Antioxidant Activity of Ethyl Acetate Extract of Amomum villosum Lour.

[Objectives]To study the antioxidant activity of the ethyl acetate extract of Amomum villosum Lour.[Methods]The removal rate of chelated iron ions,hydroxyl radicals,superoxide anion free radicals and DPPH free radicals by the ethyl acetate extract of A.villosum Lour.was determined by UV spectrophotometer.[Results]0.5000μg/mL ethyl acetate extract of A.villosum Lour.had the strongest ability to chelate with ferrous ions and to remove hydroxyl radicals,superoxide anion free radicals and DPPH free radicals.The ability to chelate with ferrous ions was 95.14%,and the removal rate of the above free radicals was 86.217%,81.44%,and 85.16%.[Conclusions]The ethyl acetate extract of A.villosum Lour.had a strong antioxidant effect,and its antioxidant capacity was related to the sample concentration,which provides a theoretical basis for its application in the development of antioxidant skin care products.

阳春砂(Amomum villosum Lour.)农家种热科2性状初报

【目的】阳春砂(Amomum villosum Lour.)是砂仁中的佳品,具有悠久的药用历史。通过系统分析阳春砂农家种热科2特征性状,为阳春砂品种识别和生产种植提供依据。【方法】对阳春砂农家种热科2植株、花、果实等性状进行跟踪调查研究与对比分析。【结果】热科2阳春砂植株高大粗壮,株高2.84 m,茎粗13.12 mm;叶片大而厚,叶长43.52 cm、宽6.46 cm,外侧叶缘波状不明显、叶面具平行于主脉的突起线,叶舌长椭圆形、棕色、长0.95 mm;花序“曲线型”,花梗长9.70 mm,小花数9.3朵,唇瓣长2.24 mm、宽2.39 mm,中脉黄色而染紫红色、顶端黄色外延、基部紫红色“八”字型痂状斑;果实扁圆形、褐色,内含种子23.2粒,千粒质量14.14 g,种子团挥发油体积分数3.6%;花期和果实成熟期晚。【结论】明确了热科2阳春砂的形态特征性状,可作为砂仁DUS测试指南的标准品种,为该品种的精准识别以及阳春砂优异资源保护与选育种奠定基础。

阳春砂(Amomum villosum Lour.)农家栽培种‘热科1’特征性状初报

阳春砂(Amomum villosum Lour.)是我国四大南药之一,具有极高的药用价值。我国阳春砂栽培种源混杂、退化严重等问题制约着其品质与药效的提升,亟待加强优质资源的筛选与改良。本研究通过调查阳春砂农家栽培种‘热科1’植物学性状,发现:阳春砂主茎高度208㎝,叶缘波状程度强且整齐,叶舌椭圆形、棕色,叶鞘绿黄色、边缘轻度棕色带;新抽芽被鲜艳棕红色膜质鳞片;花序"L"型、花梗长8.46㎝,小花数11.9朵;唇瓣中脉黄染深紫红,花粉囊覆盖处中部黄色、边缘染微红色;果实椭圆形、基部微三棱形,种子团挥发油含量3.7%,乙酸龙脑酯含量1.68%;种子褐色、不规则形状,千粒重11.42 g。本研究对阳春砂农家栽培种‘热科1’植株、花、果实与种子特性进行了详细的描述,具有鲜明的特征,作为砂仁DUS测试指南的标准品种,为识别‘热科1’品种提供科学依据,为砂仁品种保护奠定基础。

春砂仁(Amomum villosum Lour.)种子发芽的试验研究

春砂仁种植于广东、广西、福建、云南、四川等省,以广东为主要产区.生产中多采用分株繁殖的方法,由于种苗不足,大面积发展受到限制.近年不少产区试用种子繁殖,又往往因采收、贮藏、播种或长途运输等处理不当,致使种子发芽率很低.为了探讨春砂仁种子发芽的特性及其与环境之间的关系,从而找出催芽的有效措施,提高种子发芽率,我们从197s年开始,采用几种方法进行了种子发芽的比较试验.

