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Elevated pCO(2 )favours nitrate reduction in the roots of wild-type tobacco (Nicotiana tabacum cv. Gat.) and significantly alters N-metabolism in transformants lacking functional nitrate reductase in the roots.

The impact of elevated pCO(2 )on N-metabolism of hydroponically grown wild-type and transformed tobacco plants lacking root nitrate reduction was studied in order to elucidate the effects on (i) nitrate uptake, (ii) long-distance transport of N, (iii) nitrate reduction with emphasis on root-NR, and (iv) the allocation of N between the root and shoot. The findings were related to alterations of growth rates. At elevated pCO(2 )the wild type exhibited higher growth rates, which were accompanied by an increase of NO(3)(-)-uptake per plant, due to a higher root:shoot ratio. Furthermore, elevated pCO(2 )enhanced nitrate reduction in the roots of the wild type, resulting in enhanced xylem-loading of organic N (amino-N) to supply the shoot with sufficient nitrogen, and decreased phloem-transport of organic N in a basipetal direction. Transformed tobacco plants lacking root nitrate reduction were smaller than the wild type and exhibited lower growth rates. Nitrate uptake per plant was decreased in transformed plants as a consequence of an impeded root growth and, thus, a significantly decreased root:shoot ratio. Surprisingly, transformed plants showed an altered allocation of amino-N between the root and the shoot, with an increase of amino-N in the root and a substantial decrease of amino-N in the shoot. In transformed plants, xylem-loading of nitrate was increased and the roots were supplied with organic N via phloem transport. Elevated pCO(2 )increased shoot-NR, but only slightly affected the growth rates of transformed plants, whereas carbohydrates accumulated at elevated pCO(2 )as indicated by a significant increase of the C/N ratio in the leaves of transformed plants. Unexpectedly, the C/N balance and the functional equilibrium between root and shoot growth was disturbed dramatically by the loss of nitrate reduction in the root.

Carbon Dioxide↗

The uptake and partitioning of cadmium in two cultivars of potato (Solanum tuberosum L.).

The uptake and distribution of Cd in potatoes over the course of a growing season was investigated in two cultivars of potatoes that differed in tuber Cd concentration. Plants were grown in soil with supplemental Cd. The concentrations of Cd in different tissues varied greatly in the order roots>shoots>> tubers. After the initiation of tuber bulking, shoot growth ceased and the increase in total plant Cd was mostly due to accumulation in the tubers. The constancy of the Cd concentration in shoots suggested that import of Cd via the xylem must be matched by export in the phloem, which implied that Cd must have significant phloem mobility. It was found that the differences in tuber Cd between cultivars Wilwash and Kennebec were not due to differences in total uptake or growth, but to differences in Cd partitioning within the plant. This partitioning was specific to Cd and was not observed for a range of nutrient elements. Most of the differences in tuber Cd concentration between the cultivars could be accounted for by a 3-fold higher retention of Cd in the roots of cv. Wilwash. The involvement of root sequestration, and xylem and phloem pathways in the loading of Cd into tubers is considered.

Cadmium↗

Water flows in the parasitic association Rhinanthus minor/Hordeum vulgare.

Using the facultative root hemiparasite Rhinanthus minor and its host Hordeum vulgare several aspects of water relations have been measured in this parasitic association. Extraction of xylem sap by the parasite from the host's roots is facilitated by con siderably higher transpiration per leaf area in the parasite than in the host and by the fact that stomata of attached Rhinanthus were open all day and night despite extremely high ABA concentrations in the leaves. By comparison, another root hemiparasite, Melampyrum arvense, parasitizing various grasses in the field, showed normal diurnal stomatal behaviour. The abnormal behaviour of Rhinanthus stomata was not due to anatomical reasons as closure could be induced by applying high external ABA concentrations. Remarkable differences have been detected between the hydraulic conductance of barley seminal roots showing relatively low values and that of Rhinanthus seminal roots showing very high values. The latter could be related to the observed high ABA concentrations in these roots. Whole plant water uptake, transpirational losses, growth-dependent deposition, and the flows of water within the plants have been measured in singly growing Rhinanthus and Hordeum plants and in the parasitic association between the two. Water uptake, deposition and transpiration in Rhinanthus were dramatically increased after attachment to the barley host; most of the water used by the parasite was extracted as xylem sap from the host, thereby scavenging 20% of the total water taken up by the host's roots. This water uptake by the parasitized host, however, due to a parasite-induced reduction in the host's growth, was decreased by 22% as compared to non-parasitized barley. The overall changes in growth-related water deposition in the host and parasite pointed to decreased shoot growth and relatively favoured root growth in the host and to strongly favoured shoot growth in the parasite. These changes in the host became more severe, when more than one Rhinanthus was parasitizing one barley plant.

