The accumulation of accessory pigments as a function of chlorophyll a. A comparison of development and genetic control

A comparison is made between pigment accumulation in pigment-deficient genotypes of soybean ( Glycine max ) at any one sampling time (genotype control) and pigment accumulation for any one genotype as a function of development (development control). Plots of accessory chloroplast pigments against chlorophyl a show various linear and non-linear relationships that are similar whether generated by genetic differences or by the development process. These relationships are related to the formation and development of pigment-protein complexes especially the relationship of carotenoids to these complexes.     

Assessing leaf pigment content and activity with a reflectometer

This study explored reflectance indices sampled with a 'leaf reflectometer' as measures of pigment content for leaves of contrasting light history, developmental stage and functional type (herbaceous annual versus sclerophyllous evergreen). We employed three reflectance indices: a modified normalized difference vegetation index (NDVI), an index of chlorophyll content; the red/green reflectance ratio ( R _RED: R _GREEN), an index of anthocyanin content; and the change in photochemical reflectance index upon dark–light conversions (ΔPRI), an index of xanthophyll cycle pigment activity. In Helianthus annuus (sunflower), xanthophyll cycle pigment amounts were linearly related to growth light environment; leaves in full sun contained approximately twice the amount of xanthophyll cycle pigments as leaves in deep shade, and at midday a larger proportion of these pigments were in the photoprotective, de-epoxidized forms relative to shade leaves. Reflectance indices also revealed contrasting patterns of pigment development in leaves of contrasting structural types (annual versus evergreen). In H. annuus sun leaves, there was a remarkably rapid increase in amounts of both chlorophyll and xanthophyll cycle pigments along a leaf developmental sequence. This pattern contrasted with that of Quercus agrifolia (coast live oak, a sclerophyllous evergreen), which exhibited a gradual development of both chlorophyll and xanthophyll cycle pigments along with a pronounced peak of anthocyanin pigment content in newly expanding leaves. These temporal patterns of pigment development in Q. agrifolia leaves suggest that anthocyanins and xanthophyll cycle pigments serve complementary photoprotective roles during early leaf development. The results illustrate the use of reflectance indices for distinguishing divergent patterns of pigment activity in leaves of contrasting light history and functional type.     

Spectral reflectance indices and pigment functions during leaf ontogenesis in six subtropical landscape plants

Pigment combinations are regulated during leaf ontogenesis. To better understand pigment function, alterations in chlorophyll, carotenoid and anthocyanin concentrations were investigated during different leaf development stages in six subtropical landscape plants, namely Ixora chinensis Lam, Camellia japonica Linn, Eugenia oleina Wight, Mangifera indica L., Osmanthus fragrans Lowr and Saraca dives Pierre. High concentrations of anthocyanin were associated with reduced chlorophyll in juvenile leaves. As leaves developed, the photosynthetic pigments (chlorophyll and carotenoid) of all six species increased while anthocyanin concentration declined. Chlorophyll fluorescence imaging of Φ_PSII (effective quantum yield of PSII) and of NPQ (non-photochemical fluorescence quenching) and determination of electron transport rate-rapid light curve (RLC) showed that maximum ETR (leaf electron transport rate), Φ_PSII and the saturation point in RLC increased during leaf development but declined as they aged. Juvenile leaves displayed higher values of NPQ and Car/Chl ratios than leaves at other developmental stages. Leaf reflectance spectra (400–800 nm) were measured to provide an in vivo non-destructive assessment of pigments in leaves during ontogenesis. Four reflectance indices, related to pigment characters, were compared with data obtained quantitatively from biochemical analysis. The results showed that the ARI (anthocyanin reflectance index) was linearly correlated to anthocyanin concentration in juvenile leaves, while a positive correlation of Chl NDI (chlorophyll normalized difference vegetation index) to chlorophyll a concentration was species dependent. Photosynthetic reflectance index was not closely related to Car/Chl ratio, while a structural-independent pigment index was not greatly altered by leaf development or species. Accordingly, it is suggested that the high concentration of anthocyanin, higher NPQ and Car/Chl ratio in juvenile leaves are important functional responses to cope with high radiation when the photosynthetic apparatus is not fully developed. Another two leaf reflectance indices, ARI and Chl NDI, are valuable for in vivo pigment evaluation during leaf development.     

