The flower color in both F hybrids was lavender, while the BCF plant bore purple flowers. The aphid resistance and heat tolerance of MR1, MR2 and the BCF hybrid were both significantly superior to that of C. × grandiflorum 'rm20-12'. Interspecific hybridization followed by backcrossing shows clear potential for cultivar improvement in chrysanthemum. [ABSTRACT FROM AUTHOR]

MYB transcription factors are widely involved in the development of and physiological processes in plants. Here, we isolated the chrysanthemum R2R3-MYB family transcription factor CmMYB15, a homologous gene of AtMYB15. It was demonstrated that CmMYB15 expression was induced by aphids and that CmMYB15 could bind to AC elements, which usually exist in the promoter of lignin biosynthesis genes. Overexpression of CmMYB15 in chrysanthemum enhanced the resistance of aphids. Additionally, the content of lignin and the expression of several lignin biosynthesis genes increased. In summary, the results indicate that CmMYB15 regulates lignin biosynthesis genes that enhance the resistance of chrysanthemum to aphids. [ABSTRACT FROM AUTHOR]

We showed the effect of aphid infesting on the volatile organic compounds (VOCs) emitted by the infested Artemisia annua and Chrysanthemum morifolium cultivar ‘Nan nong hong feng’ plants by using headspace solid-phase microextraction (HS-SPME) method combined with gas chromatography mass spectrometry (GC–MS). In olfactometer bioassay experiment, aphids showed a preference for the odour of both healthy and infested chrysanthemum, while we found an opposite result in A. annua . Aphids tend to healthy plants compared with the infested, and the phenomenon became obvious with time. Different extracts were tested with the healthy plants and aphid infested plants. Eucalyptol, β-caryophyllene, (E)-β-farnesene, and germacrene D were released as the major constituents in both species. After aphid infesting, we observed a great increase in artemisia ketone and (E)-β-farnesene and a decrease in germacrene D in A. annua ; comparatively, eucalyptol, isoborneol and β-caryophyllene increased in chrysanthemum. Combined with the GC–MS data and olfactometer bioassay results we concluded that (E)-β-farnesene and artemisia ketone emitted from A. annua might act as a potential volatile compound to resist aphids, and the two compounds would be useful for future ecological control of aphid in chrysanthemum cultivation. [ABSTRACT FROM AUTHOR]