CO2 enrichment predisposes foliage of a eucalypt to freezing injury and reduces spring growth

Seedlings of Eucalyptus pauciflora, were grown in open-top chambers fumigated with ambient and elevated [CO₂], and were divided into two populations using 10% light transmittance screens. The aim was to separate the effects of timing of light interception, temperature and [CO₂] on plant growth. The orientation of the screens exposed plants to a similar total irradiance, but incident during either cold mornings (east-facing) or warm afternoons (west-facing). Following the first autumn freezing event elevated CO₂-grown plants had 10 times more necrotic leaf area than ambient CO₂ plants. West-facing plants had significantly greater (25% more) leaf damage and lower photochemical efficiency (F_v/F_m) in comparison with east-facing plants. Following a late spring freezing event east-facing elevated CO₂ plants suffered a greater sustained loss in F_v/F_m than west-facing elevated CO₂- and ambient CO₂-grown plants. Stomatal conductance was lower under elevated CO₂ than ambient CO₂ except during late spring, with the highest leaf temperatures occurring in west-facing plants under elevated CO₂. These higher leaf temperatures apparently interfered with cold acclimation thereby enhancing frost damage and reducing the ability to take advantage of optimal growing conditions under elevated CO₂.     

Multiphoton imaging of freezing and heating effects in plant leaves

Thermally‐induced changes in Arabidopsis thaliana leaves were investigated with a novel cryo microscope by multiphoton, fluorescence lifetime and spectral imaging as well as micro spectroscopy. Samples were excited with fs pulses in the near‐infrared range and cooled/heated in a cryogenic chamber. The results show morphological changes in the chloroplast distribution as well as a shift from chlorophyll to cell‐wall fluorescence with decreasing temperature. At temperatures below –40 °C, also second harmonic generation was observed. The measurements illustrate the suitability of multiphoton imaging to investigate thermally‐induced changes at temperatures used for cryopreservation as well as for basic investigations of thermal effects on plant tissue in general (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

叶绿素含量测定的简化

本文介绍一种避免称重、研磨、冲洗和过滤等繁琐步骤,仅将确定面积的叶片细丝用少量80%丙酮浸泡提取后便可比色计算的简便的测定叶绿素含量的方法。此法省时间、省溶剂,不受叶片含水量变化的干扰,适合于大批量叶片样品的测定,而且便于与以单位叶面积表示的光合速率联系起来分析实验结果。同时,指出一些可能影响测定结果的因素。     

杂交小麦不同发育时期叶绿素蛋白复合体的变化

杂交小麦“９０１”在不同发育时期及不同叶位,叶片色素蛋白复合体含量均高于对照品种陕２２９,尤其表现在“９０１”的旗叶,而陕２２９的倒二叶在籽粒形成后期、灌浆期,略高于“９０１”,倒三叶在籽粒发育后期至成熟期“９０１”的含量要高于陕２２９。叶绿素蛋白复合体分析结果说明,在不同发育时期、不同叶位,“９０１”叶片色素蛋白复合体中含有较高的叶绿素b,尤其表现在籽粒形成后期的旗叶上。２个品种在不同发育时期,不同时叶位之间,色素蛋白复合体组分上没有差异。仅表现在成熟期,陕２２９的倒三叶各色素带含量减少,并且消失１条叶绿素蛋白复合体带,初步认为此带是ＬＨＣＰ的寡聚体之一,而“９０１”比陕２２９晚７d倒二叶上表现出样的现象,表明“９０１”叶片色素蛋白复合体抗衰老能力很强。     

