胭脂虫寄生前后梨果仙人掌挥发性物质的变化

为了研究胭脂虫Dactylopius coccus Costa寄生前后梨果仙人掌(Opuntia ficus-indica Mill)挥发性物质的变化,以动态顶空密闭循环吸附捕集法捕集胭脂虫寄生前后梨果仙人掌的挥发物,并运用全自动热脱附-气相色谱/质谱(ATD-GC/MS)联用技术进行分析检测,用峰面积归一化法测定其相对含量。结果表明,无虫梨果仙人掌的挥发物中共检测到8种化合物,有虫的梨果仙人掌中共检测到9种化合物,化合物的种类和含量都有一定的变化;萜烯类是植食昆虫诱导的植物挥发性次生物质中的主要成分,本试验中,萜烯类在胭脂虫寄生前后的含量由23.12%降为6.98%。在无虫仙人掌的挥发物中检测出苯甲醛,含量约占1.73%,有虫的仙人掌中则未检测到;同样在有虫仙人掌挥发物中检出的其他物质如酯类(21.15%)、醇类(1.79%)和卤代芳烃类(1.51%)物质在无虫仙人掌植株中都未检测出。     

胭脂虫寄生对仙人掌生长的影响

从印榕仙人掌(Opuntia ficus-indica Mill)植株高度、新增茎片数量和新增茎片面积3个方面研究不同密度胭脂虫Dactylopius coccus Costa寄生对仙人掌生长量的影响。结果表明:1)胭脂虫寄生对寄主仙人掌有明显的抑制作用,未被寄生的仙人掌株高、茎片数量和茎片面积的增加均明显大于被寄生的仙人掌;2)随着胭脂虫寄生密度的增加,仙人掌植株生长量的增加相应减少,生长高峰期的出现也会相应推迟;3)胭脂虫的寄生密度应控制在1000头/株以下,过大会导致仙人掌的死亡。     

胭脂虫Dactylopius coccus Costa培育方法研究

【目的】为探索胭脂虫Dactylopius coccus高产培育新方法。【方法】胭脂虫的室内培养模式有采摘茎片培育模式和种植培育模式两种。本研究在采摘茎片模式下开展了培育环境构筑、平铺培育、接种方式、蔗糖培育等培育技术的研究;在种植模式下进行了接种方式、轮放培育和寄主高度控制的培育技术研究。【结果】探索出胭脂虫在两种模式下的新培育方法。【结论】在采摘茎片培育模式下,采用松针覆土保湿效果较好;平铺培育产量较高,值得推广;悬挂培育宜采用网兜内置谷草接种方式。蔗糖对胭脂虫有明显助食作用,喷洒适当浓度的蔗糖有助于提高胭脂虫的产量。在种植模式下,宜进行轮放培育,寄主植株高度控制在100 cm之内且茎片级数在4级之内,可避免植株倒伏及断离。采用纱网撒种方式,可节约接种环节中大量人力。     

胭脂虫的行为及生殖特性

对胭脂虫Dactylopius coccus Costa在产卵、孵化、固定、泌蜡、蜕皮、结茧、羽化等方面的行为特性进行观察,并就该虫在有性及无性生殖两方面进行研究。结果表明,有性生殖♀虫平均怀卵量为269粒/雌,平均产卵量232粒/雌,平均孵化率96.5%,雌雄成虫性比约为3∶1,♂蛹羽化率可达94.7%,发育起点温度为5.77℃,世代有效积温为1600.60日.度。该胭脂虫不能以无性生殖方式进行繁殖,种群的延续需要以有性生殖方式进行。     

Aetiology of Opuntia ficus-indica malformations and stunting disease

The phytoplasma aetiology of epidemic stunting of cladodes and stunted growth observed in a cactus pear plantation in Carlentini (Sicily, Italy) was investigated with graft inoculation trials and PCR/restriction fragment length polymorphism analyses of 16S ribosomal RNA (rRNA) gene and S3 ribosomal protein gene. After sequencing of the 1525‐bp from 16S rRNA gene of both naturally infected and graft‐inoculated symptomatic cactus pear samples, phytoplasma TS belonging to ribosomal subgroup 16SrII‐C were identified as aetiological agents of this worldwide spread disease.     

