Transcriptome Analysis of Inflorescence Development at the Five-Leaf Stage in Castor(Ricinus communis L.)

The yield of castor is influenced by the type of inflorescence and the proportion of female flowers.However,there are few studies on the genetic mechanism involved in the development and differentiation of castor inflorescences.In this study,we performed transcriptomic analyses of three different phenotypes of inflorescences at the five-leaf stage.In comparison to the MI(complete pistil without willow leaves),290 and 89 differentially expressed genes(DEGs)were found in the SFI(complete pistil with willow leaves)and the BI(monoecious inflorescence),respectively.Among the DEGs,104 and 88 were upregulated in the SFI and BI,respectively,compared to the MI.In addition,186 DEGs and 1 DEG were downregulated in the SFI and BI compared to the MI.Moreover,we conducted GO and KEGG enrichment analyses of the DEGs.In comparison to the MI,the SFI and BI exhibited the enrichment of functional branches in DEGs,specifically in pollen wall assembly,pollen development,and cellular component assembly involved in morphogenesis.In our study,RADL5 showed low expression levels between SFI-vs.-MI types.In addition,we found that the expression of NAC in the SFI differed from that in MI and BI,and some genes related to hormonal signaling changed their expression levels during inflorescence differentiation.These results reveal the genetic mechanism of sex genotypes in castor,which will not only guide researchers in the breeding of castor but also provide a reference for genetic research on other flowering plants.

The PcERF5 promotes anthocyanin biosynthesis in red-fleshed pear(Pyrus communis)through both activating and interacting with PcMYB transcription factors

As there is a strong interest in red-skinned pears,the molecular mechanism of anthocyanin regulation in red-skinned pears has been widely investigated;however,little is known about the molecular mechanism of anthocyanin regulation in red-fleshed pears due to limited availability of such germplasm,primarily found in European pears(Pyrus communis).In this study,based on transcriptomic analysis in red-fleshed and white-fleshed pears,we identified an ethylene response factor(ERF)from P.communis,PcERF5,of which expression level in fruit flesh was significantly correlated with anthocyanin content.We then verified the function of PcERF5 in regulating anthocyanin accumulation by genetic transformation in both pear skin and apple calli.PcERF5 regulated anthocyanin biosynthesis by different regulatory pathways.On the one hand,PcERF5 can activate the transcription of flavonoid biosynthetic genes(PcDFR,PcANS and PcUFGT)and two key transcription factors encoding genes PcMYB10 and PcMYB114.On the other hand,PcERF5 interacted with PcMYB10 to form the ERF5-MYB10 protein complex that enhanced the transcriptional activation of PcERF5 on its target genes.Our results suggested that PcERF5 functioned as a transcriptional activator in regulating anthocyanin biosynthesis,which provides new insights into the regulatory mechanism of anthocyanin biosynthesis.This new knowledge will provide guidance for molecular breeding of red-fleshed pear.

Proteomic Analyses of Three Inflorescence Styles of Castor(Ricinus communis L.)at Different Developmental Stages

Castor(Ricinus communis L.)is one of ten oil crops in the world and has complex inflorescence styles.Generally,castor has three inflorescence types:single female inflorescence(SiFF),standard female inflorescence(StFF)and bisexual inflorescence(BF).StFF is realized as a restorer line and as a maintainer line,which was applied to castor hybrid breeding.However,the developmental mechanism of the three inflorescences is not clear.Therefore,we used proteomic techniques to analyze different inflorescence styles.A total of 72 diferentially abundant protein species(DAPs)were detected.These DAPs are primarily involved in carbon and energy metabolism and carbon fixation in the photosynthetic organism pathway.The results showed that DAPs are involved in photosynthesis to control the distribution of imported carbohydrates and exported photoassimilates and thus affect the inflorescence development of castor.In addition,these DAPs are also involved in cysteine and methionine metabolism.Quantitative real-time PCR(qRT-PCR)results demonstrated that the proteomics data collected in this study were reliable.Our findings indicate that the carbon cycle and amino acid metabolism influence the inflorescence development of castor.

