Anti-Melanogenesis Activity of Supercritical Carbon Dioxide Extract from Perilla frutescens Seeds

Perilla frutescens seed (PFS) oil is reported to inhibit skin photoaging;however, its effect on melanogenesis has not yet been investigated. Herein, we tested the anti-melanogenesis activity of an oil-based extract from PFS with supercritical carbon dioxide (scCO2). In a cell culture system, B16 mouse melanoma cells were treated with the PFS scCO2 extract and other samples. The PFS scCO2 extract decreased melanin production by approximately 90% in B16 mouse melanoma cells without cytotoxicity at 100 μg/mL. This effect was greater than that of the well-known melanogenesis inhibitor, kojic acid. Although a hexane-extracted PFS oil and a squeezed PFS oil also decreased melanin production in the B16 cells, the inhibitory effect of the PFS scCO2 extract was higher than both of these. Chemical analysis of the PFS scCO2 extract and squeezed PFS oil showed that almost 90% of the components of both oils were α-linolenic acid, linoleic acid, and oleic acid. Furthermore, the ratio of those three fatty acids across both samples was almost the same. When the three fatty acids were mixed in the same ratio as in the PFS scCO2 extract, the IC50 of the mixture for melanin production in B16 melanoma cells was identical to that of the PFS scCO2 extract. However, the IC50 of the squeezed PFS oil was approximately 6.6 times higher than that of the mixture. Although those fatty acids are the main inhibitory ingredients against melanin production in all of the extracts, some factor(s) in the squeezed PFS reduce their affinity with the cells. These results indicated that the PFS scCO2 extract could be a superior melanogenesis inhibitor. Although its main ingredients are probably the same as those of the squeezed PFS oil, it is necessary to extract with scCO2 for stronger anti-melanogenesis activity.

GC-MS analysis of volatile compounds of Perilla frutescens Britton var. Japonica accessions: morphological and seasonal variability

Objective: To investigate the composition of volatile compounds in the different accessions of Perilla frutescens(P. frutescens) collected from various habitats of China and Japan. Methods: In the present study, the essential oil from the leaves of P. frutescens cultivars from China and Japan was extracted by hydro-distillation and the chemical composition and concentration of the volatile components present in the oils were determined by gas chromatography–mass spectrometry(GC–MS) analysis. Results: Among the volatile components, the major proportion was of perilla ketone, which was followed by elemicin and beta-caryophyllene in the Chinese Perilla cultivars. The main component in the oil extracted from the Japanese accessions was myristicin, which was followed by perilla ketone and beta-caryophyllene. We could distinguish seven chemotypes, namely the perilla ketone(PK) type, perilla ketone, myristicin(PM) type, perilla ketone, unknown(PU) type, perilla ketone, beta-caryophyllene, myristicine(PB) type, perilla ketone, myristicin, unknown(PMU) type, perilla ketone, elemicine, myristicin, beta-caryophyllene(PEMB) type, and the perilla ketone, limonene, betacryophyllene, myristicin(L) type. Most of the accessions possessed higher essential oil content before the flowering time than at the flowering stage. The average plant height, leaf length, leaf width of the Chinese accessions was higher than those of the Japanese accessions. Conclusion:The results revealed that the harvest time and geographical origin caused polymorphisms in the essential oil composition and morphological traits in the Perilla accessions originating from China and Japan. Therefore, these chemotypes with desirable characters might be useful for industrial exploitation and for determining the harvest time.

Studies on In Vitro Flowering and Fruiting of Perilla frutescens

Influences of PGR, sucrose, and ammonium nitrate on in vitro flowering and fruiting from cotyledon explants of P. frutescens were studied. The regenerated shoots at 2-4. cm from cotyledon explants on MS medium supplemented with 0.5 mg L^-1 BA and 1.0 mg L^-1IAA were excised and transferred to MS medium containing 30 g L^-1 sucrose, 8.25 g L^-1 ammonium nitrate, and 1.0 mg L^-1 BA. Following 40 d of culture, 86.2% of them flowered and set seeds. These seeds were germinable and developed into flowering plants in the fields. This study provides a simple system for rapid breeding of P. frutescens and studying the physiological mechanism of flowering of plants.

Effects of Plant Density on the Economic Yield and Agronomic Characters of Perilla frutescens L.

