Transformation of a CAM plant, the facultative halophyte Mesembryanthemum crystallinum by Agrobacterium tumefaciens

This study is the first to demonstrate that a foreign DNA can be delivered into cells of facultative halophyte crassulacean acid metabolism (CAM) plant, Mesembryanthemum crystallinum L. with Agrobacterium tumefaciens. This plant can be induced to change from C3 to CAM by drought and various stresses, and is a good model to study the environment stress on metabolism not only at whole plant but also at cell level. The β-glucuronidase (GUS) and kanamycin (Km) resistance genes were introduced into this plant. The average successful rate of transformation was about 54.5% with root tissue or 53.0% with hypocotyl tissue. Based on the resistance to Km, polymerase chain reaction (PCR) detection and GUS expression, transformation with Agrobacterium tumefaciens was successful for introducing foreign genes into M. crystallinum. This revised version was published online in June 2006 with corrections to the Cover Date.     

屋顶绿化植物佛甲草对温度梯度的生理生态适应

以屋顶生长的佛甲草为材料,通过光照培养箱进行不同温度条件处理,分别测量了叶片的CO2交换、叶绿素含量、叶绿素荧光参数以及植株不同部位的碳同位素比率变化(δ13 C).结果表明:持续高温/低温、较大的昼夜温差和叶表面风力的条件下,佛甲草为适应环境变化,光合会由C3代谢途径转变成景天酸代谢途径(CAM),是兼性CAM植物.短期降温会使叶片光系统Ⅱ(PSⅡ)发生不可逆失活,光合能力下降;复水后有助于PSⅡ的恢复和重建,而干旱天气会减缓恢复过程;在不利温度环境中生长的佛甲草老叶掉落较多,剩余叶片的叶绿素含量和Fv/Fm值增高,光合能力提高.δ13C测定结果显示,高温使嫩叶气孔导度降低,对成熟叶片气孔导度影响小,佛甲草茎杆虽然含有叶绿素,但没有明显的光合作用.     

Molecular Evidence for the Occurrence of H+-Transporting V-ATPase Subunit D and Two Different Forms of Subunit E in Leaves of the Obligate CAM Species Kalanchoë daigremontiana

Membrane proteins of purified tonoplast vesicles from leaves of Kalanchoë daigremontiana Hamet et Perrier were solubilized by the non-ionic detergent Triton X-114 and subsequently separated by MonoQ® anion-exchange chromatography. Special attention was given to the range of molecular masses around 30 kDa comprising the central stalk subunit peptides of the H⁺-transporting V-ATPase. Three polypeptides of apparent molecular masses of 32, 33 and 34 kDa were separated. Proteolytic fragments were obtained by trypsin digestion. Analysis by matrix-assisted laser desorption ionization (MALDI) mass spectrometry of tryptic fragments of the 32 and 33 kDa peptides and protein data- bank comparisons showed that they are two different forms of subunit E. N-terminal amino acid sequencing of tryptic fragments of the 34 kDa peptide showed that it is subunit D. This work provides for the first time unequivocal molecular evidence that the central stalk of the V-ATPase of the obligate CAM plant K. daigremontiana includes subunit D and different forms of subunit E.     

石斛属植物光合碳同化途径地理分异研究进展

系统总结了我国石斛属植物光合作用现状,结合石斛属植物光合作用碳同化途径研究,概括了兼性景天酸代谢植物的碳同化路径特征,分析了景天酸代谢表达程度与生态环境的关系,进而归纳出石斛属植物在光合作用碳同化途径方面存在地理分异。最后指出了石斛属植物系统发育研究领域存在的问题,提出从系统发育视角,采用分子系统学技术来研究石斛属植物光合碳同化途径地理分异的生物学机制。     

Day-night changes of citric-acid levels in crassulacean acid metabolism: phenomenon and ecophysiological significance

Abstract A reappraisal is offered of old and new observations of substantial day-night changes of citric-acid levels in crassulacean acid metabolism (CAM). In contrast to malic acid, the biochemical consequences and the ecophysiological significance of nocturnal accumulation of citric acid in CAM have not received due attention. Considerations show that citric-acid accumulation does not provide a means for nocturnal storage of CO₂. It may, however, serve carbon retention by internal recycling and sustain the water budget affording a vacuolar osmoticum. Since citric-acid accumulation energetically is considerably more favourable than malic-acid accumulation, this may have important ecophysiological implications. The questions raised by these reflections can and need to be tackled experimentally.     

