远志养分吸收规律及其与药效成分积累的关系

【目的】 远志 (Polygala tenuifolia Willd) 是多年生草本植物,以根入药,是我国重要的大宗药材之一。研究远志的营养元素吸收规律及其与药效成分积累的关系,为远志人工栽培中科学施肥及专用肥料的研制提供理论依据。 【方法】 以3年生远志为研究对象,2017年3月26日幼苗全部返青,从返青后40天 (5月4日) 开始,每隔20 天田间采集植株样一次,分别测定其根部和地上部干物质量,氮磷钾等8个营养元素含量以及根中3类药效成分 (总皂苷、总黄酮和总酚) 的含量。 【结果】 远志地上部干物质积累在整个生育期内呈先升高后下降的趋势,根部干物质积累呈直线上升趋势。返青后40～140 天地上部干物质积累量大于根,返青后 160～195天生长中心转移到根,根的干物质积累量大于地上部。远志植株对氮素的最大吸收时期在返青后100～140天,吸收量为18.77 g/m2,对磷、钾的最大吸收时期在返青后140 天,吸收量分别为3.07 g/m2和3.36 g/m2。远志根中总皂苷、总黄酮、总酚累积量的趋势基本一致,均为先升高后降低,累积量均在返青后180天达到最大值。总皂苷积累量出现两次峰值,分别在返青后140天 (6.28 g/m2) 和180天 (6.43 g/m2),总酚累积量也出现两次峰值,分别在返青后80天 (2.37 g/m2) 和180天 (4.31 g/m2),总黄酮积累量只有一次峰值,出现在返青后180天 (2.35 g/m2)。根药效成分经济产量在返青后180天达到最高。总黄酮含量与根中锌元素含量呈显著正相关 (P < 0.05);总酚含量与根中锌元素含量呈极显著正相关 ( P < 0.01)。 【结论】 返青后140～195天是远志根干物质积累最快的时期。远志氮营养最大效率期在返青后100～140天,磷、钾营养最大效率期在返青后140天。在远志专用肥料配制中应保证锌元素的供给,以满足远志生长发育的需求,从而有效提高药效成分的积累。远志根秋季采挖的最佳时间是返青后180天 (9月25日)。      

Dammarane-type glycosides from steamed notoginseng

Notoginseng, the root of Panax notoginseng (Burk.) F. H. Chen (Araliaceae), is a Chinese traditional medicine, which is used in both raw and processed forms due to their different pharmacological activities. Detailed chemical investigation on steamed notoginseng led to the isolation of 27 dammarane-type triterpenoids (1-27), including 4 new glycosides, namely, notoginsenosides ST-1-ST-3 and ST-5 (1-4), in addition to 3 other known compounds. Of the new compounds, 1-3 possess new aglycones. Their structures were elucidated on the basis of detailed analyses of their 1D and 2D NMR spectra and chemical reactions. The known compounds, koryoginsenoside-R1 (19), yesanchinoside D (20), 6'-O-acetylginsenoside Rg3 (24), and 3beta,6alpha,12beta-trihydroxydammar-20(21),24-diene ( 25), were isolated from notoginseng for the first time.     

Chemical and genetic assessment of variability in commercial Radix Astragali (Astragalus spp.) by ion trap LC-MS and nuclear ribosomal DNA barcoding sequence analyses

Radix Astragali (Huangqi) has been demonstrated to have a wide range of immunopotentiating effects and has been used as an adjuvant medicine during cancer therapy. Identity issues in the collection of Radix Astragali exist because many sympatric species of Astragalus occur in the northern regions of China. In order to assess the quality, purity, and uniformity of commercial Radix Astragali, 44 samples were purchased from herbal stores in Hong Kong and New York City. The main constituents, including four isoflavonoids and three saponins, were quantitatively determined by liquid chromatography mass spectrometry (LC-MS). There was significant sample-to-sample variability in the amounts of the saponins and isoflavonoids measured. Furthermore, DNA barcoding utilizing the variable nuclear ITS spacer regions of the 44 purchased Radix Astragali samples were sequenced, aligned and compared. Eight polymorphic point mutations were identified which separated the Radix Astragali samples into three groups. These results indicate that the chemical and genetic variability that exists among Radix Astragali medicinal products is still a consistency and quality issue for this herbal. Two-way ANOVA analysis showed significant effects on the contents of the seven tested compounds when both phylogenetic and geographic (i.e., point of purchase) factors were considered. Therefore, chemical profiles determined by LC-MS and DNA profiles in ITS spacer domains could serve as barcode markers for quality control of Radix Astragali.     