Comparison of Some Characteristics Between Phragmites communis and Its Salt Tolerant Variant

R5002_12, a salt tolerant line of Phragmites communis Trin., which was obtained from ethyl methane sulfonate (EMS) treated callus selected under saline stress, was compared with its wild line in respect to their molecular biological, physiological and biochemical characterizations. Five arbitrary primers were screened which showed differences in DNA amplified polymorphism between the variant and its wild line. Some new proteins appeared in the salt tolerant plant under salt stress. Electrophoresis of peroxidase and esterase also showed some differences in isozyme expression between them. The chlorophyll content of the variant was higher than that of the original variety, whether the plants were under salt stress or not.

鸭跖草Commelina communis中差异表达cDNA片段的克隆与分析

一种新的基因克隆技术称为抑制消减杂交技术(SSH)用于研究生长于2种生态环境(古铜矿山和正常土壤)中鸭跖草Commelina communis的基因表达差异。分别以Cu矿山鸭跖草作为检测子,非Cu矿山鸭跖草作为驱赶子,建立了生长于Cu矿山生态环境中鸭跖草的差异表达cDNA文库,此cDNA文库代表在Cu矿山鸭跖草中特异表达的cDNA。通过反式Northern杂交筛选出3个阳性克隆进行测序。序列分析表明其中一个cDNA为CaM-like基因。进一步对此基因进行Virtual Northern分析,结果初步表明此基因在生长于古铜矿山上的鸭跖草中上调表达。

Inorganic Elements in Kernel of Amygdalus communis L. Measured Using ICP-OES Method

[Objective] The aim was to study on distribution of inorganic elements in kernel of Amygdalus communis L., providing reference for quality evaluation of A. communis L. species. [Method] Totally 26 species of inorganic elements in kernel, including Al, B, Be, Ca, Co, Cu, Fe, Mg, Mn, Mo, Na, Ni, P, Pb, Si, Sn, Sr, Ti, Zn, Cd, As, Se, V, Hg, Cr and K were measured with inductively coupled plasma emission spectrum （ICP-OES） and principal components analysis （PCA）. [Result] A. communis L. of different species and in different factories showed a similar curve in content of inorganic elements; absolute contents of the elements differed significantly. In addition, the accumulated variance contribution of five principle factors achieved as high as 84.371% and the variance contribution made by the first three factors accounted for 67.546%, proving that Fe, Ti, Pb, Na, Se, Cu, Mo, K, Zn, Ni, Ca and Sr were characteristic elements. [Conclusion] The method, which is brief, rapid and accurate, can be used for determination of inorganic elements in kernel of A. communis L., providing theoretical references for further development and utilization of A. communis L.

Study on Cu Resistance of Ricinus communis L. Callus

[Objective] The article studies the growth and Cu absorption of Ricinus communis L. callus under Cu stress. [Method] CuSO4.5H20 solutions with different Cu concentrations were added to callus subculture medium; callus was inoculated and Cu resistance index of callus was worked out. Cu content in callus was deter- mined with the method of Varian AA240FS. [Result] With the Cu concentration at 60 mg/L, the growth of callus was inhibited, its Cu resistance index was only 33.87%. With the Cu concentration at 40 mg/L, callus was faint yellow in color, and grew rapidly with its Cu resistance index at 61.29%. Such high level resistance could remain the same after six week after continuous subculture. In the 4t week of culture, Cu resistance index in treatments with Cu concentrations at 10, 20, 30, 40 mg/L was higher than that in the 3rd week, and the content of Cu in callus of the treatments was 0.33, 0.54, 1.16, 1.40 mg/g respectively. [Conclusion] Cu con- centration at 40 mg/L in culture medium can be the threshold for selecting Cu re- sistance R. communis callus.

Exploration of larvicidal and adult emergence inhibition activities of Ricinus communis seed extract against three potential mosquito vectors in Kolkata,India

