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Jia-yi Wang, Jian-shuang Shen, Mengmeng Gu, Jia Wang, Tang-ren Cheng, Hui-tang Pan and Qi-xiang Zhang

Yellow-leafed cultivars usually do not grow as vigorous as their green-leafed counterparts, which affect their use in landscapes. To breed Forsythia cultivars with both yellow leaves and vigorous growth, crosses between F. ‘Courtaneur’ (♀) and Forsythia koreana ‘Suwon Gold’ (♂) were conducted, and 52 F1 hybrid progenies with different leaf colors (green, chartreuse, and yellow) were obtained. The progenies were categorized into three groups [Yellow Group (YG), Chartreuse Group (CG), and Green Group (GG)] based on leaf colors. The growth index (GI) and the number of branches and leaves of YG progenies were significantly lower at 2%, 35%, and 34% of GG progenies. As the leaves changed from green to chartreuse and to yellow, chlorophyll content, leaf thickness, and chlorophyll fluorescence parameters decreased and the chloroplast structures were disintegrated gradually, which influenced the leaf photosynthetic activity and led to weak growth. Compared with yellow-leafed progenies, the leaf chlorophyll content and leaf thickness of chartreuse-leafed progenies were significantly higher at 71% and 9%. The chloroplast structure of stroma lamella of chartreuse-leafed progenies was relatively intact. Carboxylation efficiency (CE), photochemical efficiency of PS II (F v/F m), and the number of branches and leaves of GG progenies were significantly higher than YG progenies; however, they have no significant difference with CG progenies. The results were promising for breeding new forsythia cultivars from moderate growth and chartreuse leaves.

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Sheng Xu, Mingmin Jiang, Jiangyan Fu, Lijian Liang, Bing Xia and Ren Wang

From a field experiment, the changes in morphophysiological characters and antioxidant enzyme activities were studied in two Lycoris species (Lycoris radiata and Lycoris aurea) subjected to 16 days of water deficit stress. With the increase of water deficit stress processing time, leaf relative water content (RWC), membrane stability index (MSI), net photosynthesis (Pn), stomatal conductance (g S), transpiration rate (E), and chlorophyll (Chl) content decreased in both studied species. The water use efficiency (WUE) showed an opposite tendency between the two species under water deficit stress, where WUE of L. aurea decreased moderately and WUE of L. aurea increased somehow. Intercellular CO2 concentration (C i) in L. aurea and L. radiata decreased in respond to water deficit stress at early stages of stress treatment, then increased throughout the rest of the stress period, and reached levels higher than those in well-watered plants at the end of the treatment. In addition, there was a significant increment in soluble sugar content and proline accumulation under water deficit stress in both species, and L. radiata showed a much more accumulation. The activity of superoxide dismutase (SOD), guaiacol peroxidase (POD), and ascorbate peroxidase (APX) increased in both plants subjected to water deficit stress while declined as the stress time increased. In L. aurea, catalase (CAT) showed a sustained increment, but it responded later and after a transient increase declined again in L. radiata under water deficit stress. In conclusion, L. radiata was more tolerant to water deficit stress than L. aurea as evidenced by its relatively higher water status, higher levels of proline, soluble sugar and pigments, and stronger photoprotection. Moreover, relatively higher antioxidant enzyme activities and lower levels of thiobarbituric acid reactive substances (TBARS) in L. radiata were also associated with its better protection against water deficit stress-induced oxidative damage.

