两个品种烟草叶片发育过程中几种光合参数变化的比较

比较烟草2个品种‘NC89’和‘JYH’叶片发育过程中几个光合参数变化的结果表明,烟草叶片发育过程中光合速率变化表现为上升期、高值持续期（APD）和速降期,叶绿素含量变化经历上升期、相对稳定期（RSP）和速降期。光合功能衰退过程中,核酮糖．1,5-二磷酸羧化酶（RuBPCase）活性比电子传递活性下降快。可逆衰退阶段的2个品种类囊体膜多肽组分和‘NC89’的核酮糖-1,5-二磷酸羧化酶加氧酶（Rubisco）大亚基基本上无变化;不可逆衰退阶段的2个品种类囊体膜多肽组分、Rubisco大小亚基均快速降解,尤其是光系统Ⅱ（PSⅡ）复合体和Rubisco小亚基。‘JYH’的叶龄为10-40d的叶中各光合参数与‘NC89’的差别不大,但‘JYH’的光合功能期短,光合功能衰退过程中光合电子传递与碳同化失衡较严重,光合功能衰退比‘NC89’早而迅速。     

Short-term damage-induced increases in tobacco alkaloids protect plants

Summary Leaf damage significantly increases the alkaloid content in undamaged leaves on damaged field-grown wild tobacco plants. Although field-grown pot-bound plants fail to exhibit the same damage-induced increase in alkaloid content, the ability to respond to leaf damage is restored 6 days after removing plants from their pots. Freshly hatched Manduca sexta larvae reared individually in the laboratory on the high-alkaloid foliage of damaged plants released from their pots gain less weight and eat less (57.2% and 45.7% of controls, respectively) than larvae fed low-alkaloid foliage from undamaged released plants. Moreover, larvae grow equally well on the foliage of damaged and undamaged pot-bound plants. The higher chlorophyll contents characteristic of damaged released plants did not negate the effects of the increased alkaloid contents on larval growth. Undamaged leaves from undamaged field-grown plants stem-fed nicotine solutions had elevated leaf nicotine and nornicotine contents. Larvae reared on these artificially induced leaves gain only 38.5% of the weight gained by larvae reared on low-alkaloid foliage. These results demonstrate that damage-induced increases in leaf alkaloids protect induced foliage from attack and are sufficient to explain the decreased growth of caterpillars on the foliage of damaged plants.     

The Only African Wild Tobacco,Nicotiana africana: Alkaloid Content and the Effect of Herbivory

Herbivory in some Nicotiana species is known to induce alkaloid production. This study examined herbivore-induced defenses in the nornicotine-rich African tobacco N. africana, the only Nicotiana species indigenous to Africa. We tested the predictions that: 1) N. africana will have high constitutive levels of leaf, flower and nectar alkaloids; 2) leaf herbivory by the African bollworm Helicoverpa armigera will induce increased alkaloid levels in leaves, flowers and nectar; and 3) increased alkaloid concentrations in herbivore-damaged plants will negatively affect larval growth. We grew N. africana in large pots in a greenhouse and exposed flowering plants to densities of one, three and six fourth-instar larvae of H. armigera, for four days. Leaves, flowers and nectar were analyzed for nicotine, nornicotine and anabasine. The principal leaf alkaloid was nornicotine (mean: 28 µg/g dry mass) followed by anabasine (4.9 µg/g) and nicotine (0.6 µg/g). Nornicotine was found in low quantities in the flowers, but no nicotine or anabasine were recorded. The nectar contained none of the alkaloids measured. Larval growth was reduced when leaves of flowering plants were exposed to six larvae. As predicted by the optimal defense theory, herbivory had a localized effect and caused an increase in nornicotine concentrations in both undamaged top leaves of herbivore damaged plants and herbivore damaged leaves exposed to one and three larvae. The nicotine concentration increased in damaged compared to undamaged middle leaves. The nornicotine concentration was lower in damaged leaves of plants exposed to six compared to three larvae, suggesting that N. africana rather invests in new growth as opposed to protecting older leaves under severe attack. The results indicate that the nornicotine-rich N. africana will be unattractive to herbivores and more so when damaged, but that potential pollinators will be unaffected because the nectar remains alkaloid-free even after herbivory.     