Objective:To determine the larvicidal and adult emergence inhibition activities of castor （Ricinus communis） seed extract against three potential mosquito vectors Anopheles stephensi（An. slephensi）,Culex quinquefasciatus（Cx.quinquefasciatus） and Aedes albopictus（Ae.albopictus） in India.Methods:The R.communis seed extract was tested,employing WHO procedure,against fourth larval instars of the three mosquito species for 24 h and larval mortalities were recorded at various concentrations（2-64μg/mL）:the 24 h LC50 values of the R.communis seed extract were determined following Probit analysis.The larval killing,antipupation and adult emergence inhibition rates of the test extract,using a single concentration of 2μLC50,were studied at different time periods（24-72 h）:the extract toxicity was tested against a fish.Oreochromis niloticus（O.niloticus）.Results:The R.communis seed extract exhibited larvicidal effects with 100%killing activities at concentrations 32-64μg/mL,and with LC50 values 7.10.11.64 and 16.84μg/mL for Cx.quinquefasciatus,An.stephensi and Ae.albopictus larvae,respectively. When the larvae were treated with the extract at a single concentration of 2×LC50,significant differences were observed,compared to control groups,in rate of pupation（P【0.001） as well as in adult formation（P【0.001）.Conclusions:The present findings suggest that the R.communis seed extract provided an excellent potential for controlling An.stephensi,Cx.quinquefasciatus and Ae. albopictus mosquito vectors.

Commelina communis L.: Copper Hyperaccumulator Found in Anhui Province of China

Commelina communis L. growing over some new copper mining wastelands at Bijiashan, Tongling City of Anhui Province, China, was found to be a copper hyperaccumulator. Its copper concentrations were 2707-6159 (4439±2434) mg kg-1, 369-831 (731±142) mg kg-1, and 429-587 (547±57) mg kg-1, respectively, in the roots, stems, and leaves. The soils supporting the growth of the species had a copper concentration ranging from 4620 to 5020 mg kg-1 and averaging 4835±262 mg kg-1, suggesting that the species could not only grow on heavily copper-contaminated soils but also accumulate extraordinarily high concentration of copper. Thus, it shows great potential in the phytoremediation of copper-contaminated soils,the restoration of mined land, geochemical prospecting, and the study of environmental pollution changes.

Antimicrobial potential of Ricinus communis leaf extracts in different solvents against pathogenic bacterial and fungal strains

Objective:To investigate the in vitro antimicrobial activities of the leaf extract in different solvents viz.,methanol,ethanol and water extracts of the selected plant Ricinus communis.Methods:Ager well diffusion method and agar tube dilution method were carried out to perform the antibacterial and antifungal activity of methanol,ethanol and aqueous extracts.Results:Metbanol leaf extracts were found to be more active against Gram positive bacteria(Bacillus subtilis:ATCC 6059 and Staphylococcus aureus:ATCC 6538)as well as Gram negative bacteria(Pseudomonas aeruginosa:ATCC 7221 and Klebsiella pneumoniae)than ethanol and aqueous leaf extracts.Antifungal activity of methanol and aqueous leaf extracts were also carded out against selected fungal strains as Aspergillus fumigatus and Aspergillus flavus.Methanolic as well as aqueous leaf extracts of Ricinus communis were effective in inhibiting the fungal growth.Conclusions:The efficient antibacterial and antifungai activity of Ricinus communis from the present investigation revealed that the methanol leaf extracts of the selected plant have significant potential to inhibit the growth of pathogenic bacterial and fungal strains than ethanol and aqueous leaf extracts.

In vitro control of Staphylococcus aureus (NCTC 6571) and Escherichia coli (ATCC 25922) by Ricinus communis L.

Objective:To evaluate antibacterial activity of hot and cold ethanol and methanol leaf extracts of Ricinus communis L(R.communis)against Staphylococcus aureus(S.aureus)(NCTC 6571)and Escherichia coli(E.coli)(ATCC 25922).Methods:Leaf powder of R.communis L.was extracted with hot(in Soxhlet)and cold ethanol and methanol,separately.The antibacterial activity of the extracts was determined by agar well diffusion and macro broth dilution methods.The extracts were also subjected to phytochemical analysis.Results:All the four test extracts showed inhibition on both S.aureus and E.coli.Hot and cold ethanol extracts revealed significantly(P<0.05)higher inhibition on S.aureus than methanol extracts,and the hot ethanol extract had the lowest minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)values(5 mg/mL and 10 mg/mL,respectively).E.coli was highly inhibited by hot extracts of both ethanol and methanol with the MIC and MBC of 40 mg/mL and 80 mg/mL,respectively.Phytochemical analysis revealed the presence of saponins,cardiac glycosides,tannins,flavonoids and terpenoids in all test extracts.Conclusions:This study demonstrates that the hot and cold methanol and ethanol extracts are potential sources for control of S.aureus and E.coli.Especially,the hot and cold extracts of ethanol are more inhibitive against 5.aureus even at lower concentration.Further study is needed to identify the specific bioactive compounds,their mode of action and their nontoxic nature in vivo condition.

