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Huang J.-C.,CAS Kunming Institute of Botany | Zhong Y.-J.,University of Hong Kong | Zhong Y.-J.,Guangdong Academy of Agricultural Sciences | Liu J.,University of Hong Kong | And 2 more authors.
Metabolic Engineering | Year: 2013

Dietary carotenoids have been shown to be beneficial to health by decreasing the risk of many diseases. Attempts to enhance carotenoids in food crops have been successful although higher plants appear to resist big changes of carotenoid biosynthesis by metabolic engineering. Here we report the generation of a more nutritious tomato by modifying the intrinsic carotenes to astaxanthin, a high-value ketocarotenoid rarely found in plants. This was achieved by co-expression of the algal β-carotene ketolase from Chlamydomonas reinhardtii and β-carotene hydroxylase from Haematococcus pluvialis, a unique pair of enzymes identified to co-operate perfectly in converting β-carotene to astaxanthin by functional complementation in Escherichia coli. Expression of the two enzymes in tomato up-regulated most intrinsic carotenogenic genes, and efficiently directed carbon flux into carotenoids, leading to massive accumulations of mostly free astaxanthin in leaves (3.12. mg/g) but esterified astaxanthin in fruits (16.1. mg/g) and a 16-fold increase of total carotenoid capacity therein without affecting the plant normal growth and development. This study opened up the possibility of employing crop plants as green factories for economical production of astaxanthin. © 2013 Elsevier Inc.


Wang S.,CAS Institute of Genetics and Developmental Biology | Wang S.,South China Agricultural University | Li S.,CAS Institute of Genetics and Developmental Biology | Liu Q.,CAS Institute of Genetics and Developmental Biology | And 9 more authors.
Nature Genetics | Year: 2015

The deployment of heterosis in the form of hybrid rice varieties has boosted grain yield, but grain quality improvement still remains a challenge. Here we show that a quantitative trait locus for rice grain quality, qGW7, reflects allelic variation of GW7, a gene encoding a TONNEAU1-recruiting motif protein with similarity to C-terminal motifs of the human centrosomal protein CAP350. Upregulation of GW7 expression was correlated with the production of more slender grains, as a result of increased cell division in the longitudinal direction and decreased cell division in the transverse direction. OsSPL16 (GW8), an SBP-domain transcription factor that regulates grain width, bound directly to the GW7 promoter and repressed its expression. The presence of a semidominant GW7 TFA allele from tropical japonica rice was associated with higher grain quality without the yield penalty imposed by the Basmati gw8 allele. Manipulation of the OsSPL16-GW7 module thus represents a new strategy to simultaneously improve rice yield and grain quality. © 2015 Nature America, Inc.


Zhang M.W.,Cornell University | Zhang M.W.,Guangdong Academy of Agricultural Sciences | Zhang R.F.,Guangdong Academy of Agricultural Sciences | Zhang F.X.,Guangdong Academy of Agricultural Sciences | Liu R.H.,Cornell University
Journal of Agricultural and Food Chemistry | Year: 2010