簇毛麦(Dasypyrum villosum)GA20ox基因的分离和鉴定

通过RT-PCR技术克隆了簇毛麦(Dasypyrum villosum)赤霉素生物合成关键酶GA20ox基因的全长序列,命名为DvGA20ox,GenBank登录号为EU142949.该基因全长1080bp,推测编码359个氨基酸的蛋白质,具有植物GA20ox基因的典型保守结构域LPWKET和NYYPXCQKP.该基因推导编码蛋白质与小麦、大麦和黑麦草GA20ox蛋白的同源性分别为98%、97%和91%.该序列重组到原核表达载体pET-32a(+)获得的重组子pET-32a(+)-DvGA20ox转化大肠杆菌BL21pLysS后,IPTG诱导表达,SDS-PAGE分析表明,DvGA20ox基因在大肠杆菌中获得了高效表达,融合蛋白相对分子质量为55×103,与理论值相符.结果为深入研究DvGA20ox蛋白的结构与功能以及该基因与株高发育的关系奠定了基础.

簇毛麦(Dasypyrumvillosum)矮秆突变体的鉴定

为给簇毛麦杂交后代群体中自然突变产生的矮秆突变体在小麦遗传改良中合理有效的利用提供理论依据,对该矮秆突变体的生物学及遗传学特性进行了初步研究。该突变体在苗期与对照植株无明显差异,在成熟期突变体株高30 cm,约为对照株高的1/3。突变株茎的倒一、二节间(最上部的两个茎节)长度分别为对照植株长度的1/9和1/6。细胞学观察表明,突变体的茎节细胞纵向延长受到抑制。此外,该突变株还表现出部分雄性不育,自然结实率低。花粉活力检测发现,突变株可育花粉只有20%,在外施GA3后花粉育性可得到恢复,推测该突变体为赤霉素敏感型矮秆突变体。遗传分析表明,矮秆突变性状受一个隐性基因控制。

簇毛麦(Dasypyrum villosum L.)中一个γ-醇溶蛋白基因序列的研究

目的:研究簇毛麦中一个新的γ-醇溶蛋白基因序列及结构特点,为小麦品质育种和该类基因的进化分析提供资料。方法:利用基因组PCR技术从簇毛麦基因组中克隆到1个γ-醇溶蛋白基因(Dv-γ)的全编码序列,利用生物信息学研究其序列结构特点。结果:该序列与已知的γ-醇溶蛋白有着很高的相似性。推导的氨基酸序列包含5个典型的结构域,其中含有8个保守半胱氨酸残基,高变重复区Ⅱ中的重复单元与其他物种来源基因有较大区别。γ-醇溶蛋白中常见的乳糜泄抗原决定簇在其内部也有发现。进化分析表明,此基因与亚家族Ⅰ中的γ-醇溶蛋白基因有着较近的亲缘关系。结论:获得了一个新的γ-醇溶蛋白基因。

Genetic behavior of Triticum aestivum–Dasypyrum villosum translocation chromosomes T6V#4S·6DL and T6V#2S·6AL carrying powdery mildew resistance

T6V#2S·6AL and T6V#4S·6DL translocation chromosomes developed from the cross of wheat and different Dasypyrum villosum accessions have good powdery mildew （PM） resistance, but their pairing and pyramiding behavior remains unclear. Results in this study indicated that the pairing frequency rate of the two differently originated 6VS chromosomes in their F1 hybrid was 18.9% according to genomic in situ hybridization （GISH）; the PM resistance plants in the F2 generation from the cross between T6V#4S·6DL translocation line Pm97033 and its PM susceptible wheat variety Wan7107 was fewer than expected. However, the ratio of the resistant vs. the susceptible plants of 15：1 in the F2 generation derived from the cross between the two translocation lines of T6V#2S·6AL and T6V#4S·6DL fitted well. Plants segregation ratio （homozygous：heterozygous：lacking） revealed by molecular marker for T6V#4S·6DL or T6V#2S·6AL in their F2 populations fitted the expected values of 1：2：1 well, inferring that the pairing of the two alien chromosome arms facilitates the transmission of T6V#4S·6DL from the F1 to the F2 generation. A quadrivalent was also observed in 21% of pollen mother cells （PMCs） of homozygote plants containing the two pairs of translocated chromosomes. The chromosome pairing between 6V#2S and 6V#4S indicates that it will be possible to obtain recombinants and clarify if the PM resistance determinant on one alien chromosome arm is different from that on the other.

Pm67, a new powdery mildew resistance gene transferred from Dasypyrum villosum chromosome 1V to common wheat(Triticum aestivum L.)

Powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici(Bgt), is a global disease that poses a serious threat to wheat production. To explore additional resistance gene, a wheatDasypyrum villosum 1 V#5(1 D) disomic substitution line NAU1813(2 n = 42) with high level of seedling resistance to powdery mildew was used to generate the recombination between chromosomes 1 V#5 and1 D. Four introgression lines, including t1 VS#5 ditelosomic addition line NAU1815, t1 VL#5 ditelosomic addition line NAU1816, homozygous T1 DL·1 VS#5 translocation line NAU1817, and homozygous T1 DS·1 VL#5 translocation line NAU1818 were developed from the selfing progenies of 1 V#5 and 1 D double monosomic line that derived from F1 hybrids of NAU1813/NAU0686. All of them were characterized by fluorescence in situ hybridization, genomic in situ hybridization, 1 V-specific markers analysis, and powdery mildew tests at different developmental stages. A new powdery mildew resistance gene named Pm67 was physically located in the terminal bin(FL 0.70–1.00) of 1 VS#5. Lines with Pm67 exhibited seedling stage immunity and tissue-differentiated reactions at adult plant stage. The sheaths, stems, and spikes of the Pm67 line were still immune, but the leaves showed a low degree of susceptibility.Microscopic observation showed that most penetration attempts were stopped in association with papillae on the sheath, and colonies cannot form conidia on the susceptible leaf of Pm67 line at adult plant stage, suggesting that the defence layers of the Pm67 line is tissue-differentiated. Thus, the T1 DL·1 VS#5 translocation line NAU1817 provides a new germplasm in wheat breeding for improvement of powdery mildew resistance.

Development of EST-PCR markers specific to the long arm of chromosome 6V of Dasypyrum villosum

Expressed sequence tags-derived polymerase chain reaction(EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provide the evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers specific for chromosome 6 VL of Dasypyrum villosum, 297 primer pairs were designed based on the expressed sequence tags(EST) sequences, which were previously mapped in different bins of the long arms of wheat homoeologous 6 AL, 6 BL, and 6 DL. By using the Triticum aestivum, D. villosum, T. durum-D. villosum amphiploid, and T. aestivum-D. villosum alien chromosome lines involving chromosome 6 V, it was found that 32(10.77%) primers could amplify specific bands for chromosome 6 V, and 31 could be allocated to chromosome arm 6 VL. These 6 VL specific markers provided efficient tools for the characterization of structural variation involving the chromosome 6 VL in common wheat background as well as for the selection of useful genes located on 6 VL in breeding programs.

The Influence of Altitude and Latitude on Breeding of Amomum tsaoko (Zingiberaceae)

Objective: To explore the effect of altitude and latitude on breeding of Amomum tsaoko Crevost et Lemaire, a flexistylous ginger, which fruit is used as common materia medica and a food condiment. Methods: The 7 populations were selected randomly from the three floristic zones of Yunnan. Adult plants and infructescences were chosen randomly to gain flower number and fructification percentage per inflorescence, and seed number per fruit. All date was analyzed by SPSS (13.0 version). Results: As A. tsaoko was distributed (or transplanted) from a habitat at lower latitude and/or altitude to a site of higher latitude and/or altitude, the flower number per inflorescence increased, on the contrary, the fructification percentage per inflorescence decreased. The competition for reproductive resource was beneficial to increase flower number and seed production. Conclusions: The habitats in south of the tropic of cancer were favorable to the reproduction of A. tsaoko, which reproductive costs were lower and harvest was higher. Increasing flower number per inflorescence may be a strategy to promote the plant to distribute into alpine habitats for both female and male reproductive success.

Tsaokoin,a New Bicyclic Nonane from Amomum tsao-ko

7α-Hydroxy-8β,9β-H- cis- bicyclo[4,3,0]non-4-ene-4-aldehyde, a nonane compound with novel skeleton, named tsaokoin, was isolated from the fruits of Amomum tsao-ko. Its structure was established on the basis of spectral analysis.

Introgression of sharp eyespot resistance from Dasypyrum villosum chromosome 2VL into bread wheat

Wheat sharp eyespot, a stem disease caused by the soilborne fungus Rhizoctonia cerealis van der Hoeven,has become a threat to wheat production worldwide. Exploiting resistance resources from wild relatives of wheat is a promising strategy for controlling this disease. In this study, a new wheat–Dasypyrum villosum T2DS·2V#4L translocation line in the background of Chinese Spring(CS) showed stable resistance to R. cerealis. Introgression of the T2DS·2V#4L chromosome into wheat cultivar Aikang 58 by backcrossing produced a marked increase in sharp eyespot resistance in NIL-T2DS·2V#4L in comparison with NILT2DS·2DL, and no detrimental effects of 2V#4L on agronomic traits were observed in the BC2F2, BC2F2:3,and BC2F2:4generations. Flow-sorted sequencing of 2V#4L yielded 384.3 Mb of assembled sequence, and8836 genes were predicted of which 6154 had orthologs in at least one of the 2AL, 2BL, and 2DL arms of CS, whereas 1549 genes were unique to 2V#4L. About 100,000 SNPs were detected in genes of 2V#4L and2DL in 10 sequenced bread wheat cultivars. A Kompetitive Allele Specific Polymerase chain reaction and30 conserved ortholog sequence markers were developed to trace the 2V#4L chromatin in wheat backgrounds. T2DS·2V#4L compensating translocation lines represent novel germplasm with sharp eyespot resistance and the markers will allow rapid detection in breeding programs.