湖北恩施烟夜蛾幼虫田间分布格局的地统计学分析

烟夜蛾以幼虫取食烟叶,给烟叶生产带来巨大损失.运用地统计学的方法,研究了湖北恩施烟区烟夜蛾幼虫的空间结构和发生动态.结果表明： 恩施烟区烟夜蛾幼虫在烟叶移栽后40 d开始为害,至烟叶成熟前期达到高峰;球形 指数套合模型为烟夜蛾幼虫田间分布的主要模型,属聚集〖JP2〗分布;空间变异度C/(C₀+C)均大于0.75,说明其结构性变异较大,随机性变异较小;烟夜蛾幼虫在烟叶旺长期进行了较大迁移,之后其数量趋于稳定,烟叶成熟期尤其稳定.     

Variation of Alkaloid Content in Erythroxylum coca Leaves from Leaf Bud to Leaf Drop

A sequential study describing the content (%) of alkaloids inleaves of Erythroxylum coca var. coca Lam. from leaf bud developmentto leaf drop has not previously been conducted. The length oftime the leaf resides on the plant and its concurrent metabolicactivity also has not been defined. In the present study, cocaine,methyl ecgonine, hygrine, tropinone, trans -cinnamoylcocaine,cis-cinnamoylcocaine, tropacocaine and cuscohygrine were monitoredto determine: (1) their content and patterns of accumulationfrom leaf bud to leaf drop; (2) the time leaves resided on theplant; and (3) whether leaves were metabolically active untilleaf drop. E. coca plants were grown in a controlled environmentfor 37 months. Leaves similar in chronological age were extractedand analysed for alkaloid content by gas chromatography (GC)and gas chromatography/mass spectrometry (GC/MS). Cocaine washighest in 7 d old rolled leaves (0·75%) and declinedto 0·39% at leaf drop. Cocaine, methyl ecgonine, hygrine,tropinone, trans -cinnamoylcocaine, cis-cinnamoylcocaine, cuscohygrineand tropacocaine content in 35 d old (mature) leaves was 0·61,0·59, 0·68, 0·08, 0·31, 0·55,0·52, and 0·05%. respectively. Cocaine, methylecgonine, hygrine, cis -cinnamoylcocaine, and cuscohygrine displayeda gradual decline from week 2 to week 36 of leaf duration. Tropinoneand tropacocaine were the least abundant of the alkaloids monitored.Cis-cinnamoylcocaine content exceeded cocaine at week 12, 16,and weeks 19 to 23 of leaf duration. Trans -cinnamoylcocainewas highest in rolled leaves (week 1) and in expanded leavesafter week 30. The monitored alkaloids appeared to be part ofthe metabolically active pool of the leaf. Leaves remained onthe E. coca plants for 36 weeks.Copyright 1994, 1999 AcademicPress Cocaine, methyl ecgonine, hygrine, tropinone, trans-cinnamoylcocaine, cis-cinnamoylcocaine, cusco-hygrine, tropacocaine, leaf bud, rolled leaves, expanded leaves, alkaloids, patterns, fluctuation, Erythroxylum coca var. coca, E, coca     

STRUCTURAL CHANGES IN POPULUS DELTOIDES TERMINAL BUDS AND IN THE VASCULAR TRANSITION ZONE OF THE STEMS DURING DORMANCY INDUCTION

Morphological and anatomical changes in shoots of vigorously growing cottonwood plants (Populus deltoides Bartr.) were studied during dormancy induction in 8-hr short days (SD) and in control plants grown in 18-hr long days (LD). Pronounced structural changes occurred in terminal buds after 4 wk and full dormancy was achieved in 7 wk of SD. Leaf expansion ceased after 5 wk of SD as foliage leaves matured to the terminal bud base at leaf plastochron index 0 (LPI 0). Within the bud, total leaf length (lamina + petiole) decreased and stipule length increased progressively each week; thus, the ratio total leaf length/stipule length decreased rapidly, especially at the position of incipient bud-scale leaves LPI - 1 and LPI - 2. These bud-scale leaves were fully developed by wk 6 and were derived from enlarged stipules and aborted laminae. The full complement of primordia within the bud at the start of SD eventually matured as foliage leaves and the first bud-scale leaf (LPI - 1) was initiated immediately following transfer to SD. Acropetal advance of the primary-secondary vascular transition zone (TZ) was associated with leaf maturation. However, it did not advance throughout the entire vascular cylinder as in LD, but only in those leaf traces serving mature leaves beneath the terminal bud. In both LD and SD treatments the same linear relationship was maintained between LPI of the TZ and LPI of the most recently matured leaf; both parameters simultaneously increased in LD and decreased in SD. Thus, the relationship between leaf maturation and advance of the TZ was maintained irrespective of environment.     