不同甘蔗品种叶片气孔对水分胁迫的响应

干旱是甘蔗面临最主要的环境胁迫之一,为了解不同甘蔗品种在干旱胁迫时的气孔响应,该研究以F172、GT21、YT93/159和 YL6四个抗旱性有显著差异的甘蔗品种为材料,采用桶栽,在伸长期进行四种不同程度的干旱胁迫(不浇水)处理：土壤持水量在①65％~70％为轻度干旱;②45％~50％为中度干旱;③25％~30％为重度干旱;④以土壤含水量为75％为对照(CK).检测不同品种不同处理甘蔗的叶片相对持水量变化,并利用扫描电镜技术观察甘蔗叶片下表皮气孔特性.结果表明：在干旱胁迫下,四个甘蔗品种叶片气孔导度急剧下降,重度干旱时耐旱性强的 F172和 GT21的气孔导度低于耐旱性弱的 YT93/159和 YL6的;复水后3 d,F172和 GT21的气孔导度上升至82．07和88．85 mmol·m-2·s-1,而 YT93/159和 YL6的仅有18．88和33．08 mmol·m-2·s-1.干旱还导致气孔下陷、闭合,气孔器的长、宽明显减小,且品种间气孔器长度变化差异显著;干旱胁迫下气孔密度增大,尤以耐旱性最强的 F172在重度干旱时达到显著差异.重度干旱时 F172与GT21的气孔闭合百分比是 YT93/159和 YL6近3~4倍.在水分胁迫下,叶片相对含水量降低,但 F172和GT21在重度干旱时仍可以保持相对较高的含水量,其它两个品种相对较低,尤以 YT93/159的最低.在复水后叶片含水量都有所恢复.这些研究结果表明不同甘蔗品种抗旱能力与叶片气孔特性和含水量密切相关.     

土壤水分胁迫对不同耐旱性春小麦品种叶片色素含量的影响

研究选用耐旱性强的“定西24”和“高原671”与耐旱性弱的“高原448”和“青春533”春小麦品种,在盆栽条件下比较了不同耐旱性品种在拔节后经不同程度水分胁迫后叶片的多种生理参数：叶片相对含水量、净光合速率、叶绿素a、叶绿素b、叶绿素总量和类胡萝卜素含量。结果表明：在土壤相对含水量75％、55％、40％和30％这4个等级上,这些生理指标均随土壤相对含水量的降低而下降,耐旱性强的品种降低慢,且降低幅度小。在水分胁迫下,4个品种叶片的叶绿素a、叶绿素b和叶绿素总量下降,证明此时光合器官的生理功能遭到破坏;在轻度水分胁迫下,耐旱性强的品种定西24和高原671叶片的类胡萝卜素含量上升;在严重水分胁迫下,4个品种的叶片类胡萝卜素含量明显下降,可能此时自由基积累超过了抗氧化剂的清除能力,从而使细胞受到损伤。根据本研究结果,可以将轻度水分胁迫下叶片类胡萝卜素含量升高作为春小麦品种耐旱性的选择指标之一。     

Regulation of phytochrome mRNA abundance in green oat leaves

Abstract. Avena sativa L. (oat) seedings were grown 4 d in continuous white light followed by 3 d in darkness. Probes derived from an oat phytochrome cDNA clone (pAP 3.2) were used in slot blot analyses to measure the abundance of phytochrome mRNA in the distinct etiolated and green portions of the leaves produced by these seedlings. Both the green and etiolated portions accumulated phytochrome mRNA to a level of about 85% of the etiolated seedling level. Subsequent experiments with similar seedlings showed that both the green and etiolated portions were capable of inducing a dramatic decline in phytochrome mRNA abundance in response to a saturating red light pulse. Despite the ability of green portions of oat leaves to accumulate phytochrome mRNA and to down-regulate phytochrome mRNA abundance in response to light, no substantial variation in phytochrome mRNA abundance was observed in green oat seedlings maintained on a 12-h day/12-h night cycle. In the same oat seedlings, the abundance of chlorophyll a/b binding protein mRNA fluctuated dramatically during the day/night cycle.     

Inhibition of photosynthesis in Nicotiana tabacum leaves treated with tabtoxin and its relation to pigment loss

Tabtoxin is the chlorosis-inducing dipeptide toxin produced by Pseudomonas syringae pv. tabaci , causal agent of the wildfire disease of Nicotiana tabacum L. cv. White Burley. In plant tissue it is hydrolysed to yield the amino acid tabtoxinine β-lactam, which inactivates glutamine synthetase (EC 6.3.1.2). This study sought to identify those physiological alterations caused by the inactivation of glutamine synthetase and which were involved in the development of the chlorotic symptom. In tobacco ( Nicotiana tabacum L. cv. White Burley) leaves injected with concentrations of taboxin that caused the inactivation of glutamine synthetase, photosynthesis was rapidly inhibited when the tissue was illuminated. This was closely followed by losses in leaf pigments, particularly chlorophyll a , and the ratio of chlorophyll a/b declined, indicating photochemical destruction. The rapid inhibition of photosynthesis in green tissue was due to reduced quantum efficiency, indicating a lesion in the light reactions. In chlorotic tissue, photosynthesis was limited by the chlorophyll level. The results indicated that the inhibition of photosynthesis may be instrumental in development of the chlorotic symptoms.     