Antinociceptive activity of Ricinus communis L.leaves

Objective:To evaluate the antinociceptive activity of the methanol extract of Ricinus communis leaves(MRCL).Methods:Antinociceptive activity was evaluated using acetic acid induced writhing test,formalin induced paw licking and tail immersion method in mice at doses of 100,125 and 130 mg/kg bw.Results:The results indicated that MRCL exhibited considerable antinociceptive activity against three classical models of pain in mice.Preliminary phytochemical analysis suggested the presence of saponin,steroids and alkaloids.Conclusions:It can be concluded that MRCL possesses antinociceptive potential that may be due to saponin,steroids and alkaloids in it.

Transesterified Chinese Spicehush (Lindera communis) seed oil as a biodiesel fuel

Studies were carried out on the transesterification of the Chinese Spicehush （Lindera communis） seed （LCS） oil with methanol for the production of biodiesel. The methyl esters of LCS oil were compared with soybean esters to determine biodiesel fuel performance and properties. The fatty acids content in the seed kernel oil （LKO） and coat oil （LCO） were quantified by gas chromatography and mass spectrometry method （C-C-MS）. LKO contains 95.1% saturated acids （capric, lauric, myristic and palmitic） and 2.3% unsaturated acids （oleic）. LCO contains 24.6% saturated acids （palmitic and stearic） and 73.6% unsaturated acids （oleic, linoleic and palmitoleic）. The kinematic viscosity, cold filter plugging point, cloud point, flash point and cetane index were determined. The methyl esters of LKO had a higher kinematic viscosity and a lower cold filter plugging point value, so it has better performance in cold weather. The biodiesel of LCS oil has fuel properties within the limits prescribed by American （ASTM D 6751-02） standards, except for a slightly lower flash point of LKO biodiesel than that prescribed by these standards （130℃）. Thus, LKO and LCO biodiesel have great potential to be used on a large scale as fuel for diesel engines.

<i>Wrinkled</i>1 (WRI1) Homologs, AP2-Type Transcription Factors Involving Master Regulation of Seed Storage Oil Synthesis in Castor Bean (<i>Ricinus communis</i>L.)

Among APETALA2 (AP2)-type plant specific transcription factor family, WRINKLED1 (WRI1), has appeared to be a master gene transcriptionally regulating a set of carbon metabolism- and fatty acid synthesis (FAS)-related genes responsible for seed specific triacylglycerols (TAGs) storage in oil plants. B3 type transcription factors, such as ABI3 and FUS3, are known to be involved in seed development, such as seed storage protein synthesis and maturation. Based on the recent whole genome sequence data of castor bean (Ricinus communis L.), putative WRI1 homologs (RcWRI1, RcWRI2) specifically expressed in castor bean seed have been identified by comparing organ specific expression profiles among seed development-related transcription factors, seed storage specific genes (Ricin, RcOleosin) and a set of FAS genes including genes for sucrose synthase (RcSUS2), biotin carboxyl carrier protein (a subunit of acetyl-CoA carboxylase, RcBCCP2) and ketoacyl-acyl carrier protein synthase (RcKAS1). Immunoreactive signals with WRI1, FUS3 and ABI5-related polypeptides were also detected in seed specifically, consistent with the expression profiles of seed development-related genes. The WRI1 binding consensus sites, [CnTnG](n)(7)[CG], designated as the AW-box, were found at the promoter region of RcBCCP2 and RcKAS1. Thus, RcWRI1 possibly play a pivotal role in seed specific TAGs storage during seed development by directly activating FAS -related genes.