Characterization of carbon metabolism in Opuntia ficus-indica Mill. exhibiting the idling mode of Crassulacean acid metabolism

Upon transfer from well-watered conditions to total drought, long-day-grown cladodes of Opuntia ficus-indica Mill. shift from full Crassulacean acid metabolism (CAM) to CAM-idling. Experiments using ¹⁴C-tracers were conducted in order to characterize the carbon-flow pattern in cladodes under both physiological situations. Tracer was applied by ¹⁴CO₂ fumigations and NaH¹⁴CO₃ injections during the day-night cycle. The results showed that behind the closed stomata, mesophyll cells of CAM-idling plants retained their full capacity to metabolize CO₂ in light and in darkness. Upon the induction of CAM-idling the level of the capacity of phosphoenolpyruvate carboxylase (EC 4.1.1.31) was maintained. By contrast, malate pools decreased, displaying finally only a small or no day-night oscillation. The capacity of NADP-malic enzyme (EC 1.1.1.40) decreased in parallel with the reduction in malate pools. Differences in the labelling patterns, as influenced by the mode of tracer application, are discussed.Abbreviations CAM Crassulacean acid metabolism - PEP-Case phosphoenolpyruvate carboxylase     

Genetic transformation of prickly-pear cactus (Opuntia ficus-indica) by Agrobacterium tumefaciens

A system for genetic transformation of an elite prickly pear cactus (Opuntia ficus-indica L., cultivar Villa Nueva) by Agrobacterium tumefaciens was developed. Beginning with direct bacterial infection by using a hypodermic syringe to the meristematic tissue termed areoles, transgenic plants were obtained by selection with 100 mg l⁻¹ kanamycin. Transient and stable GUS activities were monitored on kanamycin-resistant shoots and regenerated plants, respectively. Genetic transformation of regenerated plants growing under selection was demonstrated by PCR and Southern blot analysis; transgene copy number in the genome of transgenic plants ranged from two to six, while the transformation frequency obtained by the system reported here was of 3.2%. This method may be useful for routine transformation and introduction of several important genes in prickly pear cactus.     

Drought-induced changes in soil contact and hydraulic conductivity for roots of Opuntia ficus-indica with and without rhizosheaths

Water movement between roots and soil can be limited by incomplete root–soil contact, such as that caused by air gaps due to root shrinkage, and can also be influenced by rhizosheaths, composed of soil particles bound together by root exudates and root hairs. The possible occurrence of air gaps between the roots and the soil and their consequences for the hydraulic conductivity of the root–soil pathway were therefore investigated for the cactus t Opuntia ficus-indica, which has two distinct root regions: a younger, distal region where rhizosheaths occur, and an older, proximal region where roots are bare. Resin-embedded sections of roots in soil were examined microscopically to determine root–soil contact for container-grown plants kept moist for 21 days, kept moist and vibrated to eliminate air gaps, droughted for 21 days, or droughted and vibrated. During drought, roots shrank radially by 30% and root–soil contact in the bare root region of nonvibrated containers was reduced from 81% to 31%. For the sheathed region, the hydraulic conductivity of the rhizosheath was the least limiting factor and the root hydraulic conductivity was the most limiting; for the bare root region, the hydraulic conductivity of the soil was the least limiting factor and the hydraulic conductivity of the root–soil air gap was the most limiting. The rhizosheath, by virtually eliminating root–soil air gaps, facilitated water uptake in moist soil. In the bare root region, the extremely low hydraulic conductivity of the root–soil air gap during drought helped limit water loss from roots to a drier soil.     

