Chemical composition and biological activities of Portulaca oleracea

Portulaca oleracea(P.oleracea)is a traditional Chinese herbal that has the effects of soothing the liver and clearing collaterals,strengthening the spleen and stomach,moistening the intestines and detoxifying.With the development of the pharmaceutical industry,the medicinal value of P.oleracea is becoming increasingly prominent.It is commonly used in clinical practice for clearing heat and detoxifying,inhibiting bacteria,preventing and treating diseases such as hypertension,coronary heart disease,and cerebral infarction,indicating its broad application prospects.This study reviews the chemical components and pharmacological activities of P.oleracea in recent decades.

花椰菜(Brassica oleracea var.botrytis)抗黑腐病差异表达cDNA片段的克隆及功能的初步研究

采用花椰菜抗黑腐病近等基因系C731（感病系）和C712（抗病系）作为材料，利用cDNA—AFLP技术，研究了花椰菜黑腐病抗性在黑腐病菌侵染和非侵染条件下基因表达的情况．获得了两个阳性克隆M2和M6．Northern杂交和点杂交进一步证明M2是组成型表达的cDNA片段，只是在病菌侵染与非侵染条件下表达丰度不同．而M6是差异表达的cDNA片段．Southern杂交表明M6 cDNA片段在花椰菜基因组中是单拷贝序列．随后的序列分析发现M6cDNA片段与拟南芥1号染色体的BACF19P19中66665～66813bp有84％同源性，该片段编码拟南芥的2A6蛋白的部分序列．推测的氨基酸序列与拟南芥的2A6蛋白部分序列有91％同源性．用H2O2作为外源分子胁迫处理的初步功能分析发现：M6cDNA片段受H2O2诱导，在诱导早期16～24h高度表达，其在叶片中的积累呈逐渐上升趋势．根据序列分析和初步的功能分析的结果推测：M6cDNA片段可能就是编码花椰菜中类2A6蛋白的部分基因片段．是参与花椰菜抗病反应信号途径的相关调控基因片段．

花椰菜(Brassica oleracea var.botrytis)黑腐病抗性基因同源序列分离及克隆的研究

通过从 NBS 保守序列设计简并引物 PCR 的方法,以花椰菜(Brassica oleracea vat.botrytis)抗、感黑腐病的近等基因系为材料,分离得到 NBS-LRR 型抗性基因同源序列,并获得1个克隆,命名为 RGA330-7.Southern 杂交表明,该片段在近等基因系中存在明显的多态性,且该片段在抗黑腐病基因位点至少存在3个以上类似 RGA330-7的同源拷贝.序列分析结果认为该克隆与 NBS-LRR 型抗性基因的部分 CDSs 有很高的同源性,说明该片段属于 NBS-LRR 型.系统进化分析该序列与甘蓝型油菜的2个抗病同源序列归为一类,很可能这3个不同来源的抗性基因同源序列同属于一种抗性基因家族.因此推测该序列与花椰菜抗黑腐病基因紧密相关,为进一步克隆花椰菜抗黑腐病基因提供了可靠的候选基因,对分子标记辅助抗性育种具有重要意义。

根癌农杆菌介导白细胞介素-4基因转化甘蓝(Brassica oleracea L.)研究

利用根癌农杆菌介导法，以白细胞介素-4（interleukin-4，IL-4）基因转化中甘11号甘蓝。本研究对可能影响il-4转化率的外植体类型、外植体的预培养时间、农杆菌的侵染时间以及外植体与农杆菌的共培养时间进行了优化。试验结果表明：il-4基因对带1-2mm子叶柄的子叶的转化率显著高于下胚轴；带柄子叶在预培养1d、侵染3min、共培养1d时，转化效果最好，转化率达23．3%;下胚轴在预培养1-3d，侵染3～5min，共培养1-2d时，转化效果较好，转化率最高可达13．3％。经PCR检测及PCR-Southem杂交，初步证明目的基因il-4已经转入甘蓝的再生植株。转il-4基因甘蓝的PCR阳性再生植株已经开花并产生了后代。

与花椰菜(Brassica oleracea ssp.botrytis)抗黑腐病基因连锁的RAPD标记

利用 RAPD技术 ,在花椰菜 ( Brassica oleracea ssp.botrytis)抗感黑腐病的近等位基因系之间进行 1 0碱基随机引物的筛选 .34 5条 RAPD引物扩增的产物表明 ,有 7条引物在近等基因系中呈多态性 .进一步的重复实验表明 ,只有引物 OP2 2 4能够产生稳定的多态性 .为了确定扩增产物 OP2 2 4/1 6 0 0与抗黑腐病基因的连锁程度 ,利用 F2分离群体进行检测 ,结果表明 :花椰菜种质 C71 2抗黑腐病生理种 BJ的抗性由一对显性基因控制 .RAPD标记OP2 2 4/1 6 0 0与抗病基因连锁 ,其遗传距离为 ( 4 .5± 1 .37) c

