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Thomas S.C. Li, K.E. Bedford and P.L. Sholberg

Traditionally, American ginseng (Panax quinquefolium L.) seeds are stratified for 18 to 22 months, before seeding, in a sandbox buried outdoors in late August or early September. Uncontrolled fluctuating temperature and moisture levels and the presence of pathogenic organisms in the seed box can cause seeds to sprout prematurely, rot, dry out and die. A study was initiated to shorten the lengthy stratification period, and to increase seed viability and percentage of germination by stratifying seeds indoors under a controlled environment. Seeds were subjected to various periods of warm [15 or 20 °C (59 or 68 °F)] and cold [2 °C (35.6 °F)] temperature stratification regimes in growth chambers. Embryo growth and viability, and seed moisture content were tested periodically during stratification. The best warm regime for embryo development, seed viability and germination after subsequent cold treatment was 15 °C (59 °F). The first “split” seeds, indicating incipient germination, were observed after 3 months of warm [15 °C (59 °F)] and 4 months of cold [2 °C (35.6 °F)] treatment, when average embryo length reached 6 mm (0.24 inch). Greenhouse germination of stratified seeds was as high as 80%. The results from this study indicate that good germination is possible when ginseng seeds are stratified indoors under a controlled environment and seeds can be made to germinate at any time of the year.

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Jiao Chen, De-bao Yuan, Chao-zheng Wang, Yi-xing Li, Fen-fang Li, Ke-qian Hong and Wang-jin Lu

Many reports indicate that an abundance of really interesting new gene (RING) play key roles in regulating defense responses against abiotic and biotic stresses in plants. In this study, the cloning and functional characterization of a RING gene, MaRING2, in banana (Musa acuminata) fruit are reported. MaRING2 belongs to the NEP1-interacting protein (NIP) RING-H2 finger protein family. Gene expression profiles revealed that MaRING2 was cold responsive and induced by abscisic acid (ABA) treatment during cold storage. In this study, the MaRING2 under control of the Cauliflower mosaic virus 35S (CaMV 35S) promoter was transformed to tobacco (Nicotiana benthamiana) using agrobacterium (Agrobacterium tumefaciens)-mediated transformation. The resultant MaRING2-overexpressing transgenic plants (35S:MaRING2) exhibited significantly increased tolerance to low temperatures and were hypersensitive to exogenous ABA in terms of germination and early seedling growth. In addition, overexpression of MaRING2 enhanced the expression of stress-responsive genes under normal (before cold stress) or cold conditions. These results demonstrate the biological role of MaRING2 in conferring cold tolerance. Taken together, these results suggest that MaRING2, a C3H2C3-type RING protein, is a positive regulator of the ABA-dependent stress response.

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Dilinuer Shalimu, Ke Li, Carol C. Baskin, Jerry M. Baskin and Yujun Liu

Pomegranate is an important fruit crop cultivated in many countries, and development of new cultivars depends on the plant breeders being able to produce plants from seeds. Poor quality and low yield of cultivars are widespread problems that greatly restrict development of the pomegranate industry. Our purpose was to gain a better understanding of the seed dormancy-breaking and germination requirements of four cultivars of pomegranate from Xinjiang Province, China, which would be useful in improving old cultivars and developing new ones. Fresh pomegranate seeds incubated on moist filter paper imbibed water, but they germinated to only 16% to 20%. Sulfuric acid scarification, cold stratification, and warm followed by cold stratification significantly increased germination percentages. Seeds soaked in concentrated H2SO4 for 40 minutes followed by cold stratification for 2 months germinated to 65%, and those warm stratified for 1–3 months followed by cold stratification for 2 months germinated to 75% to 80%. Seeds of pomegranate have nondeep physiological dormancy (PD).

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D. Ke, M. Mateos, J. Siriphanich, C. Li and A.A. Kader

Leaf tissues of crisphead lettuce (Lactuca sativa L.) were kept in air or in air enriched with 5% to 20% CO at OC for 2 to 9 days followed by transfer to air or to CO2 2enhanced atmospheres at 20C for 1 day to study the mode of CO2 action on metabolism of organic and amino acids. The 20% <ch> treatment caused a decrease in intracellular pH, which alone or, in combination with, other CO2 effects, inhibited succinate dehydrogenase but activated glutamate decarboxylase. This resulted in an accumulation of succinate and γ-aminobutyrate and a reduction in concentrations of malate and glutamate. Elevated CO2 atmospheres did not affect other organic and amino acids. These effects of CO2 were influenced by temperature and concentration/duration of exposure to CO2, while type of tissue (green vs. white) and cultivars of lettuce generally had no influence.

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Hailin Shen, Zhendong Liu, Ke Yan, Liren Zou, Jinghui Wen, Yinshan Guo, Kun Li and Xiuwu Guo

Amur grape (Vitis amurensis) is a dioecious species. To elucidate the time of and reason for pistil abortion in male amur grape from the perspective of cytology, we observed the sections of pistil of a male line during its development using optical and transmission electron microscopes. The abnormity in the morphology of nucellar cell and the development of various organelles appeared before the abnormity of functional megaspore mitosis. Programmed cell death (PCD) of the nucellar cells might be an important reason for mitosis disorder, leading to the abortion of pistil in male flower. However, the abortion can be eliminated by forchlorfenuron treatment, resulting in the recovery of functional pistil in male amur grape. This study provides cytological information on the gender conversion mechanism in male amur grape, which can promote gender determination studies in Vitis species.

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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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Ji Tian, Ke-ting Li, Shi-ya Zhang, Jie Zhang, Ting-ting Song, Yong-jun Zhu and Yun-cong Yao

Anthocyanins are protective pigments that accumulate in plant organs such as fruits and leaves, and are nutritionally valuable components of the human diet. The MYB10 transcription factor (TF) plays an important role in regulating anthocyanin biosynthesis in Malus crabapple leaves. However, little is known about how the promoter regulates McMYB10 expression and influences the substantial variation in leaf anthocyanin accumulation and coloration that is observed in different crabapple cultivars. In this study, we analyzed leaf coloration, anthocyanin levels, and the expression levels of McMYB10 in the leaves of 15 crabapple cultivars with three leaf colors at various development stages, and showed that the expression of McMYB10 correlates positively with anthocyanin accumulation. We also examined the relationship between the number of R6 and R1 elements in the McMYB10 promoters of the different cultivars and the pigmentation of the new buds of spring-red cultivars, as well as the methylation level of the McMYB10 promoters at different development stages in three representative crabapple cultivars. The ratio of R6/R1 minisatellites in the promoters correlated with the color and anthocyanin accumulation in new crabapple buds, and we concluded that the differences in promoter structure and methylation level of the McMYB10 promoters coordinately affect the leaf color of crabapple cultivars.