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Liwang Liu, Guang Liu, Yiqin Gong, Wenhao Dai, Yan Wang, Fanmin Yu and Yunying Ren

Four molecular marker systems—RAPD (random amplified polymorphic DNA), ISSR (intersimple sequence repeat), SRAP (sequence-related amplified polymorphism), and SSR (simple sequence repeat)—were used to evaluate seed genetic purity of a hybrid cabbage cultivar ‘Zaoxia 16’. Genetic relationships of the F1 hybrids and their parents were analyzed with 157 RAPD primers, 54 ISSR primers, 84 SRAP primer combinations, and 44 SSR primers. Three RAPD primers (NAURP2006, NAURP2020, and NAURP2031), two ISSR primers (NAUISR1058 and NAUISR1062), one SRAP primer combination (NAUSR04/NAURS05), and two SSR primers (NAUSSR1011 and NAUSSR1031), which produced male and female parent-specific markers simultaneously, were selected for testing the genetic purity of the F1 seeds. A total of 210 ‘Zaoxia 16’ hybrid individuals were investigated with these eight selected primers. Of these, 12 appeared to be false hybrids. Nine of the 12 putative false hybrids, confirmed with all eight primers, exhibited similar banding patterns to the female parent, suggesting that they could be derived from selfing of the female parent. The results were in accordance with those from field evaluations. This study showed that RAPD, ISSR, SRAP, and SSR markers are highly efficient and reproducible for genetic purity testing of cabbage commercial hybrid seeds.

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Ren-Huang Wang, Yu-Mei Hsu, Duane P. Bartholomew, Subbiyan Maruthasalam and Chin-Ho Lin

In Taiwan, the major yield constraint in pineapple cultivation is natural flowering, which occurs when daylengths are shorter and nights are cooler. This natural (precocious) flowering increases the cost of cultivation and reduces the percentage of fruits of marketable size. Two field experiments were conducted to evaluate the inhibitory potential of aviglycine [(S)-trans-2-amino-4-(2 aminoethoxy)-3-butenoic acid hydrochloride, AVG] on natural flowering of ‘Tainon 17’ pineapple plants during the 2003 to 2004 and 2004 to 2005 cropping seasons. In the 2003 to 2004 season, bolting in the control exceeded 80% on 2 Mar. 2004, whereas no bolting was observed in the treatments. Inhibition of bolting by aviglycine (AVG) was dependent on the concentration and frequency of application. Bolting was less than 40% when plants were treated in Nov. and Dec. 2003 with 500 mg·L−1 of AVG four times at 15-day intervals or with five applications made at 10-day intervals. For the 2004 to 2005 season, bolting of plants treated with 250 or 375 mg·L−1 AVG was delayed 4 weeks relative to the control, whereas bolting was delayed 7 weeks by four or five applications of 500 mg·L−1 of AVG applied at 10- or 15-day intervals. Both experiments showed that four to five applications of 500 mg·L−1 of AVG at 10- or 15-day intervals delayed inflorescence emergence relative to the control for the duration of the treatments. We assume control was maintained for 1 to 2 weeks after treatments stopped. Based on these results, the date AVG treatments stop can be used to estimate the duration of delay in flowering. AVG inhibits ethylene biosynthesis and has the potential to be effectively used to delay or completely control the problem of precocious flowering and associated crop losses in pineapple.

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Shiow Wang, Ren-tian Feng, Linda Bowman, Ross Penhallegon and Min Ding

The effects of lingonberry (Vaccinium vitis-idaea L.) extracts on activator protein-1 (AP-1), nuclear factor-kappaB (NF-κB), and mitogen-activated protein kinases (MAPKs) were evaluated. Pretreatment of JB6 P+ mouse epidermal cells with lingonberry extracts produced a dose-dependent inhibition of AP-1 and NF-κB induced by either 12-O-tetradecanoylphorbol-13-acetate (TPA) or ultraviolet-B (UVB) light. Lingonberry extracts blocked UVB-induced phosphorylation of MAPK family members ERK1, ERK2, and p38, but not JNK. Lingonberry extracts also prevented TPA-induced phosphorylation of ERK1 and ERK2. Results of soft agar assays indicated that lingonberry extracts suppressed TPA-induced neoplastic transformation of JB6 P+

cells in a dose-dependent manner. Lingonberry extracts also induced the apoptosis of human leukemia HL-60 cells in a dose-independent manner. These results suggest that ERK1 and ERK2 may be inhibited by lingonberries, which results in suppression of AP-1 and neoplastic transformation in JB6 P+ cells and causes cancer cell death by an apoptotic mechanism in human leukemia HL-60 cells.