恶性杂草马齿苋(Portulaca oleracea)种子萌发特性的研究

为了明确恶性杂草马齿苋种子的萌发特性,本文研究了温度、光照、pH、水势、盐分及土层深度对其种子萌发的影响。结果表明:马齿苋种子在光照L∥D=14h∥10h、温度白天32℃,晚上28℃的变温条件下萌发率最高,为92%。马齿苋种子萌发适宜的酸碱度范围较宽,在5<pH<10范围内,60%以上的种子均可萌发。马齿苋种子萌发率在水势为0～-0.8MPa范围内从89.17%降低至43.33%,即马齿苋种子在较干旱的环境下仍可以萌发。马齿苋种子耐盐性较强,当NaCl浓度为160mmol/L时,其种子萌发率仍高达53.33%。当马齿苋种子位于土壤表层时,其萌发率最高,达90.83%,土层深度的增加会严重抑制其萌发生长。说明马齿苋种子的萌发受环境影响较大,掌握各种环境因子对其种子的影响,采取并制定有效的综合管理措施以遏制其进一步危害叶菜类蔬菜。

马齿苋Portulaca oleracea的化学成分

从马齿苋(Portulaca oleracea)乙酸乙酯提取物中分离得到4个化合物,经MS和NMR等波谱技术分析,并结合文献对照,确定其结构分别为谷甾-4-烯-3-醇(1),谷甾-5-烯-3-醇(2),friedelan-4-αmethyl-3-βOH(3)和β-胡萝卜甙(4),其中化合物1,2,4为首次从该类植物中分离.初步评价了化合物3的抗肿瘤活性.

Study on the Relationship Between the Ploidy Level of Microspore-Derived Plants and the Number of Chloroplast in Stomatal Guard Cells in Brassica oleracea

The relationship between the ploidy level of microspore-derived plants and chloroplast number in stomatal guard cells was studied in cabbage, broccoli, and Chinese kale. In the experiment, distribution statistics analysis and t-test were used to perform statistical analysis on chloroplast number of different ploidy level in those stomatal guard cells mentioned above, and morphology identifying and chromosome counting were used to test accuracy of counting chloroplast number in stomatal guard cells. The chloroplast average number in stomatal guard cells was very similar among the different leaf positions on the same plant and among significantly among the different ploidy the different locations in the same stoma in the same variety. All the leaf, while the chloroplast number varied distributions of the chloroplast number in different ploidy stoma were normal distribution fitted. A correlation has been established between ploidy and chloroplast number in the stomatal guard cells. In every single stoma of microspore-derived plants, the chloroplast number for a haploid should not be more than 10, diploids 11 to 15, and polyploids more than 15. The accuracy of this method for identification of different ploidy plants was 93.93%. Furthermore, the accuracy of this method was reliable and did not vary with the plants growth conditions. Therefore, the chromosome ploidy of plants derived from microspore culture in cabbage, broccoli, and Chinese kale can be identified by simply counting the chloroplast number in stomatal guard cells.

Plant Male Sterility Induced by Anti-Gene CYP86MF in Brassica oleracea var. italica

An anti-gene CYP86MF was introduced into hypocotyls of broccoli （Brassica oleracea L.var. italica Plenck） with Agrobacterium tumefaciens, and the transgenic plants were obtained by kanamycin selection. The results of PCR, Southern blot and Northern blot indicated that the anti-CYP86MF has been integrated into chromosome of the transgenic plant. And also, plants with hypogenetic stamina or ungerminated pollen were observed. The transgenic male sterility plant could fructify via artificial pollination with normal pollen. Thus it was proved that the pistil of male sterility plant was normally developed, and the sterility originated from anti-CYP86MF.

