Biolistic transfer and expression of a uidA reporter gene in different tissues of Allium sativum L.

The biolistic particle delivery system was used to transfer the uidA gene into different garlic tissues, including regenerable calli. These tissues showed a high endogenous nuclease activity preventing exogenous DNA expression. After submerging the tissues for 24 h in a 2 mm solution of the nuclease inhibitor aurintricarboxylic acid, transient expression of the uidA gene was detected. Furthermore, vacuum infiltration of aurintricarboxylic acid into the garlic tissues significantly increased the percentage of explants showing transient expression. Among four plasmids tested, pDE4 containing the CaMV35S-uidA construct produced the best results. Received: 16 May 1997 / Revision received: 26 August 1997 / Accepted: 25 September 1997     

Variazioni cariologiche e chimiche in Allium sativum L.

Abstract

Karyological and chemical variations in Allium sativum L. — Results about a research on the karyotypes of three Italian strains of Allium sativum are reported. The lack of sexual reproduction in Allium sativum favours the formation of strains regarded as clones; each of them represents an isolated genetic system. The anatomy and the chemical composition of the essence was studied to improve the understanding of the variations in the different strains comprised in this species. Two of the clones examined (Tuscan and Sicilian) belong to U type and one (Piedmontese) to the H type, according to Hrubý and Konvi?ka's classification based on the morpho-physiological characters. In this study evidence shows that: a) The three clones do not present anatomical differences. b) The morpho-physiological characters are strictly correlated with the chemical composition of the essence; in fact the gas-chromatographic study of the two U type clone essences shows the same qualitative composition, each chemical component being present more or less in the same percentage. The essence of Piedmontese clone, is qualitatively and quantitatively different from the others. c) The number and the general morphology of the chromosomes of the three root tip karyotypes agree with the literature, nevertheless the karyotypes sometimes show differences in the two satellited pairs of chromosomes. The Tuscan karyotype (U type) is comparable with the OU type of Konvi?ka and Levan. The Piedmontese karyotype (H type) is comparable with the OH karyotype of the same authors; instead the Sicilian karyotype is very different from all karyotypes in literature and from the Tuscan karyotype belonging to the same U type because of the lack of the pseudosatellite in a partner of pair no. 6. Research evidences that, for the time being, it is impossible to use only chromosome morphology to establish the clone types: nevertheless their chemical characteristics seem to correspond to their morpho-physiological features.     

大蒜分生组织培养脱病毒和快速繁殖技术

采用茎尖分生组织培养技术,获得了大蒜（Ａｌｌｉｕｍ ｓａｔｉｖｕｍ Ｌ．）的无病毒试管苗。通过基本培养基和激素配比试验,筛选出最佳的培养基组成,进行脱病毒苗的快速繁殖。结果表明：诱导愈伤组织的最适培养基为：ＭＳ＋ＢＡ０．２ｍｇ／Ｌ＋ＮＡＡ０．５ｍｇ／Ｌ,月生长率达１２．７０倍;诱导丛生芽的最适培养基为：Ｂ５＋ＢＡ０．５ｍｇ／Ｌ＋ＩＡＡ０．２ｍｇ／Ｌ,丛生芽繁殖系数高达２５．５倍,技术上达到了快速繁     

Analysis of morphological variability in the Indian germplasm of Allium sativum L.

Allium sativum germplasm collected from different parts of India was subjected to critical analysis of 14 easily noticeable morphological traits. Variability encountered in quantitative as well as qualitative, bulb and plant traits was documented. While analysing this variability, an attempt was made to highlight the genotype-environment interaction, its impact on the expression of morphological traits in this apomict and evaluate the different collections for intraspecific phylogeny.     

Establishment of embryogenic cell suspension cultures of garlic (Allium sativum L.), plant regeneration and biochemical analyses

Embryogenic cell suspension cultures of garlic (Allium sativum L.) were initiated in liquid medium from friable embryogenic tissue. The optimal parameters for culture maintenance were: (1) an initial cell density of 1–4% (v/v); (2) medium renewal every 14 days and subculturing every 28 days; (3) a low 2,4-dichlorophenoxyacetic acid concentration (0.1–0.3 mg/l). Cultures regenerated during a 14-month period. The cell suspension cultures differentiated embryos following transfer to a semi-solid embryo induction medium, with histological studies confirming and characterising the embryogenic nature of the process. Forty percent of these embryos converted into plantlets, which produced micro bulbs in vitro. The composition of the sulphur compounds of the micro bulbs obtained from cell suspension embryo-derived plantlets differed slightly from those produced by in vitro shoot proliferation-derived plantlets, but after two cycles of multiplication in the field these differences had disappeared.     

Effect of thioglycolic acid on Allium sativum L. root meristems]

Thioglycolic acid is, in the Allium test, somewhat less cytotoxic than mercaptoethanol but the characteristics of cytotoxicity are the same in both cases : the mitoclasic effect cannot be separated from a strong mitodepressive effect and is quickly followed by a lethal effect.     