Root growth, developmental changes in the apex, and hydraulic conductivity for Opuntia ficus-indica during drought

Developmental changes in the root apex and accompanying changes in lateral root growth and root hydraulic conductivity were examined for Opuntia ficus-indica (L.) Miller during rapid drying, as occurs for roots near the soil surface, and more gradual drying, as occurs in deeper soil layers. During 7 d of rapid drying (in containers with a 3-cm depth of vermiculite), the rate of root growth decreased sharply and most root apices died; such a determinate pattern of root growth was not due to meristem exhaustion but rather to meristem mortality after 3 d of drying. The length of the meristem, the duration of the cell division cycle, and the length of the elongation zone were unchanged during rapid drying. During 14 d of gradual drying (in containers with a 6-cm depth of vermiculite), root mortality was relatively low; the length of the elongation zone decreased by 70%, the number of meristematic cells decreased 30%, and the duration of the cell cycle increased by 36%. Root hydraulic conductivity ( L _P) decreased to one half during both drying treatments; L _P was restored by 2 d of rewetting owing to the emergence of lateral roots following rapid drying and to renewed apical elongation following gradual drying. Thus, in response to drought, the apical meristems of roots of O. ficus-indica near the surface die, whereas deeper in the substrate cell division and elongation in root apices continue. Water uptake in response to rainfall in the field can be enhanced by lateral root proliferation near the soil surface and additionally by resumption of apical growth for deeper roots.     

Young daughter cladodes affect CO2 uptake by mother cladodes of Opuntia ficus-indica

BACKGROUND AND AIMS: Drought damages cultivated C3, C4 and CAM plants in the semi-arid lands of central Mexico. Drought damage to Opuntia is common when mother cladodes, planted during the dry spring season, develop young daughter cladodes that behave like C3 plants, with daytime stomatal opening and water loss. In contrast, wild Opuntia are less affected because daughter cladodes do not develop on them under extreme drought conditions. The main objective of this work is to evaluate the effects of the number of daughter cladodes on gas exchange parameters of mother cladodes of Opuntia ficus-indica exposed to varying soil water contents. METHODS: Rates of net CO2 uptake, stomatal conductance, intercellular CO2 concentration, chlorophyll content and relative water content were measured in mature mother cladodes with a variable number of daughter cladodes growing in spring under dry and wet conditions. KEY RESULTS: Daily carbon gain by mother cladodes was reduced as the number of daughter cladodes increased to eight, especially during drought. This was accompanied by decreased mother cladode relative water content, suggesting movement of water from mother to daughter cladodes. CO2 assimilation was most affected in phase IV of CAM (late afternoon net CO2 uptake) by the combined effects of daughter cladodes and drought. Rainfall raised the soil water content, decreasing the effects of daughter cladodes on net CO2 uptake by mother cladodes. CONCLUSIONS: Daughter cladodes significantly hasten the effects of drought on mother cladodes by competition for the water supply and thus decrease daily carbon gain by mother cladodes, mainly by inhibiting phase IV of CAM.     

Opuntia ficus-indica cladodes as feedstock for ethanol production by Kluyveromyces marxianus and Saccharomyces cerevisiae

The feasibility of ethanol production using an enzymatic hydrolysate of pretreated cladodes of Opuntia ficus-indica (prickly pear cactus) as carbohydrate feedstock was investigated, including a comprehensive chemical analysis of the cladode biomass and the effects of limited aeration on the fermentation profiles and sugar utilization. The low xylose and negligible mannose content of the cladode biomass used in this study suggested that the hemicellulose structure of the O. ficus-indica cladode was atypical of hardwood or softwood hemicelluloses. Separate hydrolysis and fermentation and simultaneous saccharification and fermentation procedures using Kluyveromyces marxianus and Saccharomyces cerevisiae at 40 and 35 °C, respectively, gave similar ethanol yields under non-aerated conditions. In oxygen-limited cultures K. marxianus exhibited almost double the ethanol productivity compared to non-aerated cultures, although after sugar depletion utilization of the produced ethanol was evident. Ethanol concentrations of up to 19.5 and 20.6 g l^?1 were obtained with K. marxianus and S. cerevisiae, respectively, representing 66 and 70 % of the theoretical yield on total sugars in the hydrolysate. Because of the low xylan content of the cladode biomass, a yeast capable of xylose fermentation might not be a prerequisite for ethanol production. K. marxianus, therefore, has potential as an alternative to S. cerevisiae for bioethanol production. However, the relatively low concentration of fermentable sugars in the O. ficus-indica cladode hydrolysate presents a technical constraint for commercial exploitation.     

