Extraction of seed oil from Diospyros lotus optimized using response surface methodology

Oil from seeds of Diospyros lotus was extracted using a conventional method with two different solvents:hexane and petroleum ether. A central composite design with response surface methodology were used to optimize the process. A second-order polynomial equation was employed, and ANOVA was applied to evaluate the impact of various operating parameters including extraction temperature(x_1; 44.9–70.1 °C), extraction time(x_2;5.0–10.0 h) and solvent to solid ratio(x_3;11.6–28.4 mL g^(-1)), on oil yield. Experiments to validate the model showed decent conformity between predicted and actual values. Extraction conditions for optimal oil yield were 61 °C, 8.75 h extraction duration and 19.25 mL g^(-1) solvent to solid ratio. Under these conditions, the oil yield was predicted to be 5.1340%. Oil samples obtained were then analyzed using gas chromatography. The fatty acid composition revealed the major fatty acids to be oleic acid(C18:1) and linoleic acid(C18:2). The analysis of oil also demonstrated a decent ratio between omega-3 and omega-6 fatty acids. The structure of seeds was imaged using scanning electron microscopy. Oil quality was analyzed thermogravimetrically and by Fourier transform infrared spectroscopy. The assigned nutritional features of the D. lotus oil suggested that it can be used as an edible oil in pharmaceutical and food industry in the future.

Comparative investigation of the free radical scavenging potential and anticancer property of Diospyros blancoi(Ebenaceae)

Objective: To investigate the comparative effects of Diospyros blancoi(Ebenaceae)leaves(DBL), root bark(DBRB) and stem bark(DBSB) on free radicals and cancer.Methods: The polyphenol contents, antioxidant and free radical scavenging properties were determined using standard spectrophotometric methods. Cytotoxicity and anticancer activities were performed on brine shrimp nauplii and Ehrlich ascite carcinoma cells, respectively.Results: Among the extracts, DBSB showed the highest total antioxidant capacity and reducing capacity on ferrous ion. Based on 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radical scavenging activities, DBSB showed(95.760 ± 0.343)% and(67.460 ± 2.641)%scavenging with IC50of(3.10 ± 0.17) and(50.00 ± 3.11) mg/m L, respectively. The IC50 values of standard butylated hydroxytoluene and catechin(CA) for 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radicals were(8.50 ± 0.25) and(75.00 ± 0.14) mg/m L,respectively suggesting that DBSB had a significant(P < 0.05) radical scavenging activity than standards. In lipid peroxidation inhibition assay, the inhibitory activity of the extracts and the standard was in the following order: DBSB > DBRB > CA > DBL.Also, the phenolic [(139.91 ± 3.924) mg gallic acid equivalent/g] and flavonoid contents[(412.00 ± 16.70) mg catechin equivalent/g)] of DBSB were higher than that of other extracts. In addition, the DBSB showed the moderate cytotoxic and anticancer properties.Conclusions: Our results indicate that Diospyros blancoi stem bark had the significant highest antioxidant and free radical scavenging properties as well as moderate anticancer activity.Hence, we assume that the anticancer activity of this plant can be, at least in part, attributed to its content in phenolic compounds as well as its significant free radical scavenging properties.

Effects of two medicinal plants Psidium guajava L. (Myrtaceae) and Diospyros mespiliformis L. (Ebenaceae) leaf extracts on rat skeletal muscle cells in primary culture

Crude decoction, aqueous and ethanolic extracts of two medicinal plants （Psidiura guajava and Diospyros raespiliformis）, widely used in the central plateau of Burkina Faso to treat many diseases were evaluated for their antagonistic effects on caffeine induced calcium release from sarcoplasmic reticulum of rat skeletal muscle cells. These different extracts showed a decrease of caffeine induced calcium release in a dose dependent manner. Comparison of the results showed that Psidiura guajava leaf extracts are more active than extracts of Diospyros mespiliformis and that crude decoctions show better inhibitory activity. The observed results could explaine their use as antihypertensive and antidiarrhoeal agents in traditional medicine, by inhibiting intracellular calcium release.

