黄连副产物中生物碱含量测定及提取工艺研究

目的:用HPLC对黄连生产副产物的生物碱含量进行测定,探究黄连生物碱在叶柄和须根中的分布。选择副产物生物碱的最佳提取工艺回收利用生物碱。方法:采用HPLC法建立黄连生物碱的含量测定方法,结合HPLC指纹图谱对黄连生产副产物进行生物碱含量测定。采用单因素试验法,分别对提取酸度、料液比和提取次数进行试验,分析并选取最佳提取条件。结果:黄连生产副产物中黄连生物碱的含量非常可观,其中黄连须含有大量的黄连生物碱,值得进一步开发利用。提取条件为:H2SO4浓度1.5%;料液比1∶15;提取时间1 h。结论:从黄连生物碱含量、资源的综合及可持续利用角度考虑,黄连生产副产物有很好的商业利用前景。     

不同理化因子对黄连愈伤组织生长和生物碱合成的影响

古蔺野连（又称串珠连Ｃｏｐｔｉｓｇｕｌｉｎｅｎｓｉｓ）为黄连属植物中的一个野生珍稀品种,只生长在我国西南少数山区,其根茎可作中药黄连,含有与商品黄连相同的小檗碱等化学成分^[1].作者在对黄连细胞培养研究中,通过目视法和TLC比较,从古蔺野连的愈伤组织中初步筛选到了生物量增长较快、生物碱合成能力较为稳定的无性系H292,进行了其生物量及生物碱含量的动态测定。本文报道植物激素、温度、光照等对黄连愈伤组织生长和生物碱合成能力的影响。     

乌头须根总生物碱提取工艺的考察

目的:考察乌头须根中总生物碱的最佳提取工艺条件。方法:酸碱滴定法测定乌头须根中总生物碱含量,以总生物碱提取率为指标,采用L9(3)~4正交实验法筛选乌头须根总生物碱的最佳提取工艺。结果:乌头须根总生物碱含量为1.094%,影响提取的主次因素为:乙醇浓度>提取次数>提取时间>乙醇用量;优选得到的最佳提取工艺为A_3B_1C_3D_3,即以8倍量80%的乙醇提取3次,每次1.5小时。结论:乌头须根总生物碱含量较高,提取工艺条件稳定、经济、可行。     

不同红掌品种的叶片、叶柄和茎段愈伤组织的诱导及植株再生

红掌幼嫩叶片表面灭菌的最佳方法是：0．01％KMnO4 10min,0．05％链霉素、0．05％制菌霉素和0．05％头孢唑林钠各灭菌20min,然后用75％乙醇30s和HgCl2 2min,污染率为零。Anthurrium adraeanum “Rosetta“和A.adraeanum “Oilcloth-flower”愈伤组织诱导率和芽分化率显著高于A.adraeanum “Atlanta”.A.adraeanum “Fantasis”,A.adraeanum“Afrikanerin”。不同品种茎段、叶柄、叶片的愈伤组织诱导率和芽分化率有明显差异,大多数品种是幼茎段＞叶柄＞叶片。幼苗移栽在草煤或血竭渣基质中,成活率可达97％。     

银杏愈伤组织的形成及其中黄酮类化合物的产生

单一激素种类对银杏叶片,叶柄和幼茎愈伤组织的诱导中以NAA的效果最佳,2,4－D次之,6－BA最差,除胚乳外,胚,幼苗的胚根,子叶,幼茎,叶片和叶柄,以及成年树的嫩茎,叶片和叶柄各外植体在本试验条件下都能诱发愈伤组织,其中胚,子叶和叶柄的愈伤组织形成频率均可达到100．0％,叶片和幼茎在光照下的愈伤组织诱导频率比黑暗中的略高,而叶柄和胚根则相反,MS和DCR两种培养基都适合银杏幼苗叶片及叶柄愈伤组织的诱导,两者之间不存显著性差异,测得光照培养的3个组织系（ST1,ST2,ST3)中均含银杏黄酮甙元槲皮素,山柰素和异鼠李素,总含量分别为干重的0．35％,0．29％和0．14％,而黑暗中培养的这3个愈伤组织系则没有银杏黄酮的产生。     

