Exogenously-sourced ethylene increases stomatal conductance, photosynthesis, and growth under optimal and deficient nitrogen fertilization in mustard

In order to ascertain the stomatal and photosynthetic responses of mustard to ethylene under varying N availability, photosynthetic characteristics of mustard grown with optimal (80 mg N kg(-1) soil) or low (40 mg N kg(-1) soil) N were studied after the application of an ethylene-releasing compound, ethephon (2-chloroethyl phosphonic acid) at 40 days after sowing (DAS). The availability of N influenced ethylene evolution and affected stomatal conductance and photosynthesis. The effect of ethylene was smaller under deficient N where plants contained higher glucose (Glc) sensitivity, despite high ethylene evolution even in the absence of ethephon, potentially because the plants were less sensitive to ethylene per se. Ethephon application at each level of N increased ethylene and decreased Glc sensitivity, which increased photosynthesis via its effect on the photosynthetic machinery and effects on stomatal conductance. Plants grown with sufficient-N and treated with 200 μl l(-1) ethephon exhibited optimal ethylene, the greatest stomatal conductance and photosynthesis, and growth. These plants made maximum use of available N and exhibited the highest nitrogen-use efficiency (NUE).     

不同光强下高锰对黄瓜光合作用特性的影响

采用营养液培养的方法,研究了不同光强下高锰对黄瓜植株生长、叶绿素含量、叶绿素荧光参数和光合作用的影响.结果表明,高锰处理抑制了黄瓜植株的生长,与弱光处理相比强光下抑制幅度更加显著.强光下,高锰处理显著降低叶绿素含量,但降低光强却增加其含量.强光下,高锰处理显著降低原初光能转换效率(F_v/F_m)、光合电子传递量子效率(ΦPSII)和光化学猝灭系数(qP);弱光下,高锰处理对F_v/F_m和qP无显著影响.高锰处理使净光合速率(P_n)和气孔导度(G_s)下降.尤其是在强光下下降幅度更大.高锰处理使细胞间CO₂浓度(C_i)在强光下升高,而在弱光下则下降.与C_i相反,高锰处理使气孔限制值(L_s)在强光下下降,而在弱光下上升.因此,强光下高锰胁迫使净光合速率下降可能是由非气孔限制引起的,而弱光下高锰处理使净光合速率下降可能是由气孔因子限制引起的.     

The contribution of photosynthesis to the red light response of stomatal conductance

To determine the contribution of photosynthesis on stomatal conductance, we contrasted the stomatal red light response of wild-type tobacco (Nicotiana tabacum 'W38') with that of plants impaired in photosynthesis by antisense reductions in the content of either cytochrome b(6)f complex (anti-b/f plants) or Rubisco (anti-SSU plants). Both transgenic genotypes showed a lowered content of the antisense target proteins in guard cells as well as in the mesophyll. In the anti-b/f plants, CO(2) assimilation rates were proportional to leaf cytochrome b(6)f content, but there was little effect on stomatal conductance and the rate of stomatal opening. To compare the relationship between photosynthesis and stomatal conductance, wild-type plants and anti-SSU plants were grown at 30 and 300 micromol photon m(-2) s(-1) irradiance (low light and medium light [ML], respectively). Growth in ML increased CO(2) assimilation rates and stomatal conductance in both genotypes. Despite the significantly lower CO(2) assimilation rate in the anti-SSU plants, the differences in stomatal conductance between the genotypes were nonsignificant at either growth irradiance. Irrespective of plant genotype, stomatal density in the two leaf surfaces was 2-fold higher in ML-grown plants than in low-light-grown plants and conductance normalized to stomatal density was unaffected by growth irradiance. We conclude that the red light response of stomatal conductance is independent of the concurrent photosynthetic rate of the guard cells or of that of the underlying mesophyll. Furthermore, we suggest that the correlation of photosynthetic capacity and stomatal conductance observed under different light environments is caused by signals largely independent of photosynthesis.     