Assessment of biochemical and antioxidant diversities in a shallot germplasm collection from Vietnam and its surrounding countries

The shallot is considered an important genetic resource for Allium breeding programs because, compared to the bulb onion, the shallot contains higher levels of several functional chemical compounds. However, there may be differences in content, composition and beneficial activity among shallot landraces. This study was carried out to characterize the differences in phenolic, quercetin, sugar, S-alk(en)yl-l-cystein sulfoxide (ACSO), and saponin contents and antioxidant capacities of a shallot germplasm including 31 strains derived from different regions of Vietnam and six other countries. A wide variation was observed in the quantitative analyses of the chemical contents. Shallots with high contents of polyphenols, saponins, and quercetins were found in the south of Vietnam and other low-latitude countries. Meanwhile, those possessing fairly high ACSO and sugar contents were observed in the north of Vietnam. Qualitative analysis of saponins via thin layer chromatography did not show clear variation among shallot strains, but polymorphism was observed between the shallot and other Allium species, such as A. roylei. The principal component analysis could clearly discriminate shallot strains by their geographical origins. All shallot strains showed potent antioxidant activities in a DPPH assay. The highest antioxidant capacity was in the strains possessing relatively high contents of polyphenol, quercetin, and saponin. Significant correlations were found between antioxidant capacity (IC ₅₀ ^?1 ) and four groups of chemical compounds (polyphenols, quercetins, saponins, and ACSOs) (r = 0.40–0.59). A strong correlation was observed between IC ₅₀ ^?1 and quercetin contents (r = 0.59, p < 0.01). The six Fusarium-inoculated shallot strains seemed to be adequately resistant against disease, and the levels of resistance may be related to the saponin content in the bulb tissues.     

基于红外光谱和化学计量学的紫金牛药材质量快速鉴定研究

本文利用FTIR技术分别检测不同产地紫金牛及其易混淆植物的化学成分的光谱特征,并通过主成分分析模型以及主成分载荷因子等化学计量学方法,对各样本进行鉴别,找出各测试样本化学组成的差异。结果表明：f1）两种紫金牛在3378cm^-1、2923cm^-1、1735cm^-1、1621cm^-1、1444cm^-1、1373cm^-1、1321cm^-1、1238cm^-1、1153cm^-1、1033cm^-1及776cm^-1附近均有吸收峰。（21由于融水县和永福县两地相距最近,在PCA检测模型二维散点图上,这两产地样品的距离也最近,恭城县与融水县及广西兴安县相距较远,故在散点图上的距离也较远;广西兴安县的月月红是紫金牛属植物,但不属于紫金牛种,所以,在散点图上的距离也就偏离各产地紫金牛。（3）提取PCA模型的载荷因子,分析表明,在种问比较,样品月月红中脂类、苯醌、黄酮及皂苷类成分的含量较紫金牛样品丰富。在紫金牛种内比较,广西永福县的样品香豆素、酮脂及皂苷类成分的含量比其它产地高,相反地,广西恭城药市的样本中尤其是酮脂及皂苷类成分的含量最低;但是,广西恭城县的紫金牛中香豆素、苯醌、甾醇,黄酮类等成分,较其它产地丰富。     

