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Yau-Wen Yang, Pai-Yean Tai and Ying Chen

There are two evolutionary pathways in the genus of Brassica, one is rapa/oleracea lineage and the other is nigra lineage. Based on the morphological characteristics and nuclear RAPD or RFLP markers, genus Raphanus was thought more closely related to nigra lineage than to rapa/oleracea lineage (Song et al., 1990; Thormann et al., 1994). RFLP data of both chloroplast and mitochondria revealed that Raphanus is more closely related to rapa/oleracea lineage (Palmer and Herbon, 1988; Warwick and Black, 1991; Pradhan et al., 1992). We have previously demonstrated that Raphanus sativus is more closely related to nigra lineage using nuclear intergenic spacer between 5S rDNA and internal transcribed spacer region between 18S and 25S rDNA. In this study, we analyzed DNA sequences from different regions of chloroplast and showed that Raphanus sativus was closely related to rapa/oleracea lineage than to nigra lineage. These results suggest that Raphanus sativus is a hybrid between B. rapa/oleracea and B nigra lineages as proposed by Song et al (1990). The split time between these two lineages and the divergent time of Raphanus was also determined based on these chloroplast DNA sequences.

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Ting-Ting Li, Zhi-Rong Li, Kang-Di Hu, Lan-Ying Hu, Xiao-Yan Chen, Yan-Hong Li, Ying Yang, Feng Yang and Hua Zhang

Kiwifruit (Actinidia deliciosa) is a typical climacteric fruit, and its ripening is closely associated with ethylene. In this study, we present evidence that H2S alleviated ethylene-induced ripening and senescence of kiwifruit. Kiwifruit were fumigated with ethylene released from 0.4 g·L−1 ethephon solution or H2S with 1 mm sodium hydrosulfide (NaHS) as the donor or in combination. Fumigation with ethylene was found to accelerate kiwifruit ripening and H2S treatment effectively alleviated ethylene-induced fruit softening in parallel with attenuated activity of polygalacturonase (PG) and amylase. Ethylene + H2S treatment also maintained higher levels of ascorbic acid, titratable acid, starch, soluble protein, and reducing sugar compared with ethylene group, whereas suppressed the increase in chlorophyll and carotenoid. Kiwifruit ripening and senescence under ethylene treatment was accompanied by elevation in reactive oxygen species (ROS) levels, including H2O2 and superoxide anion and malondialdehyde (MDA), but combined treatment of ethylene plus H2S alleviated oxidative stress in fruit. Furthermore, the activities of antioxidative enzymes catalase (CAT) and ascorbate peroxidase (APX) were increased by ethylene + H2S treatment in comparison with ethylene alone, whereas the activities of lipoxygenase (LOX) and polyphenol oxidase (PPO) were attenuated by H2S treatment. Further investigations showed that H2S repressed the expression of ethylene synthesis-related genes AdSAM, AdACS1, AdACS2, AdACO2, and AdACO3 and cysteine protease genes, such as AdCP1 and AdCP3. Taken together, our findings suggest that H2S alleviates kiwifruit ripening and senescence by antagonizing the effect of ethylene through reduction of oxidative stress and inhibition of ethylene synthesis pathway.

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Wen-ji Xu, Feng-yang Yu, Qing-xiang Jia, Gang-jun Luo and Xiao-ying Bi

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Jie Fu, Qiaoyan Xiang, Xianbao Zeng, Mei Yang, Ying Wang and Yanling Liu

To assess the genetic diversity among lotus (Nelumbo) accessions and evaluate the correlation between genetic variation and morphological classification, we sampled 138 accessions: two of N. lutea, 112 of N. nucifera, 17 of hybrids between N. nucifera and N. lutea, and seven Japanese cultivars. The 11 selected combinations of amplified fragment length polymorphism (AFLP) primers produced 138 polymorphic loci, and the percentage of polymorphism was 28.7%. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram clustered all the accessions into two groups: Group I comprised N. lutea and its hybrids with N. nucifera; Group II included N. nucifera and its hybrids with N. lutea and Japanese cultivars. Population structure analysis identified four main clusters: N. lutea clustered mainly in C1, whereas N. nucifera clustered in C2, C3, and C4, which was consistent with the UPGMA and principal coordinate analysis results. The Japanese cultivars were related more closely to N. nucifera (genetic similarity coefficient = 0.74) than to N. lutea (0.46); hence, the Japanese cultivars can be classified as N. nucifera. Moreover, rhizome lotuses formed a separate subclade, whereas seed lotuses were interspersed among flower lotuses, which demonstrated that rhizome lotuses were distinct from flower and seed lotuses. Plant size, flower color, and other morphological criteria used commonly to classify lotuses were correlated with genetic variation to a certain extent but not sufficiently for accurate classification. It appears that it is necessary to use both DNA markers and morphological characteristics to classify lotus species and cultivars.

