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Rongcai Yuan and Jianguo Li

Effects of naphthaleneacetic acid (NAA), aminoethoxyvinylglycine (AVG), and sprayable 1-methylcyclopropene (1-MCP) alone or in combination on fruit ethylene production, preharvest fruit drop, fruit quality, and fruit maturation were examined in ‘Delicious’ apples (Malus ×domestica Borkh.). 1-MCP and AVG + NAA, when applied 15 days before anticipated harvest (DBAH) for untreated control trees, more effectively delayed preharvest fruit drop than AVG or NAA used alone. However, there was no significant difference in ethylene production between fruit treated with 1-MCP or AVG + NAA and those treated by AVG. Two applications of NAA increased fruit ethylene production and fruit softening, whereas AVG inhibited NAA-enhanced fruit ethylene production and fruit softening. There was no significant difference in fruit ethylene production, fruit firmness, and fruit drop control between one and two applications of 1-MCP. The concentrations of 1-MCP did not affect the efficacy of 1-MCP when applied 15 DBAH, but high concentration of 1-MCP more effectively delayed preharvest fruit drop than low concentration of 1-MCP when applied 7 DBAH. Both AVG and 1-MCP suppressed expression of 1-aminocyclopropane-1-carboxylate (ACC) synthase gene MdACS1, ACC oxidase gene MdACO1, and polygalacturonase gene MdPG1 in fruit. Expression of ACS5A and MdACO1 but not MdACS1 in fruit abscission zones was decreased by AVG and 1-MCP. 1-MCP more effectively suppressed expression of MdPG2 in fruit abscission zones than AVG alone.

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Jianguo Li, Hong Zhu and Rongcai Yuan

The expression of genes for ethylene biosynthesis, ethylene perception, and cell wall degradation in the fruit cortex and fruit abscission zone (FAZ) was examined in relation to preharvest fruit abscission (PFA) and fruit ripening in ‘Golden Delicious’ and ‘Fuji’ apple (Malus ×domestica Borkh.). PFA, fruit ethylene production, and fruit softening increased rapidly during fruit ripening in ‘Golden Delicious’ apples, whereas no PFA, little fruit ethylene, and gradual fruit softening were recorded in ‘Fuji’ apples. The transcript levels of 1-aminocyclopropane-1-carboxylate (ACC) synthase genes, MdACS1, MdACS3, and MdACS5A, increased rapidly in the fruit cortex of ‘Golden Delicious’ apples during ripening, but not in ‘Fuji’ apples. However, only the level of MdACS5A mRNA was up-regulated in the FAZ of ‘Golden Delicious’ apples. The transcript level of ACC oxidase gene, MdACO1, increased in the fruit cortex for both cultivars but increased only in the FAZ of ‘Golden Delicious’ apples. Expression of the ethylene receptor genes, MdETR1, MdETR2, MdERS1, and MdERS2, increased in the fruit cortex for both cultivars, but only MdETR2 and MdERS2 increased in the FAZ of ‘Golden Delicious’ apples. The transcript levels of MdPG2, a polygalacturonase gene (PG), and MdEG1, a β-1,4-glucanase gene, markedly increased only in the FAZ of ‘Golden Delicious’ apples, whereas only MdPG1 rapidly increased in the fruit cortex of ‘Golden Delicious’ apples. Our results suggested that MdACS5A, MdACO1, MdPG2, and MdEG1 in the FAZ might be related to the difference in PFA between these two cultivars, whereas MdACS1 and MdPG1 were associated with fruit softening.

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Longyi Yuan, Yang Gao and Deying Li

Petroleum-based spills on turfgrass often occur during lawn care maintenance. Damage caused by diesel and hydraulic fluid is particularly difficult to correct. The objective of this study was to compare the effectiveness of combining mulching with remediation for reseeding spilled areas in lawns. Diesel and hydraulic fluid were applied to plots at a rate of 15 L·m−2. Immediately after the spill treatments, two liquid humic amendments and an activated flowable charcoal were applied at a volume rate of 8 L·m−2, respectively, with tap water/dishwashing detergent used as a control. Nitrate nitrogen was added to each remediation treatment to facilitate remediation. The spilled areas were reseeded with perennial ryegrass (Lolium perenne) and then mulched with biochar, peat pellets, and paper pellets, respectively. At 6 weeks after seeding, humic amendment 1 and activated charcoal showed better turf quality than humic amendment 2. Peat pellet mulching presented better turf quality than other mulching methods. Reseeding perennial ryegrass and mulching with peat pellets after remediation with either humic amendment 1 or activated charcoal resulted in acceptable turf quality 6 weeks after diesel and hydraulic fluid spills. Therefore, this reestablishment method is recommended as a practical way to deal with diesel or hydraulic fluid spills in cool-season turfgrasses.

