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Identification of polysaccharides from pericarp tissues of litchi (Litchi chinensis Sonn.) fruit in relation to their antioxidant activities.

A large number of polysaccharides are present in the pericarp tissues of harvested litchi fruits. A DEAE Sepharose fast-flow anion-exchange column and a Sephadex G-50 gel-permeation column were used to isolate and purify the major polysaccharides from litchi fruit pericarp tissues. Antioxidant activities of these major polysaccharide components were also evaluated. An aqueous extract of the polysaccharides from litchi fruit pericarp tissues was chromatographed on a DEAE anion-exchange column to yield two fractions. The largest amount of the polysaccharide fraction was subjected to further purification by gel filtration on Sephadex G-50. The purified product was a neutral polysaccharide, with a molecular weight of 14 kDa, comprised mainly of 65.6% mannose, 33.0% galactose and 1.4% arabinose. Analysis by Smith degradation indicated that there were 8.7% of (1-->2)-glycosidic linkages, 83.3% of (1-->3)-glycosidic linkages and 8.0% of (1-->6)-glycosidic linkages in the polysaccharide. Furthermore, different polysaccharide fractions extracted and purified from litchi fruit pericarp tissues exhibited strong antioxidant activities. Among these fractions, the purified polysaccharide had the highest antioxidant activity and should be explored as a novel potential antioxidant.

Antioxidants↗

[Ecological control effects of Litchi chinensis-Desmodium intortum complex plant ecosystem on litchi pests].

An investigation on the community structure and dynamics of litchi pests and their natural enemies in constructed Litchi chinensis-Desmodium intortum complex plant ecosystem and single L. chinensis ecosystem showed that the total amount of litchi pests in the complex plant ecosystem was 61.27% of that in the single ecosystem in whole year, and only 50.45% in May, the key time for fruit development, which suggested that there was an interaction between D. intortum and L. chinensis. D. intortum and L. chinensis had a few common pests, but many common natural enemies. D. intortum florescence in winter provided shelter and substitutive food for the natural enemies of pests to survive in the extreme environmental conditions in winter. L. chinensis florescence was on the heel of D. intortum florescence, which provided better conditions for the natural enemies to survive and multiply. During florescence and fruit development stages of L. chinensis (from March to June), the predator/prey ratio in complex plant system was 4.22, 2.34, 2.2 and 20.63 times of that in single plant system in March, April, May and June, respectively, indicating the good control effect on pests of L. chinensis.

Animals↗

Immunomodulatory and anticancer activities of flavonoids extracted from litchi (Litchi chinensis Sonn) pericarp.

The litchi pericarp extract was subjected to partition by hexane, ethyl acetate and water. Epicatechin, proanthocyanidin B2 and proanthocyanidin B4 were isolated and purified from the ethyl acetate fraction by reverse-phase high performance liquid chromatography. The immunomodulatory activities of epicatechin, proanthocyanidin B2, proanthocyanidin B4 and the ethyl acetate fraction were examined using proliferation of mouse splenocytes. The results showed all these samples had much higher stimulatory effects on splenocyte proliferation than that of the reference, rutin. Epicatechin and the ethyl acetate fraction showed a significantly (P<0.05) stimulatory effect when the concentration was up to 12.5 micro g/ml. Proanthocyanidin B2 and proanthocyanidin B4 exhibited little lower stimulatory effects than epicatechin and the ethyl acetate fraction. The anti-breast cancer activities of epicatechin, proanthocyanidin B2, proanthocyanidin B4 and the ethyl acetate fraction were also evaluated. Epicatechin and proanthocyanidin B2 had lower cytotoxicities to human breast cancer cell MCF-7 and human embryolic lung fibroblast than paclitaxel.

Acetates↗

[Development of SSR markers in Litchi( Litchi chinensis)].