半夏茎尖培养及块茎的品质改良

离体培养半夏Ｐｉｎｅｌｌｉａ ｔｅｒｎａｔａ 茎尖获得小植株。用其叶柄作外植体在ＭＳ添加２,４－Ｄ 和ＫＴ的培养基上诱导出愈伤组织,继代培养中挑选出一种表面呈颗粒状、易分散、生长快的愈伤组织。在ＭＳ添加ＫＴ０．５ ｍ ｇ／Ｌ, ＮＡＡ ０．２ ｍ ｇ／Ｌ的培养基上,叶柄可直接分化形成小植株,愈伤组织通过形成小块茎的途径产生完整植株。块茎顶端芽分化过程中,先形成的叶原基或幼叶总是覆盖着后面的叶原基而出现一种依次叠套的特殊结构。液体浅层培养对试管苗的增殖速率比固体培养快１ 倍。叶柄和愈伤组织的小植株分化率均在７０％ 以上,移栽成活率为１００％ 。试管苗的块茎产量（鲜重）比对照（用小块茎繁殖）的净块茎产量（鲜重）高１０３％ 。试管苗块茎的总生物碱含量为０．３４４％ ,野生和人工栽培半夏块茎的总生物碱含量分别为０．２６４％ 和０．２０３％ 。     

酶浸提取方法对士的宁产率的影响

目的：研究了纤维素酶在提取生物碱过程中的应用。方法：采用酶浸法和氯仿法两种不同的工艺提取马钱子生物总碱,高效液相色谱法（HPLC）测定了马钱子生物总碱中士的宁的含量。结果：酶浸法提取士的宁和氯仿法提取士的宁的含量分别为1．83％、1．32％;酶浸法和氯仿法提取马钱子生物总碱的产率分别为：2．85％、1．86％。     

苦豆子中生物碱的分布及其在生长期内的含量变化

采用高效液相仪测定苦豆子植株不同部位和不同生长期间的生物碱含量。结果表明：苦豆子植株不同部位的总生物碱含量为21.346-41mg．g-1（DW）,以种子中为最高,花、根、果皮、根状茎、叶片、地上茎次之,复叶轴中最低;苦参碱含量为0．55-1．30m2．g-1（DW）,含量由高到低依次为种子〉花〉根〉果皮〉根状茎〉叶片〉地上茎〉复叶轴;氧化苦参碱和氧化槐果碱含量分别为3．76～12．29mg．g-1（DW）和3．36—13．33mg·g-1（DW）,这2种生物碱含量由高到低依次为种子〉花〉根〉果皮〉根状茎〉地上茎〉复叶轴〉叶片：根、根状茎、地上茎、叶中的苦参碱、氧化苦参碱和氧化槐果碱含量在7月份达到峰值;总生物碱含量在8月达到峰值。     

野生水芹与旱芹的营养成分比较分析

为研究野生水芹的营养价值,以湖北野生水芹和旱芹为材料,对新鲜叶片和叶柄的主要营养成分进行了比较分析。结果表明:两种材料叶片和叶柄的淀粉、粗纤维和维生素B1含量的差异不明显,游离氨基酸和维生素B2含量比旱芹低,但野生水芹含有丰富的蛋白质和钙,其叶柄中可溶性糖和磷的含量分别达29.70 mg/g和0.77mg/g,分别比旱芹高57.23%和37.50%;其叶片维生素C、铁含量分别达0.19 mg/g和0.35 mg/g,比旱芹分别高137.50%和191.67%,整株黄酮类物质含量是旱芹含量的2倍。这说明野生水芹具有较高的食用价值,是一种适合作为涝渍地区保护性开发的野生蔬菜资源。     

半夏组织培养诱导胚状体的正交试验

采用茎尖组织培养技术获得了半夏(Pinellia ternata(Thunb．)Breit．]的完整试管苗,并进行了半夏叶不同部位诱导胚状体和愈伤组织的试验;以生长率和总生物碱含量作为指标,对不同激素配比进行了正交试验,筛选出了最佳的培养基组成。结果表明：诱导胚状体和愈伤组织的最佳外植体是半夏的叶柄基部,诱导率分别为43．6％和100．0％,月生长率达24．43倍;最佳培养基为MS 1．0mg／L6-BA 0．1mg／L IAA。研究结果为半夏人工种子的进一步研究奠定了基础。     