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Longyi Yuan, Deying Li, Yang Gao and Wenjing Xiao

Deicing salts often are applied to sidewalks and roadways to enhance pedestrian and driving safety during freezing weather. For example, in eastern North Dakota, average annual snow days and amount are 29 days and 40 inches, respectively. This study was conducted in Fargo, ND, to investigate the population dynamics of turfgrass mixtures composed of kentucky bluegrass [KB (Poa pratensis)], creeping red fescue [RF (Festuca rubra)], and alkaligrass [ALK (Puccinellia sp.)] with the goal of optimizing turf quality by selecting seed ratios containing these species in home lawn mixtures and subject to frequent applications of deicing salts. A total of 21 mixtures were generated based on simplex-lattice design with KB, ALK, and RF contributing to 0%, 20%, 40%, 60%, 80%, and 100% of their respective full-seeding rate of 150, 150, and 300 lb/acre, respectively, after pure live seed (PLS) adjustment. The mixtures were tested at annual deicing salt rates of 0, 160, 320 lb/acre, which represent typical application. The results showed that the botanical component of the stands of grasses shifted over a 2-year period for all salt levels. Despite the good salinity tolerance of ALK reported elsewhere, it did not contribute to the improvement of turf quality in mixtures receiving deicing salts at 320 lb/acre per year. Therefore, ALK is not recommended for lawn, but mixing KB and RF in 48% and 52% of their respective full-seeding rates was recommended for areas adjacent to deicing salt applications.

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Jinhong Yuan, Man Xu, Wei Duan, Peige Fan and Shaohua Li

The responses of photosynthesis, chlorophyll fluorescence, and de-epoxidation state of the xanthophyll cycle pigments (DEPS) of micropropagated apple trees (Malus ×domestica) were investigated under whole-root water stress (WRS) and half-root water stress (HRS) induced by polyethylene glycol 6000 to simulate whole and partial root zone drying. Compared with control plants without water stress, plants under WRS and HRS exhibited reduced leaf net photosynthetic rate (Pn) and stomatal conductance (g S) with a greater reduction in WRS than in HRS plants. However, intercellular CO2 concentration (Ci) increased under WRS as water stress was prolonged, signifying a non-stomatal limitation of Pn. Regarding HRS, decreased Pn was mainly the result of a stomatal limitation explained by a relatively low Ci. Changes in photosynthesis and chlorophyll parameters indicate that severe and slight damage occurred to the photosynthetic apparatus of WRS and HRS leaves, respectively, starting at Day 3 after initiating water stress. This damage was not evident on the donor side but was expressed as a reduced capacity of the acceptor side of the photosystem II reaction centers. To prevent damage from excess light, the DEPS of WRS leaf increased. Decreased g S could explain reduced water use under an irrigation strategy of partial root zone drying in fruit trees.

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Li Xu, Suzhen Huang, Yulin Han and Haiyan Yuan

An improved three-stage protocol for plant regeneration via somatic embryogenesis of the horticulturally important plant Iris germanica L. was developed using shoot apex segments as explants. At the first stage of the experiment, 60% of callus was obtained from shoot apex segments of I. germanica on Murashige and Skoog’s (MS) medium supplemented with 4.52 μm 2,4-dichloropheoxyacetic acid (2,4-D) and 0.44 μm 6-benzyladenine (6-BA). When nonembryogenic calli were subcultured on MS medium with 11.31 μm 2,4-D and 0.44 μm 6-BA, maximum frequency of embryogenic callus (66.0%) was obtained. At the second stage, the treatment of 9% (w/v) sucrose resulted in the optimum somatic embryo (SE) formation (70.0%). More than 90.0% of SEs germinated with bipolar structure and regenerated into plantlets on plant growth regulator-(PGR)free MS medium during the third stage. Regenerated plantlets were successfully acclimatized in greenhouse environment with little somaclonal variation. Histological study showed that somatic embryogenesis stages were asynchronous and SEs developed from the surface and inner tissue of embryogenic calli.

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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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Yuanyuan Miao, Qiaosheng Guo, Zaibiao Zhu, Xiaohua Yang, Changlin Wang, Yuan Sun and Li Liu