Changes in the photosynthetic pigments in bean leaves during the first photoperiod of greening and the subsequent dark-phase. Comparison between old (10-d-old) leaves and young (2-d-old) leaves

Chlorophyll and carotenoid variations of 2-d-old and 10-d-old bean leaves (Phaseolus vulgaris var Red Kidney) were analyzed by HPLC during the first photoperiod of greening (16 h light + 8 h dark). The HPLC method used is suitable for the separation of cis- and trans-carotenoid isomers, Pchlide a and Chlide a as well as their esters. The main results are (1) before illumination the composition of the carotenoid pool is similar at the two developmental stages; (2) non-illuminated 2-d-old leaves are devoid of Pchlide a ester; (3) chlorophyll and carotenoid accumulation in 2-d-old leaves presented a lag phase twice longer than observed in 10-d-old ones; (4) Chlide a seems directly esterified to Chl a in 2-d-old leaves whereas esterification requires four steps in 10-d-old leaves and, (5) the kinetics of Chl and carotenoid accumulation are different at the two investigated developmental stages.     

Overexpression of chlorophyllide a oxygenase (CAO) enlarges the antenna size of photosystem II in Arabidopsis thaliana

The light-harvesting efficiency of a photosystem is thought to be largely dependent on its photosynthetic antenna size. It has been suggested that antenna size is controlled by the biosynthesis of chlorophyll b. To verify this hypothesis, we overexpressed the enzyme for chlorophyll b biosynthesis, chlorophyllide a oxygenase (CAO), in Arabidopsis thaliana by transforming the plant with cDNA for CAO under the control of the 35S cauliflower mosaic virus promoter. In the early de-etiolation phase, when the intrinsic CAO expression is very low, the chlorophyll a: b ratio was drastically decreased from 28 to 7.3, indicating that enhancement of chlorophyll b biosynthesis had been successfully achieved. We made the following observations in full-green rosette leaves of transgenic plants. (1) The chlorophyll a : b ratio was reduced from 2.85 to 2.65. (2) The ratio of the peripheral light-harvesting complexes (LHCII) to the core antenna complex (CPa) resolved with the green-gel system increased by 20%. (3) The ratio of the light-harvesting complex II apoproteins (LHCP) to 47-kDa chlorophyll a protein (CP47), which was estimated by the results of immunoblotting, increased by 40%. These results indicated that the antenna size increased by at least 10-20% in transgenic plants, suggesting that chlorophyll b biosynthesis controls antenna size. To the best of our knowledge, this is the first report on enlargement of the antenna size by genetic manipulations.     

Contrasting photo- and thermoperiod-induced changes in abscisic acid and lipid contents in leaves of mungbean seedlings

Seedlings of Phaseolus aureus ROXB were grown under 12/12 h light/dark cycles with the light period at 32.5°C and darkness at 10°C (normal conditions N) or with light at 10°C and darkness at 32.5°C (inverse conditions, I). I-conditions affected the level of chlorophyll and carotenoids (very low), monogalactosyldiacylgycerol (low) and phosphatidylinositol (high) in the leaves. Leaves of I-seedlings showed a sharp and durable decline of relative water content during the low temperature phase. For the N-seedlings, loss of water was restricted to the end of this period. The loss of water was accompanied by visible symptoms of wilting at specific times of day. Although the pigment content remained nearly unchanged, ABA content of leaves of both N-and I-seedlings increased during water stress. Upon return to the warm period, ABA level continued to increase after the leaves had regained turgor, this 'after stress'increase being more pronounced in the leaves of I-seedlings. Exogenous application of ABA induced a slight increase in the content of phospholipids in N- and I-leaves and a decrease in free fatty acids, whereas monogalactosyldiacylglycerol content was significantly reduced in N-leaves after application of ABA. Upon transfer of I-plants to 20°C for 12 h during the light period, pigment and chloroplastic lipid content increased rapidly whereas upon a further exposure to 10°C in light, pigments and especially monogalactosyldiacylglycerol were lost. The control of pigment and lipid metabolism and the role of ABA during chilling stress are discussed.     