Use of Dactylopius coccus Costa (Hemiptera: Coccoidea) Pigments in the Detection of Larvae and Pupae of Plutella xylostella (L.) (Lepidoptera: Yponomeutidae)

Carmines obtained from the dye of Dactylopius coccus Costa (Hemiptera: Coccoidea) were used for the detection of larvae and pupae of Plutella xylostella (L.) (Lepidoptera: Yponomeutidae) in broccoli inflorescences. Larvae were dyed with carmine II and red cochineal, while the majority of the dyes, with the exception of carmine III and the aqueous extract, were suitable to dye pupae. In the broccoli lumps exposed to the dyes, only the verge of the stems were actually dyed, right in the position where the incision took place, an appropriate characteristic for implementing this technique for commercial use.     

Changes in Osmotic Pressure and Mucilage during Low-Temperature Acclimation of Opuntia ficus-indica

Opuntia ficus-indica, a Crassulacean acid metabolism plant cultivated for its fruits and cladodes, was used to examine chemical and physiological events accompanying low-temperature acclimation. Changes in osmotic pressure, water content, low molecular weight solutes, and extracellular mucilage were monitored in the photosynthetic chlorenchyma and the water-storage parenchyma when plants maintained at day/night air temperatures of 30/20°C were shifted to 10/0°C. An increase in osmotic pressure of 0.13 megapascal occurred after 13 days at 10/0°C. Synthesis of glucose, fructose, and glycerol accounted for most of the observed increase in osmotic pressure during the low-temperature acclimation. Extracellular mucilage and the relative apoplastic water content increased by 24 and 10%, respectively, during exposure to low temperatures. These increases apparently favor the extracellular nucleation of ice closer to the equilibrium freezing temperature for plants at 10/0°C, which could make the cellular dehydration more gradual and less damaging. Nuclear magnetic resonance studies helped elucidate the cellular processes during ice formation, such as those revealed by changes in the relaxation times of two water fractions in the chlorenchyma. The latter results suggested a restricted mobility of intracellular water and an increased mobility of extracellular water for plants at 10/0°C compared with those at 30/20°C. Increased mobility of extracellular water could facilitate extracellular ice growth and thus delay the potentially lethal intracellular freezing during low-temperature acclimation.     

Carbon flow and carbohydrate metabolism during sink-to-source transition for developing cladodes of Opuntia ficus-indica

Daughter cladodes (flattened stem segments) of Opuntia ficus-indica (L.) Miller at 14-18 d after appearance on the underlying basal cladodes were sinks, requiring carbohydrate import for growth. Import stopped at 25-36 d, and the daughter cladodes became sources at 27-28 d. The activities of Rubisco, PEPCase, and sucrose-Pi synthase as well as the chlorophyll content at 14 d were not less than those at 28 d, suggesting that photosynthetic or sucrose synthesis capacity was not limiting carbon assimilation for sink cladodes. Sucrose synthase (SS) activity was three times higher than that of alkaline invertase, indicating that SS is the major enzyme for cytoplasmic sucrose hydrolysis. The SS activity was correlated with cladode growth, the highest activity coinciding with the highest growth rate. The sink-to-source transition for daughter cladodes was correlated with increases in malate and H⁺ concentrations in the vacuoles of chlorenchyma cells, with 5-fold higher nocturnal malate production and 10-fold higher H⁺ concentration in 28- than in 14-d-old daughter cladodes. The vacuolar H⁺ increase during cladode development would lower cytoplasmic pH, which may trigger metabolic events affecting the sink-to-source transition.     