Influence of Bemisia argentifolii (Homoptera: Aleyrodidae) infestation and squash silverleaf disorder on zucchini seedling growth

We investigated the effect of different levels of infestation by whiteflies, Bemisia argentifolii Bellows & Perring, on the growth and pigment concentrations of seedlings of zucchini, Cucurbita pepo L., that differed in their tolerance to squash silverleaf disorder. Genetically similar sister lines that were either tolerant (ZUC76-SLR) or susceptible (ZUC61) to silverleaf disorder exhibited reduced plant height, internode length, plant dry weight, and petiole length in response to whitefly feeding. Similar plant growth responses to whitefly feeding were observed despite that the foliage of ZUC61 silvered severely, whereas the foliage of ZUC76-SLR showed no silvering in a greenhouse experiment conducted in the spring and showed only minimal silvering in a similar greenhouse experiment conducted in the fall. In plants of both sister lines infested with 50 pairs of whiteflies and their progeny, petioles, but not the leaf blades, of uninfested leaves had reduced chlorophyll content. In another experiment, two different genetic sources of tolerance to silverleaf disorder (ZUC33-SLR/PMR and ZUC76-SLR) and a commercial silverleaf-susceptible zucchini hybrid ('Zucchini Elite') responded similarly to whitefly feeding, except the tolerant genotypes did not exhibit leaf silvering. All genotypes, silverleaf tolerant or not, had reduced dry weight, plant height, and internode length that became more pronounced as whitefly infestation increased. All genotypes had reduced levels of chlorophylls and carotenoids in uninfested young leaf blades and petioles from infested plants. Petioles, however, were more affected by feeding than leaf blades, showing a 66% reduction in chlorophylls a+b and carotenoids at the lowest infestation level (30 pairs of whitefly and their progeny), whereas pigments in leaf blades declined more slowly in response to whitefly feeding density, averaging 14-15% less at the highest infestation level (90 pairs of whitefly and their progeny). We conclude that tolerance to silverleaf disorder does not prevent stunting in zucchini seedlings nor does it protect against the systemic loss of photosynthetic and protoprotectant pigments induced by feeding of B. argentifolii whiteflies.     

广西壮族传统利用的食用色素植物

广西壮族自治区位于我国南疆,属亚热带区域,地形地貌复杂多样,植物资源极为丰富,有高等植物约     

Functional plasticity of photosynthetic apparatus and its resistance to photoinhibition in <Emphasis Type="Italic">Plantago media</Emphasis>

Morphological and functional characteristics of Plantago media L. leaves were compared for plants growing at different light regimes on limestone outcrops in Southern Timan (62°45′N, 55°49′E). The plants grown in open areas under exposure to full sunlight had small leaves with low pigment content and high specific leaf weight; these leaves exhibited high photosynthetic capacity and elevated water use efficiency at high irradiance. The maximum photochemical activity of photosystem II (F _v/F _m) in leaves of sun plants remained at the level of about 0.8 throughout the day. The photosynthetic apparatus of sun plants was resistant to excess photosynthetically active radiation, mostly due to non-photochemical quenching of chlorophyll fluorescence (qN). This quenching was promoted by elevated deepoxiation of violaxanthin cycle pigments. Accumulation of zeaxanthin, a photoprotective pigment in sun plant leaves was observed already in the morning hours. The plant leaves grown in the shade of dense herbage were significantly larger than the sun leaves, with pigment content 1.5–2.0 times greater than in sun leaves; these leaves had low qN values and did not need extensive deepoxidation of violaxanthin cycle pigments. The data reveal the morphophysiological plasticity of plantain plants in relation to lighting regime. Environmental conditions can facilitate the formation of the ecotype with photosynthetic apparatus resistant to photoinhibition. Owing to this adjustment, hoary plantain plants are capable of surviving in ecotopes with high insolation.     

Plastochron index for detecting juvenility and deciding the components of maturity period in cowpea

Growth of nine field grown cowpea genotypes was characterized using plastochron index. Attainment of the final plastochron index was considered as a point of completion of a vegetative phase. The population under study completed a vegetative phase within 50 d and exhibited flower initiation in 55 d. The gap between completion of the vegetative phase and flower initiation indicated the prevalence of juvenility in cowpea. The maturity period was found to be a sum of the periods for plastochron completion, lag I, pod development and lag II. Lag I and Lag-II periods existed due to variation in the relative sensitivity of successively formed leaves to the normal inductive conditions. A high genotypic variation for the two lag periods indicated a scope for designing an efficient plant architecture of cowpea either for grain or for vegetable purpose.     