Inter-and Intra-specific Genetic Diversity in Diospyros Using SCoT and IRAP Markers

The genus Diospyros contains more than 500 species, among them persimmon(Diospyros kaki Thunb.) is the most economically important species. However, the inter-and intra-specific genetic diversity remains unknown due to historic cultivation, domestication and introduction in different regions of China. In this study, we investigated the genetic diversity and relationship among 268 Diospyros accessions from different regions in China using start codon-targeted(SCo T) and inter-retrotransposon-amplified polymorphism(IRAP) markers. Among 268 samples,232 accessions were D. kaki varieties and 36 were closely related species, that is, D. lotus(21), D. glaucifolia(1), D. virginiana(5), D. kaki Thunb.var. silvestris(3), D. oleifera(5) and D. deyangnsis(1). In our study, a total of 90 and 97 polymorphic alleles were obtained from nine SCo T and nine IRAP markers, respectively, and these markers displayed rich polymorphisms for evaluation of the genetic diversity among 268 samples. A study on the relationship, genetic structure and analysis of molecular variance(AMOVA) showed that inter-and intra-specific gene exchanges were frequent among/within the samples from five regions. Moreover, the genetic distance between the Southwest China(SWC) and the other regions was relatively far;the samples from North China(NC), East China(EC), South and Central China(SCC), Northwest China(NWC) regions had a relatively close relationship. The unweighted pair group method with arithmetic means(UPGMA) and population structure analysis indicated that persimmons within geographic regions were clustered together, which revealed that the samples from various regions exhibited gene exchange, and the geographical proximity of persimmons resulted in a more frequent exchange. In addition, the persimmon had a close relationship with D. kaki var. silvestris, D. deyangnsis and D. oleifera species, and exhibited a far distance with respect to D. virginiana. These SCo T and IRAP markers will be helpful for understanding of the inter-and intra-specific genetic diversity of Diospyros germplasms, which play an important role in the germplasm efficient conservation, identification and utilization for Diospyros resources.

In vivo sedative and muscle relaxants activity of Diospyros lotus L

Objective: To evaluate the sedative effect of Diospyros lotus L(D. lotus) extract in mice using the open field and Rota rod tests.Methods: For the sedative and muscle relaxants activities of extract/fractions of the plant, invivo open field and phenobarbitone-induced sleeping time were used, while the Roda rod test was employed in animals for the assessment of muscle relaxant activity.Results: Results from this investigation revealed that the extracts of D. lotus have exhibited significant sedative effect in mice(45.98%) at 100 mg/kg i.p. When the extract was partitioned with different solvents, the n-hexane fraction was inactive whereas the chloroform fraction was the most active with 82.67% sedative effect at 50 and 100 mg/kg i.p. On the other hand,the ethyl acetate and n-butanol fractions displayed significant sedative effects(55.65% and40.87%, respectively) at 100 mg/kg i.p. Among the tested extract/fractions, only chloroform and ethyl acetate fractions showed significant(P < 0.05) muscle relaxant activity in the Rota rod test.Conclusions: In short, our study provided scientific background to the traditional uses of D.lotus as sedative.

Investigation of the Phytoconstituents and Antioxidant Activity of <i>Diospyros malabarica</i>Fruit Extracts