Abscisic Acid↗

Hydraulic efficiency and safety of leader shoots and twigs in Norway spruce growing at the alpine timberline.

Xylem within trees varies in its hydraulic efficiency and safety. Trees at the alpine timberline were expected to exhibit a hydraulic architecture protecting the leader shoot from winter embolism. Hydraulic and related anatomical parameters were compared as well as seasonal courses of winter embolism in leader shoots and twigs of Norway spruce trees growing at 2000 m. Leader shoots had a 1.4-fold higher specific hydraulic conductivity (ks) as well as a 4.9-fold higher leaf specific conductivity (kl) than side twigs. Vulnerability to drought-induced embolism was lower in leader shoots with a 50% loss of conductivity occurring at a water potential (Psi 50) 0.7 MPa lower than in twigs. Higher ks and kl were related to 1.2-fold wider tracheid diameters in leader shoots. Lower vulnerability corresponded to smaller pit dimensions but not to wood density. High ks and kl reflect the hydraulic dominance of the leader shoot, which is important for its water supply during summer. Low vulnerability protects the leader shoot from embolism during the winter season. In field measurements at the timberline during the winter of 2001/2002, conductivity losses of up to 56% were observed only in twigs while leader shoots showed little or no embolism. Results demonstrate that leader shoot xylem is both hydraulically efficient and safe.

Altitude↗

Interaction between two auxin-resistant mutants and their effects on lateral root formation in rice (Oryza sativa L.).

Since root elongation is very sensitive to auxin, screening for reduced inhibition in root elongation has been an important method for the detection of auxin-resistant mutants. Two recessive auxin-resistant lines of rice (Oryza sativa L. ssp. indica cv. IR8), arm1 and arm2, have been isolated by screening for resistance to 2,4-dichlorophenoxyacetic acid (2,4-D). arm1 displays a variety of morphological defects including reduced lateral root formation, increased seminal root elongation, reduced root diameter, and impaired xylem development in roots, while the arm2 phenotype is almost similar to wild-type IR8 except for a slightly reduced lateral root formation, impaired xylem development in roots and an enhanced plant height. Although the growth of arm2 roots exhibited a resistance to 2,4-D, it was sensitive to 1-naphthaleneacetic acid (NAA) as the wild type. At the same time, the arm2 roots showed a reduced [14C]2,4-D uptake while uptake of [3H]NAA was normal, suggesting that the resistance to 2,4-D of arm2 roots is due to a defect in 2,4-D uptake. To investigate the possible interaction between arm1 and arm2 genes, a double mutant has been constructed. The roots of arm1 arm2 double mutant were more resistant to 2,4-D and formed fewer lateral roots than those of either single mutant, suggesting that the two genes show synergistic effects with respect to both auxin response and lateral root formation. By contrast, all these mutants displayed the normal gravitropic response in roots, as did the wild-type plants. Taken together, Arm1 and Arm2 genes seem to function in different processes in the auxin-response pathways leading to lateral root formation.

2,4-Dichlorophenoxyacetic Acid↗

Rapid and tissue-specific changes in ABA and in growth rate in response to salinity in barley leaves.

The addition of 100 mM NaCl to the root medium of barley plants caused the rapid cessation of elongation of the growing leaf three, followed by a sudden resumption of growth during the following hour. The idea that resumption of growth is preceded and mediated by rapid and tissue-specific changes in ABA concentration and by changes in transpiration was tested. Leaf elongation velocity was recorded continuously using linear variable displacement transducers (LVDT), ABA was determined by immunoassay, and transpiration and stomatal conductivity were measured gravimetrically and by porometry, respectively. Within 10 min following addition of salt, ABA increased 6-fold in the distal portion of the leaf elongation zone; in the proximal portion, ABA accumulated with a delay. In the portion of the growing blade that had emerged ABA increased 3-fold and remained elevated during the following 20 min. This preceded a decrease in transpiration and stomatal conductivity, which, in turn, coincided with growth resumption. Twenty hours following the addition of salt, the ABA concentrations had returned to the level before stress. Leaf elongation velocity was still reduced. It is concluded that NaCl causes a rapid increase in ABA in the transpiring portion of the growing leaf. This leads to a decrease in transpiration. As a result, xylem water potential is expected to rise. The moment that the water potential gradient between the xylem and the peripheral cells in the growth zone favours water uptake again into the latter, leaf elongation resumes. The results suggest that ABA causes different responses in different leaf regions, all aimed at promoting the resumption of leaf growth.