Perilla frutescens L. is a traditional medicinal plant in china. In recent years, with more and more attention is paid to the nutritional and health values of Perilla frutescens L, it has become a potential medicinal and edible crop. Two Per- ilia frutescens cultivars, Qisu No.1 （oil type） and Qisu No.3 （leaf and seed-using type）, bred by Rapeseed Research Institute of Guizhou Academy of Agricultural Sciences, were used as test material for studying the effects of different planting density on the economic yield and agronomic characters of Perilla frutescens L. under moderate fertility level. The results showed the variation tendency of agro- nomic characters with the change of planting density was similar for the two Perilla frutescens cultivars. Among them, the plant height, main-stem nods, first effective branch length, first effective branch height, spikes per plant, and yield per plant were negatively related to the planting density; and main spike length and main- spike grains were positively related to the planting density. The effects of planting density on kilo-grain weight were not significant. The yield of the two cuitivars all increased first and then decreased with the increase of planting density. The re- gression analysis of planting density versus yield of Perilla frutescens L. was per- formed for obtaining the regression equations （Qisu No.l： y=54.046 5＋96.777 lx- 45.719 8x2, r=0.999 4; Qisu No.3：y=38.717 6＋42.111 3x-16.757 7x2, r=0.999 8）. Based on the regression equations, the optimal planting density for Qisu No.1 was 158 745 plants/hm2 with peak yield of 1 578.90 kg/hm2, and for Qisu No.3 was 188 745 plants/hm2 with peak yield of 977.55 kg/hm2. Under the optimal planting density, Qisu No.3 could also harvest 3 000 kg/hm2 of Perilla frutescens dry leaves and 2 400 kg/hm2 of Perilla frutescens stems. This research would provide an important reference for the promotion, high-yield cultivation and comprehensive utilization of Perilla frutescens L.

Perilla frutescens:A traditional medicine and food homologous plant

Perilla frutescens,an annual herb of the Labiatae family,has been cultivated in China for more than 2000 years.P.frutescens is the one of the first medicinal and edible plant published by the Ministry of Health.Its leaves,stems and seeds can be used as medicine and edible food.Because of the abundant nutrients and bioactive components in this plant,P.frutescens has been studied extensively in medicine,food,health care and chemical fields with great prospects for development.This paper reviews the cultivation history,chemical compositions and pharmacological activities of P.frutescens,which provides a reference for the development and utilization of P.frutescens resources.

Comparative transcriptome and co-expression analysis reveal key genes involved in leaf margin serration in Perilla frutescens

Objective:In this study,we aimed to identify the genes involved in leaf margin serration in Perilla frutescens.P.frutescens(Family:Lamiaceae)is widely grown in Asian countries.Perilla leaf is the medicinal part stipulated in the Chinese Pharmacopoeia.There are mainly two types of perilla leaves:one with serrated leaf margin which is the phenotype described in the pharmacopoeia and the other with smooth leaf margin.Methods:Transcriptome sequencing,co-expression analysis,and qRT-PCR analysis of six perilla tissues sampled from two different phenotypes(serrated and smooth leaves)were performed.Results:Forty-three differentially expressed genes(DEGs),which may potentially regulate leaf shape,were identified through de novo transcriptome sequencing between the two groups.Genes involved in leaf shape regulation were identified.Simultaneously,we validated five DEGs by qRT-PCR,and the results were consistent with the transcriptome data.In addition,1186 transcription factors(TFs)belonging to 45 TF families were identified.Moreover,the co-expression network of DEGs was constructed.Conclusion:The study identified the key genes that control leaf shape by comparing the transcriptomes.Our findings also provide basic data for further exploring P.frutescens,which can help study the mechanism of leaf shape development and molecular breeding.

紫苏侧枝扦插技术研究

通过对紫苏(Perilla frutescens)侧枝的处理及生根粉浓度的设计,研究不同处理对紫苏侧枝扦插的效果,确定适合紫苏侧枝扦插的最优组合。结果表明,不同扦插处理在成活率、生根数量、生根长度等方面存在差异,处理6即留取侧枝1对叶片,浸泡生根粉浓度为500 mg/L时成活率最高,达到83.3%,其生根数量、生根长度等指标极显著高于其他处理。

带固相萃取功能索氏提取装置提取紫苏挥发性成分的方法分析

设计了一种带固相萃取功能的索氏提取装置,并采用该装置提取分析紫苏(Perilla frutescens)挥发性成分。结果表明,紫苏中鉴定出75种挥发性组分,用气相色谱峰面积归一法确定其各组分相对质量含量占总挥发性组分的95.64%,其中主要成分为紫苏酮(56.16%)、石竹烯(8.38%)、异脱氢紫苏酮(8.11%)、石竹烯氧化物(4.03%)等。具有香味的成分有45种,占总挥发性组分的88.66%。该装置于密闭环境进行加热萃取,集萃取、净化和浓缩为一体,中途不需要转移样品,大大简化了样品前处理操作,有效缩短了样品前处理周期,为紫苏挥发性及半挥发性成分提供了快速、可靠的分析方法。

