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  • Author or Editor: Cheng Li x
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To determine the cause of zonal chlorosis of `Honeycrisp' apple leaves, we compared CO2 assimilation, carbohydrate metabolism, xanthophyll cycle and the antioxidant system between chlorotic leaves and normal leaves. Chlorotic leaves accumulated higher levels of non-structural carbohydrates, particularly starch, sorbitol, sucrose, and fructose at both dusk and predawn, and no difference was found in total non-structural carbohydrates between predawn and dusk. CO2 assimilation and the key enzymes in the Calvin cycle, ribulose 1,5-bisphosphate carboxylase/oxygenase, NADP-glyceraldehyde-3-phosphate dehydrogenase, phosphoribulokinase, stromal fructose-1,6-bisphosphatase, and enzymes in starch and sorbitol synthesis, ADP-glucose pyrophosphorylase, cytosolic fructose-1,6-bisphosphatase, and aldose 6-phosphate reductase were significantly lower in chlorotic leaves than in normal leaves. However, sucrose phosphate synthase activity was higher in chlorotic leaves. Thermal dissipation of excitation energy was enhanced in chlorotic leaves under full sun, lowering the efficiency of excitation energy transfer to PSII reaction centers. This was accompanied by a corresponding increase in both xanthophyll cycle pool size (on a chlorophyll basis) and conversion of violaxanthin to antheraxanthin and zeaxanthin. The antioxidant system was up-regulated in chlorotic leaves in response to the increased generation of reactive oxygen species. These findings support the hypothesis that phloem loading and/or transport is partially or completely blocked in chlorotic leaves, and that excessive accumulation of non-structural carbohydrates may cause feedback suppression of CO2 assimilation via direct interference with chloroplast function and/or indirect repression of photosynthetic enzymes.

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To determine the cause of a characteristic zonal chlorosis of `Honeycrisp' apple (Malus ×domestica Borkh.) leaves, we compared CO2 assimilation, carbohydrate metabolism, the xanthophyll cycle and the antioxidant system between chlorotic leaves and normal leaves. Chlorotic leaves accumulated higher levels of nonstructural carbohydrates, particularly starch, sorbitol, sucrose, and fructose at both dusk and predawn, and no difference was found in total nonstructural carbohydrates between predawn and dusk. This indicates that carbon export was inhibited in chlorotic leaves. CO2 assimilation and the key enzymes in the Calvin cycle, ribulose 1,5-bisphosphate carboxylase/oxygenase, NADP-glyceraldehyde-3-phosphate dehydrogenase, phosphoribulokinase, stromal fructose-1,6-bisphosphatase, and the key enzymes in starch and sorbitol synthesis, ADP-glucose pyrophosphorylase, cytosolic fructose-1,6-bisphosphatase, and aldose 6-phosphate reductase were significantly lower in chlorotic leaves than in normal leaves. However, sucrose phosphate synthase activity was higher in chlorotic leaves. In response to a reduced demand for photosynthetic electron transport, thermal dissipation of excitation energy (measured as nonphotochemical quenching of chlorophyll fluorescence) was enhanced in chlorotic leaves under full sun, lowering the efficiency of excitation energy transfer to PSII reaction centers. This was accompanied by a corresponding increase in both xanthophyll cycle pool size (on a chlorophyll basis) and conversion of violaxanthin to antheraxanthin and zeaxanthin. The antioxidant system, including superoxide dismutase and ascorbate peroxidase and the ascorbate pool and glutathione pool, was up-regulated in chlorotic leaves in response to the increased generation of reactive oxygen species via photoreduction of oxygen. These findings support the hypothesis that phloem loading and/or transport is partially or completely blocked in chlorotic leaves, and that excessive accumulation of nonstructural carbohydrates may cause feedback suppression of CO2 assimilation via direct interference with chloroplast function and/or indirect repression of photosynthetic enzymes.

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One-year-old grapevines (Vitis labrusca L. `Concord') were supplied twice weekly for 5 weeks with 0, 5, 10, 15, or 20 mm nitrogen (N) in a modified Hoagland's solution to generate a wide range of leaf N status. Both light-saturated CO2 assimilation at ambient CO2 and at saturating CO2 increased curvilinearly as leaf N increased. Although stomatal conductance showed a similar response to leaf N as CO2 assimilation, calculated intercellular CO2 concentrations decreased. On a leaf area basis, activities of key enzymes in the Calvin cycle, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), and key enzymes in sucrose and starch synthesis, fructose-1,6-bisphosphatase (FBPase), sucrose phosphate synthase (SPS), and ADP-glucose pyrophosphorylase (AGPase), increased linearly with increasing leaf N content. When expressed on a leaf N basis, activities of the Calvin cycle enzymes increased with increasing leaf N, whereas activities of FBPase, SPS, and AGPase did not show significant change. As leaf N increased, concentrations of glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), and 3-phosphoglycerate (PGA) increased curvilinearly. The ratio of G6P/F6P remained unchanged over the leaf N range except for a significant drop at the lowest leaf N. Concentrations of glucose, fructose, and sucrose at dusk increased linearly with increasing leaf N, and there was no difference between predawn and dusk measurements. As leaf N increased, starch concentration increased linearly at dusk, but decreased linearly at predawn. The calculated carbon export from starch degradation during the night increased with increasing leaf N. These results showed that 1) grapes leaves accumulated less soluble carbohydrates under N-limitation; 2) the elevated starch level in low N leaves at predawn was the result of the reduced carbon export from starch degradation during the night; and 3) the reduced capacity of CO2 assimilation in low N leaves was caused by the coordinated decreases in the activities of key enzymes involved in CO2 assimilation as a result of direct N limitation, not by the indirect feedback repression of CO2 assimilation via sugar accumulation.

