Genome-wide identification, expression and functional analysis of the core cell cycle gene family during the early somatic embryogenesis of Dimocarpus longan Lour.

Core cell cycle genes (CCCs) are essential regulators of cell cycle operation. In this study, a total of 69 CCCs family members, including 37 CYCs, 20 CDKs, five E2F/DPs, three KRPs, two RBs, one CKS and one Wee1, were identified from the longan genome. Phylogenetic and motifs analysis showed the evolutionary conservation of CCCs. Transcriptome dataset showed that CCCs had various expression patterns during longan early somatic embryogenesis (SE). Either CKS or CYCD3;2 silencing increased the expression of RB-E2F pathway genes, and the silencing of CYCD3;2 might induce the process of apoptosis in longan embryogenic callus (EC) cells. In addition, The qRT-PCR results showed that the expression levels of CDKG2, CYCD3;2, CYCT1;2, CKS and KRP1 were elevated by ABA, 2,4-D and PEG4000 treatments, while CDKG2 and CYCT1;2 were inhibited by NaCl treatment. In conclusion, our study provided valuable information for understanding the characterization and biological functions of longan CCCs.

Effects of hydrogen peroxide treatment on pulp breakdown, softening, and cell wall polysaccharide metabolism in fresh longan fruit.

Longan (Dimocarpus longan Lour.) is prone to pulp softening and pulp breakdown, leading to a loss of its nutrients including polysaccharides. ROS is one main factor affecting fruit quality. This work intended to explicate the influences of hydrogen peroxide, acting as a ROS, on pulp softening, pulp breakdown, and cell wall polysaccharides metabolism in longan fruit during storage. Contrasted to the control group, hydrogen peroxide-treated samples exhibited lower firmness, lower amounts of CWM, ISP, CSP, hemicellulose and cellulose, but higher breakdown index, WSP amount, expression levels of DlPG, DlPE, Dlß-Gal, DlCx and DlXET and activities of their corresponding enzymes (PG, PE, ß-Gal, Cx, XET). These results suggested that hydrogen peroxide reduced longan pulp firmness due to the increased gene expression levels and enzymes activities related to cell wall polysaccharide degradation to boost their decomposition, thereby led to the accelerated pulp softening and the expedited pulp breakdown of harvested longans.

Effects of chitosan treatment on the storability and quality properties of longan fruit during storage.

Effects of various concentrations of Kadozan (chitosan) treatment on storability and quality properties of harvested 'Fuyan' longans were investigated. Compared to the control samples, Kadozan treated-longans displayed lower fruit respiration rate, lower pericarp cell membrane permeability, pericarp browning index, pulp breakdown index, fruit disease index, and weight loss, but higher rate of commercially acceptable fruit, higher levels of pericarp chlorophyll, carotenoid, anthocyanin, flavonoid and total phenolics, higher amounts of pulp total soluble sugar, sucrose, total soluble solids, and vitamin C. These results revealed Kadozan treatment could increase storability and retain better quality of harvested longan fruit. Among different concentrations of Kadozan, the dilution of 1:500 (VKadozan: VKadozan + Water) showed the best results in storability and maintained the best quality of longans during storage. These findings demonstrated that Kadozan could be a facile and eco-friendly postharvest handling approach for increasing storability and lengthening shelf-life of harvested 'Fuyan' longan fruit.

The role of ROS-induced change of respiratory metabolism in pulp breakdown development of longan fruit during storage.

Compared to the control longans, hydrogen peroxide (H2O2)-treated longans exhibited higher index of pulp breakdown, higher fruit respiration rate, higher activities of pulp phosphohexose isomerase (PGI), succinate dehydrogenase (SDH), cytochrome C oxidase (CCO), ascorbic acid oxidase (AAO) and polyphenol oxidase (PPO), but lower activity of pulp nicotinamide adenine dinucleotide kinase (NADK). H2O2-treated longans also exhibited lower total activities of pulp glucose-6-phosphate dehydrogenase (G-6-PDH) and 6-phosphogluconate dehydrogenase (6-PGDH), lower levels of pulp NADP(H), but higher levels of pulp NAD(H). These data indicated that H2O2-stimulated longan pulp breakdown was owing to a decreased proportion of pentose phosphate pathway (PPP), the increased proportions of Embden-Meyerhof-Parnas pathway (EMP), tricarboxylic acid (TCA) cycle and cytochrome pathway (CCP) in total respiratory pathways. These findings further revealed that H2O2 could enhance respiration rate, and thus accelerate pulp breakdown occurrence and shorten the shelf life of longan fruit.