Diospyros malabarica (Deshi Gab) belonging to the family “Ebenaceae” grows well in the humid tropical climate of Bangladesh. In order to investigate the phytoconstituents both qualitatively and quantitatively, the seed and flesh extracts of D. malabarica were prepared using two polar solvents (i.e., water and 70% ethanol) and a nonpolar solvent (i.e., hexane). The maximum yield was obtained for aqueous and ethanolic seed extracts indicating that most of the phytoconstituents present in D. malabarica fruit are polar. The qualitative phytochemical analysis of the extracts revealed the presence of diverse amount of phytoconstituents in the extracts. On the other hand, the quantitative phytochemical analysis for phenols, tannins, flavonoids, alkaloids, saponin, proteins, reducing sugar and vitamin C revealed that the maximum amount of phenols, tannins, flavonoids and reducing sugar were present in aqueous seed extract. However, the maximum amount of total protein and vitamin C was found in ethanolic seed extract. D. malabarica seed powder contained more amount of alkaloids (i.e., 13.6%) than that of flesh (i.e., 3.4%). However, the content of saponin was more in flesh extract (0.74%) when compared to that of seed extract (0.42%). In vitro antioxidant properties of the extracts were also investigated by DPPH free radical scavenging assay, FRAP assay and reducing power assay. The highest antioxidant activity was observed for aqueous seed extract followed by the ethanolic seed, ethanolic flesh, and aqueous flesh extracts. The lowest antioxidant activity was obtained for hexane flesh extract. Furthermore, the lowest (i.e., 44.70 μg/ml) as well as the highest (2359.66 μg/ml) IC50 values were obtained for aqueous seed and hexane flesh extracts, respectively.

Diospyros lycioides Desf:Review of its botany, medicinal uses,pharmacological activities and phytochemistry

Diospyros lycioides Desf.(D. lycioides) is traditionally used as herbal medicine against various human and animal ailments in tropical Africa. The present paper reviewed information on botany, medicinal uses, phytochemistry and pharmacological activities of D. lycioides. This review was compiled using scientific literature from electronic search engine such as PubMed,Scopus, ScienceDirect, Springerlink, BioMed Central, Scielo, Medline and Science domain.Additional literatures were obtained from book chapters, books, dissertations, websites and other scientific publications. D. lycioides is used as traditional medicine in 50% of the countries where the species is native in tropical Africa. This study recorded 22 medicinal uses of D.lycioides which included abdominal pains, infertility in women, sexually transmitted infections,and used as chewing sticks(or mouthwash), toothbrushes and ethnoveterinary medicine. D.lycioides extracts demonstrated anti-adhesive, anti-inflammatory, antimetastatic, antioxidant,antifungal, antiproliferative, mutagenicity and antibacterial activities. Future research should focus on the pharmacological properties, phytochemistry, clinical trials and pharmacokinetics of D. lycioides which will enhance the therapeutic potential of the species.

Effect ofDiospyros kakienriched extender on cattle bull sperm parameters and conception rate

Objective:To explore the effect of Diospyros kaki on cattle spermatozoa during chilling and cryopreservation.Methods: Five milliliter of blended Persimmon (Diospyros kaki) flesh was added to 45 mL TCF to obtain 10% stock solution. Kaki enriched extender (KEE) was prepared by adding to TCF in concentrations 0.0/5.0 mL (control, 0%), 0.5/4.5 mL (1%), 1/4 mL (2%), 1.5/3.5 mL (3%), 2.0/3.0 mL (4%), 2.5/2.5 mL (5%), 3.0/2.0 mL (6%), 3.5/1.5 mL (7%), 4.0/1.0 mL (8%), 4.5/0.5 mL (9%) and 5.0/0.0 mL (10%) to obtain a final volume 5 mL in each tube. Whole egg yolk was added to each tube to obtain KEE with 20% egg yolk (KEEY), all tubes were centrifuged to get rid of debris. Semen was added to the supernatants in other tubes. Extended semen was subjected to evaluation (motility, alive sperm and intact sperm membrane (HOST) %) in both chilled and cryopreserved semen. Conception rate was carried out.Results:Sperm motility was significantly (P<0.0001) kept high after 11 d of chilling with the concentration 1%, 2%, 3%, 4%, 5% as compared to the control (41.67±1.67, 41.67±1.67, 40.00±0.00, 41.67±1.67 and 41.67±1.67, respectively) and also non-significantly kept high at the other concentrations up to 9 d of chilling. Addition of KEE had significantly (P<0.0033) improved post thawing sperm motility % with the concentrations 1, 2, 3, 4, 5 and 6% as compared to the control (51.67±5.27, 55.00±3.16, 48.33±1.05, 45.00±3.96, 57.00±2.50, 55.00±5.00 and 43.33±5.11 respectively).While the other concentrations exhibit no effect. Addition of KEE maintained alive sperm%, abnormalities% and % of intact spermatozoa membranes (HOST %) as good as the control with all concentrations of kaki used in our study. The conception rate upon using frozen semen in insemination showed higher conception rate in concentrations of 2%, 4% and 6 % KEE in cattle.Conclusion: It could be concluded that some concentrations ofDiospyros kaki improved bull semen quality post-chilling and post-freezing.