Genome-wide identification,evolutionary selection,and genetic variation of DNA methylation-related genes in Brassica rapa and Brassica oleracea

DNA methylation plays an important role in plant growth and development,and in regulating the activity of transposable elements(TEs).Research on DNA methylation-related(DMR)genes has been reported in Arabidopsis,but little research on DMR genes has been reported in Brassica rapa and Brassica oleracea,the genomes of which exhibit significant differences in TE content.In this study,we identified 78 and 77 DMR genes in Brassica rapa and Brassica oleracea,respectively.Detailed analysis revealed that the numbers of DMR genes in different DMR pathways varied in B.rapa and B.oleracea.The evolutionary selection pressure of DMR genes in B.rapa and B.oleracea was compared,and the DMR genes showed differential evolution between these two species.The nucleotide diversity(π)and selective sweep(Tajima’s D)revealed footprints of selection in the B.rapa and B.oleracea populations.Transcriptome analysis showed that most DMR genes exhibited similar expression characteristics in B.rapa and B.oleracea.This study dissects the evolutionary differences and genetic variations of the DMR genes in B.rapa and B.oleracea,and will provide valuable resources for future research on the divergent evolution of DNA methylation between B.rapa and B.oleracea.

A Co-Dominant Marker BoE332 Applied to Marker-Assisted Selection of Homozygous Male-Sterile Plants in Cabbage (Brassica oleracea var.capitata L.)

The dominant genic male sterility （DGMS） gene CDMs399-3 derived from a spontaneous mutation in the line 79-399-3 of spring cabbage （Brassica oleracea var. capitata L.）, has been successfully applied in hybrid seed production of several cabbage cultivars in China. During the development of dominant male sterility lines in cabbage, the conventional identification of homozygous male-sterile plants （CDMs399-3/CDMs399-3） is a laborious and time-consuming process. For marker-assisted selection （MAS） of the gene CDMs399-3 transferred into key spring cabbage line 397, expressed sequence tag-simple sequence repeats （EST-SSR） and SSR technology were used to identify markers that were linked to CDMs399-3 based on method of bulked segregant analysis （BSA）. By screening a set of 978 EST-SSRs and 395 SSRs, a marker BoE332 linked to the CDMs399-3 at a distance of 3.6 cM in the genetic background of cabbage line 397 were identified. 7 homozygons male-sterile plants in population P1170 with 20 plants were obtained finally via MAS of BoE332. Thus, BoE332 will greatly facilitate the transferring of the gene CDMs399-3 into the key spring cabbage line 397 and improve the application of DGMS in cabbage hybrid breeding.

Association Analysis and Identification of SNP Markers for Stemphylium Leaf Spot (Stemphylium botryosum f. sp. spinacia) Resistance in Spinach (Spinacia oleracea)

Stemphylium leaf spot, caused by Stemphylium botryosum f. sp. spinacia, is an important fungal disease of spinach (Spinacia oleracea L.). The aim of this study was to conduct association analysis to identify single nucleotide polymorphism (SNP) markers associated with Stemphylium leaf spot resistance in spinach. A total of 273 spinach genotypes, including 265 accessions from the USDA spinach germplasm collection and eight commercial cultivars, were used in this study. Phenotyping for Stemphylium leaf spot resistance was evaluated in greenhouse;genotyping was conducted using genotyping by sequencing (GBS) with 787 SNPs;and single marker regression, general linear model, and mixed linear model were used for association analysis of Stemphylium leaf spot. Spinach genotypes showed a skewed distribution for Stemphylium leaf spot resistance, with a range from 0.2% to 23.5% disease severity, suggesting that Stemphylium leaf spot resistance in spinach is a complex, quantitative trait. Association analysis indicated that eight SNP markers, AYZV02052595_115, AYZV02052595_122, AYZV02057770_10404, AYZV02129827_205, AYZV0-2152692_182, AYZV02180153_337, AYZV02225889_197, and AYZV02258563_213 were strongly associated with Stemphylium leaf spot resistance, with a Log of the Odds (LOD) of 2.5 or above. The SNP markers may provide a tool to select for Stemphylium leaf spot resistance in spinach breeding programs through marker-assisted selection (MAS).

亚马逊莓(Euterpe oleracea)美白相关成分和作用研究现状

作为棕榈科植物Euterpe属中分布最为广泛、研究与商业化程度最高的品种,Euterpe oleracea (E. oleracea)的果实(为亚马逊莓的一种)因其卓越的抗氧化性能而跻身"超级水果"之列,同时也成为国际上新的研究热点。本文综述了E. oleracea的相关成分和作用研究现状,指出其富含不饱和脂肪酸、氨基酸、维生素等营养成分,并含有大量的以酚类化合物为主的生物活性物质,例如黄酮类化合物、酚酸、花青素与原花青素等。其中,总脂肪酸、亚油酸、阿魏酸、槲皮素、原儿茶酸、原儿茶酸甲酯等成分被证实具有皮肤美白作用。对E. oleracea进行的细胞实验和动物实验发现,其果肉提取物具有抑制酪氨酸酶活性以及使细胞免受紫外线引起的氧化应激伤害等潜在的皮肤美白功效。文章建议在未来的研究中,对E. oleracea的美白成分进行进一步挖掘,并通过动物实验与临床试验对其美白功效进行验证。

Optimization of micropropagation and <i>Agrobacterium</i>-mediated gene transformation to spinach (<i>Spinacia oleracea</i>L.)