Hydrogen peroxide-induced changes in activities of membrane lipids-degrading enzymes and contents of membrane lipids composition in relation to pulp breakdown of longan fruit during storage.

This study aimed to investigate the effect of hydrogen peroxide (H2O2) on membrane lipids metabolism and its relation to pulp breakdown development of longan fruit during postharvest storage. Compared to the control longans, H2O2-treated longans showed higher pulp breakdown index, cell membrane permeability, and activities of phospholipase D (PLD), lipase and lipoxygenase (LOX). Moreover, H2O2-treated longans maintained higher levels of pulp phosphatidic acid (PA) and saturated fatty acids (SFA). However, H2O2-treated longans exhibited lower levels of pulp phosphatidylcholine (PC), phosphatidylinositol (PI) and unsaturated fatty acids (USFA), lower index of unsaturated fatty acids (IUFA), and lower ratio of USFA to SFA (U/S). These findings demonstrated that H2O2 caused the increased activities of enzymes involving in membrane lipids degradation and the accelerated decompositions of membrane USFA and phospholipids in longan pulp, which eventually triggered the destruction of the pulp cell membrane structure and the development of pulp breakdown in longans during storage.

Effect of Cd on growth, physiological response, Cd subcellular distribution and chemical forms of Koelreuteria paniculata.

Koelreuteria paniculata were cultivated in nutrient solution with different concentrations of Cd (0, 50, 150, 250 and 500 µM) and sampled after 90 days. The resistance, translocation, accumulation and stress responses in Koelreuteria paniculata were investigated by hydroponic experiments. The results showed that Koelreuteria paniculata is an efficient Cd excluder that can tolerate high concentrations of Cd (up to 150-250 µM of Cd). The concentration of Cd never exceeds 5 ppm in leaves and 10 ppm in roots. The high concentration of Cd (≥ 250 µM) had a toxic effect on K. paniculata and significantly restricted the plant growth. The accumulation ability of Cd by different plant tissues followed the sequence of roots > leaves > stems. The bioconcentration factors and translocation factors both were less than 1. Cd has the highest content in the cell wall and is migrated to soluble fractions and organelles at high concentrations. Undissolved Cd phosphate, pectates and protein-bound Cd were the predominant forms. The low concentration of Cd (≤150 µM) promoted the synthesis of soluble proteins, AsA and GSH, while high concentration of Cd clearly inhibited the physiological and biochemical process, caused membrane lipid peroxidation and severe membrane damages, and increased MDA and H2O2 contents. POD, CAT and SOD exhibited positive and effective responses to low concentration Cd stress, but could not remove the toxicity caused by high concentration Cd stress. The content of IAA, GA and ZT decreased and ABA content was significantly increased under high-concentration Cd stress.

Hydrogen peroxide-induced pericarp browning of harvested longan fruit in association with energy metabolism.

Energy metabolism of "Fuyan" longan fruit treated with hydrogen peroxide (H2O2), the most stable of the reactive oxygen, and its relationship to pericarp browning were investigated in this work. The results displayed that H2O2 significantly decreased contents of adenosine triphosphate (ATP) and adenosine diphosphate (ADP). It also inhibited activities of H+-ATPase, Ca2+-ATPase and Mg2+-ATPase in membranes of plasma, vacuole and mitochondria during the early-storage and mid-storage (except for mitochondrial membrane Mg2+-ATPase). These results gave convincing evidence that the treatment of H2O2 accelerating pericarp browning in harvested longans was due to a decrease of ATPase activity and available ATP content. This might break the ion homeostasis and the integrity of mitochondria, which might reduce energy charge and destroy the function and compartmentalization of cell membrane. These together aggravated browning incidence in pericarp of harvested longan fruit.

Hydrogen Peroxide Induced Changes in Energy Status and Respiration Metabolism of Harvested Longan Fruit in Relation to Pericarp Browning.

Energy status and respiration metabolism of "Fuyan" longan fruit treated by hydrogen peroxide (H2O2) and their relationship to pericarp browning were studied. The results displayed that H2O2 significantly increased the respiration rate, increased activities of respiratory terminal oxidases like cytochrome C oxidase (CCO) and ascorbic acid oxidase (AAO), decreased NAD kinase activity, maintained lower contents of NADP and NADPH as well as higher amounts of NAD and NADH, and accelerated the decrease of energy charge. These results gave convincing evidence that the treatment of H2O2 for accelerating longan pericarp browning was due to an increase of energy deficiency, an increase of respiratory metabolic pathways of Embden-Meyerhof pathway (EMP) and tricarboxylic acid (TCA) cycle, a decrease of pentose phosphate pathway (PPP) of respiratory pathway, and an increase of activities of respiratory terminal oxidases like CCO and AAO.

