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Jun Yu*, Romeo Toledo, Rakesh Singh, Leonard Pike, and Bhimanagouda Patil

Grapefruit seeds were studied for the extraction of limonoids using supercritical CO2 extraction (SC-CO2) technique. Limonin aglycone was successfully extracted with SC-CO2 directly from grapefruit seeds; and the limonin glycoside was extracted using SC-CO2 and ethanol as co-solvent from the spent seeds after the extraction of limonin aglycone. In an effort to optimize the extraction conditions of limonin aglycone, pressure, temperature, time effects were investigated. Various times of extraction, CO2 flow rate and the feeding modes of CO2 were also employed to obtain the highest yield of limonin aglycone. Optimal conditions to achieve the highest limonin aglycone (0.63 mg/g seeds) were 48.3 MPa, 50°C and 60 min with CO2 bottom feeding, flow rate about 5 L/min. The extraction conditions for limonin glycoside to achieve highest yield were further optimized. The highest extraction yield (0.62 mg limonin glycoside/g seeds) were at 48.3 MPa, 50°C, 30% molar fraction of ethanol (XEth =0.30) and 40 min with CO2 top feeding, flow rate about 5 L/min. The results demonstrated that supercritical CO2 extraction of limonoids from grapefruit seeds, a citrus juice industry byproduct, has practical significance for commercial production.

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Jun Yuan, Liyuan Huang, Naifu Zhou, Hui Wang, and Genhua Niu

Aluminum (Al) toxicity and phosphorus (P) deficiency are two crucial factors limiting the production of Camellia oleifera, which is grown commercially in red acidic soils in Southern China. The current study characterized the different forms of P and Al in the red acidic soils of C. oleifera plantations. Soil and plant tissue samples taken from 32 Camellia plantations across Hunan province were analyzed. Furthermore, a pot experiment with nutrient solutions of different Al and P contents was carried out to investigate P and Al uptake and their effect on C. oleifera growth. The results showed that the P content extracted by NaOH (Fe-P) was the highest in all types of soil samples (rhizosphere, 0–20 cm, and 20–40 cm zones), followed by P extracted by NH4F (Al-P), H2SO4 (Ca-P), and Na3C6H5O7 (O-P). HCl (In-Al), NH4Ac (Ha-Al), and Na4P2O7·10H2O (Or-Al) extracted Al were the main forms and accounted for 22.8%, 23.1%, and 23.8% of total Al, respectively. KCl extracted Al (Ex-Al) contents in the rhizosphere, 0–20 cm, and 20–40 cm soil zones were 4.78, 4.86, and 4.59 mg·kg−1, respectively. P contents in roots, young leaves, and old leaves were 0.80, 0.82, and 0.64 mg·kg−1, respectively. The highest Al content of 11.35 g·kg−1 was found in the old leaves, followed by roots and young leaves. Correlation analyses revealed that P in roots was positively associated with available P (AP) and Al-P in rhizosphere. P in roots and young leaves also had a positive correlation with Ex-Al, whereas Al in old leaves was positively correlated with In-Al and total Al. Significant correlations between Al-P, Ex-Al, and AP were detected. The pot experiment indicated that adding Al or P alone increased plant growth and Al or P uptake, respectively. When adding both Al and P, significant synergistic effect was found. These results suggest that Al is beneficial to C. oleifera, which may be the adaptive mechanism of C. oleifera to use insoluble Al-P in red acidic soil.

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Liyuan Huang, Jun Yuan, Hui Wang, Xiaofeng Tan, and Genhua Niu

High concentration of aluminum ion (Al3+) in acidic soil often negatively affects plant growth. To deepen understanding of the mechanisms of physiological response to Aluminum (Al) toxicity, changes in physiology and cell ultrastructure of oil tea (Camellia oleifera) were investigated under different Al levels. Oil tea plants were grown in pots filled with sand and treated with Al at 0, 0.5, 1.25, 2.0, or 4.0 mm. Results showed that Al at 0.5–2.0 mm improved plant growth, whereas Al at 4.0 mm inhibited root growth and damaged cell ultrastructure. Net photosynthetic rate (Pn), stomatal conductance (g s), transpiration rate (Tr), and photochemical efficiency increased as Al concentration increased from 0 to 2.0 mm; however, all parameters mentioned previously decreased at 4.0 mm. The activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in leaves treated with 2.0 mm Al reached the maximum, which were 29%, 63%, and 28% higher than that of control. When Al was ≤2.0 mm, the content of soluble sugar and soluble protein increased with increasing Al concentration. These results may indicate that oil tea adapted to Al stress through osmotic adjustment and through increasing antioxidant enzyme system. In summary, Al at low concentration (0.5–2.0 mm) improved growth and physiological performance, whereas 4.0 mm negatively impacted performance of oil tea.