Regulation of photosynthesis by sugars in sugarcane leaves

In sugarcane, increased sink demand has previously been shown to result in increased photosynthetic rates that are correlated with a reduction in leaf hexose concentrations. To establish whether sink limitation of photosynthesis is a result of sugar accumulation in the leaf, excision and cold-girdling techniques were used to modify leaf sugar concentrations in pot-grown sugarcane. In excised leaves that were preincubated in darkness for 3h, sucrose accumulation was reduced but accumulated again upon transfer to the light, while hexose concentrations remained lower than in controls (7.7 micromol mg(-1)FW versus 18.6 micromol mg(-1)FW hexose in controls). These results were associated with a 66% and 59% increase in photosynthetic assimilation (A) and electron transport rate (ETR), respectively, compared to controls maintained in the light. Similar increases in photosynthesis were observed when dark-treated leaves were supplied with 5mM sorbitol, but not when supplied with 5mM sucrose. Further analyses of (14)C-labeled sugars indicated rapid turnover between sucrose and hexose. Cold-girdling (5 degrees C) increased sucrose and hexose levels and resulted in a decline of photosynthetic rates over 5d (48% and 35% decline in assimilation rate and ETR, respectively). These sugar-induced changes in photosynthesis were independent of changes in stomatal conductance. This study demonstrates that the down-regulation of photosynthesis in response to culm sugar accumulation reported previously could be due to the knock-on effect of accumulation of sugar in leaf tissue, and supports the contention that hexose, rather than sucrose, is responsible for the modulation of photosynthetic activity.     

Dynamics of the enhanced emissions of monoterpenes and methyl salicylate, and decreased uptake of formaldehyde, by Quercus ilex leaves after application of jasmonic acid

Jasmonic acid (JA) is a signalling compound with a key role in both stress and development in plants, and is reported to elicit the emission of volatile organic compounds (VOCs). Here we studied the dynamics of such emissions and the linkage with photosynthetic rates and stomatal conductance. We sprayed JA on leaves of the Mediterranean tree species Quercus ilex and measured the photosynthetic rates, stomatal conductances, and emissions and uptake of VOCs using proton transfer reaction mass spectrometry and gas chromatography after a dark-light transition. Jasmonic acid treatment delayed the induction of photosynthesis and stomatal conductance by approx. 20 min, and decreased them 24 h after spraying. Indications were found of both stomatal and nonstomatal limitations of photosynthesis. Monoterpene emissions were enhanced (20-30%) after JA spraying. Jasmonic acid also increased methyl salicylate (MeSa) emissions (more than twofold) 1 h after treatment, although after 24 h this effect had disappeared. Formaldehyde foliar uptake decreased significantly 24 h after JA treatment. Both biotic and abiotic stresses can thus affect plant VOC emissions through their strong impact on JA levels. Jasmonic acid-mediated increases in monoterpene and MeSa emissions might have a protective role when confronting biotic and abiotic stresses.     

Light-emitting diodes as a light source for photosynthesis research

Light-emitting diodes (LED) can provide large fluxes of red photons and so could be used to make lightweight, efficient lighting systems for photosynthetic research. We compared photosynthesis, stomatal conductance and isoprene emission (a sensitive indicator of ATP status) from leaves of kudzu (Pueraria lobata (Willd) Ohwi.) enclosed in a leaf chamber illuminated by LEDs versus by a xenon arc lamp. Stomatal conductance was measured to determine if red LED light could sufficiently open stomata. The LEDs produced an even field of red light (peak emission 656±5 nm) over the range of 0–1500 mol m^-2 s^-1. Under ambient CO₂ the photosynthetic response to red light deviated slightly from the response measured in white light and stomatal conductance followed a similar pattern. Isoprene emission also increased with light similar to photosynthesis in white light and red light. The response of photosynthesis to CO₂ was similar under the LED and xenon arc lamps at equal photosynthetic irradiance of 1000 mol m^-2 s^-1. There was no statistical difference between the white light and red light measurements in high CO₂. Some leaves exhibited feedback inhibition of photosynthesis which was equally evident under irradiation of either lamp type. Photosynthesis research including electron transport, carbon metabolism and trace gas emission studies should benefit greatly from the increased reliability, repeatability and portability of a photosynthesis lamp based on light-emitting diodes.     