Comparisons of photosynthesis and photoinhibition in the CAM vine Hoya australis and several C3 vines growing on the coast of eastern Australia

Abstract. The CAM vine Hoya australis and three C₃ vines, Smilax australis, Ipomoea pes-caprae and Kennedta rubicunda, were studied at a site on the coast of northeastern New South Wales, Australia. The level of CAM activity (nocturnal acid accumulation) was comparable in H. australis growing in full sunlight and in deep shade. Acclimation to shade by H. australis was indicated by thinner leaves, increased chlorophyll content, decreased chlorophyll a/b ratios, lower dark respiration rates, and lower light compensation points. When growing in full sunlight H. australis exhibited reductions in photochemical efficiency, as indicated by reduced quantum yields and F_v/F_m fluorescence from PS II as well as low rates of photosynthesis at high light. Sun leaves of H. australis experienced a massive quenching of fluorescence from PS II during normal exposure to midday irradiance which was rapidly reversible under low irradiance conditions in the late afternoon. This quenching indicated a reduction in photochemical efficiency, part of which could be accounted for by an increase in non-radiative energy dissipation, while part of it was due to one or more processes not yet identified. Changes in PS II fluorescence from shade H. australis exposed to full sunlight suggest a decrease in the rate constant for photochemistry indicative of damage to the reaction centre, as well as an increase in non-radiative energy dissipation. The C₃ vine S. australis was also shade tolerant, but exhibited little evidence of photoinhibition when growing in full sunlight. Ipomoea pescaprae and K. rubicunda, both of which were apparently shade intolerant (being found only in full sunlight), possessed high quantum yields and much higher photosynthetic capacities than either H. australis or S. australis. From this study, and several others, it appears that plants possessing CAM experience photoinhibition to a greater degree than do C₃ species in full sunlight under natural conditions, which is probably exacerbated by some degree of CAM-idling.     

Comparative ecophysiology of CAM and C3 bromeliads. II. Field measurements of gas exchange of CAM bromeliads in the humid tropics

Abstract The results described represent the first detailed measurements of gas exchange of epiphytic plants with crassulacean acid metabolism (CAM) in the humid tropics. A portable steady-state CO₂ and H₂O porometer was used to measure net exchange rates of CO₂ and H₂O vapour (J_CO2, J_H2O), leaf temperature (T₁), air temperature (T_A), air relative humidity (RH) and photosynthetically active radiation (PAR) for bromeliads in the field during the dry season in February and March 1983 on the tropical island of Trinidad. Different lengths of tubing (up to 25 m) were used so that the gas exchange could be measured of bromeliads in situ in their epiphytic habitats. Derived parameters such as leaf-air water-vapour-concentration difference (Δ_w), water-vapour conductance of leaves (g) and internal CO₂ partial pressure (pⁱ_CO2) could be calculated. The particular problems of making such measurements in the humid tropics due to high relative humidities and high dew-point temperatures are discussed. The long and often broad, strap-like leaves of bromeliads are well suited for measurements with the steady-state porometer. It is shown that CAM activity varies along the length of individual leaves, and variability between different leaves is also demonstrated. The major phases of CAM, i.e. nocturnal stomalal opening, CO₂ uptake and dark fixation as malic acid (Phase I), daytime stomatal closure and light-dependent assimilation of CO₂ derived from decarboxylation of the malic acid (Phase III), and late-afternoon stomatal opening with direct light-dependent assimilation of atmospheric CO₂ (Phase IV) were all clearly shown by CAM bromeliads in situ. Their expression and magnitude depended on the environmental conditions. An early-morning peak of CO₂ uptake as is characteristic of Phase II of CAM was not detected during the night-day transition. A bromeliad intermediate between C₃ and CAM, Guzmania monostachia, showed substantial net CO₂ uptake in the early morning but no net uptake integrated over the whole of the night.     