Crocidolomia pavonana larval foraging: behavior and feeding site preferences on cabbage, Brassica oleracea

Experiments were conducted to determine feeding site preferences of Crocidolomia pavonana (Fabricius) (Lepidoptera: Crambidae) larvae within cabbage plants, Brassica oleracea L. var. capitata cv. Warrior (Brassicaceae), and to determine whether induced plant responses to herbivory affect the behavior of larvae. In the first experiment, intra-plant damage and larval distribution were recorded to account for the spreading pattern of damage and larval feeding behavior on a plant; larvae initially fed on the base of leaves and moved progressively to the bud, leaf tips were avoided. In the second experiment, larval performance (the duration of the first instar, survival to the second instar, and weight of second instars) was assessed when larvae fed on the bud, the base, and the tip of the youngest fully expanded leaf on a plant. Crocidolomia pavonana larvae performed best when they fed on bud leaf tissue and most poorly when they fed on leaf tissue at the base of leaves. In the third experiment, expression of induced resistance was tested on each of the three plant parts using a first-instar bioassay. Negative impacts on larval growth and development were not detected when larvae fed on the bud or base tissue when plants were damaged prior to the assay. However, negative effects were detected in larvae feeding on tip leaf tissue when the base of the leaf was damaged prior to the assay or if the bud tissue was damaged simultaneously with the assay. The findings indicate that resource heterogeneity for C. pavonana within-cabbage plants is determined by both the initial quality of food at a location and by subsequent induced changes as a result of larval feeding; both contribute to the feeding pattern observed in these gregarious larvae.     

Feeding behavior of bollworm and tobacco budworm (Lepidoptera: Noctuidae) larvae in mixed stands of nontransgenic and transgenic cotton expressing an insecticidal protein

Feeding behavior of third-instar bollworm, Helicoverpa zea (Boddie), and tobacco budworm, Heliothis virescens (F.), was observed in pure and mixed stands of nontransgenic and transgenic cotton (BTK), Gossypium hirsutum L., expressing an insecticidal protein CryIA(c) from a bacterium, Bacillus thuringiensis Berliner subsp. kurstaki. Five plant stands composed of BTK and non-BTK plants were evaluated; two pure stands and three mixed stands. Percentage ratios of BTK to non-BTK plants in the stands were 100:0, 75:25, 50:50, 25:75 and 0:100, respectively. In all stands with BTK plants, fewer bollworm and tobacco budworm larvae were found on BTK plants than non-BTK plants 24 h after infestation with third instars. At 48 h, significantly fewer tobacco budworm larvae, but not fewer bollworm larvae, were found on BTK plants. However, the number of larvae of either insect did not increase on non-BTK plants compared with the initial infestation density of three larvae per plant. The number of obacco budworm injured flower buds, and capsules was lower in all plant stands containing BTK plants compared with the pure stand of non-BTK at 48 h after infestation. Higher numbers of larvae on non-BTK plants were possibly the result of larval intoxication, reduced feeding, and increased plant abandonment and death on BTK plants rather than a classical feeding preference. Unexpectedly, the number of flower buds and capsules injured by bollworm and tobacco budworm when averaged per plant for all plants in a stand, differed little among the 75:25, 50:50 and 25:75 plant mixtures. These data suggest that larvae of both species frequently moved among plants, feeding indiscriminately on BTK and non-BTK plants.     

Biochemical strategy of sequestration of pyrrolizidine alkaloids by adults and larvae of chrysomelid leaf beetles