Spinach is one of the dioecious plant which is considered as a model plant in genetic and molecular studies of sex determination because of its special characteristics such as low chromosome number and short life cycle. An efficient protocol for Spinacia oleracea Agrobacterium-mediated gene transformation was developed. The leaf disks, roots, hypocotyls and cotyledons of this plant were inoculated with LBA4404. LBA4404 carrying pCAMBIA3301 binary vector with 35SCaMV gusint and 35SCaMV bar cassettes. Effects of two preparation condition (induction of vir genes and noninduction) were considered. Also effects of different number days of co-cultivation and pre-culture of explants were examined. After co-cultivation, the explants were transferred to regeneration medium containing 250 mg·L-1 Carbeniciline. Transient expression efficiency was calculated based on the number of blue spots per explants one week after inoculation. Based on the results of transient expression, stable transformation was carried out. After formation of callus the histochemical GUS assay was carried out on some parts of them and other parts were leaved for being regenerated.

Effects of Brassica oleracea extract on impaired glucose and lipid homeostasis in highfat diet-induced obese mice

Objective: To examine the effect of Brassica oleracea extract(BO) on impaired glucose and lipid homeostasis in high-fat diet(HFD)-induced obese mice. Methods: Obesity of ICR mice was induced by feeding a HFD(45 kcal% lard fat) for 16 weeks. During the last 8 weeks of study period, obese mice were additionally administered with BO(100 and 200 mg/kg/day). The metabolic parameters were determined. The gene expressions of hepatic lipogenesis were also studied. Results: After 8 weeks of treatment, BO(100 and 200 mg/kg) significantly reduced hyperglycemia and improved insulin sensitivity(P < 0.05). The serum lipid(total cholesterol, triglyceride, and non-esterified fatty acid) and hepatic triglyceride and nonesterified fatty acid were decreased(P < 0.05). The levels of insulin and leptin in serum were also decreased(P < 0.05). Moreover, the expressions of hepatic lipogenic genes including sterol regulatory element-binding protein 1 c, fatty acid synthase, and acetyl-CoA carboxylase were decreased by BO treatment(P < 0.05). Conclusions: These results suggest that BO is a new therapeutic agent for improving the homeostasis of glucose and lipid in HFD-induced obese mice probably by suppression of lipogenic genes in liver tissue.

The protoplasts isolation,culture and shoots regeneration of broccoli(Brassica oleracea var.italica)

Eight F1-hybrid cultivars of broccoli were studied.We obtained cell division,celled colonies and p-calli in 5 cultivars,roots and shoots regeneration in one cultivar.The leavesof propagated plantlets in vitro were cut into 1—2mm pieces,isolated with an enzyme solutioncontaining 2% cellulase and 1%macerase on a rotary shaker（50 rpm,21℃,3h,2500 lux light）,and purified with a 0.5M sucrose solution.The purified protoplasts were placed on a drop of 1%agarose.2—3 ml liquid medium was added around the agarose drops,and all of the cultures wereincubated at 25℃ under light（4000 lux）for 16 hours.3—5 days after isolation the cell divisionwas found.About 7 days after incubation 4 multicellular colonies were formed.After 3—5 wksome p-calli were developed.When the p-calli were 2—3 mm in diameter it was transferred to asolidified medium.Once they were developed to 1 cm in diameter they were transferred on a re-generation medium.About 5 months after incubation some roots and shoots grown from the calliwere

Interspecific Hybridization of Brassica campestris × B.oleracea Through Ovary Culture