Increased consumption of whole grains has been associated with reduced risk of developing major chronic diseases. These health benefits have been attributed in part to their unique phytochemicals. Previous studies on black rice mainly focused on anthocyanins. Little is known about the phytochemical profiles and antioxidant activities of different black rice varieties. The objective of this study was to determine the phytochemical profiles and antioxidant activity of rice bran samples from 12 diverse varieties of black rice. The free, bound, and total phenolic contents of black rice bran samples ranged from 2086 to 7043, from 221.2 to 382.7, and from 2365 to 7367 mg of gallic acid equiv/100 g of dry weight (DW), respectively. The percentage contribution of free phenolics to the total ranged from 88.2 to 95.6%. The average values of free, bound, and total phenolic contents of black rice bran were 8, 1.5, and 6 times higher than those of white rice bran, respectively (p < 0.05). The free, bound, and total flavonoid contents of black rice bran samples ranged from 3462 to 12061, from 126.7 to 386.9, and from 3596 to 12448 mg of catechin equiv/ 100 g of DW, respectively. The percentage contribution of free flavonoids to the total ranged from 96.3 to 97.6%. The average values of free, bound, and total flavonoid contents of black rice bran were 7.4, 1.9, and 6.7 times higher than those of white rice bran, respectively (p < 0.05). The free, bound, and total anthocyanin contents of black rice bran samples ranged from 1227 to 5096, from 4.89 to 8.23, and from 1231 to 5101 mg of cyanidin-3-glucoside equiv/100 g of DW, respectively. The percentage contribution of free anthocyanins to the total ranged from 99.5 to 99.9%. Cyanidin-3-glucoside, cyanidin-3-rutinoside, and peonidin-3-glucoside were detected in black rice bran samples and ranged from 736.6 to 2557, from 22.70 to 96.62, and from 100.7 to 534.2 mg/100 g of DW, respectively. The free, bound, and total antioxidant activities of black rice bran samples ranged from 476.9 to 180, from 47.91 to 79.48, and from 537.5 to 1876 μmol of Trolox equiv/g of DW, respectively. The percentage contribution of free antioxidant activity to the total ranged from 88.7 to 96.0%. The average values of free, bound, and total antioxidant activity of black rice bran were more than 8, 1.5, and 6 times higher than those of white rice bran, respectively (p< 0.05). The total antioxidant activity of black rice bran was correlated to the content of total phenolics, total flavonoids, and total anthocyanins and also was significantly correlated to the contents of cyanidin-3-glucoside, cyanidin-3-rutinoside, and peonidin-3-glucoside. These results indicate that there are significant differences in phytochemical content and antioxidant activity among the different black rice varieties. Black rice bran has higher content of phenolics, flavonoids, and anthocyanins and has higher antioxidant activity when compared to white rice bran. Interestingly, the phenolics, flavonoids, and anthocyanins of black rice bran are mainly present in free form. Knowing the phytochemical profile and antioxidant activity of black rice bran gives insights to its potential application to promote health. © 2010 American Chemical Society.


Wang L.,Cornell University | Wang L.,Guangdong Academy of Agricultural Sciences | Jiang Z.,Guangdong Academy of Agricultural Sciences | Lei X.G.,Cornell University
Free Radical Biology and Medicine | Year: 2012

We previously observed a stronger effect of knockout of Cu,Zn-superoxide dismutase (SOD1) than that of Se-dependent glutathione peroxidase 1 (GPX1) on murine body weight and glucose homeostasis. Two experiments were conducted to determine how hepatic lipid profiles and key metabolic regulators were correlated with this difference. SOD1-/- and GPX1-/- mice and their respective wild-type (WT) littermates (n=6 or 7/group, male) were fed a Se-adequate Torula yeast-sucrose diet and killed at 6 months of age to collect liver samples. In Experiment 1, fasted SOD1-/- mice displayed pyruvate intolerance and a 61 decrease (P<0.05) in liver glycogen compared with their WT littermates. The former had lower (P<0.05) activities of phosphoenolpyruvate carboxykinase, total protein phosphatase, and protein phosphatase 2A, but a higher (P<0.05) activity of glucokinase in the liver than the latter. In contrast, hepatic concentrations of total cholesterol, triglycerides, and nonesterified fatty acids were increased by 11 to 100 (P<0.05) in the SOD1-/- mice. Meanwhile, these mice had elevated (P<0.05) hepatic protein levels of sterol-regulatory element binding proteins 1 and 2, p53 MAPK, total and phosphorylated AMP-activated protein kinase α1 protein, protein tyrosine phosphatase 1B, and protein phosphatase 2B. In Experiment 2, GPX1-/- mice and their WT littermates were compared, but showed no difference in any of the measures. In conclusion, knockout of SOD1, but not GPX1, led to a decreased liver glycogen storage synchronized with pyruvate intolerance and elevated hepatic lipid profiles in adult mice. This striking comparison was possibly due to unique impacts of these two knockouts on intracellular tone of H2O2 and key regulators of liver gluconeogenesis, glycolysis, and lipogenesis. © 2012 Elsevier Inc. All rights reserved.