Inhibitory effects of propyl gallate on tyrosinase and its application in controlling pericarp browning of harvested longan fruits.

Tyrosinase (EC 1.14.18.1), also known as polyphenol oxidase (PPO), is a key enzyme in pigment biosynthesis of organisms. The inhibitory effects of propyl gallate on the activity of mushroom tyrosinase and effects of propyl gallate on pericarp browning of harvested longan fruits in relation to phenolic metabolism were investigated. The results showed that propyl gallate could potently inhibit diphenolase activity of tyrosinase. The inhibitor concentration leading to 50% activity lost (IC50) was determined to be 0.685 mM. Kinetic analyses showed that propyl gallate was a reversible and mixed type inhibitor on this enzyme. The inhibition constants (K(IS) and K(I)) were determined to be 2.135 and 0.661 mM, respectively. Furthermore, the results also showed that propyl gallate treatment inhibited activities of PPO and POD in pericarp of harvested longan fruits, and maintained higher contents of total phenol and flavonoid of longan pericarp. Moreover, propyl gallate treatment also delayed the increases of browning index and browning degree in pericarp of harvested longan fruits. Therefore, application of propyl gallate may be a promising method for inhibiting tyrosinase activity, controlling pericarp browning, and extending shelf life of harvested longan fruits.

[Effects of exogenous nitric oxide on physiological characteristics of longan (Dimocarpus longana) seedlings under acid rain stress].

This paper studied the effects of exogenous nitric oxide donor sodium nitroprusside (SNP) on the chlorophyll content, antioxidant enzyme activities, and osmotic regulation substances of longan (Dimocarpus longana 'Fuyan') seedlings under acid rain (pH 3.0) stress. Under the acid rain stress, the seedling leaf superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities and chlorophyll, soluble protein and soluble sugar contents decreased obviously, while the leaf malondialdedyde content had a remarkable increase, suggesting the toxic effect of the acid rain on the seedlings. Exogenous nitric oxide had dual nature on the physiological characteristics of longan seedlings under acid rain stress. Applying 0.1-0.5 mmol x L(-1) of SNP improved the SOD, POD and CAT activities and the chlorophyll, soluble protein and soluble sugar contents significantly, and decreased the malondialdedyde content. Low concentrations SNP reduced the oxidative damage caused by the acid rain stress, and 0.5 mmol x L(-1) of SNP had the best effect. Under the application of 0.5 mmol x L(-1) of SNP, the total chlorophyll, soluble protein, and soluble sugar contents and the SOD, POD and CAT activities increased by 76.0%, 107.0%, 216.1%, 150. 0%, 350.9% and 97.1%, respectively, and the malondialdedyde content decreased by 46.4%. It was suggested that low concentration (0.1-0.5 mmol x L(-1)) SNP could alleviate the toxic effect of acid rain stress on longan seedlings via activating the leaf antioxidant enzyme activities and reducing oxidative stress, while high concentration SNP (1.0 mmol x L(-1)) lowered the mitigation effect.

Effect of a saprophytic fungus on the growth and the lead uptake, translocation and immobilization in Dodonaea viscosa.

Phytoremediation is a feasible alternative to remediate soils polluted with toxic elements, which can be enhanced by manipulating plant-microbe interactions. Regarding this, free-living saprophytic fungi that interact beneficially with roots have been scarcely studied. Thus, the aim of this study was to assess the effect of a saprophytic fungus, Lewia sp., on the plant growth and the ability of Dodonaea viscosa to phytoaccumulate or phytostabilize soluble and insoluble sources of lead in a solid support. The growth of D. viscosa was influenced by both Pb and Lewia sp. While seedlings exposed to Pb showed a decrease in biomass production, in seedlings grown without Pb the biomass was stimulated by Lewia sp. The fungus strongly stimulated the weight-to-length ratio in roots. Regardless of the treatment, D. viscosa accumulated 4.4-6.5 times more Pb in roots than in shoots, conducting to low translocation factors (< 0.2). The presence of Lewia sp. significantly improved Pb accumulation, achieving high bioconcentration factors (> 22), which was attributed to an increased bioavailability and uptake of Pb due to the fungus. This study demonstrated that Lewia sp. could improve Pb-phytostabilization by D. viscosa in soils polluted with soluble and insoluble forms of Pb.

Investigation of plant hormone level changes in shoot tips of longan (Dimocarpus longan Lour.) treated with potassium chlorate by liquid chromatography-electrospray ionization mass spectrometry.