Tracer feeding experiments with (14)C-labeled senecionine and senecionine N-oxide were carried out to identify the biochemical mechanisms of pyrrolizidine alkaloid sequestration in the alkaloid-adapted leaf beetle Oreina cacaliae (Chrysomelidae). The taxonomically closely related mint beetle (Chrysolina coerulans) which in its life history never faces pyrrolizidine alkaloids was chosen as a 'biochemically naive' control. In C. coerulans ingestion of the two tracers resulted in a transient occurrence of low levels of radioactivity in the hemolymph (1-5% of radioactivity fed). With both tracers, up to 90% of the radioactivity recovered from the hemolymph was senecionine. This indicates reduction of the alkaloid N-oxide in the gut. Adults and larvae of O. cacaliae sequester ingested senecionine N-oxide almost unchanged in their bodies (up to 95% of sequestered total radioactivity), whereas the tertiary alkaloid is converted into a polar metabolite (up to 90% of total sequestered radioactivity). This polar metabolite, which accumulates in the hemolymph and body, was identified by LC/MS analysis as an alkaloid glycoside, most likely senecionine O-glucoside. The following mechanism of alkaloid sequestration in O. cacaliae is suggested to have developed during the evolutionary adaptation of O. cacaliae to its alkaloid containing host plant: (i) suppression of the gut specific reduction of the alkaloid N-oxides, (ii) efficient uptake of the alkaloid N-oxides, and (iii) detoxification of the tertiary alkaloids by O-glucosylation. The biochemical mechanisms of sequestration of pyrrolizidine alkaloid N-oxides in Chysomelidae leaf beetles and Lepidoptera are compared with respect to toxicity, safe storage and defensive role of the alkaloids.     

Utilization of Photosynthetically Fixed 14CO2 into Alkaloids in Relation to Primary Metabolites in Developing Leaves of Catharanthus roseus

Partitioning of current photosynthates towards primary metabolites and its simultaneous incorporation in leaf alkaloids was investigated in developing leaves of medicinally important Catharanthus roseus. Of the total ¹⁴CO₂ assimilated, the leaves at positions 1–6 fixed 8, 22, 25, 19, 13, and 8 %, respectively, and stem 3 %. Leaf fresh mass, chlorophyll content, and CO₂ exchange rate increased up to the third leaf. The total alkaloid content was highest in young actively growing leaves, which declined with age. Total ¹⁴C fixed and its content in ethanol soluble fraction increased up to the third leaf and then declined. The ¹⁴C content in primary metabolites such as sugars and organic acids was also highest in the 3^rd leaf. The utilization of ¹⁴C assimilates into alkaloids was maximum in youngest leaf which declined with leaf age. Hence the capacity to synthesize alkaloids was highest in young growing leaves and metabolites from photosynthetic pathway were most efficiently utilized and incorporated into alkaloid biosynthetic pathway by young growing leaves.     

The expression in tobacco plants of Aedes aegypti Trypsin Modulating Oostatic Factor (Aea-TMOF) alters growth and development of the tobacco budworm,Heliothis virescens

The production and characterisation of transgenic tobacco plantsexpressing a precursor of a regulatory peptide from Aedesaegypti (Trypsin Modulating and Oostatic Factor, Aea-TMOF) whichinterferes with the development of tobacco budworm larvae is described. Tobaccoplants were transformed with a synthetic gene containing 6 TMOF units spaced bydibasic residues, Arg-Arg, as potential post-translational cleavage sites.Peptide extracts from transgenic plants had TMOF activity and inhibitedin vitro the biosynthesis of serine proteases. Thisactivity was consistently present in T1 plants and absent in control plants.Tobacco budworm larvae, fed with transgenic leaves showed a reduced growth ratecompared to those fed with control plants. The low rather than acute toxicityofthis low impact gene is discussed in the context of alternative integrated pestmanagement strategies.     

Food preference and performance of the larvae of a specialist herbivore: variation among and within host-plant populations

Specialist herbivores are suggested to be unaffected by or attracted to the defense compounds of their host-plants, and can even prefer higher levels of certain chemicals. Abrostola asclepiadis is a specialist herbivore whose larvae feed on the leaves of Vincetoxicum hirundinaria, which contains toxic alkaloids and is unpalatable to most generalist herbivores. The food choice, leaf consumption and growth of A. asclepiadis larvae were studied to determine whether there is variation among and within host-plant populations in their suitability for this specialist herbivore. There was significant variation in food preference and leaf consumption among host-plant populations, but no differences were found in larval growth and feeding on different host-plant populations. A. asclepiadis larvae preferred host-plant populations with higher alkaloid concentrations, but did not consume more leaf material from plants originating from such populations in a no-choice experiment. There was also some variation in food preference of larvae among host-plant individuals belonging to the same population, suggesting that there was variability in leaf chemistry also within populations. Such variation in larval preference among host-plant genotypes and populations may create potential for coevolutionary dynamics in a spatial mosaic.     