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Waltram Ravelombola, Jun Qin, Yuejin Weng, Beiquan Mou, and Ainong Shi

Little has been done with respect to breeding for salt-tolerant cowpea (Vigna unguiculata) cultivars despite of salt stress being a growing threat to cowpea production. Seedling stage is one the most susceptible stages to salt stress in cowpea. Establishing a streamlined methodology for rapidly screening a large number of genotypes will significantly contribute toward enhancing cowpea breeding for salt tolerance. Therefore, the objective of this study was to establish and validate a simple approach for salt tolerance evaluation in cowpea seedlings. A total of 30 genotypes including two controls (PI582468, a salt-tolerant genotype, and PI255774, a salt-sensitive genotype) were greenhouse-grown under 0 mm and 200 mm NaCl. A total of 14 above-ground traits were evaluated. Results revealed: (1) significant differences were observed in average number of dead plants per pot, leaf injury scores, relative salt tolerance (RST) for chlorophyll, plant height, and leaf and stem biomass among the 30 genotypes; (2) all PI255774 plants were completely dead, whereas those of PI582438 were fully green after 2 weeks of salt stress, which validated this methodology; (3) RST for chlorophyll content was highly correlated with number of dead plants and leaf injury scores; (4) RST for leaf biomass was moderately correlated with number of dead plants and leaf injury scores; and (5) RST in plant height was poorly correlated with number of dead plants and leaf injury scores Therefore, less number of dead plants per pot, high chlorophyll content, and less leaf injury scores were good criteria for salt tolerance evaluation in cowpea. This study provided a simple methodology and suggested straightforward criteria to evaluate salt tolerance at seedling stage in cowpea.

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Shisheng Li, Qiufang Xiong, Jingcai Li, Yuanping Fang, and Jun Xiang

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Jun Matsumoto, Hideyuki Goto, Yasutaka Kano, Akira Kikuchi, Hideaki Ueda, and Yuta Nakatsubo

To determine the relationship among cell size, acid invertase (AI) activity, sucrose phosphate synthase (SPS) activity, and sucrose accumulation in melon (Cucumis melo L.) during early development [from 6 to 16 days after anthesis (DAA)], fruit were heated at night to a minimum of 20 °C. Cells of heated fruit were larger than those of control fruit at 16 DAA but smaller at 50 DAA. AI activity was lower and SPS activity was higher in heated than in control fruit up to 26 DAA. Sucrose, glucose, and fructose contents at 26 and 50 DAA were higher in heated than in control fruit. Heating caused cells to reach mature size earlier than those of control fruit, and maturity was accompanied by earlier decline in AI activity and an earlier increase in SPS activity that promoted soluble sugar accumulation.

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Jing Mao, Hongliang Xu, Caixia Guo, Jun Tong, Yanfang Dong, Dongyun Xu, Fazhi Chen, and Yuan Zhou

Although tolerance to high temperature is crucial to the summer survival of Iris germanica cultivars in subtropical areas, few physiological studies have been conducted on this topic previously. To remedy this, this study explored the physiological response and expression of heat shock factor in four I. germanica cultivars with varying levels of thermotolerance. The plants’ respective degrees of high-temperature tolerance were evaluated by measuring the ratio and area of withered leaves under stress. Several physiological responses to high temperatures were investigated, including effects on chlorophyll, antioxidant enzymes, proline, and soluble protein content in the leaves of four cultivars. CaCl2 was sprayed on ‘Gold Boy’ and ‘Royal Crusades’ considered being sensitive to high temperatures to study if Ca2+ could improve the tolerance, and LaCl3 was sprayed on ‘Music Box’ and ‘Galamadrid’ with better high-temperature tolerance to test if calcium ion blocker could decrease their tolerance. Heat shock factor genes were partially cloned according to the conserved region sequence, and expression changes to high-temperature stress with CaCl2 or LaCl3 treatments were thoroughly analyzed. Results showed that high temperature is the primary reason for large areas of leaf withering. The ratio and area of withered leaves on ‘Music Box’ and ‘Galamadrid’ were smaller than ‘Gold Boy’ and ‘Royal Crusades’. CaCl2 slowed the degradation of chlorophyll content and increased proline and soluble protein in ‘Gold Boy’ and ‘Royal Crusades’ but had no significant effect on activating peroxidase or superoxide to improve high-temperature tolerance. Genetic expression of heat shock factor in ‘Gold Boy’ and ‘Royal Crusades’ was upregulated by Ca2+ at later stages of leaf damage under high-temperature stress. LaCl3 down-regulated the physiological parameters and expression level of heat shock factor in ‘Music Box’ and ‘Galamadrid’. These results suggest that different I. germanica cultivars have varying high-temperature tolerance and furthermore that Ca2+ regulates their physiological indicators and expression level of heat shock factor under stress.