三七收获后不同年限土壤微生物代谢多样性分析

采用Biolog技术,对云南省砚山县的盘龙、阿猛、干河3个乡镇进行了三七历年种植地的调查研究,以未种植过三七的土壤为对照,研究三七收获后1~6 a不同年限对土壤养分、土壤微生物对碳源利用以及土壤微生物多样性的影响。结果表明,反映土壤微生物活性的平均颜色变化率(AWCD)并未随年限增加呈现明显变化规律;微生物培养96 h活性旺盛。对Biolog板31种碳源吸光度值聚类分析表明,3个乡镇所取三七收获后1 a和6 a土壤微生物碳源利用均可聚为一类,其土壤微生物碳源利用特征相似,其余不同年限土壤微生物碳源利用聚类并未出现一定规律;与未种植过三七的土壤相比,三七收获后1 a、6 a土壤微生物均对碳水化合物类、聚合物类、羧酸类和酚类碳源的利用分别高出25.97%~55.59%和53.14%~65.68%;随着三七收获后年限的增加,土壤微生物对碳水化合物、氨基酸类、羧酸类碳源利用呈收获后2 a升高、4 a和5 a下降趋势。收获三七后1~6 a土壤与未种植过三七的土壤相比,氮、磷、钾及有机质含量差异均不显著。除干河乡外,阿猛乡和盘龙乡收获三七后1~6 a土壤微生物与未种植过三七的相比,Shannon-Wiener指数、丰富度指数、Simpson指数均值总体差异不明显。试验说明三七轮作至少需6 a以上时间,土壤微生物对不同碳源的优势利用可反映出三七连作障碍与土壤微生物群落结构差异有密切联系。     

基于形态性状和指标成分的远志遗传多样性分析及综合评价

为了揭示远志种质资源遗传多样性地理分布的特点和种质资源群体间的遗传关系,并筛选出相对合理的评价指标,本研究利用形态性状数据以及3个主要指标成分(远志皂苷、远志𠮿酮Ⅲ和3,6'-二芥子酰基蔗糖)含量,对省内11个不同来源的26份和省外7个不同来源的16份三年生远志进行遗传多样性分析。研究发现远志在形态性状以及指标成分含量上具有明显差异,变异系数和遗传多样性指数分别在13%~49%、19%~30%和0.7~1.7、0.9~1.4之间;远志𠮿酮Ⅲ含量与花色深度呈极显著正相关性,与株高呈显著正相关性,与木芯粗呈极显著负相关性;3,6'-二芥子酰基蔗糖含量与细叶远志皂苷含量呈显著正相关性,与株高呈极显著负相关性;细叶远志皂苷含量与茎分支数呈显著负相关性。主成分分析将15个指标简化为4个因子,累计贡献率达67.764%;形态性状和指标成分含量的聚类分析结果均将远志分为三大类群,但各类群成员并不一致,利用综合评价F值筛选出8份优良种质;各种质的综合得分范围为-1.15~1.23;利用逐步回归分析筛选到远志种质评价的7个关键性指标,分别为茎分支、叶色、花色、根重、木芯重、远志𠮿酮Ⅲ含量和3,6'-二芥子酰基蔗糖含量。综上所述,参试远志种质资源主要形态性状和指标成分的遗传多样性较为丰富,变异幅度较大。本研究从42份种质资源中筛选出8份综合表现良好的优异单株,可用于进一步开发和利用,为远志种质创新利用、品种改良提供了理论依据。     

Triterpene saponins from aerial parts of Medicago arabica L

Eight major triterpene saponins have been isolated from the aerial parts of Medicago arabica and their structures elucidated by FAB-MS and NMR analysis. Three of them are new compounds and are identified as 3-O-(alpha-L-arabinopyranoside) bayogenin, 3-O-(alpha-L-arabinopyranosyl), 28-O-(beta-D-glucopyranoside) bayogenin, and 3-O-[alpha-L-arabinopyranosyl(1-->2)-beta-D-glucuronopyranosyl], 28-O-beta-D-glucopyranoside 2-beta-hydroxyoleanolic acid. Two saponins, identified as 3-O-(alpha-L-arabinopyranoside) hederagenin and 3-O-(alpha-L-arabinopyranosyl), 28-O-(beta-D-glucopyranoside) hederagenin are known compounds but not previously reported as saponin constituents of Medicago species, while three other saponins, being mono- and bidesmosides of hederagenin, have been previously isolated from roots of M. sativa.     