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Ji-Lian Zheng, Lan-Ying Hu, Kang-Di Hu, Jun Wu, Feng Yang and Hua Zhang

Hydrogen sulfide (H2S) has been identified as a multifunctional signaling molecule in plants. Here, we show that H2S delayed postharvest senescence of fresh-cut apples (Malus ×pumila) in a dose-dependent manner. Exogenous H2S application maintained significantly higher levels of ascorbic acid, flavonoids, total phenolics, reducing sugars and soluble proteins, and lower levels of free amino acids in apple slices compared with controls. Further investigations showed that H2S significantly reduced the accumulation of superoxide radicals, hydrogen peroxide (H2O2) and malondialdehyde (MDA). Apple fruits fumigated with H2S contained significantly higher activities of ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol peroxidase (POD) and superoxide dismutase (SOD), and lower activities of lipoxygenase (LOX), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and protease relative to controls. H2S also upregulated MdDHAR expression and downregulated the expression of MdLOX2, MdPG1, MdPPO, MdACO1, MdERS1, and MdETR1 in postharvest apple tissue. The present study indicates that H2S was involved in delaying postharvest senescence of apples by acting as an antioxidant and by regulating senescence-related gene expression.

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Yu Bai, Ying Zhou, Xiaoqing Tang, Yu Wang, Fangquan Wang and Jie Yang

The appropriate timing of bolting and flowering is one of the keys to the reproductive success of Isatis indigotica. Several flowering regulatory pathways have been reported in plant species, but we know little about flowering regulatory in I. indigotica. In the present study, we performed RNA-seq and annotated I. indigotica transcriptome using RNA from five tissues (leaves, roots, flowers, fruit, and stems). Illumina sequencing generated 149,907,857 high-quality clean reads and 124,508 unigenes were assembled from the sequenced reads. Of these unigenes, 88,064 were functionally annotated by BLAST searches against the public protein databases. Functional classification and annotation assigned 55,991 and 23,072 unigenes to 52 gene ontology (GO) terms and 25 clusters of orthologous group (COG) categories, respectively. A total of 19,927 unigenes were assigned to 124 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and 80 candidate genes related to plant circadian rhythm were identified. We also identified a number of differentially expressed genes (DEG) and 91 potential bolting and flowering-related genes from the RNA-seq data. This study is the first to identify bolting and flowering-related genes based on transcriptome sequencing and assembly in I. indigotica. The results provide foundations for the exploration of flowering pathways in I. indigotica and investigations of the molecular mechanisms of bolting and flowering in Brassicaceae plants.

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Ying Qu, Xue Bai, Yajun Zhu, Rui Qi, Geng Tian, Yang Wang, Yonghua Li and Kaiming Zhang

Leaves of Begonia semperflorens accumulate anthocyanins and turn red under low temperature (LT). In the present work, LT increased H2O2 content and superoxide anions production rate, causing significant increases in the activities of enzymes and contents of reduced components involved in the ascorbate-glutathione cycle (AsA-GSH cycle). As a result, LT-exposed seedlings increased the expression of genes involved in anthocyanin biosynthesis, and accumulated anthocyanin. Based on LT condition, application of N,N'-dimethylthiourea (DMTU) decreased reactive oxygen species (ROS) content, and unbalanced the AsA-GSH-controlled redox homeostasis. As a result, seedlings in the LT + DMTU group did not accumulate anthocyanin. Our results suggest that ROS may act as an important inducer in LT-induced anthocyanin biosynthesis.

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Wei Hu, Ju-Hua Liu, Xiao-Ying Yang, Jian-Bin Zhang, Cai-Hong Jia, Mei-Ying Li, Bi-Yu Xu and Zhi-Qiang Jin

The banana, a typical climacteric fruit, undergoes a postharvest ripening process followed by a burst in ethylene production that signals the beginning of the climacteric period. Postharvest ripening plays an important role in improving the quality of the fruit as well as limiting its shelf life. To investigate the role of glutamate decarboxylase (GAD) in climacteric ethylene biosynthesis and fruit ripening in postharvest banana, a GAD gene was isolated from banana, designated MuGAD. Coincidently with climacteric ethylene production, MuGAD expression as well as the expression of the genes encoding the Musa 1-aminocyclopropane-1-carboxylate synthase (MaACS1) and Musa 1-aminocyclopropane-1-carboxylate oxidase (MaACO1) greatly increased during natural ripening and in ethylene-treated banana. Moreover, ethylene biosynthesis, ripening progress, and MuGAD, MaACS1, and MaACO1 expression were enhanced by exogenous ethylene application and inhibited by 1-methylcyclopropene (1-MCP). Taken together, our results suggested that MuGAD is involved in the fruit ripening process in postharvest banana.