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Ji-Yu Zhang, Zhong-Ren Guo, Rui Zhang, Yong-Rong Li, Lin Cao, You-Wang Liang and Li-Bin Huang

This study examined the ability to vegetatively propagate 1-year-old pecan (Carya illinoinensis) through the rooting of hardwood cuttings. Cuttings were treated with varying concentrations of different auxins and different combinations of media and ambient temperatures. Under different temperature conditions, all auxin treatments induced the rooting of cuttings but did not promote sprouting. The effectiveness of the induction of adventitious roots was as follows: 1-naphthalene acetic acid (NAA) > indole 3-butyric acid > indole 3-acetic acid. The base of the parent shoot treated by NAA at a concentration of 0.09%, planted in substrate with bottom heat was the most effective, with 82% rooting, 8.3 roots/cutting and root lengths of 7.3 cm. These findings suggested that auxin and substrate/air temperature differences are both indispensable in the process of adventitious roots formation in pecan. This study revealed that the propagation of hardwood cuttings derived from branches of 1-year-old pecan is possible.

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Ni Jia, Qing-Yan Shu, Dan-Hua Wang, Liang-Sheng Wang, Zheng-An Liu, Hong-Xu Ren, Yan-Jun Xu, Dai-Ke Tian and Kenneth Michael Tilt

Petal anthocyanins were systematically identified and characterized by high-performance liquid chromatography (HPLC)–electrospray ionization–mass spectrometry (MS) coupled with diode array detection among nine wild herbaceous peony (Paeonia L.) species (15 accessions). Individual anthocyanins were identified according to the HPLC retention time, elution order, MS fragmentation patterns, and by comparison with authentic standards and published data. Six main anthocyanins, including peonidin-3,5-di-O-glucoside, peonidin-3-O-glucoside-5-O-arabinoside (Pn3G5Ara), peonidin-3-O-glucoside, pelargonidin-3,5-di-O-glucoside, cyanidin-3,5-di-O-glucoside, and cyanidin-3-O-glucoside (Cy3G), were detected. In addition to the well-known major anthocyanins, some minor anthocyanins were identified in herbaceous peony species for the first time. Detection of the unique anthocyanins cyanidin-3-O-glucoside-5-O-galactoside and pelargonidin-3-O-glucoside-5-O-galactoside in both Paeonia anomala L. and P. anomala ssp. veitchii (Lynch) D.Y. Hong & K.Y. Pan indicated these two species should belong to the same taxon. Pn3G5Ara was found only in European wild species and subspecies suggesting different metabolic pathways between European and Chinese accessions. Anthocyanins conjugated with galactose and arabinose were observed in the genus Paeonia for the first time. The North American species, Paeonia tenuifolia L., had high Cy3G content in flower petals. This anthocyanin composition is distinct from the anthocyanin composition in Asian and European species and possibly is responsible for the vivid red coloration in flowers.

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Zhuang-Zhuang Liu, Tao Chen, Fang-Ren Peng, You-Wang Liang, Peng-Peng Tan, Zheng-Hai Mo, Fan Cao, Yang-Juan Shang, Rui Zhang and Yong-Rong Li