Contribution to the study of heterogeneity in the leaves of a spring wheat plant

The effect of the individual leaf blades of spring wheat on the dry matter of stalks, chaff and grain (caryopses), of spikes and total overground part, was studied. In the experimental plants the individual leaf blades were detached according to the scheme given, at the beginning of shooting, (A), at the beginning of earing (B), and at the beginning of flowering (C). The dry matter (fresh weight) of the stalk was least decreased if either the lowest or the uppermost leaf blade was severed during the developmental phase of shooting. The dry as well as the fresh weights of chaff were least affected in those plants where the leaf blade was removed during the developmental phase of flowering. Both the dry and fresh weights of caryopses were least decreased if either the lowest or the uppermost leaf blade was removed during the developmental phase of flowering. The dry weight as well as the relative water content of chaff and ear grains were most decreased following removal of leaf blades during the developmental phase of shooting. The relative water content of chaff, grains and ears was most decreased following removal of developed leaf blades during the developmental phase of earing. It was confirmed that in addition to the photosynthetic activity of leaves the photosynthesis of other parts of the stem system (stalk internodes, ear awns etc.) participated in the production of total dry matter of experimental plants. The photosynthetic activity of leaf blades was particularly high up to the earing phase, while subsequently the photosynthesis of extrafoliar area (stalk internodes and ears) predominated. In spite of this, participation of the total leaf area is very high in the formation of grain dry matter (over 50%), as well as of the total dry matter of plant (over 80%).     

Etiology of a Root Rot Disease Complex of Alstroemeria in Alberta, Canada

Fusarium oxysporum, Pythiu-m ultimum, and Rhizoctonia solani were isolated from the basal stems of diseased alstroemeria showing symptoms of dark brown stripes along leaf margins, leaf chlorosis, plant wilting, browning or rotting of basal stem, rhizome, and storage and fibrous roots. The pathogen isolated most frequently was Fusarium spp. (40.5 % of plants examined). Pythium spp. and R. solani were isolated less frequently (5.5 % and 6.8 % of plants examined, respectively). F. oxysporum caused the highest mortality in alstroemeria when rhizomes were grown in unsterilized soil-less mix medium. This is the first report in North America of a root-rot disease complex affecting alstroemeria.     

玉竹的组织培养与快速繁殖

以玉竹[Polygonatum odoratum (Mill.) Druce]根状茎、叶片和茎段为外植体,于附加不同激素配比的MS培养基中诱导愈伤组织、不定芽和不定根,探讨增殖培养和植株再生的条件.结果表明,叶片和茎段外植体诱导愈伤组织和芽的分化率很低;而根状茎外植体易于培养,有较高的诱导率和增殖倍数,其愈伤组织、不定芽和不定根的诱导率分别可达87%、90%和99%以上.适宜根状茎外植体愈伤组织诱导的培养基为MS+1.0 mg/L 6-BA+0.5 mg/L NAA,有利于增殖和丛生芽分化的培养基为MS+2.0 mg/L 6-BA+0.5 mg/L IBA和MS+3.0 mg/L 6-BA+0.1 mg/L NAA,而1/2MS+3.0～5.0 mg/L NAA适宜诱导试管苗生根培养.试管苗的移栽成活率可达85%以上.     

洪湖野菰及其化学成分分析

野菰(Zizania latifolia)是湖北省洪湖中优势水生维管束植物,其群落占全湖355平方公里面积的127平方公里。茎和叶的年生物量为4379克鲜重/平方米,全湖总年产量121700吨干重,目前未被利用。野菰各器官的蛋白质和氨基酸含量分别以百分干重表示:根,7.0和4.76;根状茎,11.3和8.85;茎,9.5和7.15;嫩茎梢,22.4和16.53;叶,16.8和14.61。500克干叶的必需氨基酸含量接近100克干重草鱼幼鱼背肌的必需氨基酸含量。脂肪:叶中3.4～4.2,茎中2.2;粗纤维:叶中26.8～28,茎中24.2;灰分:叶中10.0,茎中5.8。菰茎含可溶性糖类30％以上,其中葡萄糖,果糖、蔗糖和麦芽糖是主要成分。结果表明野菰是一种优质饲料。     

Boron Deficiency Induced Changes in Translocation of 14CO2-Photosynthate Into Primary Metabolites in Relation to Essential Oil and Curcumin Accumulation in Turmeric (Curcuma longa L.)