Effects of addition of external nitric oxide on the allocation of photosynthetic electron flux in Rumex K-1 leaves under osmotic shock

Photosynthetic electron flux allocation, stomatal conductance, and the activities of key enzymes involved in photosynthesis were investigated in Rumex K-1 leaves to better understand the role of nitric oxide (NO) in photoprotection under osmotic stress caused by polyethylene glycol. Gas exchange and chlorophyll fluorescence were measured simultaneously with a portable photosynthesis system integrated with a pulse modulated fluorometer to calculate allocation of photosynthetic electron fluxes. Osmotic stress decreased stomatal conductance, photosynthetic carbon assimilation, and nitrate assimilation, increased Mehler reaction, and resulted in photoinhibition. Addition of external NO enhanced the stomatal conductance, photosynthetic rate, activities of glutamine synthetase and nitrate reductase, and reduced Mehler reaction and photoinhibition. These results demonstrated that osmotic stress reduced CO₂ assimilation, decreasing the use of excited energy via CO₂ assimilation which caused significant photoinhibition. Improving stomatal conductance by the addition of external NO enhanced the use of excited energy via CO₂ assimilation. As a result, less excited energy was allocated to Mehler reaction, which reduced production of reactive oxygen species via this pathway. We suppose that Mehler reaction is not promoted unless photosynthesis and nitrogen metabolism are prominently inhibited.     

Increased stomatal conductance induces rapid changes to photosynthetic rate in response to naturally fluctuating light conditions in rice

A close correlation between stomatal conductance and the steady-state photosynthetic rate has been observed for diverse plant species under various environmental conditions. However, it remains unclear whether stomatal conductance is a major limiting factor for the photosynthetic rate under naturally fluctuating light conditions. We analysed a SLAC1 knockout rice line to examine the role of stomatal conductance in photosynthetic responses to fluctuating light. SLAC1 encodes a stomatal anion channel that regulates stomatal closure. Long exposures to weak light before treatments with strong light increased the photosynthetic induction time required for plants to reach a steady-state photosynthetic rate and also induced stomatal limitation of photosynthesis by restricting the diffusion of CO₂ into leaves. The slac1 mutant exhibited a significantly higher rate of stomatal opening after an increase in irradiance than wild-type plants, leading to a higher rate of photosynthetic induction. Under natural conditions, in which irradiance levels are highly variable, the stomata of the slac1 mutant remained open to ensure efficient photosynthetic reaction. These observations reveal that stomatal conductance is important for regulating photosynthesis in rice plants in the natural environment with fluctuating light.     

Stomatal limitation of photosynthesis as affected by water stress and CO2 concentration

A water stress effect on photosynthesis and transpiration of wheat seedlings at 50-500 µmol(CO2) mol-1 was measured in an open gas exchange system. The limitation of photosynthesis by stomatal conductance was quantified by a stomatal control coefficient of the net photosynthetic rate. The stomatal control coefficient increased linearly as the water potential of root media decreased to -1 MPa, and it decreased with increasing CO2 concentration.     

Relationships between leaf conductance to CO2 diffusion and photosynthesis in micropropagated grapevine plants, before and after ex vitro acclimatization

In vitro-cultured plants typically show a low photosynthetic activity, which is considered detrimental to subsequent ex vitro acclimatization. Studies conducted so far have approached this problem by analysing the biochemical and photochemical aspects of photosynthesis, while very little attention has been paid to the role of leaf conductance to CO(2) diffusion, which often represents an important constraint to CO(2) assimilation in naturally grown plants. Mesophyll conductance, in particular, has never been determined in in vitro plants, and no information exists as to whether it represents a limitation to carbon assimilation during in vitro growth and subsequent ex vitro acclimatization. In this study, by means of simultaneous gas exchange and chlorophyll fluorescence measurements, the stomatal and mesophyll conductance to CO(2) diffusion were assessed in in vitro-cultured plants of the grapevine rootstock '41B' (Vitis vinifera 'Chasselas'xVitis berlandieri), prior to and after ex vitro acclimatization. Their impact on electron transport rate partitioning and on limitation of potential net assimilation rate was analysed. In vitro plants had a high stomatal conductance, 155 versus 50 mmol m(-2) s(-1) in acclimatized plants, which ensured a higher CO(2) concentration in the chloroplasts, and a 7% higher electron flow to the carbon reduction pathway. The high stomatal conductance was counterbalanced by a low mesophyll conductance, 43 versus 285 mmol m(-2) s(-1), which accounted for a 14.5% estimated relative limitation to photosynthesis against 2.1% estimated in acclimatized plants. It was concluded that mesophyll conductance represents an important limitation for in vitro plant photosynthesis, and that in acclimatization studies the correct comparison of photosynthetic activity between in vitro and acclimatized plants must take into account the contribution of both stomatal and mesophyll conductance.     