紫苏杂种F_(1)的SSR标记鉴定及性状分析

鉴定真实的F1杂种是开展紫苏杂交育种和构建遗传学研究群体的基础。本研究以‘冀紫1号’为母本,‘冀紫2号’为父本配制杂交组合,利用简单重复序列(simple sequence repeat,SSR)标记进行F_(1)杂种真伪性鉴定,分析了亲本和真杂种的农艺性状和生理生化指标。结果表明从29对SSR引物中筛选出1对扩增稳定、条带清晰、能够区分双亲及杂交种的引物ZS-14,利用该引物共鉴定出46株真实杂种。杂种植株在株高、叶长、叶宽、叶片重量和可溶性糖等性状上综合了双亲的优势。本研究结果为紫苏杂种真伪性鉴定及杂交育种提供了指导。

Smashing Tissue Extraction and GC Analysis of Active Fatty Acids from Oil Cake of Perilla Seeds

Objective To optimize the extraction technology of perilla seeds oil from the oil cake of perilla seeds(OCPS)by using the contents of active fatty acids as evaluation standard.Methods The fatty acids were extracted from OCPS,the residue of perilla seeds after cold-press,by smashing tissue extraction(STE),the new technology selected through comparing with classical leaching extraction(LE),Soxhlet extraction(SE),ultrasonic extraction(UE),and supercritical-CO2 fluid extraction(SFE).For optimized condition of STE,orthogonal test was designed and completed.The contents of five fatty acids in extracted oil and OCPS were determined by GC.Results The optimized extraction parameters were smashing for 1.5 min under extraction power of 150 W and 1:6 of the material/solvent ratio.The contents of five fatty acids in the oils extracted by five techniques from OCPS and determined by GC were as follows:α-linolenic acid(41.12%-51.81%),linoleic acid(15.38%-16.43%),oleic acid (18.93%-27.28%),stearic acid(2.56%-4.01%),and palmitic acid(7.38%-10.77%).Conclusion The results show that STE is the most efficient technology with the highest yield(LE:0.57%;SE:1.03%;UE:0.61%;SFE:0.80%;STE: 1.17%)and shortest time(LE:720 min;SE:360 min;UE:30 min;SFE:120 min;STE:1.5 min)among five tested extraction technologies.It is first reported using STE to extract herbal oil enriched with active fatty acids.

紫苏SNP分子标记开发及遗传多样性分析

紫苏(Perilla frutescens(L.))是一种药食兼用型的植物,具有丰富的营养价值和药用价值,本研究利用特异性位点扩增片段测序技术(specific-locus amplified fragment sequencing,SLAF-seq)对30份紫苏种质资源进行了分子标记开发,以‘日本晴’(水稻)为对照,测序获得紫苏全基因组范围内的SNP分子标记。通过测序共获得106.92 Mb Reads数据,各样本Reads数据在1742153~9815872之间,样本平均测序质量值Q30为95.91%;平均GC含量为39.16%。通过生物信息学分析,本研究获得406599个SLAF标签,样本的平均测序深度为19.13倍,其中多态性的SLAF标签共有140387个,共得到419223个群体SNP标记。利用开发的SNP分子标记将30份紫苏种质资源分为2组,紫苏SNP分子标记的研究可为紫苏的遗传图谱构建、种质资源鉴定以及农艺性状的关联分析提供理论基础。

苏子种质资源的遗传多样性分析

为明确苏子种质资源亲缘关系,本研究以收集的44份苏子种质资源为材料,采用SSR、AFLP和SRAP分子标记对其进行遗传多样性和聚类分析。结果表明,SSR、SRAP和AFLP引物的平均多态性分别为63.6%、85.2%和92.6%,44份种质间的遗传相似系数为0.717~0.967(平均为0.864);在遗传相似系数0.762处,44份苏子材料可分为2大类,黄陵苏子和玉泉苏子聚为第Ⅰ类,其余42份材料聚为第Ⅱ类;在相似系数0.84处,可将第二类分为4个亚类。从主成分分析二维图结果来看,44份材料分成5个组,黄陵苏子和玉泉苏子为一组,与其它几组距离较远。本研究结果说明这些苏子种质之间遗传多样性较低,但部分材料间存在较大遗传差异,其中黄陵苏子和玉泉苏子与其他42份材料亲缘关系较远。本研究综合采用了3种标记进行苏子遗传多样性的评价,可为优异种质的创新利用提供理论参考,也为后续大规模的评价鉴定提供依据。