四种珍稀濒危石斛属植物光合特性及叶片显微结构的比较研究

为探究珍稀濒危石斛属植物的光合特性及其叶片显微结构特征,该研究对滇桂石斛（Dendrobium scoriarum）、喇叭唇石斛（D.lituiflorum）、罗河石斛（D.lohohense）和钩状石斛（D.aduncum） 4种石斛属植物的净光合速率（P_n）日变化、光响应曲线、CO₂响应曲线、叶绿素含量和叶片显微结构等指标进行了测定。结果表明：（1）滇桂石斛和钩状石斛白天P_n为正,上午8:00—10:00出现最大峰值,夜间P_n低于白天,存在P_n为正的情况;喇叭唇石斛和罗河石斛白天P_n日变化呈双峰曲线,白天P_n为正,夜间P_n为负值。（2） 4种石斛属植物中,喇叭唇石斛的光合能力最强,罗河石斛的光适应范围最宽。（3）喇叭唇石斛的初始羧化效率（α）、潜在最大净光合速率(A_max)和光呼吸速率（R_p）显著性（P<0.05）大于其他3种石斛。（4）滇桂石斛和钩状石斛的叶片厚度（LT）大于喇叭唇石斛和罗河石斛,而气孔密度（SD）则呈现相反趋势。（5）喇叭唇石斛的叶绿素总量（Chl）、叶绿素a（Chla）和叶绿素b（Chlb）的含量均显著性（P<0.05）大于其他3种石斛。（6）叶肉厚度（MT）与最大净光合速率(P_max)呈显著性（P<0.05）负相关;SD与P_max呈极显著性（P<0.01）正相关,与A_max呈显著性（P<0.05）正相关;Chl总量与P_max、A_max呈显著性（P<0.05）正相关。综上认为,较厚的叶肉组织和低气孔密度可能是4种石斛属植物应对弱光环境以及附生于树干或生长于岩石缺水环境的适应机制,叶肉厚度、气孔密度及叶绿素总含量是影响4种石斛光合能力大小的重要因素,在引种栽培过程中营造适宜的光照环境、适当增加CO₂浓度有利于4种石斛属植物的生长。该研究结果为石斛属植物种质资源保育和引种栽培提供了参考。     

Diel water movement between parenchyma and chlorenchyma of two desert CAM plants under dry and wet conditions

Abstract. Electric-circuit analogue models of the water relations of crassulacean acid metabolism (CAM) succulents such as Agave deserti and Ferocactus acanthodes have predicted diel movement of water between the water-storage parenchyma and the photo-synthetic chlorenchyma. Injection of tritiated water into either tissue in the laboratory confirmed substantial and bidirectional water movements, especially under conditions of wet soil. For A. deserti , water movement from the water-storage parenchyma to the chlorenchyma increased at night as the chlorenchyma osmotic pressure increased. Although nocturnal osmotic pressure increases and transpiration for both species were minimal in the field under dry conditions, diel changes in the deuterium: hydrogen ratio (expressed as ΔD) were similar for the water-storage parenchyma and the chlorenchyma. Such indication of [substantial mixing of water between the tissues over a 24-h cycle was more evident under wet conditions in the field. For A. deserti , ΔD then increased by 32%o from the afternoon to midnight and was essentially identical in the water-storage parenchyma and the chlorenchyma. For F. acanthodes , the diel changes in ΔD were one-third those of A. deserti , and ΔD was always slightly higher for the chlorenchyma than for the water-storage parenchyma, apparently reflecting the lower surface-to-volume ratio of A. deserti. In summary, data obtained using radioactive and stable isotopes strongly supported model predictions concerning diel cycles of internal water distribution for these CAM species.     

Comparative ecophysiology of CAM and C3 bromeliads. I. The ecology of the Bromeliaceae in Trinidad

Abstract This article deals with the physiological ecology of the Bromeliaceae, a large neotropical family containing both terrestrial and epiphytic forms, as well as many species with crassulacean acid metabolism (CAM). The article is in two parts. In the first, we review what is known of the occurrence of CAM and C₃ species in the Bromeliaceae. The photosynthetic pathways are discussed in the context of the major taxonomic divisions within the family and the great diversity of bromeliad life-forms. Of the three subfamilies, the Pitcairnioideae contain both C₃ and CAM species and are essentially all terrestrial. In contrast, the Tillandsioideae are entirely epiphytic or saxicolous, with CAM species being restricted to the genus Tillandsia, And in the Bromelioideae all species show CAM, but terrestrial and epiphytic forms are found in about equal numbers. The evidence suggests that both CAM and the epiphytic habit arose more than once in the family's evolutionary history. In the second part we consider the photosynthetic ecology of the various bromeliad life-forms in more detail using the specific example of Trinidad (West Indies). CAM bromeliads tend to be centred on the drier regions of the island and C₃ forms on the wetter areas. However, at any one site there is a marked vertical stratification of species within the forest profile. Based on the known habitat preferences of the bromeliads, six contrasting sites were selected for field studies in Trinidad. These ranged from arid coastal scrub to montane rain forest, the vegetational and climatic characteristics of which are described here. The constancy of δ¹³C values (carbon-isotope ratios) for individual CAM species in these markedly different habitats emphasized the need for ecophysiological studies to characterize environmental effects on CO₂ assimilation and transpiration. The following papers in this series present the results of a comparative investigation of gas exchange and leaf water relations of CAM and C₃ bromeliads in situ at the various sites.     