灌水频率和施肥量对三七有效成分积累与发病率的影响

为探明灌水频率和施肥量对三七有效成分积累及发病率的影响。以2年生三七为试验对象,设置W1（3 d灌1次水）、W2（5 d灌1次水）、W3（7 d灌一次水）、W4（9 d灌1次水）4个灌水频率水平;F1（75 kg/hm2）、F2（150 kg/hm2）2个施肥水平;采用完全组合设计,共8个处理。结果表明,随着灌水频率的增加,三七光合速率、蒸腾速率、叶水势均呈先增加后减小趋势,在花期达到峰值,且W2处理均最大。不同处理的株高、茎粗、叶长及叶宽变化趋势大致相同且均在营养生长期大幅增加,在果期达到最大。W3F2处理下有效成分积累量最大（1084.93 mg/kg）,特别是与W1F2相比,W3F2处理三七有效成分积累量极显著增加了61.10%。此外,随着灌水频率的增加,三七根腐病、黑斑病在高频率灌水W1处理条件下发病率最高（P<0.05）,分别为15.79%、3.41%;干叶病发病率最低,仅为2.11%。综上所述,微喷灌施肥条件下低频灌水W3（7 d灌1次水）和高肥F2（150 kg/hm2）组合处理适宜三七叶片生长,且三七根中有效成分积累多,发病率低。本研究对优化三七水肥管理,实现三七绿色可持续发展具有重要理论及指导意义。     

化肥和生物有机肥配施对鲜食型甘薯块根产量、品质及土壤肥力的影响

  【目的】  研究生物有机肥和化肥对鲜食型甘薯干物质积累、产量品质及土壤肥力的影响,为鲜食型甘薯的优质高效生产提供技术支撑。  【方法】  2018和2019年,以鲜食型主栽品种‘济薯26’为供试品种,在山东济南丘陵区进行了两年定位田间试验。设置不施肥(对照)、单施化肥、单施生物有机肥、化肥生物有机肥配施处理。在移栽后50、100和150天,取样分析植株干物质积累和分配。于成熟期,调查分析块根产量及品质,以及0—20 cm土层土壤肥力指标。  【结果】  与单施化肥相比,单施生物有机肥和化肥生物有机肥配合处理均显著提高了两年度移栽后150天甘薯植株干重和块根干重,块根产量以化肥配合生物有机肥处理最高,单施生物有机肥次之,均显著高于单施化肥。与单施化肥相比,单施生物有机肥可显著提高两个年度土壤脱氢酶和蔗糖酶活性,显著提高第二年度土壤脲酶和碱性磷酸酶活性及土壤速效磷、速效钾和有机质含量,促进干物质向块根分配,显著提高了第二年度块根还原糖、可溶性糖和维生素C含量。与施用生物有机肥相比,化肥生物有机肥配施显著提高了两个年度移栽后100天和150天植株干重和块根干重,显著提高第二年度土壤碱性磷酸酶和脱氢酶活性,提高土壤碱解氮和速效磷含量,还原糖和可溶性糖含量无显著变化,但显著增加类胡萝卜素和维生素C含量。相关分析表明,块根品质指标与土壤有机质、速效磷和速效钾含量显著正相关,与土壤酶活性亦有显著正相关关系。  【结论】  连续进行化肥与生物有机肥配施,可显著提高土壤碱性磷酸酶和脱氢酶活性,增加土壤碱解氮和速效磷含量,促进甘薯植株和块根干物质的持续积累,获得高产,同时保持较高的块根糖含量和维生素C含量,是提高鲜食型甘薯产量和品质的有效措施。     

Genetic diversity analysis of hulless barley from Shangri-la region revealed by SSR and AFLP markers

For adding the hulless barley resources of Shangri-la region to the global barley resource library, a basic work was done by us to assess their genetic diversity of this region. The genetic diversity of 60 hulless barley samples collected from three counties in Shangri-la region of Yunnan Province, were studied using SSR (simple sequence repeats) and AFLP (amplified fragment length polymorphism) markers. A total of 70 alleles were detected for 19 pairs of SSR primers, and 525 band containing 464 polymorphic bands were revealed for 5 pairs of AFLP primers. The value of polymorphism information content (PIC) ranged from 0.03 to 0.86 for SSR primers. The total numbers of alleles were 51, 55, 43 in three populations and the polymorphic bands were 188, 205 and 141. The genetic distances and genetic identity among the three populations showed their close relationship. The gene diversity among populations relative to the total population diversity (Gst) was 0.13 for SSR markers and 0.02 for AFLP markers and indicated that just 13 and 2% variations were among populations, respectively. The UPGMA cluster analysis revealed that all of the samples grouped randomly rather than clustered into distinct groups corresponding to their populations, row types and spring/fall types. We concluded that there was high genetic diversity in the population of Shangri-la region and the formation of diversity was related to complex environment and inhabitants’ traditional practices.     