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Paeonia ostii ‘Feng Dan’ is an economically important, multipurpose woody plant in terms of its medical, ornamental, and oil values; however, there is a noticeable contradiction between the increasing demands and the lack of excellent germplasm resources because of traditional breeding and propagation approaches. In vitro embryo culture is an attractive option for this issue. This study presents a protocol for in vitro immature embryo culture in P. ostii ‘Feng Dan’, which involves two steps: 1) immature seeds at 30 days after anthesis (DAA) (cellularization stage of endosperm, proembryo stage) or after being cultured in vitro for cotyledon embryo formation (upward micropyle with placenta was the best inoculation method with the highest ratio of seed with cotyledon embryo of 66.67%); and 2) seedling establishment was realized within 7 months via embryo (at 40 DAA or after) germination, shoot induction, rooting, and acclimatization. The multiplication potential was increased with embryo maturity. This protocol provides an available reference for embryo rescue and propagation of tree peony and will be beneficial to shortening the breeding cycle.

Open Access

Nucleotide sequences of the second intron of the nuclear LEAFY gene (FLint2) and trnL-trnF region of the chloroplast genome were used to analyze the phylogenetic relationships among eight wild mandarins (Citrus reticulata Blanco), 19 mandarin landraces, and 19 related species of Citrus L. Forty-six trnL-trnF sequences and 111 FLint2 sequences were obtained from 46 ingroup accessions, with an average length of 1059.7 and 776.7 bp respectively. Phylogeny reconstructions were conducted separately for these two data sets using maximum parsimony and maximum likelihood criteria. Monophyly of mandarins was supported by both of these data sets, and in this clade, most mandarin landraces formed an unresolved polytomy, whereas ‘Jiangyong 1’, ‘Chongyi A2’, ‘Chongyi A1’ (or ‘Jiangyong 4’ in FLint2 data), and ‘Daoxian 1’ wild mandarins formed a subclade. ‘Mangshan A1’ and ‘Daoxian 5’ wild mandarins were sisters to this mandarin clade. A hybrid origin of five mandarin landraces and several mandarin-related species was suggested as a plausible hypothesis to explain the incongruence between the FLint2 and trnL-trnF data sets.

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Sea buckthorn (Hippophae rhamnoides) is an ecologically and economically valuable species that has been widely cultivated as a new berry crop rich in nutritional and medicinal compounds. RNA Sequencing (RNA-Seq) simple sequence repeat (SSR) markers were developed to evaluate the genetic relationships among 91 plants of 31 cultivars from two subspecies, mongolica and sinensis, as well as intraspecific hybrids between them. A total of 7540 RNA-Seq SSRs were identified as potential molecular markers, in which AG/CT (27.57%) was the most abundant unit type. AT/AT (9.93%), and AAG/CTT (11.95%) are the other main repeat motifs. A total of 110 primer pairs were randomly selected for validation of amplification. Seventeen SSR loci, located in genes encoding metabolic processes and cellulose synthases, were identified to be polymorphic among different sea buckthorn cultivars. These SSR loci generated 48 alleles, ranging from 2 to 5 per locus. Cluster analysis based on the proportion of shared alleles and unweighted pair group method with arithmetic average (UPGMA) algorithm divided all the genotypes into two main groups, with all of the ssp. sinensis cultivars (native to China) and hybrids in one group and ssp. mongolica cultivars (introduced from Russia) in the other group, which was in good agreement with their taxonomic classification. The RNA-Seq SSRs developed in this study have a potential use in the conservation of sea buckthorn germplasm and marker-assisted breeding (MAB).

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Twenty-day-old `Bush Blue Lake 47' common bean plants grown in a growth chamber at 25 days/22C night and a 12-hour photoperiod regime were foliar sprayed with 0.5% GLK-8903 including 0.05% Tween-20. After 24 hours of treatment, plants were chilled in a cold room (4C day/night, 12 hours of light). After 3 days of chilling, leaves of untreated controls were injured, as visually characterized by leaf wilting, whereas leaves of the GLK-8903-treated plants still retained turgor. During chilling, the activity of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) decreased. GLK-8903 treatment had no effect on SOD and POD activities; however, the CAT activity was reduced significantly after GLK-8903 treatment either at 25 or at 4C. During chilling, the content of malondialdehyde, a decomposition product of phospholipid peroxidation, increased in treated plants and untreated controls, with increased content significantly lower in the former compared with the latter. The GLK-8903 per se and total lipid extracted from GLK-8903-treated plants were able to reduce the linoleic acid oxidation in vitro. The mechanism by which GLK-8903 alleviates chilling injury in bean plants is discussed.