Photosynthetic Characteristics of the Palisade Mesophyll and Spongy Mesophyll in the CAM/C4 Intermediate Plant,Peperomia camptotricha*

Several photosynthetic parameters were examined in the different tissue layers of leaves from the recently characterized crassulacean acid metabolism/C₄ intermediate plant, Peperomia camptotricha (Nishio and Ting, 1987). Light appears to control the development of certain photosynthetic characteristics within the tissue layers, while factors other than light seem to dictate others. Analysis of the chlorophyll content (including P700) and chlorophyll-proteins indicated that more light harvesting chlorophylls were associated with reaction centers in the tissues that were shaded by overlying tissue. Electron transport activity and chlorophyll-protein analysis indicated that the ratio of photosystem I to photosystem II in the spongy mesophyll (the abaxial tissue type and hence most shaded) was elevated relative to the overlying chlorophyll-rich median palisade mesophyll. The elevated photosystem I relative to photosystem II in the spongy mesophyll of Peperomia camptotricha may be related to C₄ metabolism and an increased requirement for ATP.     

佛甲草(Sedum lineare)在干旱条件下的气体交换

在干旱条件下,测定了广州市1、4、5和年期屋顶绿化植物佛甲草叶片的气体交换参数、相对含水量(RWC)和比叶重(LMA),探讨了佛甲草在屋顶绿化中的生理生态适应性.结果表明不同生长期佛甲草叶片的RWC和LMA相对较低,分别在81.79%～87.58%和0.059～0.072 kg·m^-2之间,RWC和LMA在不同生长期的差异不显著(P>0.05).不同生长期佛甲草的气体交换参数CO₂同化速率(A)、气孔导度(g_s)和蒸腾速率(E)均呈现相似的日变化格型,A、g_s和E的日均值差异不显著(P>0.05).A、g_s和E之间的日变化呈明显的正相关关系(P<0.01).根据植物的RWC和气体交换参数的特征,推断佛甲草属兼性CAM植物,不同生长期佛甲草的主要生理生态指标相似.     

Daily fluctuations of titratable acidity, content of organic acids (malate and citrate) and soluble sugars of varieties and wild relatives of Ananas comosus L. growing under natural tropical conditions

The genus Ananas has its centre of origin in northern South America. In this area, several varieties of Ananas comosus are widely cultivated, and a number of wild species are found growing under variable conditions of light intensity, soil fertility and water availability. Here we report detailed daily courses of titratable acidity, and malate, citrate and free-sugars content of several cultivated varieties of A. comosus and of A. ananassoides, a closely related species growing on granitic rock-outcrops in southern Venezuela. Day-night oscillations of both malate and citrate were detected in plants growing under full sun, but malate was by far the most important organic anion associated with CAM performance in ail populations sampled. Fructose was the dominant compound in the neutral fraction, but only sucrose showed a consistent inverse relation with the cycle of titratable acidity. The diel oscillations of free sugars measured were not always enough to account for the amount of organic anions accumulated during the night. Plants cultivated under shady conditions always showed a lower night-time increase in titratable acidity and organic acids, and also smaller oscillations in the amount of free sugars than sun exposed plants. In all populations growing under full sun, osmolality increased during the night, but it was not always possible to explain these changes on the basis of variations in molar concentrations of organic acids and sugars. Besides, no diel variations in the cations K⁺, Ca²⁺ and Mg²⁺ were detected. K⁺ was always the dominant cation (K/Ca ratios ～ 19), while Mg²⁺ was always higher than Ca²⁺ (Mg/Ca ～ 2).     

不同碱度条件下中华水韭昼夜CO2气体交换的特征

 运用pH-drift的方法研究了在不同碱度条件下中华水韭(Isoetes sinensis)的沉水叶片昼夜CO₂吸收的特征。结果表明中华水韭的沉水叶片具有昼夜吸收水中CO₂的能力, 而不具备利用水中的HCO ₃^–的能力, 进一步证明了水生植物中华水韭的光合碳同化途径具有景天酸代谢(CAM)的特征。中华水韭沉水叶片光照条件下对水中CO₂的吸收速率在一定的浓度范围内正相关于水中的CO₂浓度。光照条件下, 中华水韭的pH-drift实验的pH补偿点分别为(8.1±0.3)和(7.9±0.1) mmol·L^–1, 最终[CT]/Alk值为(1.009±0.01)和(1.022±0.004)。碱度对中华水韭夜晚CO2的吸收速率有显著的影响(F = 38.73, p < 0.000 1)。总碱度1.70 mmol·L^–1溶液中的中华水韭沉水叶片在相对较低的CO₂浓度(0.04±0.001 mmol·L^–1)水平下即表现出对CO₂的净吸收。调查了野外一处中华水韭沉水种群的生境pH值及CO₂浓度的昼夜变化, 发现水体碱度约为1.59 mmol·L^–1, 一昼夜的pH值波动不大, 平均为(6.1±0.04), 昼夜CO₂浓度存在波动, 午夜水中的CO₂浓度是午后的近3倍。