Research progress on chemical constituents and pharmacological activities of Amomum longiligulare T.L.Wu

In order to develop and utilize the tropical medicinal resource Amomum(A.)longiligulare T.L.Wu more reasonably and effectively,this paper summarizes the research progress in chemical constituents and pharmacological activities of A.longiligulare by consulting the literatures,so as to provide a certain theoretical basis for exploring its material basis.The chemical constituents of A.longiligulare mainly include volatile oil,diphenylheptane and flavonoids,which have good efficacy in anti-peptic ulcer,as well as antioxidant,analgesic and other pharmacological activities.This paper briefly discusses the reasons for the differences in the composition of volatile oil studied by different scholars.By summarizing its pharmacological activities,it is found that its various pharmacological activities may be the basis of its anti-peptic ulcer.

Chemical constituents and biological activities of essential oils of Amomum genus(Zingiberaceae)

Amomum Roxb.includes the aromatic and medicinal plants native to tropical and subtropical Asia belonging to the family Zingiberaceae.Members of Amomum genus have been used for a long time in traditional medicine for the treatment of throat trouble,congestion of lungs,inflammation of eyelids,and digestive disorders,etc.Amomum essential oils have been studied for their chemical profiles in which limonene,allo-aromadendrene,1,8-cineole,camphor,farnesyl acetate,α-pinene,β-pinene,caryophyllene,camphene,D-camphor,santolina triene,methyl chavicol,bornyl acetate,β-elemene,δ-3-carene,etc.were the major compounds.Furthermore,the oils extracted from Amomum plants have been reported to possess antimicrobial,antioxidant,insecticidal,larvicidal,cytotoxic,anti-scabies,and anti-inflammatory activities.This review focuses on the chemical constituents and biological activities of the essential oils isolated from the different plant parts of Amomum plants.The objective of the present review is to highlight therapeutic potentials and provide evidence for future medicinal applications of these species of genus Amomum.

Physical Land Suitability Assessment for the Large Cardamom Amomum subulatum Roxb.Cultivation in Hills of Kathmandu Valley

Nepal is one of the top three exporters of larger cardamom （Amomum subulatum Roxb.） in the world market contributing largely to the Nepalese economy. Sub-Himalayan range of hilly area of eastern Nepal is popular for large cardamom cultivation. Being similar phytogeographical and ecological setting, cultivation has begun in hills of Kathmandu Valley however land suitability needs to be evaluated with reference to specific need of the crop. Kathmandu is a valley with alluvial bottom and altitudinal range between 1200 and 2730m above sea level having cool to warm temperate climatic range. Varieties of aspects and slope gradient have potentiality for the large cardamom cultivation. Aim of the study was set to categorize available land area in the range of suitability for large cardamon cultivation on the basis of physical characteristics of the land and ecological need of crop. Land evaluation methodology has been designed along with site specific amendment in FAO framework （1976）. Land use map in scale of 1：25 000 was used in geographic information system environment. Suitability was done through map overlaying methods. Aim of the study was set to categorize available land area in the range of suitability categories for large cardamom cultivation based on physical characteristics of the land matching with ecological need of crop. Analysis of data shows that altitudinal range at uplands of Kathmandu Valley and annual average rainfall and temperature are found to be within suitability range for Amomum subulaturn cultivation. Sandy loam soil, slightly acidic in nature with gentle sloping land of about 5-8° provides highly suitable condition. Result showed that more than one-third of total land area above 1800 m of altitude is suitable. Selectively, of the total area, 5% are north facing slope, 13% gentle slopes are highly suitable. 16% lands with north east orientation are moderately suitable. Most of the land areas which are marginaly suitabile for vegetable and cereal crops are highly suitable for large cardamom. Hills of northern boundary hills found to be suitable due to more rainfall distribution pattern compared to southern boundary hills. Therefore, land evaluation also identifies limitations and input requirements for sustainable production.