The stolon is the main asexual reproductive organ of the medicinal plant Tulipa edulis and has special morphology. In the greenhouse experiment presented herein, the dynamic changes in carbohydrates and related enzymes, proteins, and endogenous hormones of stolons during T. edulis stolon formation were investigated. The results showed that the content of total soluble sugar, sucrose, reducing sugar, fructose, and starch were all significantly enhanced in the middle period when stolon emerged and maintained at relatively high levels until the later period of stolon formation, while protein content decreased during stolon formation. The activities of amylase (AMY), sucrose phosphate synthase (SPS), and sucrose synthase (SS) peaked in the initial period and were negatively correlated with soluble sugars. However, adenosine diphosphoglucose pyrophosphorylase (AGPase) activity increased as stolon formation progressed, and the changes in soluble starch synthase (SSS), granule-bound starch synthase (GBSS) activities presented a single peak, reaching their maximums in the middle period. AGPase, SSS, and GBSS activities were all positively related to starch content. Moreover, quantitative real-time polymerase chain reaction (qRT-PCR) verified the changes in SS and SSS activities via the expression levels of the SS, SSSI, and SSSII genes. The gibberellin (GA) and zeatin riboside (ZR) content attained their maximum in the initial period of stolon formation. Indole-3-acetic acid (IAA) and abscisic acid (ABA) remained at high levels during the initial and middle period and decreased significantly during the later period of stolon formation, inversely to the ratio of ABA:IAA. Analysis of the physiological changes in T. edulis stolon indicated that the accumulation of soluble sugars and starch via various enzymes, a high level of IAA and a low ABA to IAA ratio mainly contributed to stolon development of T. edulis. This paper explored carbohydrate levels and endogenous hormones profiles during stolon formation, which provided the theory basis for further regulating stolon growth of T. edulis.

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Wei Li, Rongcai Yuan, Jacqueline K. Burns, L.W. Timmer and Kuang-Ren Chung

Colletotrichum acutatum J. H. Simmonds infects citrus flower petals, causing brownish lesions, young fruit drop, production of persistent calyces, and leaf distortion. This suggests that hormones may be involved in symptom development. To identify the types of hormones, cDNA clones encoding proteins related to ethylene and jasmonate (JA) biosynthesis, indole-3-acetic acid (IAA) regulation, cell-wall modification, signal transduction, or fruit ripening were used to examine differential gene expressions in calamondin (Citrus madurensis Lour) and/or `Valencia' sweet orange (Citrus sinensis Osbeck) after C. acutatum infection. Northern-blot analyses revealed that the genes encoding 1-aminocyclopropane-1-carboxylate (ACC) oxidase and 12-oxophytodienoate required for ethylene and JA biosynthesis, respectively, were highly up-regulated in both citrus species. Both gene transcripts increased markedly in petals, young fruit and stigmas, but not in calyces. The transcripts of the genes encoding IAA glucose transferase and auxin-responsive GH3-like protein, but not IAA amino acid hydrolyase, also markedly increased in both species 5 days after inoculation. The expansin and chitinase genes were slightly up-regulated, whereas the senescence-induced nuclease and ß-galactosidase genes were down-regulated in calamondin. No differential expression of transcripts was detected for the genes encoding expansin, polygalacturonase, and serine-threonine kinase in sweet orange. As compared to the water controls, infection of C. acutatum increased ethylene and IAA levels by 3- and 140-fold. In contrast, abscisic acid (ABA) levels were not significantly changed. Collectively, the results indicate that infection by C. acutatum of citrus flowers triggered differential gene expressions, mainly associated with IAA, ethylene, and JA production and regulation, and increased hormone concentrations, consistent with the hypothesis of the involvement of phytohormones in postbloom fruit drop.

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Ze Li, Xiaofeng Tan, Zhiming Liu, Qing Lin, Lin Zhang, Jun Yuan, Yanling Zeng and Lingli Wu

Camellia oleifera Abel. is one of four major woody oil plants in the world. The objective of the current study was to evaluate the effect of different plant growth regulators (PGRs) and concentrations on direct organogenesis using cotyledonary nodes, hypocotyls, and radicle explants. High induction frequency of adventitious shoots were obtained from cotyledonary nodes, hypocotyls, and radicle explants (85.2%, 73.6%, and 41.0%, respectively) when cultured on half-strength Murashige and Skoog (1/2 MS) medium containing 2.0 mg·L−1 6-benzylaminopurine (BA) and 0.1 mg·L−1 indole-3-acetic acid (IAA). Microshoots from cotyledonary nodes, hypocotyls, and radicle explants were then transferred to 1/2 MS medium containing 2.0 mg·L−1 BA and 0.05 mg·L−1 indole-3-butyric acid (IBA) for shoot multiplication, resulting in 6.9 shoots per explant. The shoots were transferred to Woody Plant Medium (WPM) supplemented with various α-naphthalene acetic acid (NAA) and gibberellic acid (GA3) for shoot elongation. The mean length of shoots and the number of leaves per shoot were 3.7 and 6.6 cm, respectively, in WPM supplemented with 0.5 mg·L−1 NAA and 3.0 mg·L−1 GA3. The highest rooting of shoots (90.2%) or the number of roots per shoot (7.2) was obtained when elongated microshoots were transferred to 1/2 MS medium supplemented with 3.5% perlite, 1.0 mg·L−1 IBA and 2.0 mg·L−1 NAA. The rooted plantlets were successfully acclimatized in the greenhouse with a survival rate of 90.0%. The in vitro plant regeneration procedure described in this study is beneficial for mass propagation and improvement of C. oleifera through genetic engineering.