中国大蒜(Allium sativum L.)18个品种的酯酶同工酶多态性分析

采用聚丙烯酰胺凝胶电泳(PAGE)分离了中国大蒜3个生态型(低温反应敏感型、低温反应中间型和低温反应迟钝型)中18个较典型的品种的酯酶同工酶,并用排序分析法对18个品种的亲缘关系进行了分析,将18个品种分为3个变种群：1．“苏联”蒜变种群(var．Russia),2．吉木萨尔白皮蒜变种群(var．：Jimusaer。),3．中国内陆大蒜变种群(var．China)。其中中国内陆大蒜变种群又可分为5个品种群：①关中蒜品种群,②西北蒜品种群,③西南蒜品种群,④云贵蒜品种群,⑤华东蒜品种群。实验结果初步证实,大蒜生态型不能完全等同于基因型,酯酶同工酶的变化可能更能说明大蒜的亲缘进化关系。     

Antifungal saponins from bulbs of garlic,Allium sativum L. var. Voghiera

A bioassay-guided phytochemical analysis of the polar extract from the bulbs of garlic, Allium sativum L., var. Voghiera, typical of Voghiera, Ferrara (Italy), allowed the isolation of ten furostanol saponins; voghieroside A1/A2 and voghieroside B1/B2, based on the rare agapanthagenin aglycone; voghieroside C1/C2, based on agigenin aglycone; and voghieroside D1/D2 and E1/E2, based on gitogenin aglycone. In addition, we found two known spirostanol saponins, agigenin 3-O-trisaccharide and gitogenin 3-O-tetrasaccharide. The chemical structures of the isolated compounds were established through a combination of extensive nuclear magnetic resonance, mass spectrometry and chemical analyses. High concentrations of two eugenol diglycosides were also found for the first time in Allium spp. The isolated compounds were evaluated for their antimicrobial activity towards two fungal species, the air-borne pathogen Botrytis cinerea and the antagonistic fungus Trichoderma harzianum.     

Measurement of the inorganic pyrophosphate in tissues of Pisum sativum L.

Purified pyrophosphate: fructose 6-phosphate 1-phosphotransferase (EC 2.7.1.90) was used to measure the inorganic pyrophosphate in unfractionated extracts of tissues of Pisum sativum L. The fructose 1,6-bisphosphate produced by the above enzyme was measured by coupling to NADH oxidation via aldolase (EC 4.1.2.13), triosephosphate isomerase (EC 5.3.1.1) and glycerol-3-phosphate dehydrogenase (EC 1.1.1.8). Amounts of pyrophosphate as low as 1 nmol could be measured. The contents of pyrophosphate in the developing embryo of pea, and in the apical 2 cm of the roots, were appreciable; 9.4 and 8.9 nmol g^-1 fresh weight, respectively. The possibility that pyrophosphate acts in vivo as an energy source for pyrophosphate: fructose 6-phosphate 1-phosphotransferase and for UDPglucose pyrophosphorylase (EC 2.7.7.9) is considered.     

大蒜种质资源对蒜蛆的抗性评价

就大蒜种质资源对蒜蛆的抗性进行了虫圃田间鉴定,并对抗性与大蒜主要植物学性状和大蒜辣素含量的相关性进行了分析。结果表明:52份材料的感虫指数分布在7.14~90.38之间,种质资源间抗虫性差异达到了显著水平;聚类分析并结合感虫指数将52份种质资源分为高抗、抗、中抗、中感、感、高感6个类别,其中高抗和抗性材料分别为4份和8份。相关分析表明:感虫指数与植株的7个形态数量性状的相关性均未达到显著水平,而与鳞茎的鳞芽背宽、鳞茎的大蒜辣素含量均呈极显著的负相关,即抗性与二者显著正相关。表明大蒜辣素含量越高,大蒜对蒜蛆的抗性越强;从鳞芽背宽较宽和高大蒜辣素含量的大蒜资源中筛选抗蒜蛆的种质可能性更大。     

Role of garlic (Allium sativum L.) in human and plant diseases

A resurgence of interest in garlic due to recent revelations of its beneficial effects in the treatment of various human and plant diseases and also due to validation of claims made in traditional systems of medicine has resulted a plethora of publications on different aspects of garlic in recent years. Chemical constituents of garlic and their variations on the methods of isolation have been discussed in the present review. Effect of garlic and its constituents against various human and plant pathogenic and saprophytic microorganisms has also been reviewed.     

Flavor formation in tissue cultures of garlic (Allium sativum L.)

Differentiated and undifferentiated cultures of garlic (Allium sativum L.) were analyzed for the study of flavor formation in cultures. Attempts were made to correlate alliin content with free and bound amino acid contents and with enzymes like phenylanine ammonialyase (E.C. 4.1.1.5) and alliin-lyase (E.C.4.4.1.4) which play important roles in formation of the flavor percursor alliin.It was observed that in differentiating cultures showing shoot formation, there is an increase in alliin content as well as in free and bound amino acid contents. Corresponding to this there was also an increase in the activity of phenylalanine ammonialyase in differentiating cultures. Alliin-lyase activity was found to be significantly different in differentiating and undifferentiated cultures. The significance of these results is discussed.     