Antisense inhibition of the iron-sulphur subunit of succinate dehydrogenase enhances photosynthesis and growth in tomato via an organic acid-mediated effect on stomatal aperture

Transgenic tomato (Solanum lycopersicum) plants expressing a fragment of the Sl SDH2-2 gene encoding the iron sulfur subunit of the succinate dehydrogenase protein complex in the antisense orientation under the control of the 35S promoter exhibit an enhanced rate of photosynthesis. The rate of the tricarboxylic acid (TCA) cycle was reduced in these transformants, and there were changes in the levels of metabolites associated with the TCA cycle. Furthermore, in comparison to wild-type plants, carbon dioxide assimilation was enhanced by up to 25% in the transgenic plants under ambient conditions, and mature plants were characterized by an increased biomass. Analysis of additional photosynthetic parameters revealed that the rate of transpiration and stomatal conductance were markedly elevated in the transgenic plants. The transformants displayed a strongly enhanced assimilation rate under both ambient and suboptimal environmental conditions, as well as an elevated maximal stomatal aperture. By contrast, when the Sl SDH2-2 gene was repressed by antisense RNA in a guard cell-specific manner, changes in neither stomatal aperture nor photosynthesis were observed. The data obtained are discussed in the context of the role of TCA cycle intermediates both generally with respect to photosynthetic metabolism and specifically with respect to their role in the regulation of stomatal aperture.     

外源硫对镉胁迫下马齿苋光合性状和矿质元素吸收的影响

从光合反应系统揭示外源硫(S)诱导马齿苋镉(Cd)耐受性的生理机制,为外源S缓解重金属毒害提供理论依据.采用营养液培养,研究外源S供体(NH4)2SO4对100 mg/L Cd胁迫下马齿苋叶片光合色素、光合特性、叶绿素荧光参数和矿质营养元素的影响.结果表明,Cd胁迫可显著降低马齿苋叶片中叶绿素a和叶绿素b含量;净光合速率、蒸腾速率、气孔导度均显著降低,而胞间二氧化碳浓度上升,表明非气孔因素是Cd胁迫诱导马齿苋光合抑制的主要因素;同时,PSⅡ实际光化学效率(ФPSII)、电子传递效率(J)、化学猝灭系数(qP)显著下降,而非化学猝灭系数(qN)显著上升,表明Cd胁迫影响马齿苋PSⅡ反应系统的正常运行.外施400 mg/L(NH4)2SO4显著提高马齿苋叶片叶绿素a含量、叶绿素b含量和叶绿素a/b比值,增强马齿苋叶片光合作用和PSⅡ原初光化学反应量子效率.对5种与光反应系统密切相关的矿质元素含量进行分析发现,Cd处理显著增加马齿苋叶片中的Ca和Fe含量,显著抑制马齿苋对Mg、Mn和Cu的吸收.Cd胁迫下马齿苋叶片的变黄与Mg、Mn的亏缺有关,而与Fe缺乏无关;添加外源S可显著提升马齿苋叶片中Ca、Mg、Fe、Cu和Mn含量,从而增强Cd胁迫下马齿苋叶片的PSII反应系统功能.     

A coupled model of photosynthesis,stomatal conductance and transpiration for a rose leaf (Rosa hybrida L.)

The following three models were combined to predict simultaneously photosynthesis, stomatal conductance, transpiration and leaf temperature of a rose leaf: the biochemical model of photosynthesis of Farquhar, von Caemmerer and Berry (1980, Planta 149: 78-90), the stomatal conductance model of Ball, Woodrow and Berry (In: Biggens J, ed. Progress in photosynthesis research. The Netherlands: Martinus Nijhoff Publishers), and an energy balance model. The photosynthetic parameters: maximum carboxylation rate, potential rate of electron transport and rate of triose phosphate utilization, and their temperature dependence were determined using gas exchange data of fully expanded, young, sunlit leaves. The stomatal conductance model was calibrated independently. Prediction of net photosynthesis by the coupled model agreed well with the validation data, but the model tended to underestimate rates of stomatal conductance and transpiration. The coupled model developed in this study can be used to assist growers making environmental control decisions in glasshouse production.     