Abscisic Acid↗

Abscisic acid (ABA) flows from Hordeum vulgare to the hemiparasite Rhinanthus minor and the influence of infection on host and parasite abscisic acid relations.

Using the facultative root hemiparasite Rhinanthus minor and Hordeum vulgare as a host, the flows, depositions, and metabolism of abscisic acid (ABA) within the host, within the parasite, and between host and parasite have been studied. When the plants were supplied with 5 mM NO(3)(-), there were weak or no effects of parasitism on ABA flows, biosynthesis, and ABA degradation in barley. However, ABA deposition was significantly affected in the leaf laminae (3-fold) and in the leaf sheath (2.4-fold), but not in roots. Dramatic changes in ABA flows, metabolism, and deposition on a per plant basis, however, have been observed in Rhinanthus. Biosynthesis in the roots was 12-fold higher after attachment, resulting in 14-fold higher ABA flows in the xylem. A large portion of this ABA was metabolized, a small portion was deposited. Phloem flows of ABA were increased 13-fold after attachment. The concentrations of ABA in tissues and transport fluids were higher in attached Rhinanthus by an order of magnitude than in host tissues and xylem sap. The same tendency was also found in a comparison between single Rhinanthus and unparasitized barley. As compared with 5 mM NO(3)(-), lower NO(3)(-) or 1 mM NH(4)(+) supply doubled the ABA concentrations in barley leaf laminae, while having only small or non-significant effects in the other organs. The possible function of ABA for the parasite is discussed.

Abscisic Acid↗

Root-derived cytokinins as long-distance signals for NO3--induced stimulation of leaf growth.

Leaf growth of many plant species shows rapid changes in response to alterations of the form and the level of N supply. In hydroponically-grown tomato (Lycopersicon esculentum L.), leaf growth was rapidly stimulated by NO(3)(-) application to NH(4)(+) precultured plants, while NH(4)(+) supply or complete N deprivation to NO(3)(-) precultured plants resulted in a rapid inhibition of leaf growth. Just 10 microM NO(3)(-) supply was sufficient to stimulate leaf growth to the same extent as 2 mM. Furthermore, continuous NO(3)(-) supply induced an oscillation of leaf growth rate with a 48 h interval. Since changes in NO(3)(-) levels in the xylem exudate and leaves did not correlate with NO(3)(-)-induced alterations of leaf growth rate, additional signals such as phytohormones may be involved. Levels of a known inhibitor of leaf growth, abscisic acid (ABA), did not consistently correspond to leaf growth rates in wild-type plants. Moreover, leaf growth of the ABA-deficient tomato mutant flacca was inhibited by NH(4)(+) without an increase in ABA concentration and was stimulated by NO(3)(-) despite its excessive ethylene production. These findings suggest that neither ABA nor ethylene are directly involved in the effects of N form on leaf growth. However, under all experimental conditions, stimulation of leaf growth by NO(3)(-) was consistently associated with increased concentration of the physiologically active forms of cytokinins, zeatin and zeatin riboside, in the xylem exudate. This indicates a major role for cytokinins as long-distance signals mediating the shoot response to NO(3)(-) perception in roots.

Abscisic Acid↗

GAL4-GFP enhancer trap lines for genetic manipulation of lateral root development in Arabidopsis thaliana.

Lateral root development occurs throughout the life of the plant and is responsible for the plasticity of the root system. In Arabidopsis thaliana, lateral root founder cells originate from pericycle cells adjacent to xylem poles. In order to study the mechanisms of lateral root development, a population of Arabidopsis GAL4-GFP enhancer trap lines were screened and two lines were isolated with GAL4 expression in root xylem-pole pericycle cells (J0121), i.e. in cells competent to become lateral root founder cells, and in young lateral root primordia (J0192). These two enhancer trap lines are very useful tools with which to study the molecular and cellular bases of lateral root development using targeted gene expression. These lines were used for genetic ablation experiments by targeting the expression of a toxin-encoding gene. Moreover, the molecular bases of the enhancer trap expression pattern were characterized. These results suggest that the lateral-root-specific GAL4 expression pattern in J0192 is due to a strong enhancer in the promoter of the LOB-domain protein gene LBD16.

Arabidopsis↗

Mucilage production during the incompatible interaction between Orobanche crenata and Vicia sativa.

