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Huai-Fu Fan, Wen Chen, Zhou Yu and Chang-Xia Du

Salt stress reduces the fresh weight, dry weight, and relative growth rate of cucumber (Cucumis sativus) seedlings and results in serious quality loss in cucumber production. Our previous study indicated that the netting-associated peroxidase (CsaNAPOD) protein in cucumber seedling roots was induced by salt stress. Here, we amplified the coding sequence of CsaNAPOD from a cDNA isolated from the roots of cucumber seedlings. Sequence analysis indicated that the coding sequence of CsaNAPOD is 1035 bp, encoding a deduced protein of 344 amino acids, with a predicated molecular weight of 37.2 kD and theoretical isoelectric point of 5.64. The deduced amino acid sequence of CsaNAPOD showed high sequence similarity to peroxidases (PODs) from other plant species. Moreover, CsaNAPOD possesses the typical sequence structures of class III PODs and indicated that CsaNAPOD belongs to this subfamily. CsaNAPOD was highly expressed in the roots and was weakly expressed in the stems and leaves of cucumber seedlings. Salt stress significantly increased the expression of CsaNAPOD in the leaves during the entire experimental period compared with the control, and the expression of CsaNAPOD in roots was reduced at 6 hours and induced at 48 and 72 hours by salt treatment. In stems, the expression of CsaNAPOD declined at 48 and 72 hours as a result of the salt treatment compared with the control. These results indicate that the expression of CsaNAPOD responded to salt stress in cucumber seedlings, and the expression patterns under salt stress in different tissues were not identical. Our research suggests that CsaNAPOD may have potential function during the plant response to salt stress.

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Ju Ding, Kai Shi, Yan-Hong Zhou and Jing-Quan Yu

Root and foliar applications of 24-epibrassinolide (EBL), an immobile phytohormone with antistress activity, were evaluated for their effects on reducing fusarium wilt and their influence on antioxidant and phenolic metabolism in roots of cucumber plants (Cucumis sativus L. cv. Jinyan No. 4). EBL pretreatment significantly reduced disease severity together with improved plant growth and reduced losses in biomass regardless of application methods. EBL treatments significantly reduced pathogen-induced accumulation of reactive oxygen species (ROS), flavonoids, and phenolic compounds, activities of defense-related and ROS-scavenging enzymes. The enzymes included superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, catalase as well as phenylalanine ammonia-lyase and polyphenoloxidase. There was no apparent difference between two application methods used. EBL applications triggered a slight increase in H2O2 concentration followed by increases in the transcript levels of WRKY transcription factor and defense-related genes. This study demonstrated that EBL enhanced resistance to fusarium wilt by a novel mechanism that was not related to its active transport or increase in antioxidant system.

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Roger J. Sauve, Suping Zhou, Yingchun Yu and Wolfram George Schmid

A randomly amplified polymorphic DNA (RAPD) technique was used to identify and determine the phylogenetic relationships of 37 hosta accessions representing the major subgenera, sections and groups in the genus Hosta. Results of this study show that RAPD markers were able to differentiate not only the main groups, whose plants shared many genetic traits, but also cultivars within a species. Some accessions were identified by a single primer while others had high intercross linkage and required many markers for their separation. The phylogenetic clustering showed that H. plantaginea, the only night-blooming species, and H. ventricosa, the only known natural tetraploid, are unique and should be classified separately. The four species in the subgenus Bryocles, section Lamellatae H. venusta, H. minor, H. capitata, and H. nakaiana have very low genetic similarity since they do not share many amplified fragments. The other accessions were classified into four main clusters; cluster 1: H. venusta, H. tardiva, H. pycnophylla, H. tsushimensis `Ogon', H. montana, H. tibae, H. montana f. macrophylla, H. kikutii `Kikutii', H. longissima `Longifolia', H. rectifolia `Rectifolia', H. takahashii and H.`Undulata'; cluster 2: H. laevigata, H. sieboldiana, H. pycnophylla × H. longipes f. latifolia, H. longipes `Urajiro' and H. ibukiensis; cluster 3: H. capitata, H. kikutii `Polyneuron', H. nigrescens, H. kikutii `Yakusimensis', H. pachyscapa, H. kikutii `Caput-Avis', H. longipes f. latifolia, H. hypoleuca, H. okamotoi, H. densa and H. takiensis; and cluster 4: H. aequinoctiiantha, H. rupifraga, H. `Amanuma', H. minor and H. kikutii `Densa'.