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Lu Zhang, You-biao Hu, Hua-sen Wang, Sheng-jun Feng, and Yu-ting Zhang

Plant growth and development are determined by complex exogenous and endogenous cues. A plant follows several temporally distinct developmental stages, including embryonic, vegetative, and reproductive. The vegetative stage, which is usually the longest stage, can be subdivided into juvenile and adult phases. The transition from the juvenile to the adult phase, also called the vegetative phase change, is characterized by anatomical, morphological, and physiological changes in the vegetative parts of the shoot. Recent studies in several systems have identified the genetic temporal mechanisms of this process, which is regulated by an endogenous age cue (i.e., microRNA156/157) and its targeted genes (i.e., Squamosa promoter binding protein-box transcription factors). This review summarizes the recent advances in the study of the underlying regulatory mechanisms of vegetative phase change. This review also describes the modes of miRNA action and the functions of their targeted genes in this highly conserved developmental process.

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Shuang Jiang, Haishan An, Xiaoqing Wang, Chunhui Shi, Jun Luo, and Yuanwen Teng

Simple sequence repeats (SSRs) are widely used in cultivar identification, genetic relationship analysis, and quantitative trait locus mapping. Currently, the selection of hybrid progeny plants in molecular marker-assisted breeding mostly relies on SSR markers because of their ease of operation. In Pyrus, a large number of SSR markers have been developed previously. The method to identify polymorphic SSRs quickly is still lacking in cultivated as well as wild pear species. We present a large number of polymorphic SSRs identified using a quick in silico approach applied across 30 cultivated and wild accessions from Pyrus species. A total of 49,147 SSR loci were identified in Pyrus, and their genotypes were evaluated by whole-genome resequencing data of 30 Pyrus accessions. The results show that most SSR loci were dinucleotide repeat motifs located in intergenic regions. The genotypes of all SSR loci were revealed in all accessions. A total of 23,209 loci were detected, with more than one genotype in all Pyrus accessions. We selected 702 highly polymorphic SSR loci to characterize the pear accessions with an average polymorphism information content value of 0.67, suggesting that these SSR loci were highly polymorphic. The genetic relationship of Pyrus species in the neighbor-joining (NJ) tree and population structure showed a clear division between the oriental and occidental accessions. The population structure split all oriental pears into two groups: cultivars and wild accessions. These new findings of the polymorphic SSR loci in this study are valuable for selecting appropriate markers in molecular marker-assisted breeding in Pyrus.

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Xinjing Qu, Hui Wang, Ming Chen, Jiao Liao, Jun Yuan, and Genhua Niu

Oil tea (Camellia oleifera) is an important edible oil tree. However, its growth and yield are strongly limited by drought. This study investigated the physiological and metabolic responses of two common oil tea cultivars, Huajin and Changlin53, to moderate and severe drought stress. Based on the photosynthetic and physiological indices, ‘Changlin53’ may be more tolerant to drought than ‘Huajin’. A total of 41 key metabolites induced by drought stress, including 12 amino acids, 12 organic acids, 10 carbohydrates, 3 fatty acids, and 4 phenols, have been identified by liquid chromatography-mass spectrometry. Under moderate drought stress, the contents of carbohydrates, amino acids, and some organic acids in ‘Changlin53’ were significantly increased; however, under severe drought stress, the contents of soluble sugars were decreased and the synthesis ability of amino acids and organic acids were enhanced. The glutamic acid–mediated proline biosynthesis pathway and salicylic acid synthesis were continuously upregulated in ‘Changlin53’ under moderate and severe drought stress, which could regulate osmotic pressure and maintain intracellular environmental stability. Under moderate drought stress, the contents of monosaccharides, amino acids, and organic acids increased in ‘Huajin’ leaves. Furthermore, the shikimic acid–mediated secondary metabolite synthesis pathway was weakened. More secondary metabolites were used to increase glycolysis and tricarboxylic acid cycle to accelerate energy production and to enhance the glutamic acid–mediated proline biosynthesis pathway, which are necessary to increase osmotic regulation. Under severe drought stress, the contents of carbohydrates, organic acids, and some amino acids were significantly decreased in ‘Huajin’ leaves, indicating serious damage. These results deepened our understanding of the mechanisms involved in oil tea drought tolerance, which will help improve water management of oil tea seedlings.