Isolation and structural elucidation of cytotoxic compounds from the root bark of Diospyros quercina (Baill.) endemic to Madagascar

Objective:To isolate and characterize the cytotoxic compounds from Diospyros quercina(Baill.)G.E.Schatz&Lowry(Ebenaceae).Methods:An ethno-botanical survey was conducted in the south of Madagascar from July to August 2010.Bio-guided fractionation assay was carried out on the root bark of Diospyros quercina,using cytotoxicity bioassay on murine P388 leukemia cell lines as model.The structures of the cytotoxic compounds were elucidated by 1D and 2D NMR spectroscopy and mass spectrometry.Results:Biological experiments resulted in the isolation of three bioactive pure compounds(named TR-21,TR-22,and TR-23)which exhibited very good in vitro cytotoxic activities with the IC_(50)values of(0.017 5±0.0060)μg/mL,(0.089±0.005)μg/mL and(1.027±0.070)μg/mL respectively.Thus,they support the claims of traditional healers and suggest the possible correlation between the chemical composition of this plant and its wide use in Malagasy folk medicine to treat cancer.Conclusions:The ability of isolated compounds in this study to inhibit cell growth may represent a rational explanation for the use of Diospyros quercina root bark in treating cancer by Malagasy traditional healers.Further studies are,therefore,necessary to evaluate the in vivo antineoplastic activity of these cytotoxic compounds as effective anticancer drugs.

Research on Dyeing Process for Polyamide with Aqueous Extracted from the Fruits of Diospyros Mollis

In this study, polyamide fabrics were dyed with aqueous extracts from Diospyros Mollis fruits. The aqueous extracts from Diospyros Mollis fruits were extracted with water solution at room temperature and a ratio of 1：5 for Diospyros Mollis to water. The influences of temperature in range of 50-100 ℃, time in range of 30-80 minutes and a ratio of aqueous extracts to water in range of 3/1-1/11 were investigated. The results showed that the aqueous extracts from Diospyros Mollis fruits could dye polyamide fabrics to have black color with color strength and fastness properties to be very good level of 4-5.

Phytochemical Profile, Antioxidant, and Anti-Inflammatory Activities, Safety of Use and Spasmolytic Effects of Aqueous Decoction Extract of Diospyros mespiliformis Leaves Hochst. ex A. DC. (Ebenaceae) on the Isolated Duodenum of Rat