A total of 100 SSR sequences were isolated and cloned by means of SAM (Selectively Amplified Microsatellite)techniques and another one was obtained by searching the NCBI and EMBL databases. There were 89 SSR sequences used for design of special primers. As a result, the primers were designed at 82 loci from 71 fragments. Forty-one special primers were synthesized, pairing with 5'anchored degenerate SSR primer, to detect 39 SSR loci. Fifteen of them amplified the corresponding SSR sequences and Other 11 SSR primer pairs amplified non-expected fragments. In the end, 21 polymorphic primer pairs were selected from 26 primer pairs which amplified clear and robust DNA fragment by using the genome DNA of 37 litchi germplasm materials, and 22 locus-specific SSR markers were obtained.

Base Sequence↗

Distribution changes of calcium and programmed cell death in the pistil of litchi (Litchi chinensis Sonn.) flower during its development.

Potassium pyroantimonate precipitation method was used for investigating calcium distribution and cell ultrastructure change during development of pistils of litchi male and female flower. The results showed that at the megasporocyte stage of female flowers, calcium precipitates was located mainly at cell wall and intercellular space of inner integument near the micropyle and style cells, and to a lesser extent in vacuoles. Vascular tissues also contained much calcium precipitates. In inner integument cells near the micropyle of male flowers, the vacuole contained most of the calcium precipitates. Calcium precipitates in style cell and vascular tissues of male flowers was sparse and seldom seen. After meiosis of megasporocyte, pistils of female flowers continued to grow and those of male flowers aborted. In female flowers, calcium precipitates concentration became lower and calcium precipitates was probably transported to the places for future pollen bourgeoning and fertilization. Cell wall calcium precipitates concentration increased in the inner integument cells near the micropyle. Calcium precipitates concentration increased from topper style cells to lower ones. In male flowers, inner integument cells near the micropyle underwent the programmed cell death (PCD): flow of calcium from vacuoles into nucleus might had triggered the PCD process. A continuous channel was formed between perinuclear space and cytoplasm membrane lumen, and calcium flowed freely between nuclear membrane and plasma membrane. At certain time and locations, calcium precipitates was newly appeared at some organelles like endoplasimic reticulum, mitochondria and peroxisomes. This calcium redistribution in cells might trigger and regulate the process of PCD. In male flowers, style cells containing no calcium precipitation soon began to degenerate.

Apoptosis↗

Phenolic composition of litchi fruit pericarp.

Litchi (Litchi chinensis, Sapindaceae) is a nonclimacteric subtropical fruit that, once harvested, loses its red pericarp color because of browning reactions probably involving polyphenols. Low-pressure chromatography, high-pressure liquid chromatography, UV-visible spectral analysis, mass spectrometry, and nuclear magnetic resonance studies have allowed the determination and quantification of the polyphenolic composition of litchi pericarp. Litchi skins contain significant amounts of polyphenolic compounds. The principal characteristic of the litchi skin polyphenolic compounds is their ortho-diphenolic structure, which gives them high oxidability. Four major pigments were formally identified as cyanidin 3-rutinoside, cyanidin glucoside, quercetin 3-rutinoside (rutin), and quercetin glucoside. The tannin content was characterized after the depolymerization thiolysis reaction. Tannins (polymeric proanthocyanidins) are mainly constituted with epicatechin units linked by A- and B-type bonds. The different phenolic compounds of litchi cv. Kwai Mi were quantified by HPLC. Condensed tannins were the most abundant (4 mg x g(-1) of fresh skin), followed by epicatechin and procyanidin A2 (1.7 and 0.7 mg x g(-1) of fresh pericarp, respectively). The amount of anthocyanins was found to be comparable to that of flavonols, with a value of approximately 0.4 mg x g(-1) of fresh pericarp.

Chromatography, Liquid↗

Differential expression of litchi XET genes in relation to fruit growth.