Using three varieties of Brassica campestris, Hauarad (708), Maoshan-3 (714) and Youbai (715),as the maternal plants and one variety of Brassica oleracea Jingfeng-1 (6012) as paternalplants, crosses were made to produce interspecific hybrids through ovary culture techniques.The ovaries from the cross between B. campestrisB.oleracea (7086012 and 7146012) werecultured and ovary culture was more effective in terms of obtained seeds when ovaries werecultured in vitro at 9 d after pollination (DAP). While for the cross of 7156012, it wasbetter when ovaries in vitro cultured at 12 DAP. Among three cross combinations, the cross of7146012 showed the best response and 43 seeds per ovary were obtained. Among the mediastudied, the ovaries from the cross of 7086012 cultured on MS media supplemented with 3.0 mgL-1 BA0.1 mg L-1 NAA showed better response, and its rate of seeds per ovary reached 44.0%.While the ovaries from the other two crosses (7146012 and 7156012) showed the best responsewhen cultured on B5 media supplemented with 3.0 mg L-1 BA + 0.2 mg L-1 NAA, and the rates of seedsper ovary reached 72.0 and 60.0%, respectively. All seeds obtained from the three crosscombinations were cultured on the MS media supplemented with 1.0 mg L-1 BA + 0.05 mg L-1 NAA,and the seeds from the cross of 7156012 showed the best germination response and thepercentage of germinations reached 66.7%. The regenerated plantlets were obtained from theseseedlings after cultured on the MS media supplemented with 0.05 mg L-1 NAA. Cytological studyshowed that these regenerated plants were all true hybrids of B.campestrisB.oleracea.

Genetic diversity analysis of Brassica oleracea L.by SSR

SSR analysis on genetic diversity of 30 samples was carried out. Five primers selected from 36 primers were used to amplify 30 samples in this experiment, PCR products were separated by 6% polyacrylamide gel electrophoresis, silver staining and photographed. The results of SSR were analyzed by UPGMA clustering. The results showed that a total of 21 gene alleles were detected by 5 SSR primers. The number of alleles ranged from 2 to 5 with an average of 4.2.PIC range was 0.257-0.92 1, with an average of 0.543. The average coefficient of genetic similarity of SSR markers among materials was 0.432. Some of cabbage cultivars in the experiment were divided into four groups except cultivars which come from Japan.

Effect of H_2O_2 on Growth of Collard( Brassis oleracea L. ) Seedlings Under Salt Stress

Collard variety( Brassis oleracea L. var. acephala f. tricolor Hort.) as a research material was treated with exogenous H_2O_2 and H_2O_2 scavenger dimethyl thiourea under 100 mmol/L NaC l stress. Two days later,growth rate,dry weight,fresh weight and relative water content of the plants were determined. After 6h of treatment,the activity and gene expression of three antioxidant enzymes,superoxide dismutase( SOD),catalase( CAT) and ascorbate peroxidase( APX) in plants,were measured. The results showed that the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in collard seedlings were higher in the treatment of salt stress with the addition of 0. 05 mmol/L exogenous H_2O_2 than in the simple salt stress treatment; and when endogenous H_2O_2 was removed,the growth rate,dry weight,fresh weight,relative water content,and the activity and gene expression of the three antioxidant enzymes in plant seedlings were lower than those under simple salt stress. It is speculated that under salt stress,H_2O_2 is involved in the regulation of antioxidant defense gene expression,and it might be an important regulator of salt-induced antioxidant system in collard leaves.

Isolation and Expression Profiling of a CONSTANS-Like Gene and Two FLOWERING LOCUS T-Like Genes from Spinacia oleracea L.

Spinach (Spinacia oleracea L.) develops leaf rosettes under short-day conditions, and starts reproductive growth including bolting and flowering under long-day conditions. Japanese people prefer Oriental spinach that bolts easily with a shorter photoperiod than European spinach. This is one of the main reasons that Oriental spinach is difficult to grow year-round. In order to understand spinach flowering mechanisms and obtain knowledge for spinach breeding, we isolated one CONSTANS-like (COL) and two FLOWERING LOCUS T (FT) homologs, which are key components of photoperiodic regulation of flowering time, from a Japanese cultivar. The expression of SoCOL1 showed diurnal rhythm with the highest expression at the end of the dark cycle. This diurnal rhythm is similar to the expression of BvCOL1 from sugar beet (Beta vulgaris), whose flower-promoting effect was observed when overexpressed in Arabidopsis. Phylogenetic analysis showed that SoCOL1 is the closest homolog of BvCOL1, suggesting that SoCOL1 is an ortholog of BvCOL1. SoFT1 and SoFT2 are closely related to BvFT1 and BvFT2, respectively. The expression of SoFT1 and SoFT2 were induced in advance of flower bud formation after changing the photoperiod, but the expression level of SoFT1 was much lower than SoFT2. Currently, we are speculating that SoFT2 is a flower-promoting factor of spinach, and that SoFT1 has a role in light signaling because the expression of SoFT1 showed a diurnal rhythm.