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Chen Chen, Meng-Ke Zhang, Kang-Di Hu, Ke-Ke Sun, Yan-Hong Li, Lan-Ying Hu, Xiao-Yan Chen, Ying Yang, Feng Yang, Jun Tang, He-Ping Liu and Hua Zhang

Aspergillus niger is a common pathogenic fungus causing postharvest rot of fruit and vegetable, whereas the knowledge on virulence factors is very limited. Superoxide dismutase [SOD (EC 1.15.1.10)] is an important metal enzyme in fungal defense against oxidative damage. Thus, we try to study whether Cu/Zn-SOD is a virulence factor in A. niger. Cu/Zn-SOD encoding gene sodC was deleted in A. niger [MA70.15 (wild type)] by homologous recombination. The deletion of sodC led to decreased SOD activity in A. niger, suggesting that sodC did contribute to full enzyme activity. ΔsodC strain showed normal mycelia growth and sporulation compared with wild type. However, sodC deletion markedly increased the cell’s sensitivity to intracellular superoxide anion generator menadione. Besides, spore germination under menadione and H2O2 stresses were significantly retarded in ΔsodC mutant compared with wild type. Further results showed that sodC deletion induced higher superoxide anion production and higher content of H2O2 and malondialdehyde (MDA) compared with wild type, supporting the role of SOD in metabolism of reactive oxygen species (ROS). Furthermore, ΔsodC mutant had a reduced virulence on chinese white pear (Pyrus bretschneideri) as lesion development by ΔsodC was significantly less than wild type. The determination of superoxide anion, H2O2, and MDA in A. niger-infected pear showed that chinese white pear infected with ΔsodC accumulated less superoxide anion, H2O2, and MDA compared with that of wild type A. niger, implying that ΔsodC induced an attenuated response in chinese white pear during fruit–pathogen interaction. Our results indicate that sodC gene contributes to the full virulence of A. niger during infection on fruit. Aspergillus niger is one of the most common species found in fungal communities. It is an important fermentation industrial strain and is also known to cause the most severe symptoms in fruit during long-term storage (Pel et al., 2007). Meanwhile, plants activate their signaling pathways to trigger defense responses to limit pathogen expansion. One of the earliest host responses after pathogen attack is oxidative burst, during which large quantities of ROS are generated by different host enzyme systems, such as glucose oxidase (Govrin and Levine, 2000). ROS such as singlet oxygen, superoxide anion, hydroxyl (OH), and H2O2 are released to hinder the advance of pathogens (Gara et al., 2003). ROS can react with and damage cellular molecules, such as DNA, protein, and lipids, which will limit fungal propagation in the host plant (Apel and Hirt, 2004).

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Ying Chen, Xinlu Chen, Fei Hu, Hua Yang, Li Yue, Robert N. Trigiano and Zong-Ming (Max) Cheng

Agave species are economically important plants in tropical and subtropical desert ecosystems as ornamentals as well as potential bioenergy crops. However, their relatively long life cycles and the current lack of biotechnology tools hinder their breeding. In this study, an efficient system for micropropagation was developed for Agave americana L. by using basal stems as explants and grown on a modified Murashige and Skoog medium (MSI) or a 1/2 MSI medium supplemented with various concentrations of 6-benzylaminopurine (BA) for shoot proliferation. The highest number of shoots (18.5 shoots/explant) from basal stems was obtained on MSI supplemented with 13.32 μM BA. An efficient shoot regeneration system was also developed from leaf tissues. Combinations of auxin with cytokinin, basal media, and leaf regions were optimized for shoot induction. Adventitious shoot formation from leaf segments was induced and proliferated with combination ranging of 0.54 to 2.68 μM [α-naphthaleneacetic acid (NAA)] with 8.88 to 13.32 μM (BA), and the maximum frequency (≈69%) was obtained with 2.68 μM NAA plus 13.32 μM BA. MSI medium and the basal segment of leaf affected shoot induction. The highest rooting frequency and mean number of shoots occurred in 1/2 MSI containing with 4.92 μM indole-3-butyric acid (IBA) alone (90%, 3.4) or 1.48 μM IBA plus 1.61 μM NAA (92%, 5.2). Survival of in vitro plantlets after transfer and acclimatization to ex vitro conditions was 87%. This is the first complete protocol for micropropagation of A. americana.