Growth and photosynthetic pigments responses of two varieties of Catharanthus roseus to triadimefon treatment

Triadimefon, potential fungicide cum plant-growth retardant was used in this study to investigate its effect on the growth and the photosynthetic pigment contents of two varieties of Catharanthus roseus (L.) G. Don. The plants of both varieties were subjected to 15 mg l(-1) triadimefon treatment by soil drenching 30, 45, 60, and 75 days after planting (DAP). Plants were uprooted on 90 DAP, and morphological parameters, like plant height, number of leaves, leaf area, root length and fresh and dry weights were determined. The photosynthetic pigments, like chlorophylls a and b, total chlorophyll, carotenoids, floral pigment, anthocyanin, were extracted and estimated. It was observed that plant height, number of leaves and leaf area were decreased and that root length, fresh and dry weights were increased under triadimefon treatment. The photosynthetic and floral pigments were increased under triadimefon treatment in both varieties. The results suggest that the application of this plant-growth retardant (triadimefon) has favourable effects on the reduction of plant height; it can thus be used for replacing manual hand pruning and for improving floral and vegetation colour in bedding plants like C. roseus.     

Possible involvement of red pigments in defense against mercury in Pseudomonas K-62

Abstract To study the physiological role of the red pigments in soil strain Pseudomonas K-62, we isolated a red pigment-deficient white mutant from the soil strain by treatment with mitomycin C and compared the phenotypic properties of the mutant and parent strain. The red pigments, which were classified as one of carotenoids based on their physicochemical properties, were separated into two groups, designated pigment A and B respectively on NH-Chromatorex HPLC.The crude pigments and pigment B which could react with Hg²⁺ in the wild-type Pseudomonas K-62 and its mercury-resistant plasmid-deficient strain were enhanced by the addition of Hg²⁺. The white mutant thus obtained showed a greater sensitivity to Hg²⁺ than the wild-type reddish strain despite containing the resistant plasmids. The major component in pigment B was identified by mass spectrometric analysis as 1-hydroxy-1-methoxy-1,2, 1',2',7',8'-hexahydro-ψ,ψ-caroten-4-one, a carotenoid monoketone. These results suggested that red pigments, especially pigment B, may account, at least partially, for defense against Hg²⁺ in the bacterial environments.     

Relationships between the photochemical reflectance index (PRI) and chlorophyll fluorescence parameters and plant pigment indices at different leaf growth stages

This study aimed to evaluate the photochemical reflectance index (PRI) for assessing plant photosynthetic performance throughout the plant life cycle. The relationships between PRI, chlorophyll fluorescence parameters, and leaf pigment indices in Solanum melongena L. (aubergine; eggplant) were studied using photosynthetic induction curves both in short-term (diurnal) and long-term (seasonal) periods under different light intensities. We found good correlations between PRI/non-photochemical quenching (NPQ) and PRI/electron transport rate (ETR) in the short term at the same site of a single leaf but these relationships did not hold throughout the life of the plant. In general, changes in PRI owing to NPQ or ETR variations in the short term were <20?% of those that occurred with leaf aging. Results also showed that PRI was highly correlated to plant pigments, especially chlorophyll indices measured by spectral reflectance. Moreover, relationships of steady-state PRI/ETR and steady-state PRI/photochemical yield of photosystem II (Φ(PSII)) measured at uniform light intensity at different life stages proved that overall photosynthesis capacity and steady-state PRI were better correlated through chlorophyll content than NPQ and xanthophylls. The calibrated PRI index accommodated these pigments effects and gave better correlation with NPQ and ETR than PRI. Further studies of PRI indices based on pigments other than xanthophylls, and studies on PRI mechanisms in different species are recommended.     

Longitudinal leaf gradients of UV-absorbing screening pigments in barley (Hordeum vulgare)

In vivo leaf characteristics were examined to describe longitudinal gradients of UV-absorbing screening pigments in barley. Chlorophyll fluorescence properties and in vivo absorption spectra (210–750 nm) of leaves were measured from the base to the tip. Barley leaves showed strong longitudinal gradients of chlorophyll, where chlorophyll concentration increased within the first 5–8 cm from the leaf base, and did not significantly change for the remaining part of the leaf. Fluorescence microscopy was used to localize cell wall bound screening pigments different from flavonoids, since flavonoids lack a blue-green fluorescence emission (Lichtenthaler and Schweiger 1998). Measurements of in vivo chlorophyll fluorescence indicated that the ratio of UV-absorbing screening pigments per leaf area increases from the leaf base to the tip. These gradients were confirmed by in vivo absorption spectra. It is demonstrated that leaves in the early stage of development are less protected against UV-radiation than fully developed mature leaf regions. The experiments show that measurements of in vivo chlorophyll fluorescence are ideally suited as a fast non-invasive tool to estimate the epidermal UV-transmittance in different leaf sections.     