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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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Xuhong Zhou, Xijun Mo, Yalian Jiang, Hao Zhang, Rongpei Yu, Lihua Wang, Jihua Wang and Suping Qu

The omission of second division gene (OSD1) gene plays a fundamental role in meiosis and is associated with 2n gamete formation in Arabidopsis thaliana. The objective of this work was to unravel the mechanisms leading to 2n pollen production, and isolate and analyze the expression patterns of OSD-like (OSDL) genes in carnation (Dianthus caryophyllus). We found an absence of the second meiotic division caused the formation of 2n pollen. Three homoeologous genes were cloned and labeled as OSDLa, OSDLb, and OSDLc in a diploid carnation. The cDNAs were 1180 bp for OSDLa, 1288 bp for OSDLb, and 971 bp for OSDLc. A strong similarity was found between the amino sequences of OSDLb and OSDLc. An evident feature of OSDLs proteins is the presence of D-box and MR-tail domains; however, the GxEN/KEN-box domain, which is distinct among the other plant proteins was absent. Quantitative real time polymerase chain reaction (qRT-PCR) analysis showed that OSDL genes maintain continuous expression in buds and other tissues. OSDLa has the highest expression in buds of 1.1–1.2 cm long (stage 2), and OSDLb has a high level of expression in buds of 0.9–1.0 cm long (stage 1) and stage 2 buds and ovary tissues in three carnation cultivars. The expression level of OSDLc was highest in ovaries. These expression patterns strongly suggest that OSDLs in carnation involve male meiosis and ovary development. These findings can have potential applications in fundamental polyploidization research and plant breeding programs in carnation.

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Yan-Chang Wang, Lei Zhang, Yu-Ping Man, Zuo-Zhou Li and Rui Qin

Big fruit size and nice red pigmentation combined with good flavor should be the major target for red-fleshed kiwifruit (Actinidia spp.) breeding programs. Genetic diversity and plant characteristics were evaluated on a set of kiwifruit accessions with predominantly red flesh to identify the superior individuals for further breeding or study of commercial application. The leading phenotypic characters varied widely among the accessions. Accession R reached average fruit weight ≈100 g, whereas it ranged from 43.15 to 84.71 g for the other accessions. Fruits of L and Q were flatter in shape than the others. The core volume accounted for fruit proportions ranging from 2.33% to 11.42%. ‘Chuhong’, ‘Honghua’, and K exhibited a round fruit apex, whereas most others showed a depressed apex. R, L, and Q had the highest a* values in the inner pericarp and also the most appealing visual coloration. Results revealed significantly higher soluble solid content (SSC), total sugar, and sugar/acid ratio in Q, R, and L. The 12 pairs of simple sequence repeat (SSR) markers were successfully used to characterize the genetic variability and confirm true-to-type identity for four accessions. However, the limited number of markers had no ability to discriminate among the other 11 accessions. Based on additional 28 SSRs, six of the indistinguishable accessions were confirmed to be genetically different, and three seemed to belong to the same clone vine. The results demonstrated that application of SSR data could improve the efficiency of identifying red-fleshed kiwifruit germplasm.

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Xiaotao Ding, Yuping Jiang, Dafeng Hui, Lizhong He, Danfeng Huang, Jizhu Yu and Qiang Zhou

Adequate greenhouse environmental management is very important for improving resource use efficiency and increasing vegetable yield. The objective of this study was to explore suitable climate and cultivation management for cucumber to achieve high yield and build optimal yield models in semi-closed greenhouses. A fruit cucumber cultivar Deltastar was grown over 4 years in greenhouse and weekly data of yields (mean, highest and lowest) and environmental variables, including total radiation, air temperature, relative humidity, and carbon dioxide (CO2) concentration were collected. Regression analyses were applied to develop the relationships and build best regression models of yields with environmental variables using the first 2 years of data. Data collected in years 3 and 4 were used for model validation. Results showed that total radiation, nutrient, temperature, CO2 concentration, and average nighttime relative humidity had significant correlations with cucumber yields. The best regression models fit the mean, lowest, and highest yields very well with R 2 values of 0.67, 0.66, and 0.64, respectively. Total radiation and air temperature had the most significant contributions to the variations of the yields. Our results of this study provide useful information for improving greenhouse climate management and yield forecast in semi-closed greenhouses.