On the limits of applicability of spectrophotometric and spectrofluorimetric methods for the determination of chlorophyll a/b ratio

The concentration limits for spectrophotometric and spectrofluorimetric determinations of the chlorophyll (Chl) a/b ratio in barley leaves were studied using 80% acetone extracts at room temperature. The optimum sample absorbances (at 663.2 nm – maximum of the Q_Y) band of Chl a) for the Chl a/b determination were determined. For given spectrometers and sample positions, these absorbances ranged between 0.2 and 1.0 and 0.008–0.1 for the absorption and fluorescence methods, respectively. Precision of the measurements and the distorting effects are discussed. The lower limits of both absorption and fluorescence methods depend on sensitivity of the spectrometers for the Chl b detection. The spectrophotometric determination of Chl a/b ratio at higher Chl concentrations can be distorted by the chlorophyll fluorescence signal. The extent of this distortion depends on sample-detector geometry in any given type of the spectrometer. The effect of inner filter of Chl molecules and the detection instrumental function affect the value of the upper limit for the spectrofluorimetric method. Both methods were applied to estimate the Chl a/b ratio in pigment extracts from greening barley leaves, which are characterized by a low Chl concentration and a high Chl a/b ratio at the beginning of greening process.     

Effect of PEG-6000 imposed drought stress on RNA content,relative water content (RWC), and chlorophyll content in peanut leaves and roots

Drought, one of the environmental stresses, plays crucial role in reduction in plant production on majority of agricultural fields of world, In order to evaluate drought stress on RNA content Relative water content (RWC), and chlorophyll content, Water deficit was induced by Polyethylene glycol (PEG) in peanut (Arachis hypogaea), accession number ICGV 91114. In this current study we evaluate RNA content and Relative water content (RWC) both in leaves and roots and chlorophyll content in leaf. The present study was undertaken with the aim to investigate the effect of water deficit imposed by PEG-6000, 40 old day seedlings were treated with varying concentrations of polyethylene glycol-6000 (PEG-6000; w/v-5%, 10%, 15% & 20%) for 24 h. The results showed that RNA content and Relative water content (RWC) content was significantly reduced in both leaves and roots with increased concentration of PEG, In leaves, a concentration dependent decline in chlorophyll content with increasing concentration of polyethylene glycol-6000 (PEG-6000). Reduction in chlorophyll ‘a’ level was to a greater extent than the chlorophyll ‘b’. Thus, this attributes can be used as screening tool for drought tolerance in peanut.     

干旱胁迫下菊芋各器官生物量及物质分配规律

以红皮和白皮菊芋为研究材料,通过设置不同的灌水梯度,研究了干旱胁迫对菊芋各器官生物量及物质分配规律的影响。试验结果表明,干旱胁迫降低了红皮和白皮菊芋各器官的生物量;其中,50%灌水条件下两种菊芋的总生物量、地上生物量和地下生物量显著高于10%、20%、30%和40%;红皮菊芋根冠比随着灌水量的减少呈现逐渐增加的变化趋势,白皮菊芋呈现"升-降-升"的变化趋势,且在相同水分处理条件下红皮菊芋茎叶比和根冠比均高于白皮;红皮菊芋随着灌水量的降低根系贡献率呈现逐渐增加的变化趋势,叶片贡献率、块茎贡献率、花贡献率、叶柄贡献率、茎秆贡献率均呈现逐渐降低的变化趋势,白皮菊芋块茎贡献率、花贡献率随灌水量的降低呈现逐渐降低的变化趋势,其他各器官的贡献率随灌水量降低没有规律可循。     

Inhibition of degreening in the peel of bananas ripened at tropical temperatures.

Changes in the plastid ultrastructure as revealed by thin-section electron-microscopy, chlorophyll a/b ratio, and the polypeptides of the thylakoid chlorophyll-protein complexes have been examined during the degreening of bananas (Musa AAA Group, Cavendish Subgroup) and plantains (Musa AAB Group, Plantain Subgroup) ripened at 20°C and 35°C. In bananas, where degreening is inhibited at temperatures above 24°C, ripening at the higher temperature results in a retention of thylakoid membranes, a relatively delayed breakdown in chlorophyll b, and a reduced dismantling of pigment-protein complexes. By contrast, in plantains, where degreening is complete within 4 days at both 20°C and 35°C, thylakoid membranes and their associated pigment-protein complexes are lost, and there is a rapid increase in chlorophyll a/b ratios at both ripening temperatures. It is suggested that the retention of thylakoid membranes is an important factor in the failure of Cavendish bananas to degreen when ripened at tropical temperatures, and that the degreening problem may be related to the comparatively high chlorophyll b content of the preclimacteric fruit.     