A novel role of water-soluble chlorophyll proteins in the transitory storage of chorophyllide

All chlorophyll (Chl)-binding proteins involved in photosynthesis of higher plants are hydrophobic membrane proteins integrated into the thylakoids. However, a different category of Chl-binding proteins, the so-called water-soluble Chl proteins (WSCPs), was found in members of the Brassicaceae, Polygonaceae, Chenopodiaceae, and Amaranthaceae families. WSCPs from different plant species bind Chl a and Chl b in different ratios. Some members of the WSCP family are induced after drought and heat stress as well as leaf detachment. It has been proposed that this group of proteins might have a physiological function in the Chl degradation pathway. We demonstrate here that a protein that shared sequence homology to WSCPs accumulated in etiolated barley (Hordeum vulgare) seedlings exposed to light for 2 h. The novel 22-kD protein was attached to the outer envelope of barley etiochloroplasts, and import of the 27-kD precursor was light dependent and induced after feeding the isolated plastids the tetrapyrrole precursor 5-aminolevulinic acid. HPLC analyses and spectroscopic pigment measurements of acetone-extracted pigments showed that the 22-kD protein is complexed with chlorophyllide. We propose a novel role of WSCPs as pigment carriers operating during light-induced chloroplast development.     

Pear plantlets cultured ‘in vitro’ under lime-induced chlorosis display a better adaptive strategy than quince plantlets

In many fruit trees species lime-induced chlorosis causes serious economic damage. In the last few years, the in vitro culture technique has been applied to test rootstocks for susceptibility to iron chlorosis and to study biochemical and molecular aspects of the syndrome. In this study in vitro shoot cultivation of quince rootstocks MA, BA29 and pear cv. ‘Conference’ was used to unravel the relationship between iron deficiency, presence of bicarbonate, and growth development and chloroplast pigment behaviour. Stresses were obtained by using MS medium supplemented with various concentrations of FeNaEDTA and/or KHCO₃. Substrate pH, shoot growth and development parameters, total leaf iron and photosynthetic pigment contents of plantlets were independently determined after 24 days of culture. The medium was acidified by the species under all stress conditions. Iron deficiency and bicarbonate condition led to different growth patterns and modular development among the genotypes. This reflects a different sensitivity and plastic adaptation to the elements of this stress. Iron concentration in leaves and the shoot apex of plantlets decreased in all genotypes. However, chloroplast pigments only decreased in quince plantlets under iron deficiency and bicarbonate condition while they did not change in pear plantlets. Our results demonstrate firstly that a plantlet without a root system, which is cultivated in vitro, is able to sense iron deficiency and bicarbonate enriched conditions and, consequently, activates biochemical and physiological responses. Secondly, acidification appears to be related to iron concentration in tissues of quince rootstocks and pear cultivar. Finally, chloroplast biochemical elements are strongly and differently regulated under both stress conditions and between genotypes.     

Functional role of red (retro)-carotenoids as passive light filters in the leaves of Buxus sempervirens L.: increased protection of photosynthetic tissues?

Red (retro)-carotenoids accumulate in chloroplasts of Buxus sempervirens leaves during the process of winter leaf acclimation. As a result of their irregular presence, different leaf colour phenotypes can be found simultaneously in the same location. Five different colour phenotypes (green, brown, red, orange, and yellow), with a distinct pattern of pigment distribution and concentration, have been characterized. Leaf reddening due to the presence of anthocyanins or carotenoids, is a process frequently observed in plant species under photoinhibitory situations. Two main hypotheses have been proposed to explain the function of such colour change: antioxidative protection exerted by red-coloured molecules, and green light filtering. The potential photoprotective role of red (retro-) carotenoids as light filters was tested in Buxus sempervirens leaves. In shade leaves of this species the upper (adaxial) mesophyll of the lamina was replaced by the equivalent upper part of a different colour phenotype. These hybrid leaves were exposed to a photoinhibitory treatment in order to compare the photoprotective effect exerted by adaxial parts of phenotypes with a different proportion of red (retro)-carotenoids in the lower mesophyll of a shade leaf. The results indicated that the presence of red (retro)-carotenoids in the upper mesophyll did not increase photoprotection of the lower mesophyll when compared with chlorophyll, and the best protection was achieved by an upper green layer. This was due to the fact that the extent of photoinhibition was proportional to the amount of red light transmitted by the upper mesophyll and/or to the chlorophyll pool located above. These results do not exclude a protective function of carotenoids in the upper leaf layer, but imply that, at least under the conditions of this experiment, the accumulation of red pigments in the outer leaf layers does not increase photoprotection in the lower mesophyll.     