Effect of methanol extract of Ricinus communis seed on reproduction of male rats

Aim： To investigate the effect of methanol extract of Ricinus communis seed （RCE） on male rats reproductive functions. Methods： Thirty-two male albino rats were divided into four groups. Groups 1, 2 and 3 were gavaged with 0.2 mL of 2.5% tween 80 （RCE vehicle; control） or 20 mg/（kg-d） and 40 mg/（kg-d） of RCE, respectively, for 30 days, and group 4 was also gavaged with 40 mg/（kg.d） of RCE, but was allowed a recovery periold of 30 days. Five untreated female rats were cohabited with male rats in each group from day 25 of RCE treatment for 5 days, except group 4, where cohabitation began on day 25 of the recovery period. All male rats were sacrificed 24 h after the experiments. The female rats were laparatomized on day 19 of pregnancy and the number and weight of litters were recorded. Results： There was a significant decrease （P 〈 0.01） in the weight of the reproductive organs, sperm functions and serum levels of testosterone in RCE treated rats. There was disorganization in the cytoarchitecture of the testes, disruption of the seminiferous tubules and erosion of the germinal epithelium. The number and weight of litters of rats in groups 2 and 4 decreased significantly （P 〈 0.05） but no changes were observed in group 3. RCE caused no changes in liver, kidney, heart or body weights in male rats. Conclusion： RCE has a reversible negative impact on male reproductive functions, which appears to be mediated via gonadal disruption in testosterone secretion. （Asian J Androl 2006 Jan; 8： 115-121）

Genome-wide Identification,Classification and Expression Analysis of Lhc Supergene Family in Castor Bean(Ricinus communis L.)

Light-harvesting chlorophyll a/b-binding （LHC） proteins are a group of nuclear-encoded thylakoid proteins that play a key role in plant photosynthesis and are widely involved in light harvesting, energy transfer to the reaction center, maintenance of thylakoid membrane structure, photoprotection and response to en- vironmental conditions, etc. Although/dw supergene family is well characterized in model plants such as Arabidopsis, rice and poplar, little information is available in castor bean （Ricinus communis L. ）. In this study, a genome-wide search was carried out for the first time to identify castor bean L/w genes and analyze the gene structures, biochemical properties, evolutionary relationships and expression characteristics based on the published data of castor bean genome and ESTs. According to the results, a total of 28 Rclhcs genes representing 13 gene families （ l_hca , l_hcb , Elip , Ohpl , Ohp2 , SEP1, SEP2 , SEP3 , SEP4 , SEP5 , PsbS , Rieske and FCII） and 25 subgene families were identified in castor bean genome; to be specific, 25 and 5 genes were found to have corresponding ESTs in NCBI and have al- ternative splicing isoforlns, respectively. These RcLhcs contain 0 to 9 introns and distribute on 26 of the 25 878 released scaffolds. All RcLhcs genes were found to be expressed in all examined tissues, i.e. leaf, flower, II/III stage endosperm, V/VI stage endosperm and seed, with the highest expression level in leaf tissue.

In vitro establishment of a highly effective method of castor bean (Ricinus communis L.) regeneration using shoot explants

An efficient plant regeneration protocol was established for castor bean （Ricinus communis L.）, in which 0.3 mg L-1 thidiazuron （TDZ） induced shoot clusters and increased the number of adventitious shoots from hypocotyl tissue. Our results showed that treatment under dark conditions significantly promoted the average number of shoots per explant to 37.36±4.54 （with a 6-d treatment）. Modified 1/2 Murashige and Skoog （MS） basal medium supplemented with 440 mg L-1 Ca2＋, 0.2 mg L-1 gibberellic acid and 0.1 mg L-1 TDZ significantly increased shoot elongation rates and lowered vitrification rates. Further- more, 1/2 MS media supplemented with 0.2 mg L-11-naphthaleneacetic acid induced a higher rooting rate compared with other culture conditions.

Genetic diversity and population structure of Commelina communis in China based on simple sequence repeat markers

Commelina communis（Asiatic dayflower） is a troublesome weed in China. Genetic variation of 46 C. communis populations from different collection sites in our country was investigated using 12 simple sequence repeat（SSR） primer pairs. Polymorphism analysis results showed high level of genetic diversity among these populations. The alleles（bands） were amplified by these primer pairs. The polymorphic proportion was 18.25%, and the average polymorphism information content was 0.1330. The highest effective number of alleles was 1.9915 at locus YP33, and the lowest value was 1.0000 at both loci YP25 and YP31. C. communis showed major average observed heterozygosity value（0.8655） than that of average expected heterozygosity（0.1330）. C. communis populations were divided into three groups on the basis of unweighted pair-group method with arithmetic mean cluster analysis（Dice genetic similarity coefficient=0.772） and genetic structure analysis（K=3）, and a principal coordinate analysis. The results of this study further illustrated that C. communis populations contained abundant genetic information, and the 12 SSR markers could detect the microsatellite loci of C. communis genomic DNA. These results might indicate that C. communis maintains high genetic diversity among different populations.