Xyloglucan endotransglycosylase (XET) catalyses the transglycosylation of xyloglucan, the major hemicellulose polymer, which has been thought to mediate the cross-linking of cellulose microfibrils in cellular walls and proposed to be involved in the control of cell wall relaxation. To understand the relationship between litchi fruit cracking and gene expression patterns, three XET genes from litchi fruit were identified and then examined for their expression profiles in pericarp and aril tissues at different development stages, using a cracking-resistant cultivar, 'Huaizhi', and a cracking-susceptible cultivar, 'Nuomici'. Three full-length cDNAs of 1267, 1095 and 1156 bp encoding XETs, named LcXET1, LcXET2 and LcXET3, respectively, were isolated from expanding fruit using RT-PCR and RACE-PCR (rapid amplification of cDNA ends) methods. Northern blotting analysis showed that LcXET1 mRNA accumulation occurred much earlier in aril tissues at 59 days after anthesis (DAA) than in pericarp tissues at 73 DAA in 'Nuomici'. However, it appeared at almost the same time (66 DAA) in pericarp and aril tissues in 'Huaizhi', which suggested that differential accumulation of LcXET1 in pericarp and aril tissues in 'Nuomici' and 'Huaizhi' was closely associated with fruit cracking. LcXET2 mRNA accumulation could be detected in pericarp and aril tissues throughout fruit development but exhibited a differential accumulation pattern between pericarp and aril tissues. In the aril of 'Nuomici', intensive signal bands were detectable at 59-73 DAA in rapidly expanding fruits of 'Nuomici' but only weak bands could be found in the pericarp tissues. In contrast, moderate signal bands were detectable both in pericarp and aril tissues of 'Huaizhi' fruits. Furthermore, LcXET3 showed constitutive expression in both pericarp and aril tissues of developing 'Nuomici' and 'Huaizhi' litchi fruit. In addition, differential expression patterns of three XETs genes were observed in different tissues of litchi, with only LcXET1 being fruit-specific. To further address the role of LcXET in fruit cracking, alpha-naphthalene acetic acid (NAA) was used to treat 'Nuomoci' to reduce fruit cracking. Enhanced LcXET1 mRNA accumulation appeared in pericarp while LcXET2 and LcXET3 mRNA accumulation enhanced in aril tissues in the NAA-treated fruits. Thus, LcXET1 is more likely to play a role in reducing litchi fruit cracking than LcXET2 and LcXET3.

Amino Acid Sequence↗

[Photosynthesis and free radical yield of Litchi chinensis leaves under increased CO2 partial pressure in atmosphere].

The maximum photosynthetic rate of litchi saplings leaves grown under a CO2 partial pressure of 77 +/- 5 Pa was 23% lower than that of 39.3 Pa, and slight decreases in respiration rate in light and CO2 compensation point excluding of respiration in light were observed. The maximum rates of carboxylation (Vcmax) and photosynthetic electron transport (Jmax) were decreased in saplings grown under 77 +/- 5 Pa CO2 partial pressure. It may suggest that there was a lower energy level of photosystem I(PSI) in the saplings leaves under 77 +/- 5 Pa CO2 partial pressure, and free radical yield decreased by 39% in the saplings under 77 +/- 5 Pa CO2, as comparied with that under atmospheric CO2. The percentage of infection by Peronophythora litchi increased from 1.8% under atmospheric CO2 to 9.5% under enriched CO2. It may mean that the decreasing photosynthetic and respiration metabolism would make it lower the production of O2-., and leaves were infected by P. litchi more easily. The controlling of the popularization of P. litchi for litchi plantation must be paid wide attention, as air CO2 partial pressure will increase continuously.

Carbon Dioxide↗

[A comparative study of postharvest fruit senescence in different litchi cultivars].

Among the 5 tested litchi (Litchi chinensis Sonn.) cultivars ("Huaizhi", "Guiwei", "Nuomici", "Hongmili" and "Shuijingqiu", "Nuomici" became deteriorated much faster than other cultivars while "Guiwei" fruit was the slowest in the rotting process (Fig. 1A). Fruit deterioration was accompanied by fruit desiccation (Fig. 2B), but the speed of water loss was not significantly correlated to fruit deterioration rate, indicating that it was not the key factor causing the difference in postharvest performance among cultivars. Fruit deterioration rate was significantly positively correlated to membrane leakage (Fig. 2A), suggesting the capacity to maintain membrane integrity is closely related to the shelflife of litchi. Skin browning potential, uronic acid concentration, degree of methylation of pectin and soluble Ca content in pericarp as well as total Ca content in the pulp were not significantly correlated with fruit deterioration. Content of structural Ca (water-insoluble but acetic acid-soluble calcium, membrane or wall-bound Ca), the major form of Ca in the pericarp, was negatively correlated to fruit deterioration rate (Fig. 2E). The results proved that differences in fruit desiccation rate, browning potential, Ca other than structural form were not the major cause leading to difference in postharvest performance among different cultivars. "Guiwei" being more tolerant to desiccation than other cultivars is likely associated its higher structural Ca concentration in the pericarp.