Eco-physiological studies on the response of taxodiaceous conifers to shading,with special reference to the behaviour of leaf pigments

The composition of leaf pigments was studied in 32 species of gymnosperms. In 19 species belonging to four families, both evergreen and deciduous leaves turned to reddish brown in late autumn and winter. The red pigment detected in these leaves proved to be rhodoxanthin. The colouring was due to the disappearance of chloroplast pigments and the subsequent synthesis of rhodoxanthin. Neither qualitative nor quantitative differences were recognised in the chloroplast-derived pigments of green leaves between the species with and without the ability to produce rhodoxanthin. Unlike green leaves in which ca. 10% of total carotenoid content was α-carotene, reddish brown leaves scarcely contained α-carotene. Changes in pigment composition during the reddish colouring of leaves and their regreening were studied in detail and discussed.     

不同叶色三叶海棠叶片成色色素分析

结合民族植物学和药理学的研究方法,对西双版纳地区傣族、哈尼族和基诺族等3个少数民族民间利用番石榴（Psidium guajava）、余甘子（Phyllanthus emblica）和水柳（Homonoia riparia）的传统知识进行调查研究及体外抗菌活性实验。结果表明：番石榴和余甘子在村寨中较为常见,当地少数民族将其种植于庭院中,常作为果蔬食用,食用番石榴嫩叶可缓解拉肚子的症状,治疗腹痛、腹泻。水柳生长在水边,傣族会将其叶作为腌酸鱼的配料之一。根据文献记载,番石榴、余甘子和水柳的叶部位作为药使用时,常煎水外洗,治疗皮肤瘙痒。对这3种药用植物叶部位采用80%乙醇浸泡制备的提取物进行体外抗菌实验,结果显示番石榴、余甘子和水柳3种药用植物对金黄色葡萄球菌和大肠埃希菌均有较好的抑菌和杀菌活性,其最小抑菌浓度MIC在98~390 μg·mL 1之间,最小杀菌浓度MBC在98~781 μg·mL 1之间。番石榴和水柳叶对铜绿假单胞菌有一定抑菌和杀菌活性,其MIC和MBC范围均为6 250~12 500 μg·mL 1。由此可见,这3种药用植物的民间利用具有一定的合理性和药用开发价值。     

基于高光谱遥感的小麦冠层叶片色素密度监测

作物叶片色素状况是评价植株光合效率和营养胁迫的重要指标,冠层叶片色素密度（单位土地面积叶片色素总量）的实时无损监测对作物生长诊断、产量估算及氮素管理具有重要意义。以包括不同品质类型（高蛋白、中蛋白和低蛋白）的多个小麦品种在不同施氮水平下的连续2a大田试验为基础,研究了小麦叶片色素密度与冠层高光谱参数的定量关系。结果表明,叶片色素（叶绿素a、叶绿素b、叶绿素a＋b和类胡萝卜素）密度随施氮水平增加而提高,不施氮处理的叶片色素密度随生育进程而下降,施氮处理的叶片色素密度呈单峰曲线,品种间存在明显差异。群体叶片色素密度的敏感波段主要分布在可见光区,而红边区域导数光谱表现更显著。光谱参数VOG2、VOG3、RVI（810,560）、SRE／SBE和SDr／SDb等与叶绿素密度关系较为密切,线性方程决定系数R^2均在0．858以上,而与类胡萝卜素密度关系减弱,决定系数R^2低于0．780,且参数间差异较小。经独立试验资料的检验表明,VOG2、VOG3、SRE／SBE和SDr／SDb对不同色素的估测结果较好,预测相对误差RE低于17．6％,虽然对叶绿素b的准确性稍低。总体上,光谱参数VOG2、VOG3、SRE／SBE和SDr／SDb与小麦群体叶片色素密度关系密切,特别是对叶片叶绿素a和叶绿素a＋b的密度可以进行准确可靠的实时监测。     

Leaf shape variation and herbivore consumption and performance: a case study with Ipomoea hederacea and three generalists