Relationship between chloroplast structure and O2 evolution rate of leaf discs in plants from different biotopes in South Finland

Abstract The chloroplast ultrastructure, especially the thylakoid organization, the polypeptide composition of the thylakoid membranes and photosynthetic O₂ evolution rate, chlorophyll (Chl) content and Chi a/b ratio were studied in leaves of nine plants growing in contrasting biotopes in the wild in South Finland. All the measurements were made at the beginning of the period of main growth on leaves approaching full expansion, when the CO₂-saturated O₂ evolution rate (measured at 20°C and 1500 μmol photons m^?2s^?1) was at a maximum, ranging from 19.2 to 6.9 μmol O₂ cm^?2 h^?1. Among the species, the Chi a/b ratio varied between 3.75 and 2.71. In the mesophyll chloroplasts, the ratio of the total length of appressed to non-appressed thylakoid membranes varied between 1.07 and 1.79, the number of partitions per granum varied between 2.8 and 12.0 and the grana area between 21 and 42% of the chloroplast area. There was a significant relationship between the rate of O₂ evolution of the leaf discs and the thylakoid organization in the mesophyll chloroplasts. The higher the O₂ evolution rate, the lower was the ratio of the total length of appressed to non-appressed thylakoid membranes and also the lower the grana area. Although the relationship of the photosynthetic rate with the Chi content and the Chi a/b ratio of the leaves was not as clear, a significant negative correlation existed between the Chi a/b ratio and the ratio of appressed to non-appressed thylakoid membranes, indicating lateral heterogeneity in the distribution of different Chl- protein complexes.     

海盐对绿竹叶片反射光谱及叶绿素荧光参数的影响

沿海防护林是海岸生态系统中最基本的生物资源,由于沿海地区地下水位高,土壤含盐量高,肥力低,生态环境恶劣等特点,所以优良植物种选择是加快沿海优良防护林体系建设的关键。为了探讨绿竹耐盐性以及为沿海地区防护林选择优良植物种,以2年生绿竹(Bambusa oldhamii)为材料,采用水培法进行不同浓度的海盐处理,利用Unispec-SC型单通道光纤光谱仪和非调制式叶绿素荧光仪对绿竹叶片反射光谱和叶绿素荧光参数等进行测定。结果表明:当海盐浓度小于1.2%时,绿竹叶片中叶绿素a、叶绿素b和类胡萝卜素含量,以及"三边"参数与对照无显著差异,当海盐浓度升高到1.6%时,色素与对照相比分别降低了63.2%、62.8%和47.2%(P0.01),红边位置(λr ed)和红边面积(Sred)相比对照显著减小(P0.05)。1.2%浓度的海盐处理,红边归一化指数(rNDVI)、绿度归一化指数(gNDVI)、类胡萝卜素反射指数2(CRI700)和光化学反射指数(PRI)显著降低(P0.05),与对照相比分别降低了27.3%、23.3%、19.5%和43.9%;当海盐浓度增加到1.6%时,rNDVI、改良红边比值指数(mND)、gNDVI、改良归一化差值指数(mSR700)、类胡萝卜素反射指数1(CRI550)、CRI700和PRI等参数与对照相比分别下降了42.4%、43.9%、32.6%、21.5%、47.2%、49.9%和58.5%。绿竹叶片PSⅡ最大量子产率(Fv/Fm)、电子传递的量子产额(ΦEo)、单位反应中心捕获的用于电子传递的能量(ETo/RC)、单位面积反应中心数目(RC/CS)和叶片性能指数(PIABS)等参数,1.6%海盐处理与对照相比分别降低了50.8%、28.6%、21.7%、52.1%和92.3%,单位反应中心复合体吸收的能量(ABS/RC)比对照提高了96.9%。说明绿竹具有一定的耐盐性。