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Yu Liu, Miao He, Fengli Dong, Yingjie Cai, Wenjie Gao, Yunwei Zhou, He Huang and Silan Dai

The NAC transcription factor is a peculiar kind of transcription factor in plants. Transcription factors are involved in the expression of plant genes under different conditions, and they play a crucial role in plant response to various biotic and abiotic stress. We transferred the ClNAC9 gene into Chrysanthemum grandiflora ‘niu9717’ by Agrobacterium tumefaciens–mediated transformation. The results of kanamycin-resistant screening, polymerase chain reaction (PCR) detection, and Northern blot analysis proved that the target gene had been integrated into the genome of the target plants. Wild-type (WT) plants and transgenic plants were treated with different concentrations of NaCl, NaHCO3, and drought stress, and physiological indexes, such as antioxidant system activity (superoxide dismutase, peroxidase, catalase), malondialdehyde accumulation, and leaf relative water content, were measured. We also observed changes in plant morphology. The physiological indexes’ changing range and extreme values suggested that transgenic plants’ resistance to salinity, alkali, and drought stress was significantly higher than WT plants. Transgenic plant growth was less inhibited compared with WT plants, indicating that the ClNAC9 gene increased the resistance of transgenic plants under the stress of salinization, alkalization, and drought.

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Yi Zhang, Xiao-Hui Hu, Yu Shi, Zhi-Rong Zou, Fei Yan, Yan-Yan Zhao, Hao Zhang and Jiu-Zhou Zhao

We studied the effects of exogenous spermidine (Spd) on plant growth and nitrogen metabolism in two cultivars of tomato (Solanum lycopersicum) that have differential sensitivity to mixed salinity-alkalinity stress: ‘Jinpeng Chaoguan’ (salt-tolerant) and ‘Zhongza No. 9’ (salt-sensitive). Seedling growth of both tomato cultivars was inhibited by salinity-alkalinity stress, but Spd treatment alleviated the growth reduction to some extent, especially in ‘Zhongza No. 9’. Exogenous Spd may help reduce stress-induced increases in free amino acids, ammonium (NH4 +) contents, and NADH-dependent glutamate dehydrogenase (NADH-GDH) activities; depress stress-induced decreases in soluble protein and nitrate content; and depress nitrate reductase, nitrite reductase, glutamine synthetase (GS), NADH-dependent glutamate synthase (NADH-GOGAT), glutamate oxaloacetate transaminase (GOT), and glutamate pyruvate transaminase (GPT) activities, especially for ‘Zhongza No. 9’. Based on our results, we suggest that exogenous Spd promotes the assimilation of excess toxic NH4 + by coordinating and strengthening the synergistic action of NADH-GDH, GS/NADH-GOGAT, and transamination pathways, all during saline-alkaline stress. Subsequently, NH4 + and its related enzymes (GDH, GS, GOGAT, GOT, and GPT), in vivo, are maintained in a proper and balanced state to enable mitigation of stress-resulted damages. These results suggest that exogenous Spd treatment can relieve nitrogen metabolic disturbances caused by salinity-alkalinity stress and eventually promote plant growth.

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Hai-Fang Yang, Hye-Ji Kim, Hou-Bin Chen, Jillur Rahman, Xing-Yu Lu and Bi-Yan Zhou

Litchi trees flower at the apex of terminal shoots. Flowering is affected by the maturity of terminal shoots before growth cessation occurs during the winter. In this study, we focused on changes of flowering in three important cultivars, Guiwei, Feizixiao, and Huaizhi, from Dec. 2012 to Mar. 2013 under natural winter conditions. Flowering rate, carbohydrate accumulation, and expression of the flowering-related genes were determined at three different developmental stages of terminal shoots with dark green, yellowish green and yellowish red leaves, respectively. The results showed that the total soluble sugar and starch contents in the dark green leaves were the highest, whereas those in the yellowish red leaves were the lowest. Trees with dark green terminal shoots had the highest flowering rates, whereas those with yellowish green or yellowish red shoots had relatively lower flowering rates. SPAD was highest in dark green leaves and lowest in yellowish red leaves at the start of the trial. The SPAD value of yellowish red leaves slightly increased but did not reach the levels of the dark green leaves, whereas levels of the other leaf stages remained fairly constant. Expression level of the litchi homolog FLOWERING LOCUS C (LcFLC), the floral inhibitor in yellowish red leaves, increased from 16 Jan., whereas that in dark green leaves declined to a level lower than the yellowish red leaves on 4 Feb. Expression level of the litchi homolog CONSTANTS (LcCO), the floral promoter in dark green leaves, was higher than that of yellowish red leaves before 26 Jan. Expression level of the litchi homolog FLOWERING LOCUS T 2 (LcFT2), encoding florigen, was higher in dark green leaves than in the other two leaf types. Our results suggest that terminal shoots should be matured and leaves should turn green for successful flowering. Mature leaves had higher expression levels of the floral promoter and florigen. In litchi production, leaves of the terminal shoots (potential flowering branches) should be dark green during floral induction and differentiation stages, and winter flushes should be removed or killed.