泽漆营养器官发育解剖学研究

采用石蜡切片法、半薄切片法对泽漆营养器官的发育过程进行了观察,同时对3种器官中乳汁管的分布和大小进行了分析。结果表明:泽漆根的发育类似于草本双子叶植物根的一般发育规律。初生木质部为三原型。茎的初生结构由表皮、皮层、维管束环和髓构成,其髓中有空腔,而茎的次生生长过程中维管形成层的活动短暂,仅产生少量次生维管组织,不形成周皮。叶的发育包括原分生组织、初生分生组织和成熟结构3个阶段,属于异面叶结构。泽漆乳汁管主要分布在维管束韧皮部的外侧。在3种器官中,乳汁管直径差异较大,依次为根>茎>叶。     

The involvement of sucrose,glucose and other metabolites in the synthesis of triterpenes and dopa in the laticifers of Euphorbia lathyris

A quantitative triterpene analysis was made of latex stem tissue of Euphorbia lathyris. Young plants seedlings of E. lathyris were incubated with various labelled precursors. Incorporation into triterpenes was obtained from [2-¹⁴C]mevalonic acid, [1-¹⁴C]acetate, [3-¹⁴C]pyruvate, [U-¹⁴C]sucrose, [U-¹⁴C]glucose, [U-¹⁴C]xylose, [U-¹⁴C]glyoxylate, [2,3-¹⁴C]succinic acid, [1-¹⁴C]glycerol [U-¹⁴C]serine. Both sugars tyrosine appeared to be effective precursors in DOPA synthesis inside the laticifers. Exogenously supplied mevalonic acid was only involved in triterpene synthesis outside the laticifers. GC-RC of triterpenes synthesized from [U-¹⁴C]glucose revealed the origin of these compounds in the latex. The labelled triterpenes obtained after incorporation of the other mentioned labelled precursors were only partly synthesized in the laticifers. For quantitative data on latex triterpene synthesis seedlings were incubated with [U-¹⁴C]sucrose, [U-¹⁴C]glucose, [U-¹⁴C]xylose [1-¹⁴C]acetate in the presence of increasing amounts of unlabelled substrate. From the amount of ¹⁴C incorporated into the triterpenes the amount of substrate directly involved in triterpene synthesis was calculated, as was the absolute triterpene yield. Sucrose showed the highest triterpene yield, equivalent to the daily increase of the triterpene content of growing seedlings. The possible significance of the other precursors in triterpene synthesis in the laticifers is discussed.     

甘遂乳汁管的解剖学及营养器官中二萜类化合物分布的研究

利用石蜡切片、半薄切片及组织化学的方法对甘遂各器官中乳汁管的类型、分布和大小进行了研究。结果表明,甘遂乳汁管为无节分枝型,在各器官中主要分布在维管束韧皮部的外侧或周围,此外,在根的中柱鞘薄壁细胞、叶片的叶肉组织、果实的中果皮及胚乳细胞内也有少量乳汁管的分布。甘遂乳汁管的大小在各器官中略有不同,其中茎中直径最大为39.79μm,块根中直径居中为36.90μm,叶肉组织中直径最小,仅有8.94μm;乳汁管的密度在其营养器官中差异较大,依次为叶>茎>根>块根。组织化学实验结果显示二萜类化合物在甘遂的营养器官中分布广泛。根中柱鞘和韧皮部的薄壁组织细胞,茎皮层、维管形成层和韧皮部的薄壁组织细胞,叶的叶肉细胞以及叶脉中的厚角组织和薄壁组织细胞的腔中都显示不同程度的红色,而乳汁管中乳汁的显色较深。     

番木瓜蛋白酶的免疫荧光定位

番木瓜在我国广东、广西、云南等地栽种,它的乳汁富含番木瓜蛋白酶,该酶主要应用于食品工业、酿造业及化妆品等。目前我们已完成精酶生产工艺的研究并投入了批量生产,然而该酶在果实里的分布特征及组织定位尚不清楚。我们通过幼果切片的组织学染色,观察到番木瓜的乳汁管结构,并用间接免疫荧光法初步确定了番木瓜凝乳蛋白酶的存在部位。     

Chemical Composition and Activity of Essential Oils of Carissa macrocarpa (Eckl.) A.DC. Cultivated in Tunisia and Its Anatomical Features

This is the first study investigating the chemical composition of essential oils (EOs) isolated from different tissues of Carissa macrocarpa (Eckl .) A.DC., their antimicrobial activity and the anatomical characters of the aerial organs and the fruits. The main EO components were pentadecanal and tetradecan‐1‐ol (31.9 and 16.5% in fresh leaf EO, respectively), (E)‐nerolidol and caryophyllene oxide (27.3 and 15.0% in fruit EO, respectively), linalool and hexahydrofarnesyl acetone (30.9 and 24.9% in stem EO, respectively), benzyl benzoate (24.3% in flower EO). The fruit EO was more active against Candida albicans (MIC = 0.46 mg/mL) compared to the reference antibiotic (17.66 mg/mL). Furthermore, at this concentration it inhibited all the Gram‐positive bacteria. Concerning the anatomical features, it is noteworthy to mention the presence of a large cluster of calcium oxalate crystals inside some parenchymatous cells. Large ducts corresponding to non articulated laticifers were identified in the cortex of leaf, stem and fruit pericarp. The laticifers categories and their distribution are taxonomically important to discriminate this species from others acclimated in different countries. Considering the obtained results, EOs of C. macrocarpa can be a good source of antimicrobial compounds, contributing to solve the problem of microbial resistance to antibiotics.     