Growth and alkaloid contents in leaves of Tabernaemontana pachysiphon Stapf (Apocynaceae) as influenced by light intensity,water and nutrient supply

The growth of Tabernaemontana pachysiphon (Apocynaceae) plants and the alkaloid content of leaves were investigated in the greenhouse at three levels of nutrient supply under two contrasting water and light regimes. We determined height increment, above-ground biomass production, leaf size, specific leaf weight and the content of the alkaloids apparicine, A2, isovoacangine, tubotaiwine and tubotaiwine-N-oxide. The effects of major controlling factors such as light, water and nutrient supply could be directly correlated with growth and were largely independent of each other. In contrast, leaf-alkaloid contents were influenced by interdependencies among the main factors and individually affected in a synergistic or antagonistic manner which deviated from the effects on growth. The following general trends could be identified with respect to the quantitatively predominant alkaloids apparicine, tubotaiwine and isovoacangine. Increasing nutrient supply had a positive effect on both growth and alkaloid content. Drought increased alkaloid content, but retarded growth. High light intensity lowered alkaloid content but promoted growth. We investigated the relationship between primary production and the production of secondary metabolites with respect to relative and total alkaloid content as well as in relation to the leaves' nitrogen status. Our results showed that under conditions of low nutrient supply, higher proportions of leaf nitrogen were allocated to alkaloids than at moderate or high nutrient supply. Under conditions of drought and low light, all plants allocated almost equal proportions of leaf nitrogen to alkaloids, regardless of fertiliser. Total alkaloid content per plant, however, increased with fertilisation. With respect to the N-allocation strategy, we found no indication of a trade-off between primary production and the production of secondary metabolites in this species. Rather, our results are in accordance with the carbon nutrient balance hypothesis.     

Effect of transgenic plants expressing high levels of a tobacco anionic peroxidase on the toxicity of Anagrapha falcifera nucleopolyhedrovirus to Helicoverpa zea (Lepidoptera: Noctuidae)

Wild type and corresponding transgenic tomato (Lycopersicon esculentum Miller) and two tobacco (Nicotiana spp.) plants that express high levels of a tobacco anionic peroxidase were used to determine what type of interactions occurred between peroxidase altered plant chemistry and the baculovirus Anagrapha falcifera nucleopolyhedrovirus (AfMNPV) for control of neonate corn earworms, Helicoverpa zea (Boddie). Transgenic plants expressed approximately five to 400 times higher peroxidase activity than corresponding tissues of wild type plants. The H. zea larvae typically fed 1.5 times less on transgenic compared with wild type leaf disks. There was only one experiment (of three with tomato leaves) where the larvae that fed on transgenic leaves were less susceptible to the virus based on nonoverlapping 95% confidence intervals for LC50 values. When the exposure dose was corrected for reduced feeding on the transgenic leaf disks, the insecticidal activity of the virus was not significantly different for larvae fed on transgenic versus wild type plants. Eight other experiments (with tomato and two species of tobacco) indicated either no significant effect or enhanced susceptibility (when corrected for feeding rates) to the virus of larvae fed on the transgenic leaves. These results indicate enhanced insect resistance in plants expressing high levels of a specific anionic peroxidase may be compatible with applications of AfMNPV. Potential reasons for this compatibility are discussed.     

RNA interference (RNAi)-induced suppression of nicotine demethylase activity reduces levels of a key carcinogen in cured tobacco leaves

Technologies for reducing the levels of tobacco product constituents that may contribute to unwanted health effects are desired. Target compounds include tobacco-specific nitrosamines (TSNAs), a class of compounds generated through the nitrosation of pyridine alkaloids during the curing and processing of tobacco. Studies have reported the TSNA N '-nitrosonornicotine (NNN) to be carcinogenic in laboratory animals. NNN is formed via the nitrosation of nornicotine, a secondary alkaloid produced through enzymatic N -demethylation of nicotine. Strategies to lower nornicotine levels in tobacco ( Nicotiana tabacum L.) could lead to a corresponding decrease in NNN accumulation in cured leaves. The major nicotine demethylase gene of tobacco has recently been isolated. In this study, a large-scale field trial was conducted to evaluate transgenic lines of burley tobacco carrying an RNA interference (RNAi) construct designed to inhibit the expression of this gene. Selected transgenic lines exhibited a six-fold decrease in nornicotine content relative to untransformed controls. Analysis of cured leaves revealed a commensurate decrease in NNN and total TSNAs. The inhibition of nicotine demethylase activity is an effective means of decreasing significantly the level of a key defined animal carcinogen present in tobacco products.     