Changes in leaf growth, net photosynthetic rate (P _N), incorporation pattern of photosynthetically fixed ¹⁴CO₂ in leaves 1–4 from top, roots, and rhizome, and in essential oil and curcumin contents were studied in turmeric plants grown in nutrient solution at boron (B) concentrations of 0 and 0.5 g m^-3. B deficiency resulted in decrease in leaf area, fresh and dry mass, chlorophyll (Chl) content, and P _N and total ¹⁴CO₂ incorporated at all leaf positions, the maximum effect being in young growing leaves. The incorporation of ¹⁴CO₂ declined with leaf position being maximal in the youngest leaf. B deficiency resulted in reduced accumulation of sugars, amino acids, and organic acids at all leaf positions. Translocation of the metabolites towards rhizome and roots decreased. In rhizome, the amount of amino acids increased but content of organic acids did not show any change, whereas in roots there was decrease in contents of these metabolites as a result of B deficiency. Photoassimilate partitioning to essential oil in leaf and to curcumin in rhizome decreased. Although the curcumin content of rhizome increased due to B deficiency, the overall rhizome yield and curcumin yield decreased. The influence of B deficiency on leaf area, fresh and dry masses, CO₂ exchange rate, oil content, and rhizome and curcumin yields can be ascribed to reduced photosynthate formation and translocation.     

In vitro organogenic competence of different organs and tissues of lily of the valley ‘Grandiflora of Nantes’

The in vitro organogenic competence of rhizome and flower stalk discs, flower buds and meristems of lily of the valley (Convallaria majalis L.) Grandiflora of Nantes was studied. Except for the meristems, the other organs were all capable of organogenesis. Rhizome and flower stalk discs formed roots, while flower buds formed flower bud-like structures, and it appeared very difficult to modify the type of organogenesis. After six subcultures of the flower bud-like structures, a series of developmental stages were distinguished, i.e. regeneration of flower bud-like structures first, then leaf-like organs, and finally vegetative buds. This work pointed out the organogenic competence of the rhizome and flower stalk fragments for the first time.Abbreviations BA 6-benzyladenine - 2iP N-isopentenyladenine - NAA 1-naphthaleneacetic acid     

Partitioning of 14C-Photosynthate of Leaves in Roots,Rhizome, and in Essential Oil and Curcumin in Turmeric (Curcuma longa L.)

Incorporation of photosynthetically fixed ¹⁴C was studied at different time intervals of 12, 24, and 36 h in various plant parts—leaf 1 to 4 from apex, roots, and rhizome—into primary metabolites—sugars, amino acids, and organic acids, and secondary metabolites—essential oil and curcumin—in turmeric. The youngest leaves were most active in fixing ¹⁴C at 24 h. Fixation capacity into primary metabolites decreased with leaf position and time. The primary metabolite levels in leaves were maximal in sugars and organic acids and lowest in amino acids. Roots as well as rhizome received maximum photoassimilate from leaves at 24 h; this declined with time. The maximum metabolite concentrations in the roots and rhizome were high in sugars and organic acids and least in amino acids. ¹⁴C incorporation into oil in leaf and into curcumin in rhizome was maximal at 24 h and declined with time. These studies highlight importance of time-dependent translocation of ¹⁴C-primary metabolites from leaves to roots and rhizome and their subsequent biosynthesis into secondary metabolite, curcumin, in rhizome. This might be one of factors regulating the secondary metabolite accumulation and rhizome development.     

Biomass and nutrient allocation of sawgrass and cattail along a nutrient gradient in the Florida Everglades