Wang L.,Zhejiang University | Lou G.,Zhejiang University | Ma Z.,Zhejiang University | Liu X.,Guangdong Academy of Agricultural Sciences
Food Chemistry | Year: 2011

Litchi (Litchi chinensis Sonn.) is widely accepted as a delicious fruit in China and its seeds have been commonly used in traditional Chinese medicine to relieve neuralgic pain. In the present study, chemical investigation of the 95% ethanol extract of Litchi chinensis seeds led to the isolation of four new compounds, 2α,3α-epoxy-5,7,3′,4′-tetrahydroxyflavan- (4β-8-catechin) (5), 2β,3β-epoxy-5,7,3′,4′- tetrahydroxyflavan-(4α-8-epicatechin) (7), litchiol A (9) and litchiol B (12), together with 11 known ones, 2,5-dihydroxy-hexanoic acid (1), soscopoletin (2), coumaric acid (3), protocatechuic acid (4), 2α,3α-epoxy-5,7, 3′,4′-tetrahydroxyflavan-(4β-8)-epicatechin (6), pterodontriol d-6-O-β-d-glucopyranoside (8), Narirutin (10), naringin (11), dihydrocharcone-4′-O-β-d-glucopyranoside (13), pinocembrin-7- rutinoside (14), pinocembrin-7-neohesperidoside (15). Their structures were mainly elucidated on the basis of NMR, MS, IR, CD and UV spectral evidences. Antioxidant activities of 14 compounds were determined by DPPH radical-scavenging assay and Trolox equivalent antioxidant capacity assay, and the results showed that four compounds, protocatechuic acid (4), 2α,3α-epoxy-5,7,3′,4′-tetrahydroxyflavan-(4β-8- catechin (5), 2α,3α-epoxy-5,7,3′,4′-tetrahydroxyflavan- (4β-8)-epicatechin (6), 2β,3β-epoxy-5,7,3′,4′- tetrahydroxyflavan-(4α-8)-epicatechin (7), exhibited moderate antioxidant activities. © 2010 Elsevier Ltd. All rights reserved.


Patent
Zhejiang University and Guangdong Academy of Agricultural Sciences | Date: 2015-06-04

Provided in the present invention are a separated endogenous rice gene resistant to high temperature (hereinafter referred to as the OsZFP gene for short) and a polypeptide encoded thereby, optimizing rice cells comprising the heat-resistant gene of the present invention or the polypeptide encoded thereby, and the plant cell preparation method thereof. Further provided are new methods and technologies for breeding new varieties of heat-resistant crops, comprising the related regulatory sequence for heat-resistance and a closely linked molecular marker denoting the heat-resistant gene and the sequence thereof.


Li D.M.,Guangdong Academy of Agricultural Sciences
Genetics and molecular research : GMR | Year: 2012

NAC proteins, which are plant-specific transcription factors, have been identified to play important roles in plant response to stresses and in plant development. The full-length cDNAs that encode 2 putative NAC proteins, designated as MmATAF1 and MmNAP, respectively, were cloned from Mikania micrantha by rapid amplification of cDNA ends. The full-length cDNAs of MmATAF1 and MmNAP were 1329 and 1072 bp, respectively, and they encoded deduced proteins of 260- and 278-amino acid residues, respectively. The proteins MmATAF1 and MmNAP had a calculated molecular mass of 29.81 and 32.55 kDa and a theoretical isoelectric point of 7.08 and 9.00, respectively. Nucleotide sequence data indicated that both MmATAF1 and MmNAP contained 2 introns and 3 exons and that they shared a conserved genomic organization. Multiple sequence alignments showed that MmATAF1 showed high sequence identity with ATAF1 of Arabidopsis thaliana (61%) and that MmNAP showed high sequence identity with NAP of A. thaliana (67%) and CitNAC of Citrus sinensis Osbeck (62%). Phylogenetic analysis showed that the predicted MmATAF1 and MmNAP proteins were classified into the ATAF and NAP subgroups, respectively. Transient expression analysis of onion epidermal cells indicated nuclear localization of both MmATAF1-GFP and MmNAP-GFP fusion proteins. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis indicated that MmATAF1 was expressed in all the tissues tested, but in varying abundance, while MmNAP was specifically expressed in stems, petioles, shoots, and leaves, but not in roots. The transcript levels of MmATAF1 and MmNAP in shoots and in infected stems were induced and strengthened by wounding, exogenous ZnSO(4), abscisic acid, salicylic acid, and Cuscuta campestris infection on the basis of semi-quantitative RT-PCR and real-time PCR analyses, respectively. Collectively, these results indicated that MmATAF1 and MmNAP, besides having roles in M. micrantha adaptation to C. campestris infection and abiotic stresses, also integrated signals derived from both C. campestris infection and abiotic stresses.