Host Plant Quality Factors That Influence the Growth and Development of Oxyops vitiosa, a Biological Control Agent of Melaleuca quinquenervia

The Australian weevil Oxyops vitiosa was released in 1997 in Florida as a biological control agent of Melaleuca quinquenervia. The larvae of this agent are flush-feeders, found only on the growing tips of their host. Knowledge of this restriction to feeding on the growing tips and other nutritional requirements may assist in the establishment and dispersal of this species. Therefore, O. vitiosa survival was assessed when neonates were fed M. quinquenervia leaves from branches that had dormant buds or emerging bud leaves. Additionally, the influence of leaf quality from different sites and within sites was determined by the feeding of neonates emerging bud leaves collected at three sites and from three leaf qualities (poor, intermediate, and high). Within-site leaf qualities were described in the field by leaf color and in the laboratory by percentage dry mass and nitrogen. Larval survival was lowest when fed leaves from branches that had dormant buds. Associated with this low survival were high leaf toughness and percentage dry mass. When larvae were fed emerging bud leaves, most of the variation in larval survival and performance was attributed to differences in within-site plant quality. Generally, the highest-quality leaves had relatively low percentage dry mass and high percentage nitrogen. Larval survival generally decreased when fed the poor-quality leaves, and in one site, the intermediate-quality leaves. Larvae required less time to develop to adults when fed the high-quality leaves. Development time increased in females but not in males when the larvae were fed the poor-quality leaves. Adult biomass of both females and males generally increased when the larvae were fed the high-quality leaves from two of the three sites. The results indicate that the larvae of O. vitiosa are restricted to feeding on flush foliage with low toughness. Additionally, variations in foliar percentage dry mass and nitrogen influence larval survival and performance. This knowledge benefited the development of mass-production nursery sites and the selection of suitable release sites, which facilitated the establishment of this biological control agent.     

Effect of tobacco budworm (Lepidoptera: Noctuidae) infestation level on budworm-resistant and -susceptible varieties of flue-cured tobacco in North Carolina

Field experiments were conducted from 1972 to 1978 and from 1998 to 1999 to evaluate tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae), larval feeding on flue-cured tobacco, Nicotiana tabacum (L.), yield in eastern North Carolina. In the earlier studies, using variety Coker 319, treatment plots were evaluated when either 0 or 100% of plants in a plot were infested with H. virescens larvae. Treatment differences based on actual yield loss (kilograms per hectare) were compared with estimations of yield loss based on leaf consumption and leaf loss. Results indicate actual yield loss when 100% of plants were infested was less than the corresponding estimates of yield loss. In the later experiments, two tobacco budworm-resistant lines, 'CU 263' and 'CU 370', were compared with a commercial susceptible variety, K 326, when 0, 10, 20, or 40% of plants were infested (1998) and 0, 10, 40, 75, or 100% of plants were infested (1999). Although significant increases in leaf equivalents consumed were associated with infestations exceeding the recommended threshold, differences were not detected for yield (kilograms per hectare), quality (dollars per kilogram), and value (dollars per hectare) within each tobacco line. Additionally, there was not a significant correlation between value and infestations level for any of the tobacco lines. These results provide economic support for tolerance of a higher treatment threshold. Although K 326 sustained more leaf equivalent loss than CU 263 and CU 370, the value of K 326 harvested was higher than that of CU 263 and CU 370. To justify use of resistant varieties, the combination of pest pressure and the benefit of host plant resistance must be greater than the capacity of a susceptible variety to produce competitive yields, despite sustaining significantly higher loss.     

Characterization of bright tobaccos by multivariate analysis of C CPMAS NMR spectra

Univariate and multivariate statistics were applied to characterize cured bright tobacco samples on the basis of their ¹³C CPMAS NMR spectra and leaf constituent analysis. NMR spectra were obtained for 55 samples selected from a set of 134 samples of graded bright tobacco leaves from crop year 1999. Historical leaf constituent analyses were available for total alkaloids, reducing sugars, total nitrogen, and insoluble ash. In addition, we applied HPLC to quantify the two abundant plant polyphenols, chlorogenic acid, and rutin. Principal component analysis (PCA) and partial least squares (PLS) of the NMR spectra revealed systematic relationships between groups of samples related to these substances and afforded predictive quantitative models for the analyzed constituents. Analysis of the PLS significant variables showed that leaf polysaccharides, alkaloids, and minerals are major determinants influencing the grading of cured bright tobacco leaves.