Studies of selected plant raw materials as alternative sources of triterpenes of oleanolic and ursolic acid types

The qualitative and quantitative evaluation of triterpene aglycones of saponin fractions isolated from vegetative and generative organs of three Silphium species, Silphium perfoliatum, Silphium trifoliatum, and Silphium integrifolium, as compared to materials used in the herbal industry such as Panax quinquefolium root and Calendula officinalis flower, was performed. The analyses revealed that triterpene aglycones of saponins isolated from tested Silphium and Calendula species were oleanolic acid and ursolic acid. It was found that Panax roots contained only the aglycone of oleanolic acid within the triterpene saponin group. The leaves of Silphium harvested in May were characterized by the highest content of oleanolic acid-They contained 17.03 mg/g dry weight of the triterpenic acid, on average. The seasons before flowering and at the beginning of that stage appeared to be the most efficient periods for leaf collection in reference to triterpene aglycone contents in plant yield. Moreover, it was found that inflorescences of S. trifoliatum and S. integrifolium contained oleanolic acid in amounts of 22.05 and 17.95 mg/g dry weight respectively, whereas Calendula flowers contained 20.53 mg/g dry weight. The oleanolic acid content in Panax roots was 3.15 mg/g dry weight. Ursolic acid most abundantly occurred in S. integrifolium and S. trifoliatum at concentrations of about 14.98 mg/g dry weight in leaves harvested before flowering (June) and to 15.50 mg/g dry weight in leaves collected during flowering.     

长期施氮对谷子根系内生真菌群落特征的影响

为探讨长期施氮对作物运输根和吸收根内生真菌群落特征的影响及其机制。以农田长期定位施肥试验(1995—2019年)为基础,通过高通量测序技术,研究长期5个施氮肥处理(CK、N1、N2、N1P、N2P。其中CK、N1、N2施氮量分别为0,55.2,110.4 kg/hm~2)对谷子不同功能根内生真菌群落组成和结构的影响。结果表明:CK处理中吸收根的OTUs数和真菌多样性(shannon指数和Chao 1指数)均显著高于运输根(P0.05)。门水平上,吸收根的Ascomycota和Glomeromycota丰度显著高于运输根(P0.05),但Mortierellomycota和Basidiomycota丰度显著低于运输根(P0.05)。在属水平上,优势菌属Mortierella和Bipolaris在吸收根内的相对丰度显著低于运输根(P0.05),但Fusarium和Minimedusa显著高于运输根(P0.05)。高氮施肥吸收根和运输根的OTUs数、Chao 1指数分别提高了23.62%,23.87%,其他施肥则相反。施肥运输根的shannon指数提高了7.12%～19.62%,吸收根则相反。施肥吸收根致病菌属(Bipolaris、Fusarium、Magnaporthiopsis、Microdochium)的相对丰度分别增加了52.99%,40.74%,133.06%,200.00%,运输根中抗病菌属(Minimedusa)的相对丰度提高了40.16%～97.11%。施肥谷子根系碳、氮、磷含量分别提高了2.33%～11.63%,13.30%～94.66%,3.89%～263.92%,氮磷肥配施处理的影响尤为显著(P0.05)。施肥运输根生物量提高了106.67%～336.41%,吸收根的生物量降低了35.20%～60.20%。长期施肥通过提高土壤速效磷、全磷、可溶性氮和根系碳含量而提高吸收根内生真菌丰度,通过提高根系生物量而降低运输根内生真菌丰度,低氮磷配施肥(N1P)对土壤养分含量及菌群发展影响最大。探明长期施氮肥对黄土丘陵区谷子不同功能根系内生真菌分布格局变化的影响,为作物的高产优质及农田土壤生态环境的良好构建提供理论依据。     

Phytochemistry of wild populations of Panax quinquefolius L. (North American ginseng)

A survey of the phytochemistry of Panax quinquefolius L. (North American ginseng) collected from wild populations in Ontario, Quebec, Maine, Vermont, and Wisconsin was undertaken. Reverse-phase HPLC was used to determine the natural variation of levels of ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd and their total in leaf, stem, and root of authentic wild-grown material. The totals in roots varied from 1 to 16%, with the greatest number of individual samples having 4-5% total ginsenosides. The lack of ginsenoside Rf in roots of authentic wild populations confirmed its status as a phytochemical marker differentiating American and Asian ginseng. Ten geographically isolated wild populations were collected, and several showed significant variation in the levels of major ginsenosides. There was no statistical difference in mean ginsenoside content between wild and cultivated P. quinquefolius roots at 4 years of age, suggesting there is no phytochemical justification for wild crafting. Baseline data on total ginsenoside levels for authentic wild P. quinquefolius reported here provide reference levels for quality assurance programs.     