Gu W.,Guangdong Academy of Agricultural Sciences
Wei sheng wu xue bao = Acta microbiologica Sinica | Year: 2012

The aim of this study was to screen microorganisms that could degrade rice straw. We used selective medium to screen strains and determined straw fracture tension strength, weight loss, lignocellulose decomposition rate and extracellular enzyme activity as re-screening methods after 10 days shake flask culture. We isolated two antinomycetes (A3 and A6), the highest cellulose enzyme activity of holoenzyme, beta3-Glucosidase, endonuclease and exonclease for A3 were 12.84, 6.23, 24.56 and 14.00 U/mL, and for A6 12.85, 6.53, 17.80 and 18.80 U/mL. The hemicelluloses enzyme activity was 83.05 for A3 and 52.98 U/mL for A6. Both strains belonged to Streptomyces. With 10 days' treatment, inoculated straws showed a decrease of straw fracture tension strength by 62.67% (A3) and 66.67% (A6), while weight loss of straw was 31.50% (A3) and 35.83% (A6). A3's decomposition rate of cellulose, hemicellulose and lignin was 38.73% , 33.16% and 20.68% , and 47.69% , 28.64% and 22.59% for A6. Antinomycetes A3 and A6 could degrad cellulose, hemicellulose and lignin.


Luo C.,Guangdong Academy of Agricultural Sciences
BMC genomics | Year: 2013

Hyperpigmentation of the visceral peritoneum (HVP) has recently garnered much attention in the poultry industry because of the possible risk to the health of affected animals and the damage it causes to the appearance of commercial chicken carcasses. However, the heritable characters of HVP remain unclear. The objective of this study was to investigate the genetic parameters of HVP by genome-wide association study (GWAS) in chickens. HVP was found to be influenced by genetic factors, with a heritability score of 0.33. HVP had positive genetic correlations with growth and carcass traits, such as leg muscle weight (rg = 0.34), but had negative genetic correlations with immune traits, such as the antibody response to Newcastle disease virus (rg = -0.42). The GWAS for HVP using 39,833 single nucleotide polymorphisms indicated the genetic factors associated with HVP displayed an additive effect rather than a dominance effect. In addition, we determined that three genomic regions, involving the 50.5-54.0 Mb region of chicken (Gallus gallus) chromosome 1 (GGA1), the 58.5-60.5 Mb region of GGA1, and the 10.5-12.0 Mb region of GGA20, were strongly associated (P < 6.28 × 10-7) with HVP in chickens. Variants in these regions explained >50% of additive genetic variance for HVP. This study also confirmed that expression of BMP7, which codes for a bone morphogenetic protein and is located in one of the candidate regions, was significantly higher in the visceral peritoneum of Huiyang Beard chickens with HVP than in that of chickens without pigmentation (P < 0.05). HVP is a quantitative trait with moderate heritability. Genomic variants resulting in HVP were identified on GGA1 and GGA20, and expression of the BMP7 gene appears to be upregulated in HVP-affected chickens. Findings from this study should be used as a basis for further functional validation of candidate genes involved in HVP.


Zhang E.,Guangdong Academy of Agricultural Sciences | Chen X.,Guangdong Academy of Agricultural Sciences | Liang X.,Guangdong Academy of Agricultural Sciences
Electrophoresis | Year: 2011

In this short communication, a novel protocol for resolubilization of TCA-precipitated plant proteins for 2-DE is described. Guanidine hydrochloride (Gdn-HCl) is used as an intermediate for protein solubilization in which proteins are reduced and alkylated with tributylphosphane (TBP) and 2-vinylpyridine (2-VP). The blocking of -SH groups at Cys residues can greatly improve the solubility of TCA-precipitated proteins and obtain more high-quality protein spots in the 2-DE gel. This protocol is compatible with silver stain and MS identification. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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