叶面喷施硝酸盐对丁香叶片气孔动态、光合作用的影响

以哈尔滨市绿化植物紫丁香（Syringa oblata Lindl.）为研究对象,借助便携式光合测定系统和电子显微镜,研究了叶面喷施硝酸盐（50 mmol·L^-1 NaNO₃溶液）对叶片气孔动态（气孔分布、气孔开度）和光合生理过程（光合—呼吸的光诱导、光化学效率、叶绿素含量以及细胞间隙二氧化碳浓度、气孔导度）的影响。结果表明：叶片上、下表皮均有气孔、腺毛分布,喷施处理第5 d时,叶片的气孔开度和导度、光合速率、叶绿素含量等未受到显著影响,但14 d时,气孔关闭、导度下降,光合—呼吸的光诱导过程受到显著影响。     

Gas exchange and resource-use efficiency of Leymus cinereus (Poaceae): diurnal and seasonal responses to naturally declining soil moisture

We examined factors that limit diurnal and seasonal photosynthesis in Leymus cinereus, a robust tussock grass from shrub-steppes of western North America. Data from plants in a natural stand and in experimental field plots indicate that this bunchgrass has 1) a high photosynthetic capacity, 2) high leaf nitrogen content and high nitrogen-use efficiency, 3) a steep leaf-to-air diffusion gradient for carbon dioxide, which enhances intrinsic water-use efficiency, and 4) photosynthetic tissues that tolerate severe water stress and recover quickly from moderate water stress. Midday depressions of CO₂ assimilation (A) and stomatal conductance were slight in plants with plentiful water, but marked in plants subject to moderate water stress. Midday stomatal closure in moderately stressed plants reduced intercellular carbon dioxide concentration (c_i) by ≈40 μl liter^-1. The maximum rate of A achieved during the day for severely stressed plants (predawn water potential = -4 MPa) was one-third and daily carbon gain per unit leaf area was about one-fourth that of well-watered plants. For plants in the natural stand, CO₂-saturated photosynthesis declined almost linearly with decreasing soil water availability over the growing season, whereas there was little effect on A at CO₂ ambient levels or on carboxylation efficiency until predawn water potentials reached -1.8 MPa. Nitrogen-use efficiency declined with diminishing soil moisture, but there was no seasonal change in stomatal limitation or instantaneous water-use efficiency as estimated from A vs. c_i curves at optimal leaf temperature and moderate atmospheric evaporative demand. Thus, reduced stomatal conductance in response to increased evaporative demand may increase stomatal limitation diumally, but over the growing season, stomatal limitation estimated from A vs. c_i curves is relatively constant because maximum stomatal conductance is closely tuned to the CO₂ assimilatory capacity of the mesophyll.     

植物气孔导度的机理模型

Ball-Berry气孔导度模型及其修正模型是评价植物叶片气孔调节的重要工具。该文从CO₂分子在叶片气孔中扩散这个最基本的物理过程出发, 应用物理学中的分子扩散和碰撞理论、流体力学与植物生理学等知识, 严格推导出叶片气孔导度的机理模型。利用美国Li-Cor公司生产的Li-6400光合作用测定仪控制CO₂浓度、湿度和温度, 测量了华北平原冬小麦(Triticum aestivum)的光响应数据和气孔导度数据。拟合结果表明: 推导的气孔导度机理模型较之Ball-Berry气孔导度模型和Tuzet等气孔导度模型, 能更好地描述冬小麦的气孔导度与净光合速率之间的关系。如果用气孔导度的机理模型耦合光合作用对光响应的修正模型, 则耦合模型可以很好地描述华北平原冬小麦叶片气孔导度对光强的响应曲线, 并可直接估算冬小麦的最大气孔导度和对应的饱和光强, 同时可以研究最大气孔导度是否与最大净光合速率同步的问题。拟合结果还表明: 冬小麦在30 ℃、560 μmol·mol^-1CO₂, 或在32 ℃、370 μmol·mol^-1CO₂条件下, 最大气孔导度与最大净光合速率并不同步。     

Seasonal changes in photosynthetic characteristics of Anemone raddeana,a spring-active geophyte,in the temperate region of Japan