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Abstract

Leaves of mefluidide-treated corn (Zea mays L.) plants showed significant protection from chilling injury compared with untreated, chilled controls. On a per-plant basis, leaves sprayed with 5, 10, 15, or 20 ppm mefluidide had 6%, 14%, 9%, or 12% damage, respectively, of the total leaf area after a 6-day exposure to 4°C, whereas controls had more than 60% damage. Treated plants had more leaf area, were taller, had greater fresh and dry weights, and flowered 6 days earlier than controls. Plant growth was stunted with a 20-ppm spray, although plants were protected from chilling injury. During a 6-day chilling period, a consistent level of about 10% electrolyte leakage was observed among treated plants, whereas from control tissue, electrolyte leakage increased from 10% to 40%. These results suggest that mefluidide may modulate the permeability of the plasma membrane in the chilling environments. Chemical name used: N-[2,4-dimethyl-5-[(trifluoromethyl)sulfonyl]amino]phenyl]acetamide (mefluidide).

Open Access

Own-rooted one-year-old `Concord' grapevines were fertigated twice weekly for 11 weeks with 1, 10, 20, 50, OR 100 μmol iron (Fe) from ferric ethylenediamine di (o-hydroxyphenylacetic) acid in a complete nutrient solution. As Fe supply increased, leaf total Fe content did not change, whereas active Fe (extracted by 2, 2'-dipyridyl) and total chlorophyll content increased curvilinearly. CO2 assimilation and stomatal conductance increased curvilinearly with increasing active Fe, whereas intercellular CO2 concentrations decreased linearly. Activities of key Calvin cycle enzymes, Rubisco, NADP-glyceraldehyde-3-phosphate dehydrogenase, phosphoribulokinase, stromal fructose-1,6-bisphosphatase (FBPase), and a key enzyme in sucrose synthesis, cytosolic FBPase, all increased linearly with increasing active Fe. No difference was found in the activities of ADP-glucose pyrophosphorylase and sucrose phosphate synthase of leaves between the lowest and the highest treatments, whereas slightly lower activities were observed in the middle Fe treatments. Content of 3-phosphoglycerate increased curvilinearly with increased active Fe, whereas glucose-6-phosphate and fructose-6-phosphate did not change. Glucose, fructose, sucrose, starch, and total non-structural carbohydrates at both dusk and pre-dawn increased with increasing active Fe. Carbon export from starch breakdown during the night, calculated as the difference between dusk and predawn levels, increased as active Fe increased. In conclusion, Fe limitation reduces the activities of Rubisco and other photosynthetic enzymes, and hence CO2 assimilation capacity. Fe-deficient grapevines have lower concentrations of non-structural carbohydrates in source leaves, and therefore, are source limited.

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Own-rooted 1-year-old `Concord' grapevines (Vitis labruscana Bailey) were fertigated twice weekly for 11 weeks with 1, 10, 20, 50, or 100 μm iron (Fe) from ferric ethylenediamine di (o-hydroxyphenylacetic) acid (Fe-EDDHA) in a complete nutrient solution. As Fe supply increased, leaf total Fe content did not show a significant change, whereas active Fe (extracted by 2,2′-dipyridyl) content increased curvilinearly. Chlorophyll (Chl) content increased as Fe supply increased, with a greater response at the lower Fe rates. Chl a: b ratio remained relatively constant over the range of Fe supply, except for a slight increase at the lowest Fe treatment. Both CO2 assimilation and stomatal conductance increased curvilinearly with increasing leaf active Fe, whereas intercellular CO2 concentrations decreased linearly. Activities of key enzymes in the Calvin cycle, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), stromal fructose-1,6-bisphosphatase (FBPase), and a key enzyme in sucrose synthesis, cytosolic FBPase, all increased linearly with increasing leaf active Fe. No significant difference was found in the activities of ADP-glucose pyrophosphorylase (AGPase) and sucrose phosphate synthase (SPS) of leaves between the lowest and the highest Fe treatments, whereas slightly lower activities of AGPase and SPS were observed in the other three Fe treatments. Content of 3-phosphoglycerate (PGA) increased curvilinearly with increasing leaf active Fe, whereas glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), and the ratio of G6P: F6P remained unchanged over the range of Fe supply. Concentrations of glucose, fructose, sucrose, starch, and total nonstructural carbohydrates (TNC) at both dusk and predawn increased with increasing leaf active Fe. Concentrations of starch and TNC at any given leaf active Fe content were higher at dusk than at predawn, but both glucose and fructose showed the opposite trend. No difference in sucrose concentration was found at dusk or predawn. The export of carbon from starch breakdown during the night, calculated as the difference between dusk and predawn measurements, increased as leaf active Fe content increased. The ratio of starch to sucrose at both dusk and predawn also increased with increasing leaf active Fe. In conclusion, Fe limitation reduces the activities of Rubisco and other photosynthetic enzymes, and hence CO2 assimilation capacity. Fe-deficient grapevines have lower concentrations of nonstructural carbohydrates in source leaves and, therefore, are source limited.

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