Optimization of microwave-assisted extraction (MAP) for ginseng components by response surface methodology

Response surface methodology (RSM) was applied to predict optimum conditions for microwave-assisted extraction-a MAP technology-of saponin components from ginseng roots. A central composite design was used to monitor the effect of ethanol concentration (30-90%, X(1)) and extraction time (30-270 s, X(2)) on dependent variables, such as total extract yield (Y(1)), crude saponin content (Y(2)), and saponin ratio (Y(3)), under atmospheric pressure conditions when focused microwaves were applied at an emission frequency of 2450 MHz. In MAP under pre-established conditions, correlation coefficients (R (2)) of the models for total extract yield and crude saponin were 0.9841 (p < 0.001) and 0.9704 (p < 0.01). Optimum extraction conditions were predicted for each variable as 52.6% ethanol and 224.7 s in extract yield and as 77.3% ethanol and 295.1 s in crude saponins, respectively. Estimated maximum values at predicted optimum conditions were in good agreement with experimental values.     

45Ca示踪技术研究三七提取物对细胞Ca2+内流与外溢的影响

采用45Ca同位素示踪技术,进行了三七提取物对大鼠内脏器官细胞Ca2+内流与外溢的影响研究。结果表明,从三七中提取的总皂苷和总黄酮对大鼠的心脏、肾脏、动脉平滑肌等内脏器官细胞钙通道具有一定的调控作用,既能阻滞Ca2+内流,也能促进已流入细胞内的Ca2+外溢。与总黄酮相比,总皂苷的作用更强,其调控作用可与阳性对照药物Verapamil相比。这一结果提示皂苷和黄酮提取物具有钙拮抗作用,可能是治疗冠心病的有效成分。     

NrDNA internal transcribed spacer revealed molecular diversity in strains of red seaweed <Emphasis Type="Italic">Porphyra yezoensis</Emphasis> and genetic insights for commercial breeding

Unraveling the cryptic genetic diversity and selective breeding network in various Porphyra strains is of significance for conservation and utilization of economically important nori crops, for both current and future needs. Here, we used nuclear ribosomal spacer (ITS1) region to investigate the genetic variation and intra-specific relatedness of 59 Porphyra yezoensis Ueda specimens worldwide using phylogenetic and parsimony genealogical approaches. 23 nrDNA ITS1 genotypes were revealed and clustering analysis grouped them into two distinct clades. High genetic diversity was detected in wild P. yezoensis strains from Miyagi and Hokkaido Prefectures in Japan, while the cultivated strains from China and South Korea exhibited relatively higher genetic diversity likewise, which provided crucial genetic insights for future commercial breeding of P. yezoensis on a global scale. In addition, phylogenetic study has revealed the genetic relationship of strains with unknown parentage to those with known parentage, and also ITS1 sequence pattern could be correlated with the geographic origin of P. yezoensis specimens. All these pedigree information generated from this research can be used to select parents for inter-specific or intra-specific selective breeding and cross breeding to maximize the preservation of stock resource and sustainable development of nori industry.     