Purification of the alliin lyase of garlic, Allium sativum L

1. Alliin lyase (EC 4.4.1.4) was purified up to sevenfold from garlic-bulb homogenates. The enzyme was unstable to storage at -10 degrees , particularly in dilute concentrations, but the addition of glycerol (final concentration 10%, v/v) stabilized the activity completely for at least 30 days. 2. The purified enzyme had an optimum pH for activity at 6.5. The addition of pyridoxal phosphate stimulated the reaction rate and the stimulation became more marked as the purification proceeded. 3. Hydroxylamine (10mum) and cysteine (0.5mm) inhibited the enzyme activity by more than 80%. Spectral studies indicated that cysteine reacted with pyridoxal phosphate bound to the protein. 4. The K(m) values for S-methyl-, S-ethyl-, S-propyl-, S-butyl- and S-allyl-l-cysteine sulphoxides were determined. With S-allyl-l-cysteine sulphoxide the K(m) was 6mm and the V(max.) was greater than those with the other substrates tested. 5. The thioether analogues of the substrates were competitive inhibitors for the lyase reaction. The K(i) decreased with increasing chain length of the alkyl substituent. With S-ethyl-l-cysteine sulphoxide as substrate the K(i) was 33, 8 and 5mm respectively for S-methyl-, S-ethyl- and S-propyl-l-cysteine. 6. The addition of EDTA or Mg(2+), Mn(2+), Co(2+) or Fe(2+) stimulated the reaction rate. Other bivalent cations either had no effect or gave a strong inhibition. In the presence of EDTA no further increase of activity was observed with added Mg(2+).     

大蒜产香内生真菌的分离及其挥发性成分分析

大蒜(Allium sativum L.)是百合科葱属常见的植物,富含丰富的生物活性成分和营养物质。本研究采用组织分离法从新鲜的大蒜中分离内生真菌并进行纯化,获得1株产香浓郁的菌株KLBMPAS015。采用固相微萃取固定吸附,结合气相色谱-质谱解吸附分析技术(SPME-GC-MS)对菌株KLBMPAS015的挥发成分进行分析。结果表明:该菌株代谢产物的香气成分主要为:n-丁酸2-乙基己基酯(n-butyric acid 2-ethylhexyl ester)(1.36%)、1-丁醇3-甲基-丙酸(1-butanol,3-methyl-,propanoate)(25.40%)和戊基惕各酸(iso-amyl tiglate)(65.51%)。产香真菌KLBMPAS015的戊基惕各酸含量相对较高,在天然香料生产中具有重要的潜在应用价值。     

Tankyrase activity in organs and tissues of mice

Tankyrase, one of the NAD⁺ ADP-ribosyltransferases, is a target for drugs developed for their anticancer and other pharmacological activities. We designed an assay for estimation of the inhibition or activation of the enzyme in pre-clinical studies. In mice, the highest specific activity of tankyrase was observed in thymus, spleen, pancreas, and bone marrow. In murine liver, tankyrase is active in ontogenesis and during reparative regeneration; however, the basal activity is hardly detectable in normal liver and most of other organs of adult animals. We suggest that tankyrase is a part of the tissue growth and repair machinery, while its age-dependent inhibition, when an organism stops growing, turns on phenoptosis.     

Sister chromatid exchanges in garlic (Allium sativum L.) callus cells

A technique is described for differential staining of sister chromatids and the study of sister chromatid exchanges (SCEs) in garlic (Allium sativum L.) callus cells. BrdU incorporation into newly synthesized DNA was ensured by culturing calli on medium containing 100 M BrdU+0.01 M FudR+1 M Urd. SCEs were visualized by FPG staining technique and their frequency was analysed. Mean frequency of SCEs in callus cells was higher than that in meristem root-tip cells. Using the same staining method, cell cycle time of callus cells was analysed. It was found that it ranges from 48 to 132 hrs. The method described represents a new approach in the study of genetic instability of plant cells cultured in vitro.Abbreviations BrdU 5-bromo-2-deoxyuridine - 2,4-D 2,4-dichlorophenoxyacetic acid - FPG fluorescent-plus-Giemsa - FudR 5-fluoro-2-deoxyuridine - SCE sister chromatid exchange - SSC 0.15 M NaCl + 0.015 M Na-citrate - T thymidine-containing strand of the DNA duplex - B 5-bromo-2-deoxyuridine-containing strand of the DNA duplex - Urd uridine     

Physiological responses and chromosomal aberration in root tip cells of Allium sativum L. to cerium treatments

Plant and Soil - Lower concentrations of REEs promote plant growth, whereas higher concentrations of REEs negatively affect plant growth, soil accumulation, and environmental pollution. To clarify...