Characterization and Expression Profiling of Genes Encoding the Gibberellin 3-oxidase in Dasypyrum villosum Dwarf Mutant

Gibberellin 3-oxidase catalyzes the conversion of inactive gibberellin(GA) species into GAs with biological activity and it is subjected to strict developmental controls in the life cycle of a plant. In this study, 33 gene sequences, encoding the gibberellin 3-oxidase(GA3ox) from Dasypyrum villosum and its dwarf mutant, were obtained. Each contained a 1 107 bp coding sequence(CDS) that encoded a putative protein containing 369 amino acids. The GA3ox protein showed 77% to 97% homology and shared the major conserved structural domains of GA3ox proteins with rice, sorghum bicolor, oat, barley, and wheat. Sequence alignment showed that there were 20 single nucleotide polymorphisms(SNPs) and 22 Insertion/deletions(In Dels) among these sequences, which could be divided into 2 haplotypes, haplotypes Ⅰ and Ⅱ. Haplotype Ⅰ was found in the wild type and was1 495 bp in length, and haplotype Ⅱ was found in the dwarf mutant and was 1 485 bp in length. The Q-PCR results showed that GA3ox was expressed in the leaves, roots, internodes, and stem nodes, and that there was a significant difference in the transcript level of the GA3ox between the wild type and dwarf mutant. The transcript levels of GA3ox in the leaves at the seedling stage, stem elongation stage and the heading stage, in the root and stem nodes at the stem elongation stage and in the internodes at the heading stage of the wild type, were significantly higher than those in the dwarf mutant. However, GA3ox expression in the rest of the wild type tissues at the 3 stages was slightly higher than or not different from the dwarf mutant.The results suggested that the wild type and mutant allele sequences of GA3ox in D. villosum showed 2 amino acid changes in exons and variations in the lengths of introns or the SNPs in introns, which most probably impaired the function of the enzyme,affected the GA3ox expression level, and eventually gave rise to dwarfing.

Optimization of extraction process of Amomum longiligulare T.L.Wu polysaccharides by response surface methodology

Objective:To optimize the technological conditions of ultrasonic-assisted extraction of Amomum longiligulare T.L.Wu polysaccharides and increase the yield of polysaccharides.Methods:The polysaccharide was prepared by water extraction and alcohol precipitation method,and the yield of polysaccharide was taken as an index.The effects of ultrasonic time,extraction times,ultrasonic temperature,and water-to-material ratio on polysaccharide yield were investigated through a single factor experiment.Combined experimental design and response surface analysis were used to optimize the extraction process of Amomum longiligulare T.L.Wu polysaccharides.Results:The optimum conditions of ultrasonic-assisted extraction were determined as follows:extraction time of 29 min,three extraction times,water bath extraction temperature of 68℃,water-to-material ratio of 15:1;under these conditions,the polysaccharide yield was 10.69%.Conclusion:The results are close to the predicted values of the model.This optimization test is effective and feasible,and provides a reference for the related research of Amomum longiligulare T.L.Wu.

豆蔻属Amomum药用植物资源的初步研究

目的为豆蔻属药用资源的利用和保护奠定基础。方法野外调查和文献整理。结果豆蔻属Amomum是姜科的第二大属,我国有记载的豆蔻属药用植物共20种和变种,其中,砂仁、豆蔻、草果是被《中国药典》收载的3味主流药材,其他均为民间或民族用药。砂仁、豆蔻、草果均属于芳香化湿药,砂仁的应用最广泛,用于51个药典方剂,其次是豆蔻,用于21个药典方剂,草果用于5个药典方剂。结论结合豆蔻属分子系统学研究结果和传统中药材知识,易于鉴别砂仁、豆蔻、草果3味药材,以及砂仁、草果及其混淆品的鉴别。豆蔻属药用植物的分布较为狭窄,对其资源的保护和开发利用值得重视。

Antioxidant activities of different extracts from Amomum longiligulare Fruits

Objective:To study the antioxidant activities of five extracts ofAmomi longiligulare (A. longiligulare) to determine the antioxidant fraction.Methods: The ethanol extract ofA. longiligulare was extracted by systematic solvent extraction to obtain the petroleum ether, dichloromethane, ethyl acetate, n-butanol fractions successively. Thein vitro antioxidant activities of the five extracts were evaluated by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical and hydroxyl radical (OH-) scavenging assay methods.Results: The five extracts showed different extent of antioxidant activities in vitro, especially the ethyl acetate and dichloromethane fractions.Conclusion: The difference of antioxidant activity of different extracts may be associated with the type and structure of antioxidant components in each fraction of A. longiligulare fruits.