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Jing Ma, Zheng Li, Bin Wang, Shunzhao Sui and Mingyang Li

Expansins are extracellular proteins that are involved in cell wall modifications such as cell wall disassembly, cell separation, and cell expansion. Little is known about expansin gene expression during flower development of wintersweet (Chimonanthus praecox). In the present study, an expansin gene, CpEXP1, was isolated from the wintersweet flower cDNA library through random sequencing; this gene encodes a putative protein of 257 amino acids with the essential features conserved, like in other alpha expansins. The CpEXP1 gene exhibited different transcription levels in different tissues and had a significantly higher expression in flowers than other tissues. It is strongly correlated with the development of the flower. The expression of CpEXP1 increased in the flower buds or whole flowers from Stage 1 to 4 and decreased from Stage 5 to 6 during natural opening. Ethephon (an ethylene releaser) treatment promoted cut flower senescence, whereas 1-methylcyclopropene (1-MCP) (an ethylene perception inhibitor) delayed the process of flower wilting. This result is associated with the concomitant lower transcript levels of CpEXP1 in the ethephon-treated samples as well as the steady expression in the 1-MCP-treated samples compared with that in control flowers. The studies show the interesting observation that the expression of an expansin gene CpEXP1 is correlated with the development of Chimonanthus praecox flowers, the upregulation during flower opening vs. the downregulation during senescence.

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Renwei Huang, Daofeng Liu, Min Zhao, Zhineng Li, Mingyang Li and Shunzhao Sui

Lobularia maritima (L.) Desv. is an important ornamental plant. We investigated an efficient method to induce tetraploid plants of L. maritima (L.) Desv. by treating germinating seeds and apical growing points of seedlings with a range of concentrations of colchicine for different periods of time. Examination of the ploidy level by counting chromosome numbers at metaphase confirmed that the chromosome number of diploid plants was 2n = 2x = 24, whereas 2n = 4x = 48 was observed in tetraploid plants. The morphological characteristics of the diploid and colchicine-induced tetraploid plants were compared. Increases in the size of leaves, flowers, and stomata were observed in the tetraploid plants compared with the diploids. However, the stomatal density and plant height of the tetraploid plants were lower than for the diploid plants. This study presents the first report of autotetraploid plants of L. maritima (L.) Desv., and of the successful generation of tetraploid plants with improved ornamental traits by colchicine treatment.

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Rui Li, Lu Fan, Jingdong Lin, Mingyang Li, Daofeng Liu and Shunzhao Sui

Kalanchoe (Kalanchoe blossfeldiana) is a common potted flower that is popular throughout the world. Brown spot (caused by Stemphylium lycopersici) is one of the common foliage diseases in kalanchoe. This disease tends to infect leaves of kalanchoe plants in hot and humid environments, reducing their aesthetic value. The current investigation aimed to generate mutations resistant to brown spot in ‘Mary’ kalanchoe through chemical mutagenesis followed by molecular marker identification. Putative mutants were developed by treating embryogenic calluses with ethyl methanesulfonate (EMS) at median lethal doses (LD50)–either a 0.8% concentration for 2 hours or a 1.0% concentration for 0.5 hours. Brown spot crude toxin solution was used as the selection agent to identify disease-resistant calluses during tissue culture. The optimal crude concentration (60%) was determined by soaking calluses with different concentrations of crude pathogen: 0%, 20%, 40%, 60%, and 80% (v/v). A total of 32 anti-brown spot lines were regenerated and tested for disease resistance with detached leaves. Three regenerated EMS mutant lines showed no obvious brown spot lesions on their leaves after the disease resistance assay and were subjected to polymorphism identification by start codon targeted (SCoT) molecular markers. Three (SCoT40, SCoT71, and SCoT72) of 45 selected primers were chosen to identify the mutants. This work may lay the foundation for further development of new disease-resistant cultivars of kalanchoe.