Summary Seasonal changes in the photosynthetic characteristics of intact involucral leaves of Anemone raddeana were investigated under laboratory conditions. Net photosynthesis and constant water vapor pressure deficit showed almost the same seasonal trend. They increased rapidly from mid-April immediately after unfolding of the leaves and reached the maximum in late-April, before the maximum expansion of the leaves. They retained the maximum values until early-May and then decreased toward late-May with a progress of leaf senescence. The calculated values of intercellular CO₂ concentration and relative stomatal limitation of photosynthesis showed no significant change throughout the season. The carboxylation efficiency as assessed by the initial slope of Ci-photosynthesis curve and the net photosynthesis under a high Ci regime varied seasonally in parallel with the change of the light-saturated photosynthesis. The results indicate that the seasonal changes in light-saturated net photosynthesis are not due to a change of stomatal conductance, but to a change in the photosynthetic capacity of mesophyll. Nevertheless, leaf conductance changed concomitantly with photosynthetic capacity, indicating that the seasonal change in stomatal conductance is modulated by the mesophyll photosynthetic capacity such that the intercellular CO₂ concentrations is maintained constant. The shape of light-photosynthesis curve was similar to that of sun-leaf type. The quantum yield also changed simultaneously with the photosynthetic capacity throughout the season.Contribution No. 2965 from the Institute of Low Temperature Science     

Ca2+缓解 NaCl胁迫引起的玉米光合能力下降的作用

 以‘鲁玉14号玉米’（‘Zea mays cv. Luyu 14’）为材料,研究了Ca2+ 对NaCl胁迫下玉米叶片的气孔导度、净光合速率、Mehler反应及其对超氧化物歧化酶SOD和抗坏血酸过氧化物酶APX活性的影响,探讨了Ca2+在盐胁迫中的可能作用及其作用机制。正常生长条件下外施 2～16 mmol·L－1 Ca2+造成气孔导度下降,但在盐胁迫条件下外施 2～8 mmol·L－1 Ca2+却能降低盐胁迫造成的气孔导度下降。另外,无论正常生长条件下还是盐胁迫下,依赖Mehler反应的电子传递都会因 Ca2+的加入而增强,但后者的增强程度较大。对照植株经 2～8 mmol· L－1 Ca2+处理后总电子传递明显降低,而外施 2～8 mmol·L－1 Ca2+ 对盐胁迫植株的总电子传递却有明显的促进作用。8 mmol· L－1 Ca2+显著地提高了盐胁迫下SOD和APX的活性,相比之下,Ca2+对APX活性的促进作用更加显著。Ca2+的加入明显减轻了盐胁迫对玉米的抑制作用,这主要归因于Ca2+降低了盐胁迫造成的气孔关闭,改善了光合作用,并通过促进Mehler反应一方面直接耗散过剩的激发电子避免了电子传递链的过度还原,另一方面建立跨膜的pH梯度刺激依赖叶黄素循环的热耗散来耗散过剩光能,从而缓解了盐胁迫下过剩光能对玉米造成的伤害。而Mehler反应产生的活性氧可以被活性提高的抗氧化酶所清除。     

Compensatory photosynthesis as a response to partial debudding in ezo spruce, <Emphasis Type="Italic">Picea jezoensis</Emphasis> seedlings

The effects of partial debudding on photosynthesis, stomatal conductance and nitrogen contents of 1-year-old needles and newly grown needles were studied in Picea jezoensis (Sieb. et Zucc.). Seventy-five percent of the buds of P.jezoensis seedlings were removed soon after bud break. Gas exchange was measured three times for 1-year-old needles and twice for current-year needles. The photosynthetic rates of 1-year-old needles were greater in debudded seedlings than in control seedlings, and the difference increased as the growing season progressed. This greater photosynthetic rate in debudded seedlings was accompanied by greater stomatal conductance. However, neither the photosynthetic rates nor the stomatal conductance of current-year needles differed between debudded and control seedlings after the needles had fully expanded. Debudding also had no effect on mass-based nitrogen contents in either the 1-year-old or the current-year needles. Area-based nitrogen in the 1-year-old needles did not differ between debudded and control seedlings, but was greater in debudded seedlings than in control seedlings in current-year needles. These results suggest that the enhanced photosynthetic rate is more likely a result of an increased root/leaf ratio that reduces the stomatal limitation of photosynthetic rate than a result of altered sink-source relationships or increased leaf nitrogen content.