Diospyros mespiliformis Hochst. ex A. DC. (Ebenaceae) is a multi-use plant, including for therapeutic purposes. It is used in alternative medicine in Burkina Faso to treat conjunctivitis, menorrhagia, dysentery, and diarrhea. The aim of our study was to evaluate the chemical profile, antioxidant and anti-inflammatory activities, safety of use and spasmolytic effects of the aqueous decoction of Diospyros mespiliformis leaves. Phytochemical screening by HPTLC and assay of compounds of interest were carried out. Four methods were used to assess antioxidant activity. Inhibitory activity against 15-lipoxygenase and phospholipase A2 was assessed. Acute oral toxicity of the extract was tested on female mice (NMRI). Following these tests, the extract contained bioactive compounds of interest such as flavonoids, tannins, sterols, triterpenes, and saponosides. The total phenolic and flavonoid contents of the aqueous decoctate were 70.59 ± 3.20 mg EAT/g and 31.57 ± 0.78 mg EQ/g respectively. The extract was less active than Trolox with inhibitory concentrations of 50% (IC50) for the ABTS, DPPH, FRAP, and LPO tests of 7.53 ± 0.08 μg/mL, 29.47 ± 0.06 μg/mL, 1128.83 ± 4.82 mol EAA/g, and 32.30 ± 1.60 μg/mL respectively. The extract has an anti-inflammatory effect with inhibition of phospholipase A2 compared to betamethasone. In addition, the aqueous extract produced an antispasmodic effect with Emax of 70% and 80% respectively during contractions induced by BaCl2 and ACh. Finally, this study provided basic scientific data and could justify the use of D. mespiliformis leaves in the treatment of diarrhea.

Antinociceptive Effect of Methanol Extract of Diospyros malabarica (Desr.) Kostel Leaves in Mice

Background: Diospyros malabarica (Desr.) Kostel, a small to medium-sized tree in the Ebenaceae family, is known as “Deshi Gab” in Bangladesh. Fever, diabetes, snake bite, diarrhea, biliousness, and ulcer ailments are all treated with the herb. This study’s goal was to examine in mouse models the antinociceptive properties of methanol extract of Diospyros malabarica leaves (MEDM). Methods: For the purpose of determining the antinociceptive activity in mice, five distinct pain models including hot plate, tail immersion, acetic acid-induced writhing, formalin and glutamate-induced nociception tests were used. The conventional medications were morphine sulphate (5 mg/kg, intraperitoneally) and diclofenac sodium (10 mg/kg, intraperitoneally). While the control group was expecting deionized water, MEDM was given orally at dosages of 200, 400, and 600 mg/kg (0.1 mL/mouse, orally). Results: According to the current research, MEDM strongly reflected the antinociceptive activity of all mouse models of chemical and heat-induced pain (*p < 0.05). 400 and 600 mg/kg demonstrated a considerable (*p < 0.05) ability to prolong the reaction of latency to pain in opposition to thermally produced nociception in hot plate and tail immersion tests. Inhibition levels in the acetic acid-induced writhing test were 11.57%, 37.77%, and 51.83%, respectively. The extract suppressed 20.78%, 45.48%, and 56.93% of licking during the initial stages of formalin-induced nociception. In the late phase, the extract showed higher rates of licking than the control group (13.14%, 50.28%, and 66.85%). The glutamate-induced nociception test was significantly (*p < 0.05) prevented by the plant extract. Compared to the control, it demonstrated an inhibition of licking of 22.85%, 47.32%, and 63.42%, respectively. Conclusions: It is evident that the plant extract has exceptional analgesic properties. To determine the precise processes behind antinociceptive effect and to identify the substances that produce this activity, more research is required. The study’s findings also support the long-standing use of MEDM in painful conditions.

Bioassay-guided isolation of novel and selective urease inhibitors from Diospyros lotus

Two new dimeric naphthoquinones, 5′,8′-dihydroxy-6,6′-dimethyl-7,3′-binaphthyl-1,4,1′,4′-tetraone(1; Di-naphthodiospyrol D) and 5′,8′-dihydroxy-5,8-dimethoxy-6,6′-dimethyl-7,3′-binaphthyl-1,4,1′,4′-tetraone(2; Di-naphthodiospyrol E), along with known naphthoquinones diospyrin(3) and 8-hydroxy diospyrin(4) were isolated from the chloroform fraction of extract of Diospyros lotus roots. Their structures were elucidated by advanced spectroscopic analyses, including HSQC, HMBC, NOESY, and J-resolved NMR experiments. The fractions and compounds 1-4 were evaluated for urease activity and phosphodiesterase-Ⅰ, carbonic anhydrase-Ⅱ and α-chymotrypsin enzyme inhibitory activities. Compounds 1 and 2 and their corresponding fractions showed significant and selective inhibitory effects on urease activities. The IC_(50) values of 1 and 2 were 260.4 ± 6.37 and 381.4 ± 4.80 μmol·L-1, respectively, using thiourea(IC_(50) = 21 ± 0.11 μmol·L^(-1)) as the standard inhibitor. This was the first report demonstrating that the naphthoquinones class showed urease inhibition.