酸性红壤上玉米不同部位固氮微生物群落丰度和组成特征

  【目的】   发挥微生物固氮功能可降低农田化学氮肥的投入,对于缓解土壤酸化具有重要意义。固氮微生物广泛存在于植物体的各个部位,了解酸性土壤上作物不同部位固氮微生物群落特征,为挖掘其功能潜力提供数据支撑和理论基础。   【方法】   选择性状差异较大的耐铝玉米品种先玉335和铝敏感品种Mo17为试验材料,在酸性红壤上种植1个月后,收集玉米地上部、根部和根际土壤样品,分析玉米生物量、养分含量和根际土壤基本理化性质;采用实时荧光定量PCR和高通量测序技术,以nifH基因为标靶,分析不同玉米品种、不同取样部位 (叶部、根部和根际土壤) 固氮微生物群落的丰度、多样性和结构组成。   【结果】   先玉335地上和地下生物量显著高于Mo17,但是植株氮、磷浓度和根际土壤pH、铵态氮和速效钾含量却低于Mo17。实时荧光定量PCR结果显示,nifH基因丰度在玉米不同部位间的差异程度明显大于玉米品种间的差异,其中根部丰度显著高于根际土壤和叶部,而且先玉335根部nifH基因丰度要高于Mo17。高通量测序分析显示,变形菌门 (Proteobacteria) 和蓝藻门 (Cyanobacteria) 是固氮微生物的优势门,根际土壤和根部的优势菌属是慢生根瘤菌属 (Bradyrhizobium),而叶部优势菌属是细鞘丝藻属 (Leptolyngbya),多数优势菌属相对丰度在玉米不同部位间呈现显著差异。取样部位显著影响固氮微生物群落多样性指数 (Shannon指数和OTU数量),但同一部位品种间没有明显差异,其中根际多样性指数显著高于根部,两者均高于叶部。非度量多维尺度 (NMDS) 和PERMANOVA分析显示,取样部位对固氮微生物群落结构组成的影响程度显著高于品种的影响,根际土壤、根部和叶部三者之间的群落组成差异达到显著水平,而品种间仅仅根际土壤样品差异显著。   【结论】   酸性红壤上,玉米不同部位间的固氮微生物群落丰度、多样性和结构组成差异程度显著高于品种间的差异。根部固氮微生物丰度最高,暗示固氮潜力更大。叶部固氮微生物群落组成显著不同于根际土壤和根部,且有最低的丰度和多样性,这与叶片苛刻的环境条件密切相关。     

Analysis of genetic diversity in Indian robusta coffee genepool (<Emphasis Type="Italic">Coffea canephora</Emphasis>) in comparison with a representative core collection using SSRs and AFLPs

Genetic diversity among 40 accessions (Coffea canephora) of robusta coffee genepool available in India was determined in comparison with 14 representative samples from a C. canephora core collection and three accessions of C. congensis, using amplified fragment length polymorphism (AFLP) and simple sequence repeats (SSR) markers. Both these molecular approaches were able to generate unique fingerprints for each of the accessions analysed. All the 12 SSR primers used in the present study were found polymorphic, with an average of six alleles per primer pair. Comparative analysis revealed the higher amount of diversity in representatives from a core collection than in the Indian genepool. Moreover, a total of 205 polymorphic AFLP bands were scored in all the 57 accessions analysed. The genetic relationship among 57 accessions was compared on the basis of SSR and AFLP polymorphisms. Genetic similarity dendrograms showed high correlation between the two marker systems. This study clearly established the high amount of diversity present in core samples, which is not represented in Indian genepool. Furthermore, the three accessions of C. congensis did not exhibit any significant diversity from other robusta accessions supporting the school of thought that C. congensis forms a biotype of C. canephora. The potential use of SSRs and AFLP markers in genetic diversity analysis for better ex situ management and also for exploitation of diversity in breeding programmes is discussed.     

Triterpene saponins from the roots of Medicago hybrida

Fourteen triterpene saponins (1-14) have been isolated from the roots of Medicago hybrida and their structures elucidated by FAB-MS and NMR analysis. Two of them are new compounds and were identified as hederagenin 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (7) and oleanolic acid 3-O-[beta-D-galactopyranosyl(1-->2)-beta-D-glucuronopyranosyl]-28-O-[alpha-L-rhamnopyranosyl(1-->4)-beta-D-glucopyranoside] (14). Seven saponins being mono- and bidesmosides of hederagenin (1, 5, 6, 9), one bidesmoside of bayogenin (2), and two bidesmosides of 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid (11) and oleanolic acid (13) are known compounds but not previously reported as saponin constituents of Medicago, whereas five other saponins, being mono- and bidesmosides of medicagenic acid (3, 4, 8, 10, 12), and one monodesmoside of hederagenin (8) have been previously isolated from other Medicago species. The presence of 2beta,3beta-dihydroxyolean-12-en-23-al-28-oic acid might represent an interesting intermediate in the biosynthesis of these substances.