Effects of drought on water relations and nonstructural carbohydrates in cladodes of Opuntia ficus-indica

Nocturnal acid accumulation, water content, osmotic pressure (π), and nonstructural carbohydrates were determined in the chlorenchyma and the water-storage parenchyma of Opuntia ficus-indica (L.) Miller for well-watered plants and those subjected to drought for 15 weeks. During the 15-week drought, total cladode water content decreased by 57%, the water-storage parenchyma losing a greater fraction of water than the chlorenchyma, which most likely helped maintain nocturnal acid accumulation in the latter tissue. Despite the preferential water loss from the water-storage parenchyma, it had a lower π than the chlorenchyma over the 15 weeks of drought, suggesting a substantial decrease in osmotically active solutes in the water-storage parenchyma. Also, the measured π increases of both tissues were much less than those predicted based on the loss of water during drought and the initial content of osmotically active solutes under well-watered conditions. A decrease in the amount of soluble sugars (glucose. fructose and sucrose) occurred in plants subjected to drought. accounting for 46% and 81% of the difference between the measured and the predicted increases in π of the chlorenchyma and the water-storage parenchyma. respectively. The decrease in soluble sugars was associated with an equivalenl increase in polysaccharides, presumably starch, in the water-storage parenchyma. but not in the chlorenchyma.     

Rapid propagation of tuna (Opuntia ficus-indica) and plant establishment in soil

Explants from young joints of mature plants of tuna (Opuntia ficus-indica Mill.) were cultured on Murashige and Skoog (MS) medium containing 8.8 M benzyladenine (BA) and 0.5 M naphthaleneacetic acid (NAA). Shoots produced were utilized as secondary explants. Each shoot was cut longitudinally from apex to base into two explants, and some of these explants were cut transversely into proximal and distal explants. The size and number of shoots produced was affected by size and position of the explant within its source. The shoots were rooted in vitro or ex vitro and plants were successfully established in soil from both rooting methods.Abbreviations AC activated charcoal - BA benzyladenine - IBA indolebutyric acid - MS Murashige & Skoog (1962) medium - NAA naphthaleneacetic acid     

A Stereological Analysis of the Succulent Tissue 5Opuntia ficus-indica (L.) Mill.: I. DEVELOPMENT OF MUCILAGE CELLS

A correlated stereological and developmental analysis of thesucculent tissue of Opuntia ficus-indica was carried out. Thestereological parameters related to mucilage cells were describedas a function of their distance from the apical meristem. Polynomesof the second and third degree were found to describe the developmentalprocess. Three zones of development could be distinguished.The relation of the structure of the tissue to its physiologicalfunction and mucilage secretion is discussed.     

Isolation and structure of D-xylans from pericarp seeds of Opuntia ficus-indica prickly pear fruits

Xylans were isolated from the pericarp of prickly pear seeds of Opuntia ficus-indica (OFI) by alkaline extraction, fractionated by precipitation and purified. Six fractions were obtained and characterized by sugar analysis and NMR spectroscopy. They were assumed to be (4-O-methyl-D-glucurono)-D-xylans, with 4-O-alpha-D-glucopyranosyluronic acid groups linked at C-2 of a (1-->4)-beta-D-xylan. The sugar composition and the 1H and 13C NMR spectra showed that their chemical structures were very similar, but with different proportions of D-Xyl and 4-O-Me-D-GlcA. Our results showed that, on average, the water soluble xylans have one nonreducing terminal residue of 4-O-methyl-D-glucuronic acid for every 11 to 14 xylose units, whereas in the water non-soluble xylans, xylose units can varied from 18 to 65 residues for one nonreducing terminal residue of 4-O-methyl-D-glucuronic acid.     

A Stereological Analysis of the Succulent Tissue 5Opuntia ficus-indica (L.) Mill.: II. ULTRASTRUCTURAL DEVELOPMENT OF THE MUCILAGE CELLS

Five stages of the development of the cytoplasm of mucilagecells were followed by stereological analysis and analysed togive numerical (N_v), volumetric (V_v) and surface (S_v) densitiesof organelles and membranes. A quantitative evaluation of membrane areas, membrane sites,and mucilage secretion was obtained based on morphological andhistochemical data. The developmental process from early differentiationto complete degeneration of the cytoplasm is discussed in quantitativeterms.