Number of Species and Geographical Distribution of Diospyros L.(Ebenaceae) in China

Diospyros L. is the largest, most widely distributed, and economically important genus of the family Ebenaceae. However, various researchers disagree on the number of species and geographical distribution of Diospyros in China. For the purpose of providing a scientific basis for the effective protection and rational use of Diospyros, the present study involved a comprehensive review of the literature, electronic databases, and herbaria as well as field investigations. The results showed as the following.(1) While 174 scientific names have been published for various taxa of Diospyros in China, by screening and excluding synonyms, 65 species names were considered acceptable; these account for about 12%of the total species of Diospyros globally.(2) A review of herbarium specimens documented that Diospyros occurs in 28 administrative regions of China, more than the literature records. Species richness decreases from south to north in China while Guangxi, Yunnan, Guangdong, and Hainan have the greatest number of species.(3) Among the 65 relatively reliable Diospyros species, 45 species are endemic to China and 18 species are stenochoric. Some species may have become extinct as a result of environmental degradation and the status of some germplasm remain unresolved. In brief, Chinese Diospyros includes abundant, China-specific, and widely distributed features. Nevertheless, comprehensive species delimitation and the analysis of phylogenetic relationships will still require further study.

Allometric equations for aboveground biomass estimation of Diospyros abyssinica (Hiern) F. White tree species

Introduction:Quantifying forest biomass requires the application of allometric equations which is a fundamental step.Generalized allometric equations have been applied to quantify aboveground biomass(AGB)of forests.But,adopting generalized allometric equations to quantify AGB of different forests creates uncertainty.Therefore,developing species-and sitespecific allometric equations is essential to accurately quantify the biomass.The study was aimed to develop species-specific allometric equations for Diospyros abyssinica(Hiern)F.White in Yayu Coffee Forest Biosphere Reserve using the Semi-destructive method.The vegetation types of Yayu Coffee Forest Biosphere Reserve is categorized to Moist Evergreen Montane Rainforest of Ethiopia.Results and discussion:Evaluating statistical relationships of AGB against predictor variables,eight allometric equations were formulated.AGB was regressed against trunk diameter(D),total height(H),and wood density(ρ)individually and in combination.Selection of allometric equations was employed using model performance statistics.Equations with a higher coefficient of determination(adjusted R^(2)),lower residual standard error,and Akaike information criterion(AIC)values were found best-fitted.Relationships of AGB and independent variables were found statistically significant(p<0.000).Overall,formulating species-and site-specific allometric equations is significant for accurate estimation of forest biomass and carbon stock budget.

PEG-6000模拟干旱胁迫对毛叶柿种子萌发及生长量的影响

为了解干旱胁迫下毛叶柿(Diospyros dumetorum)种子萌发情况,探究干旱对毛叶柿种子萌发期生长量的影响。以毛叶柿种子为材料,用PEG-6000溶液(0、5%、10%、15%、20%、25%)处理种子以此来模拟不同程度的干旱胁迫。结果表明:1)轻度干旱(5%~10%PEG)能促进种子相对发芽率和相对发芽势的增长,超过10%PEG浓度,种子萌发指标与对照组相比均受到不同程度的抑制。2)随着PEG浓度的增加,幼苗生长指标均值有所下降,与对照组(0PEG)相比,轻中度干旱(PEG0~10%)的胚根长和胚芽长有所增加,胚根长对干旱胁迫的响应更敏感。3)随着PEG浓度的增加,种子受胁迫程度越高,种子地下生长量呈现增加趋势,PEG浓度与根干质量和鲜质量呈极显著相关(P<0.01)。结论:干旱胁迫下毛叶柿种子通过降低相对萌发率、增加胚根鲜质量、干质量和根冠比的生长适应干旱胁迫。