Calcium↗

Litchi chinensis fatty acid diversity: occurrence of the unusual cyclopropanoic fatty acids.

Litchi chinensis (Sapindaceae) is a tree that originates from China and is cultivated for its sweet fruits all over the world in warm climates. Unusual fatty acids such as cyclopropanoic fatty acids have been identified in the seeds of Litchi. Because of their potential value for industry (as inks, cosmetics, detergents, lubricants, etc.), the variability in the relative levels of unusual fatty acids in the seeds of 28 different Litchi varieties was analysed at two locations (on Réunion Island in the Indian Ocean) and on two different harvest dates. Except for one variety, all the seeds contained cis-9,10-methylene-octadecanoic acid (C(19)CA) at a relative level of 35-48%. The only variety that contained no or only traces of C(19)CA was Groff, seeds of which were significantly much smaller than those from all other varieties.

China↗

[Effects of bagging on the fruit quality in Litchi chinensis fruit and pesticide residues in it].

Different color bags were used to cover Litchi chinensis fruit to study the effects of bagging on its quality and pesticide residues. The results showed that bagging significantly improved the fruit color. Using bagging techniques, the I and II class fruit was accounted for 57.87% to 81.57%, 30% higher than that of control, and the weight per fruit increased significantly. Among the color bags used, white bag was the best. When the end of the bag was opened, it was benefit for decease control, but not good for pest control. When the end of the bag was closed, it was good for pest control, but bad for decease control. Bagging had no effects on the taste of Litchi chinensis fruit, but might increase the residues of fenpropathrin and trichlorphon. More study should to be carried out to select the suitable pesticides accompanied with bagging techniques.

Color↗

Development, characterization and variability analysis of microsatellites in lychee (Litchi chinensis Sonn., Sapindaceae).

We report 12 microsatellites enriched in CT repeats obtained from a genomic library of the lychee ( Litchi chinensis Sonn.) cultivar Mauritius. The polymorphisms revealed by these microsatellites were evaluated in a collection of 21 lychee cultivars. A total of 59 fragments were detected with these 12 SSRs, with an average of 4.9 bands/SSR. Three primer pairs seem to amplify more than a single locus. The mean expected and observed heterozygosities over the 9 single-locus SSRs averaged 0.571 (range: 0.137-0.864) and 0.558 (range: 0.169-0.779) respectively. The total value for the probability of identity was 7.53 x 10(-5). In addition, the selected SSRs were used to amplify DNA from four longan cultivars. Eleven of the 12 SSRs produced amplification fragments in longan, and eight of these fragments were polymorphic. All except two of the products amplified from longan were the same size as those amplified from lychee, suggesting a close genetic proximity between the two species. The SSRs studied produced 22 different patterns, allowing the unambiguous identification of 16 lychee and the 4 longan cultivars studied. Discrimination was possible with just four selected microsatellites. Two groups with two and three undistinguishable cultivars were obtained, reflecting probable synonymies. Unweighted pair-group method of artimetic averages (UPGMA) cluster analysis divided the lychee cultivars studied into two main groups, one consisting of ancient cultivars and the other with more diverse recent cultivars. This is the first report of microsatellite development in the Sapindaceae, and the results demonstrate the usefulness of microsatellites for identification, similarity studies and germplasm conservation in lychee and related species.

Litchi↗

Potential anticancer activity of litchi fruit pericarp extract against hepatocellular carcinoma in vitro and in vivo.