The effect of leaf shape variation on plant-herbivore interactions has primarily been studied from the perspective of host seeking behavior. Yet for leaf shape to affect plant-herbivore coevolution, there must be reciprocal effects of leaf shape variation on herbivore consumption and performance. We investigated whether alternative leaf morphs affected the performance of three generalist insect herbivores by taking advantage of a genetic polymorphism and developmental plasticity in leaf shape in the Ivyleaf morning glory, Ipomoea hederacea. Across four experiments, we found variable support for an effect of leaf shape genotype on insects. For cabbage loopers (Trichoplusia ni) and corn earworms (Helicoverpa zea) we found opposing, non-significant trends: T. ni gained more biomass on lobed genotypes, while H. zea gained more biomass on heart-shaped genotypes. For army beetworms (Spodoptera exigua), the effects of leaf shape genotype differed depending on the age of the plants and photoperiod of growing conditions. Caterpillars feeding on tissue from older plants (95 days) grown under long day photoperiods had significantly greater consumption, dry biomass, and digestive efficiency on lobed genotypes. In contrast, there were no significant differences between heart-shaped and lobed genotypes for caterpillars feeding on tissue from younger plants (50 days) grown under short day photoperiods. For plants grown under short days, we found that S. exigua consumed significantly less leaf area when feeding on mature leaves than juvenile leaves, regardless of leaf shape genotype. Taken together, our results suggest that the effects of leaf shape variation on insect performance are likely to vary between insect species, growth conditions of the plant, and the developmental stage and age of leaves sampled. Handling editor: May Berenbaum.     

Morphophysiological differences in leaves of <Emphasis Type="Italic">Lavoisiera campos</Emphasis>-<Emphasis Type="Italic">portoana</Emphasis> (Melastomataceae) enhance higher drought tolerance in water shortage events

Lavoisiera campos-portoana Barreto (Melastomataceae) has two kinds of leaves, pubescent and glabrous, and branches may have one or both types of leaves at the same moment. The plant is endemic to high altitude rocky fields in Brazil where rainfall is very seasonal. We predicted that these two leaf types are adaptations to different regimes of water availability. In experimental conditions of drought stress, we measured relative water content (RWC), photosynthetic pigments, chlorophyll a fluorescence and osmotic potential, and we counted stomates and measured stomatal conductance on both sides of leaves and compared these between the two leaf types. Stomatal conductance and electron transport rate at a given photosynthetic photon flux were greater in pubescent leaves than in glabrous leaves, and both declined during drought stress. Excessive photon flux density in glabrous leaves was greater during stress and after rehydration. Photosynthetic pigment content and RWC did not change between leaves, and values reduced during the stress period. Both types of leaves showed osmotic adjustment capacity, which occurred earlier in glabrous ones. These morphophysiological differences highlight the adaptation strategies of this plant to withstand drought, since the glabrous portion of the plant presents a preventive behavior, while the pubescent portion only shows the same responses in more advanced stages of drought stress.     

Multivariate analysis of polypeptide synthesis in field-grown maize inbreds and hybrids

Summary A leaf disc method is described to permit the localized incorporation of ³⁵S-l-methionine into polypeptides synthesized in individual leaves of maize plants grown in the field. The method of incorporation employs minimal external manipulation of the intact leaf, is simple, repeatable, and may be used at any plant age after leaf emergence. Incorporation (cpm/g protein) in 12 leaves per plant was compared among three inbred (Oh43, W23, M14) and three F₁ hybrid (Oh43/M14, W23/M14, Oh43/W23) genotypes. The incorporation was 40% higher (hybrid versus inbred) in 9 of the 12 leaves studied. Samples from leaf 07 (7th leaf numbered from base of plant) for four inbreds (Oh43, M14, B73, Mol 7) and two pairs of reciprocal F₁ hybrids (Oh43/M14, M14/Oh43; B73/Mo17, Mo17/B73) were labelled in situ using the leaf disc method. Each cultivar was sampled at three different ages in each of 1985, 1986, and 1987. High-resolution, two-dimensional isoelectric focusing sodium-dodecyl-sulfate polyacrylamide gel electrophoresis and fluorography were used to display the polypeptides synthesized in the samples. Multivariate methods — Principal Coordinate Analysis, Cluster Analysis, and Standard Deviation Distance — were used to analyze variation and to identify trends in the variation for year, genotype, and age sampled. Our analyses disclose a hierarchy to polypeptide synthesis variation in maize leaves: differences in polypeptide synthesis are greater for year-to-year comparisons than differences due to sample age, which in turn are greater than differences for inbred versus hybrid comparisons.