君迁子组培苗生根移栽体系优化

以常规君迁子组培苗为试材,研究不同植物生长调节剂处理下组培苗的生根效果,并探索移栽后不同保湿时间对组培苗移栽成活率的影响,以期获得君迁子(Diospyros lotus L.)组培苗生根移栽的高效技术体系,并为我国柿砧木无性系扩繁提供参考依据。结果表明:在1.0 mg·L^(-1) IBA+0.5 mg·L^(-1) IAA处理下,君迁子组培苗生根效果最好,生根率可达95.56%,平均根条数为7.93,平均鲜质量0.59 g,根系活力0.99μg·g^(-1)·h^(-1),平均总根长26.57 cm,平均根系表面积10.33 cm~2,平均根体积0.32 cm~3,平均根直径可达0.61 mm;移栽后保湿培养10 d组培苗成活率最高为90.44%,移栽至大田5个月后平均株高64.32 cm,茎粗6.86 mm,可用于接穗的嫁接繁殖。

柿种质资源田间耐寒性调查分析

对国家柿种质资源圃内资源进行调查认为，2008年初的低温虽不是陕西历史上冬季的最低温度，但由于持续时间长，同期平均温度低于正常年份，是造成大部分柿资源严重受冻的直接主要原因。冻害程度与柿资源的种类、品种、嫁接采用的砧木、树体生长势的不同有很大关系。调查结果表明：涩柿品种耐寒性高于甜柿品种；我国原产的甜柿品种耐寒性大于日本甜柿品种；甜柿品种中耐寒性强弱顺序为：阳丰〉次郎〉禅寺丸〉大秋〉早秋〉西村早生；同一品种嫁、接于西村早生为中间砧上的冻害重于直接嫁接在君迁子上的；高接1～2a的资源及树势衰弱树受冻害程度高。

罗田甜柿种质资源ISSR分子标记初步鉴定

分别采集湖北省罗田县、麻城市自然分布的28、8株罗田甜柿(Diospyros kaki Thunb cv.Oriental persimmon)古树优系及小涩柿、无核、宝盖、秋焰甜柿优系为研究材料,利用简单重复序列间扩增分子标记(Inter simple sequence repeat,ISSR)技术对40个甜柿优系进行初步认证及亲缘关系分析。结果表明:1从21条ISSR引物中筛选出了9条能够扩增出清晰、稳定条带的引物,9条引物共扩增出185条谱带,其中多态性条带有185条,多态性条带比率为100%;2应用NTSYS2.10e软件对扩增结果进行非加权组平均法(Unweighted pair-group method with arithmetic means,UPGMA)聚类分析,40个甜柿优系大致被分为两大类,遗传相似系数从0.39到0.92;3罗田甜柿古树基因资源丰富,每个基因型都有其独特的指纹图谱,遗传多样性高。罗田甜柿优系间的亲缘关系与地理来源有一定的相关性,麻城市与罗田县地理位置交界区域的罗田甜柿资源相对集中在A区,其他地理位置采集优系集中在B区。分析结果可为罗田甜柿种质资源开发、基因资源保存、后期品种审定及鉴别提供理论依据。

柿EST-SSR标记在柿属植物中的通用性研究

公共数据库和柿转录组测序产生的大量EST序列为开发新的SSR标记提供了有效资源。我们在前期利用这些数据开发出112个EST-SSR标记,此次研究选用其中11个标记对柿属(Diospyros L.)15个种20份试材进行了标记通用性检测。结果表明,所有引物均能扩增出产物,多态性引物比率达100%,并且UPGMA聚类结果可靠,说明柿EST-SSR标记在近缘种中可以通用。上述通用性标记为柿属植物遗传多样性研究提供了标记资源。