Litchi fruit pericarp (LFP) extract contains significant amounts of polyphenolic compounds, and exhibits powerful antioxidative activity against fat oxidation in vitro. The purpose of this study is to confirm the anticancer activity of LFP extract against hepatocellular carcinoma in vitro and in vivo, and to elucidate the mechanism of its activity. Human hepatocellular carcinoma cell line was tested in vitro for cytotoxicity, colony formation inhibition, and cell cycle distribution through flow cytometry after treatment with water-soluble crude ethanolic extract (CEE) from LFP. Murine hepatoma bearing-mice were fed doses of 0.15, 0.3, and 0.6g/kg/day of water-soluble CEE in DH(2)O p.o. for 10 days, respectively, to test the anticancer activity and BrdU incorporation of cancer cells in vivo. LFP extract demonstrated a dose- and time-dependent inhibitory effect on cancer cell growth; IC(50) was 80microg/ml, and significantly inhibited colony formation in vitro, tumor growth and BrdU incorporation into cancer cells in vivo. The tumor inhibitory rates at doses of 0.15, 0.3, and 0.6g/kg/day were 17.31% (P>0.05), 30.77% (P<0.05), and 44.23% (P<0.01), respectively. BrdU labeled tumor cells of treated animals were 11.80+/-2.79%, and were significantly lower than that in untreated controls (23.00+/-5.42%, P<0.05). Our findings showed that LFP extract exhibited potential anticancer activity against hepatocellular carcinoma in vitro and in vivo through proliferating inhibition and apoptosis induction of cancer cells.

Animals↗

Anticancer activity of litchi fruit pericarp extract against human breast cancer in vitro and in vivo.

Litchi fruit pericarp (LFP) extract contains significant amounts of polyphenolic compounds and exhibits powerful antioxidative activity against fat oxidation in vitro. The purpose of this study is to confirm the anticancer activity of LFP extract on human breast cancer in vitro and in vivo, and to elucidate the mechanism of its activity. Human breast cancer cells were tested in vitro for cytotoxicity, colony formation inhibition, BrdU incorporation, and gene expression profiling after treatment with LFP extract. Seven nude mice bearing human breast infiltrating duct carcinoma orthotopically were tested for its anticancer activity and expression of caspase-3 in vivo by oral administration of 0.3% (0.3 mg/ml) of LFP water-soluble crude ethanolic extract (CEE) for 10 weeks. LFP extract demonstrated a dose- and time-dependent inhibitory effect on cell growth (IC(50) = 80 microg/ml), and it significantly inhibited colony formation and BrdU incorporation of human breast cancer cells. Oligonucleotide microarray analysis identified 41(1.22%) up-regulated and 129 (3.84%) down-regulated genes after LFP water-soluble CEE treatment; the predominantly up-regulated genes were involved in various biological functions including cell cycle regulation and cell proliferation, apoptosis, signal transduction and transcriptional regulation, and extracellular matrix/adhesion molecules; and down-regulated genes were mainly associated with adhesion, invasion, and malignancy of cancer cells. A 40.70% tumor mass volume reduction and significant increase of casepase-3 protein expression were observed in vivo experiment. The findings in this study suggested that LFP extract might have potential anticancer activity on both ER positive and negative breast cancers, which could be attributed, in part, to its DNA damage effect, proliferating inhibition and apoptosis induction of cancer cells through up-regulation and down-regulation of multiple genes involved in cell cycle regulation and cell proliferation, apoptosis, signal transduction and transcriptional regulation, motility and invasiveness of cancer cells; ADP-ribosyltransferase (NAD+; poly (ADP-ribose) polymerase)-like 1 (ADPRTL1), Cytochrome P450, subfamily I (CYP1A1) and Hyaluronan-mediated motility receptor (HMMR) might be the main molecular targets at which LFP water-soluble CEE acted.

Animals↗

Issatchenkia hanoiensis, a new yeast species isolated from frass of the litchi fruit borer Conopomorpha cramerella Snellen.