Cytosine methylation plays important roles in regulating gene expression and modulating agronomic traits. In this study, the fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) technique was used to study variation in cytosine methylation among seven pecan (Carya illinoinensis) cultivars at four developmental stages. In addition, phenotypic variations in the leaves of these seven cultivars were investigated. Using eight primer sets, 22,796 bands and 950 sites were detected in the pecan cultivars at four stages. Variation in cytosine methylation was observed among the pecan cultivars, with total methylation levels ranging from 51.18% to 56.58% and polymorphism rates of 82.29%, 81.73%, 78.64%, and 79.09% being recorded at the four stages. Sufficiently accompanying the polymorphism data, significant differences in phenotypic traits were also observed among the pecan cultivars, suggesting that cytosine methylation may be an important factor underlying phenotypic variation. Hypermethylation was the dominant type of methylation among the four types observed, and full methylation occurred at higher levels than did hemimethylation in the pecan genomes. Cluster analysis and principal coordinate analysis (PCoA) identified Dice coefficients ranging from 0.698 to 0.778, with an average coefficient of 0.735, and the variance contribution rates of the previous three principal coordinates were 19.6%, 19.0%, and 18.2%, respectively. Among the seven pecan cultivars, four groups were clearly classified based on a Dice coefficient of 0.75 and the previous three principal coordinates. Tracing dynamic changes in methylation status across stages revealed that methylation patterns changed at a larger proportion of CCGG sites from the 30% of final fruit-size (30%-FFS) stage to the 70%-FFS stage, with general decreases in the total methylation level, the rate of polymorphism, and specific sites being observed in each cultivar. These results demonstrated that the F-MSAP technique is a powerful tool for quantitatively detecting cytosine methylation in pecan genomes and provide a new perspective for studying many important life processes in pecan.

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Ren-jun Feng, Li-li Zhang, Jing-yi Wang, Jin-mei Luo, Ming Peng, Jun-feng Qi, Yin-don Zhang and Li-fang Lu

Cold stress is one of the most important environmental factors affecting crop growth and agricultural production. Induced changes of gene expression and metabolism are critical for plants responding and acclimating to cold stress. Banana (Musa sp.) is one of the most important food crops in the tropical and subtropical countries of the world. Banana, which originated from tropical regions, is sensitive to cold, which can result in serious losses in commercial banana production. To investigate the response of the banana to cold stress conditions, changes in protein expression were analyzed using a comparative proteomics approach. ‘Brazil’ banana (Musa acuminata AAA group) is a common banana cultivar in southern China. ‘Brazil’ banana plantlets were exposed to 5 °C for 24 hours and then total crude protein was extracted from treatment and control leaves by phenol extraction, separated with two-dimensional gel electrophoresis, and subsequently identified by mass spectrometry (MS). Out of the more than 400 protein spots reproducibly detected, only 41 protein spots exhibited a change in intensity by at least 2-fold, with 26 proteins increasing and 15 proteins decreasing expression. Of these, 28 differentially expressed proteins were identified by MS. The identified proteins, including well-known and novel cold-responsive proteins, are involved in several cellular processes, including antioxidation and antipathogen, photosynthesis, chaperones, protein synthesis, signal transduction, energy metabolism, and other cellular functions. Proteins related to antioxidation, pathogen resistance, molecular chaperones, and energy metabolism were up-regulated, and proteins related to ethylene synthesis, protein synthesis, and epigenetic modification were down-regulated in response to cold temperature treatment. The banana plantlets incubated at cold temperatures demonstrated major changes in increased reactive oxygen species (ROS) scavenging, defense against diseases, and energy supply. Increased antioxidation capability in banana was also discovered in plantain, which has greater cold tolerance than banana in response to cold stress conditions. Therefore, we hypothesized that an increased antioxidation ability could be a common characteristic of banana and plantain in response to cold stress conditions. These findings may provide a better understanding of the physiological processes of banana in response to cold stress conditions.

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Rui Zhang, Fang-Ren Peng, Dong-Liang Le, Zhuang-Zhuang Liu, Hai-Yang He, You-Wang Liang, Peng-Peng Tan, Ming-Zhuo Hao and Yong-Rong Li

Scion wood of ‘Caddo’ and ‘Desirable’ pecan (Carya illinoinensis) was grafted onto the epicotyl of 1-month-old, open-pollinated ‘Shaoxing’ pecan seedlings for evaluation as a grafting technique to reduce the time to produce grafted trees. The results showed that seedlings grafted with “base scions” had higher survival than those grafted with “terminal scions” for both ‘Caddo’ and ‘Desirable’. Also, grafting with paraffinic tape could achieve greater success rate than that with medical tape. The most ideal time to perform this grafting was late April in Nanjing, China, when pecan seedlings were about 35 days old. This study demonstrated that the technique yielded successful epicotyl grafting of >70%, and it could thus be applied in practice.