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Shunzhao Sui, Jianghui Luo, Daofeng Liu, Jing Ma, Weiting Men, Lu Fan, Yu Bai and Mingyang Li

Wintersweet (Chimonanthus praecox) is a woody garden plant with fragrant flowers, which blooms in deep winter. The vase life of fresh cut flowers is 8–9 days. We applied ethylene and 1-methylcyclopropene (1-MCP; an ethylene action inhibitor) to test the role of ethylene in flower opening and senescence. In addition, abscisic acid (ABA), gibberellic acid (GA3), two cytokinins, 6-benzylaminopurine (6-BA), and zeatin (ZT) were also applied. The expression pattern of CpSRG1, a senescence-related gene, was analyzed. Ethylene treatment accelerated flower opening and senescence, decreasing vase life by 2.1 days. It also decreased flower break strength, indicating the induction of abscission. 1-MCP slowed opening, delayed senescence, and prolonged vase life by 2.6 days. Ethylene dramatically induced the expression of the CpSRG1 gene, while 1-MCP suppressed it. ZT promoted flower opening and increased vase life by 1.6 days. It suppressed the expression of CpSRG1. 6-BA, GA3, or ABA had no significant effect on flower opening and senescence of wintersweet.

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Daofeng Liu, Jing Ma, Jianfeng Yang, Tien V. Nguyen, Huamin Liu, Renwei Huang, Shunzhao Sui and Mingyang Li

Wintersweet is a woody ornamental plant and has a long history of human cultivation. Few molecular markers have been characterized and remain scant in wintersweet. This study aimed to mine simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs) from the transcriptomic database of wintersweet. A total of 3972 SSRs and 97,060 putative SNPs/indels (92,307 SNPs and 4753 indels) were identified in this data set. This study marks the highest number of SSR and SNP markers discovered to date from wintersweet by using transcriptome sequencing data. These identified markers will provide a useful source for molecular genetic studies such as genetic diversity and characterization, association mapping, and map-based gene cloning in wintersweet.

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Chandra Thammina, Mingyang He, Litang Lu, Kaishuang Cao, Hao Yu, Yongqin Chen, Liangtao Tian, Junmei Chen, Richard McAvoy, Donna Ellis, Degang Zhao, Yuejin Wang, Xian Zhang and Yi Li

Euonymus alatus (Thunb.) Sieb., commonly known as “burning bush,” is an extremely popular landscape plant in the United States as a result of its brilliant showy red leaves in fall. However, E. alatus is also seriously invasive because of its prolific seed production and effective seed dispersal by birds. Thus, development of sterile, non-invasive, seedless triploid E. alatus is in high demand. In this article, we report successful production of triploid E. alatus using endosperm tissues as explants. In our study, ≈50% of immature endosperm explants and 14% of mature endosperm explants formed compact, green calli after culture in the dark for 8 weeks and then under light for 4 weeks on Murashige and Skoog (MS) medium supplemented with 2.2 μM BA and 2.7 μM α-naphthaleneacetic acid (NAA). Approximately 5.6% of the immature endosperm-derived calli and 13.4% of mature endosperm-derived calli initiated shoots within 8 weeks after they were cultured on MS medium with 4.4 μM benzyladenine (BA) and 0.5 μM indole-3-butyric acid (IBA). Eighty-five percent of shoots rooted after culture on woody plant medium (WPM) containing 4.9 μM IBA for 2 weeks and then on hormone-free WPM medium containing 2.0 g·L−1 activated charcoal for 4 weeks. Eight independently regenerated triploid plants have been identified. Triploid plant regeneration rates observed were 0.42% from immature endosperm explants and 0.34% from mature endosperm explants, respectively, based on the number of endosperm explants cultured. Because triploid plants cannot produce viable seeds, and thus are sterile and non-invasive, some triploid E. alatus plant lines reported here can be used to replace the currently used invasive counterparts. Chemical names used: benzyladenine (BA), indole-3-butyric acid (IBA), and α-naphthaleneacetic acid (NAA).