Phytochemical analysis,hypotensive effect and antioxidant properties of Myrtus communis L.growing in Algeria

Objective:To analyze Myrlus communis chemically and evaluate the hypotensive effects and antioxidant properties of methanol,chloroform,ethyl acetate and aqueous extracts from the leaves of this plant.Methods:Total phenolic and flavonoid contents as well as the antioxidant potential of methanol,chloroform,ethyl acetate and aqueous extracts have been investigated by using different in vitro methods.The hypotensive effects of methanol and ethyl acetate extracts were evaluated in anaesthetized rats by using the method of invasive blood pressure recording.Moreover,ethyl acetate extract was subjected to analysis by different chromatographic methods in order to identify new compounds.Results:Chemical analysis of ethyl acetate extract revealed the presence of myrecitin-3-O-α-rhamnoside.Ethyl acetate extract was found to have the highest total phenolic and total flavonoid contents with the values of 435.37 ma gallic acid equivalents/g dried weight and 130.75 mg quercetin equivalent/g dried weight,respectively.Ethyl acetate extract also exhibited the highest activity in scavenging 2.2-diphenyl-1-picrylhydrazyl.2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid),hydroxy]radical and reducing power:whereas,methanol extract exhibited higher chelating activity than ethyl acetate extract did.Chloroform was found to be strong inhibitor of lipid peroxidation in β-carotene bleaching assay(91.19%).ferric thiocyanate method(87.55%).and thiobarbituric acid method(82.59%) as compared to butylated hydroxytoluene.Intravenous administration of methanol and ethyl acetate extract(0.04 to 12 mg/kg body weight) decreased the maximum mean arterial blood pressure with values of 20.6%and 32.49%at 12 mg/kg body weight,respectively in anesthetized rats.Conclusions:This study provides a scientific basis for the use of Myruts communis in traditional medicine as hypertensive agent as well as additional resources for natural antioxidants.

In vitro antimicrobial and larvicidal properties of wild Ricinus communis L. in Mauritius

Objective: To explore the phytochemical and antimicrobial activities as well as the insecticidal properties of the different sections of Ricinus communis(castor) plant in Mauritius.Methods: Qualitative and quantitative methods were used for the determination of phytochemicals in the crude leaves, pericarp, seeds, bark and root extracts obtained by using polar and non-polar solvents. The disc diffusion and micro-dilution methods were used to evaluate the antimicrobial activities of the crude solvent extracts against 13 microorganisms. The insecticidal properties of the crude extracts on larvae of Bactrocera zonata(Diptera: Tephritidae), which caused important economic losses to local fruits were also investigated.Results: All the extracts from the different parts of the plant showed antimicrobial activity against most tested microorganisms. The polar solvents' extracts of the fully mature parts of the castor plant were active against the Pseudomonas aeruginosa(ATCC 27853),Escherichia coli(E. coli)(ATCC 25922), E. coli(0145:H28 Acc. No. CP006027.1) with inhibition zones ranging from 16 mm to 19 mm and against Bacillus cereus(ATCC11778)(B. cereus), Listeria innocua(ATCC 33090)(L. innocua). Lowest microbial inhibitory concentration was recorded for B. cereus, L. innocua, Staphylococcus aureus(ATCC 29213), E. coli(ATCC 25922) and Proteus mirabilis strain(NCTC 11938) with value of 3.2 mg/m L. The most active extract against both Gram-negative and Grampositive bacteria was extracted from the fully mature pericarps and it was the most active against E. coli(ATCC 25922), B. cereus, L. innocua and Staphylococcus aureus(ATCC 29213). In addition, the extracts obtained by using polar solvent and fully mature leaves demonstrated the strongest larvicidal activity against Bactrocera zonata(100%).Conclusions: Ricinus communis(castor) plant extracts possess larvicidal properties providing an effective eco-friendly control for fruit flies. The antimicrobial results justify the use of this plant in traditional medicine and the practice of supplementing decoctions/concoctions with conventional antibiotics.