β-1,3-Glucanase is highly-expressed in laticifers of Hevea brasiliensis

Clones encoding -1,3-glucanase have been isolated from a Hevea cDNA library prepared from the latex of Hevea brasiliensis using a probe Nicotiana plumbaginifolia cDNA encoding -1,3-glucanase, gnl. Nucleotide sequence analysis showed that a 1.2 kb Hevea cDNA encoding a basic -1,3-glucanase showed 68% nucleotide homology to gnl cDNA. Northern blot analysis using the Hevea cDNA as probe detected a mRNA of 1.3 kb which was expressed at higher levels in latex than in leaf. In situ hybridization analysis using petiole sections from Hevea localized the -1,3-glucanase mRNA to the laticifer cells. Genomic Southern analysis suggested the presence of a low-copy gene family encoding -1,3-glucanases in H. brasiliensis.     

Immunocytochemical identification of laticifers

Summary sensitive immunocytochemical method for the identification of laticifers has been developed. Frozen sections of various laticifer-bearing plant material, mounted on slides, were first flooded with the IgG fraction of rabbit anti-latex antiserum, prepared using whole latex ofAsclepias syriaca, then flooded with fluorescein-conjugated IgG fraction goat anti-rabbit IgG to visualize laticifers. Positive fluorescence was observed for laticifers in shoots and embryos ofA. syriaca andStapelia bella and embryos ofA. tuberosa. Laticifers did not fluoresce in shoots ofA. tuberosa andEuphorbia tirucalli, in embryos ofE. marginata, or in petioles ofMusa paradisiaca andCichorium intybus. Controls prepared with uninjected rabbit serum were negative (no fluorescence).     

Purification and characterization of a wound-inducible thaumatin-like protein from the latex of Carica papaya

A 22.137 kDa protein constituent of fresh latex was isolated both from the latex of regularly damaged papaya trees and from a commercially available papain preparation. The protein was purified up to apparent homogeneity and was shown to be absent in the latex of papaya trees that had never been previously mechanically injured. This suggests that the protein belongs to pathogenesis-related protein family, as expected for several other protein constituents of papaya latex. The protein was identified as a thaumatin-like protein (class 5 of the pathogenesis-related proteins) on the basis of its partial amino acid sequence. By sequence analysis of the Carica genome, three different forms of thaumatin-like protein were identified, where the latex constituent belongs to a well-known form, allowing the molecular modeling of its spatial structure. The papaya latex thaumatin-like protein was further characterized. The protein appears to be stable in the pH interval from 2 to 10 and resistant to chemical denaturation by guanidium chloride, with a of 15.2 kcal/mol and to proteolysis by the four papaya cysteine proteinases. The physiological role of this protein is discussed.     

Romaine lettuce latex deters feeding of banded cucumber beetle: a vehicle for deployment of biochemical defenses

Latex is widely found among plant species and is known to play a defensive role against certain herbivores. Two romaine lettuce, Lactuca sativa L. (Asteraceae) cultivars, 'Valmaine' (resistant) and 'Tall Guzmaine' (susceptible), were selected to study the potential of latex as a defense mechanism against the banded cucumber beetle, Diabrotica balteata LeConte (Coleoptera: Chrysomelidae). Latex from Valmaine strongly inhibited feeding of adult D. balteata compared to latex from Tall Guzmaine when applied to the surface of artificial diet. Beetles consumed significantly less diet from disks treated with Valmaine latex than they consumed from diet treated with Tall Guzmaine latex, in both choice and no-choice tests. In a choice test involving diet disks treated with Valmaine latex from young leaves vs. disks treated with latex from mature leaves, the beetles consumed significantly less diet treated with latex from young leaves. No significant difference in diet consumption was found between diets treated with latex from young and mature leaves of Tall Guzmaine in choice tests. Three solvents of differing polarity (water, methanol, and methylene chloride) were tested to extract deterrent compounds from latex; Valmaine latex extracted with water:methanol (20:80) strongly inhibited beetle feeding when applied to the surface of artificial diet. In no-choice tests, fewer beetles were observed feeding on diet treated with water:methanol (20:80) extract of Valmaine latex than on diet treated with a similar extract of Tall Guzmaine latex, resulting in significantly less consumption of the diet treated with the Valmaine latex extract. These studies suggest that moderately polar chemicals within latex may account for resistance in Valmaine to D. balteata.     

The involvement of amino acids in latex lipid synthesis in Euphorbia lathyris seedlings

The breakdown of triglycerides and proteins in the endosperm of Euphorbia lathyris was assayed in a 14 day germination period. Six days after germination, the average daily production was 2.7 μmol of amino acids. Arginine, glutamine, asparagine and glutamic acid accounted for 53% of the total amino acids. Excised cotyledons with 1 cm hypocotyls were used for amino acid uptake and their involvement in terpenoid synthesis was studied. Glutamine and aspartate were hardly involved in apolar lipid synthesis. Leucine, isoleucine, valine and threonine were mainly incorporated into the triterpenes in the laticifers. Alanine and serine were also involved in phytosterol synthesis in the adjacent tissue. In the 14 day germination period, ca3% of the daily yield of latex triterpenes may be synthesized from a variety of amino acids.