The new ascogenous yeast species Issatchenkia hanoiensis was discovered in the frass of the litchi fruit borer Conopomorpha cramerella Snellen. The yeast forms unconjugated persistent asci containing one to two roughened ascospores. The yeast has a CoQ-7 system, which is typical for the genus Issatchenkia. The closest species to I. hanoiensis as indicated by analysis of the partial ribosomal DNA large-subunit (D1/D2) sequence is the asexual species Candida pseudolambica. The two share 94.2% similarity in the sequenced region. Other species of Issatchenkia were also among the closest relatives of I. hanoiensis, the level of similarity ranging from 89.8% to 94.1%. The type culture is strain HB1.3.13=CBS 9198=NRRL Y-27509.

Animals↗

Shoot development, chlorophyll, gas exchange and carbohydrates in lychee seedlings (Litchi chinensis).

Shoot growth, chlorophyll concentrations, gas exchange and starch concentrations were studied in lychee (Litchi chinensis Sonn.) seedlings of cultivar "Wai Chee" grown in a heated greenhouse at Nambour in subtropical Australia (27 degrees S). We also examined the effects of shoot defoliation and root pruning on leaf expansion. Shoot growth showed a rhythmic cycle under constant greenhouse conditions, with a mean duration of flushing of 20 days and an interval of 10 days over three cycles. Shoots and leaves expanded in a sigmoidal pattern to about 80 mm and 500 cm(2), respectively, for each flush. Starch concentrations of the lower stem and roots decreased as the young red leaves expanded, and increased as the fully expanded leaves turned dark green. Chlorophyll concentrations and net CO(2) assimilation rate were highest in the fully expanded dark green leaves. Removing 50% of the area of each fully expanded leaf had little effect on the expansion of younger leaves, but total biomass of defoliated plants was only 60% of that of controls. In contrast, removing half the roots just before bud swelling reduced final leaf area by 80%. We conclude that the young shoot has relatively low rates of photoassimilation until the leaves are fully expanded and dark green, and depends on assimilates from elsewhere in the plant. During leaf expansion, translocation of assimilates to the shoot occurred at the expense of the roots.

Biomass↗

Effects of leaf, shoot and fruit development on photosynthesis of lychee trees (Litchi chinensis).

Changes in gas exchange with leaf age and fruit growth were determined in lychee trees (Litchi chinensis Sonn.) growing in subtropical Queensland (27 degrees S). Leaves expanded in a sigmoid pattern over 50 days during spring, with net CO2 assimilation (A) increasing from -4.1 +/- 0.9 to 8.3 +/- 0.5 micromol m-2 s-1 as the leaves changed from soft and red, to soft and light green, to hard and dark green. Over the same period, dark respiration (Rd) decreased from 5.0 +/- 0.8 to 2.0 +/- 0.1 micromol CO2 m-2 s-1. Net CO2 assimilation was above zero about 30 days after leaf emergence or when the leaves were half fully expanded. Chlorophyll concentrations increased from 0.7 +/- 0.2 mg g-1 in young red leaves to 10.3 +/- 0.7 mg g-1 in dark green leaves, along with stomatal conductance (gs, from 0.16 +/- 0.09 to 0.47 +/- 0.17 mol H2O m-2 s-1). Fruit growth was sigmoidal, with maximum values of fresh mass (29 g), dry mass (6 g) and fruit surface area (39 cm2) occurring 97 to 115 days after fruit set. Fruit CO2 exchange in the light (Rl) and dark (Rd) decreased from fruit set to fruit maturity, whether expressed on a surface area (10 to 3 micromol CO2 m-2 s-1 and 20 to 3 micromol CO2 m-2 s-1, respectively) or on a dry mass basis (24 to 2 nmol CO2 g-1 s-1 and 33 to 2 nmol CO2 g-1 s-1, respectively). Photosynthesis never exceeded respiration, however, the difference between Rl and Rd was greatest in young green fruit (4 to 8 micromol CO2 m-2 s-1). About 90% of the carbon required for fruit growth was accounted for in the dry matter of the fruit, with the remainder required for respiration. Fruit photosynthesis contributed about 3% of the total carbon requirement of the fruit over the season. Fruit growth was mainly dependent on CO2 assimilation in recently expanded dark green leaves.

Carbon Dioxide↗