The endogenous levels of indole-3-acetic acid (IAA), gibberellins (GAs), abscisic acid (ABA) and cytokinins (CKs) and their changes were investigated in shoot tips of ten longan (Dimocarpus longan Lour.) trees for off-season flowering until 60 days after potassium chlorate treatment in comparison with those of ten control (untreated) longan trees. These analytes were extracted and interfering matrices removed with a single mixed-mode solid phase extraction under optimum conditions. The recoveries at three levels of concentration were in the range of 72-112%. The endogenous plant hormones were separated and quantified by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). Detection limits based on the signal-to-noise ratio ranged from 10 ng mL(-1) for gibberellin A4 (GA4) to 200 ng mL(-1) for IAA. Within the first week after potassium chlorate treatment, dry weight (DW) amounts in the treated longan shoot tips of four gibberellins, namely: gibberellin A1(GA1), gibberellic acid (GA3), gibberellin A19 (GA19) and gibberellin A20 (GA20), were found to increase to approximately 25, 50, 20 and 60 ng g(-1) respectively, all of which were significantly higher than those of the controls. In contrast, gibberellin A8 (GA8) obtained from the treated longan was found to decrease to approximately 20 ng g(-1)DW while that of the control increased to around 80 ng g(-1)DW. Certain CKs which play a role in leaf bud induction, particularly isopentenyl adenine (iP), isopentenyl adenosine (iPR) and dihydrozeatin riboside (DHZR), were found to be present in amounts of approximately 20, 50 and 60 ng g(-1)DW in the shoot tips of the control longan. The analytical results obtained from the two-month off-season longan flowering period indicate that high GA1, GA3, GA19 and GA20 levels in the longan shoot tips contribute to flower bud induction while high levels of CKs, IAA and ABA in the control longan contribute more to the vegetative development.

[Regulation function of calcium on photosynthesis of Dimocarpus longana Lour. cv. wulongling under simulated acid rain stress].

Studies on the regulation function of calcium on photosynthesis of Dimocarpus longana under simulated acid rain stress showed that the photoreduction activity of chloroplasts was activated when the concentration of calcium ion in reaction medium ranged from 0 to 5 mmol.L-1, and peaked at the 3.5 mmol.L-1, which was 41.90% higher than that of control. Conversely, the activity of chloroplasts reduced 26.06% in the reaction medium with a concentration of 2 mmol.L-1 EGTA, as compared with the control. Both Mn2+ and Mg2+ could inhibit photoreduction activity. The photophosphorylation activity increased when the concentration of calcium ion in reaction medium ranged from 0 to 6 mmol.L-1, and peaked at the 4.5 mmol.L-1, while superoxidase dismutase (SOD) activity rose from 0 to 6 mmol.L-1 and peaked at 6 mmol.L-1. Calcium ion with the concentration of both 10 mmol.L-1 and 15 mmol.L-1 could increase the content of chlorophyll(Chl), stabilize the membrane structure of leaf discs, and reduce the membrane permeability under simulated acid rain with pH value of 3.0. The effect in 15 mmol.L-1 were better than in 10 mmol.L-1. However, the injury of acid rain to leaves was strengthened when the concentration of calcium was higher than 20 mmol.L-1. Net photosynthesis rate (Pn) rose when leaves sprayed with 15 mmol.L-1 Ca(NO3)2 before treatment of acid rain stress of pH 2.5. All of the results represented the excellent protection function of calcium on D. longana leaves under simulated acid rain.

Tunicamycin and Brefeldin A induce in plant cells a programmed cell death showing apoptotic features.

The recent identification of DAD (defender against apoptotic death) gene in plants suggests that the N-linked glycosylation of proteins could be an important control point of plant programmed cell death. In this paper we describe the effects of Tunicamycin, an inhibitor of N-linked protein glycosylation, and Brefeldin A, an inhibitor of protein trafficking from the Golgi apparatus, on sycamore (Acer pseudoplatanus L.) cell cultures. These two chemicals proved able to induce a strong acceleration of the cell death; changes in cell and nucleus morphology; an increase in DNA fragmentation, detectable by a specific immunological reaction; and the presence of oligonucleosomal-size fragments (laddering) in DNA gel electrophoresis. Moreover, Brefeldin A, but not Tunicamycin, strongly stimulated the production of hydrogen peroxide. These results indicate that also in plants chemicals interfering with the activities of endoplasmic reticulum and of Golgi apparatus strongly induce a form of programmed cell death showing apoptotic features.