Biomass and nutrient allocation in sawgrass (Cladium jamaicense Crantz) and cattail (Typha domingensis Pers.) were examined along a nutrient gradient in the Florida Everglades in 1994. This north to south nutrient gradient, created by discharging nutrient-rich agricultural runoff into the northern region of Water Conservatio ea 2A, was represented by three areas (impacted, transitional and reference). Contrasting changes of plant density and size along the gradient were found for communities of both species. For the sawgrass community, more small plants were found in ref ce areas, whereas few large plants were found in impacted areas. In contrast, for the cattail community, bigger plants were found in reference areas, and smaller plants were found in impacted areas. Both species allocated approximately 60% of their total biomass to leaves and 40% to belowground tissues. However, sawgrass biomass allocation to leaves, roots, shoot bases and rhizomes (65%, 19%, 11%, and 5%, respectively) was similar among the three areas. In contrast, cattail plants growing in referen reas showed higher root allocation (27.3%), but lower leaf allocation (51.1%) than those growing in impacted areas (14.6% and 65.8% for root and leaf allocation, respectively). Cattail had higher phosphorus concentrations than sawgrass in tissues associated with growth functions (leaves, roots, and rhizomes). In contrast, sawgrass had higher phosphorus and nitrogen concentrations than cattail in tissues primarily associated with resource storage (shoot bases). From impacted to reference areas, for sawgrass, there was a decrease of leaf TP from 605 to 248 (mg/kg), root TP from 698 to 181 (mg/kg), rhizome TP from 1,139 to 142 (mg/kg), and shoot base TP from 5,412 to 400 to (mg/kg). For cattail, leaf TP decreased from 1,175 to 556 (mg/kg), root TP de sed from 1,100 to 798 (mg/kg), rhizome TP decreased from 1390 to 380 (mg/kg), and shoot base TP decreased from 2,990 to 433 (mg/kg). N/P ratios of sawgrass in reference areas were 27, 63, 38, and 50 for leaves, roots, rhizomes, and shoot bases, respectively, whereas in impacted areas they were 11, 21, 6, and 2, respectively. The greatest TP storage was found in impacted areas. Differences in seed output, seed number, and mean seed weight were found for both species as well. Each cattail flower stalk duced approximately 105 tiny seeds (0.048 ± 0.001 mg) while each sawgrass flower stalk produced about 103 large seeds (3.13 ± 0.005 mg). These results suggest that phosphorus is a limiting resource in the Everglades and that the two species have different life history strategies. These data provide an ecological basis for making informed management and planning decisions to protect and restore the Everglades.     

SHOOT APEX ORGANIZATION AND ORIGIN OF THE RHIZOME-BORNE ROOTS AND THEIR ASSOCIATED GAPS IN DENNSTAEDTIA CICUTARIA

The shoot apex of Dennstaedtia cicutaria consists of three zones—a zone of surface initials, a zone of subsurface initials, and a cup-shaped zone that is subdivided into a peripheral region and central region. A diffuse primary thickening meristem, which is continuous with the peripheral region of the cup-shaped zone, gives rise to a broad cortex. The roots occurring on the rhizomes are initiated very near the shoot apex in the outer derivatives of the primary thickening meristem. The roots that occur on the leaf bases also differentiate from cortical cells. Eventually, those cortical cells situated between the newly formed root apical cell and the rhizome procambium (or leaf trace) differentiate into the procambium of the root trace, thus establishing procambial continuity with that of the rhizome or leaf trace. Parenchymatous root gaps are formed in the rhizome stele and leaf traces when a few of their procambial cells located directly above the juncture of the root trace procambium differentiate into parenchyma. As the rhizome procambium or leaf trace continues to elongate, the parenchyma cells of the gap randomly divide and enlarge, thus extending the gap.     

THE OCCURRENCE OF “ROOT GAPS” IN THE RHIZOME SOLENOSTELE AND LEAF TRACE OF DENNSTAEDTIA CICUTARIA

The vascular system of the rhizome axes of Dennstaedtia cicutaria consists of a solenostele with an amphicribral vascular bundle centrally located in the pith. Each leaf has a single trace which is an amphicribral vascular bundle. At each node of an axis there is a complex consisting of the main axis, leaf base, and a branch axis attached to the basiscopic margin of the leaf base. Numerous roots are present on the rhizomes and the abaxial side of the leaf bases. Parenchymatous gaps occur in the rhizome solenostele and the leaf trace directly above the departure of some of the root traces. These gaps are termed root gaps. In some instances the root gaps are confluent. However, not all of the root traces have an associated root gap. The leaf trace is inserted laterally on the main and branch axes at the node so that the acroscopic leaf trace margin anastomoses with the main axis of the vascular system and the basiscopic margin with that of the branch axis. Two leaf gaps are associated with each leaf trace, one occurring in the main axis solenostele and the other in the branch axis solenostele. The medullary bundle of each axis anastomoses with each leaf trace at its point of attachment to the rhizome solenostele. Thus, the medullary bundle forms a continuous vascular strand from leaf trace to leaf trace in any given rhizome axis.