Orobanche spp. (broomrapes) are holoparasites lacking in chlorophyll and totally dependent on their host for their supply of nutrients. O. crenata is a severe constraint to legumes cultivation and breeding for resistance remains as one of the best available methods of control. However, little is known about the basis of host resistance to broomrapes. It is a multicomponent event, and resistance based on hampering development and necrosis of broomrape tubercles has been reported. In the present work, the formation of mucilage and occlusion of host xylem vessels associated with the death of O. crenata tubercles were studied histologically. Samples of necrotic O. crenata tubercles established on resistant and susceptible vetch genotypes were collected. The samples were fixed, sectioned and stained using different procedures. The sections were observed at the light microscopy level, either under bright field, epi-fluorescence or confocal laser scanning microscopy. A higher proportion of necrotic tubercles was found on the resistant genotype and this was associated with a higher percentage of occluded vessels. Mucilage is composed mainly by carbohydrates (non-esterified pectins) and the presence of polyphenols was also detected. The mucilage and other substances composed by parasite secretions and host-degraded products was found to block host vessels and obstruct the parasite supply channel, being a quantitative defensive response against O. crenata in vetch, and probably also in other legumes and plants. The presence of foreign substances (i.e. parasite secretions) and host-degraded products (i.e. carbohydrates from cell walls) inside host vessels seems to activate this response and leads to xylem occlusion and further death of established Orobanche tubercles.

Adhesives↗

Intact plant MRI for the study of cell water relations, membrane permeability, cell-to-cell and long distance water transport.

Water content and hydraulic conductivity, including transport within cells, over membranes, cell-to-cell, and long-distance xylem and phloem transport, are strongly affected by plant water stress. By being able to measure these transport processes non-invasely in the intact plant situation in relation to the plant (cell) water balance, it will be possible explicitly or implicitly to examine many aspects of plant function, plant performance, and stress responses. Nuclear magnetic resonance imaging (MRI) techniques are now available that allow studying plant hydraulics on different length scales within intact plants. The information within MRI images can be manipulated in such a way that cell compartment size, water membrane permeability, water cell-to-cell transport, and xylem and phloem flow hydraulics are obtained in addition to anatomical information. These techniques are non-destructive and non-invasive and can be used to study the dynamics of plant water relations and water transport, for example, as a function of environmental (stress) conditions. An overview of NMR and MRI methods to measure such information is presented and hardware solutions for minimal invasive intact plant MRI are discussed.

Biological Transport↗

The expression of an aquaporin promoter from Mesembryanthemum crystallinum in tobacco.

The promoter region of the MipB gene encoding an aquaporin from Mesembryanthemum crystallinum was isolated and used in a transcriptional fusion to control uidA expression in tobacco. The sequence of the promoter was determined for 2 kb upstream of the translation initiation site. Three start sites were utilized with approximately equal frequency, located 176, 170, and 161 bases, respectively, upstream of the translation initiation site. As judged by analysis of GUS expression, promoter MipB retains its specificity in transgenic tobacco. In germinating seedlings, all cells showed GUS expression of different intensities with the strongest signals in root meristems. In older seedlings, GUS staining was observed in rapidly expanding cells--root and apical meristem, and lateral root primordia. In mature plants, strong GUS activity was located to glandular trichomes, subepidermal cells of the stem and petioles, to cells surrounding vascular tissues as well as in xylem parenchyma cells. In immature floral organs, GUS expression was strong in sepals, petals, stamen, and pistil. The intensity declined as they matured. In general, this promoter was active in rapidly expanding cells and cells with high water flux capacity, especially in the xylem parenchyma.

Aquaporins↗

Two azuki bean XTH genes, VaXTH1 and VaXTH2, with similar tissue-specific expression profiles, are differently regulated by auxin.

To gain insight into the functional diversity of the XTH (xyloglucan endotransglucosylase/hydrolase) gene family, we analyzed the expression profiles of two azuki bean genes, VaXTH1 and VaXTH2, which share a striking resemblance in their amino acid sequences. The two XTH genes exhibit essentially similar tissue-specific expression profiles, in that both mRNAs are found predominantly in the phloem fibers of growing internodes. However, their expression profiles are not identical. Whereas VaXTH1 is expressed in xylem cells in the basal part of the internode, little or no expression of VaXTH2 is found in the xylem. Furthermore, they exhibit spatially divergent RNA distribution profiles along the internode, VaXTH1 being expressed nearer to the top of the internode than VaXTH2. This indicates their temporally divergent expression profiles during development of the phloem fiber. Indole-3-acetic acid (IAA) up-regulates both of the mRNA levels. However, this effect of IAA on the VaXTH1 gene is nullified in 0.25 M mannitol, which prevents cell expansion without affecting auxin action per se. In contrast, the IAA-induced up-regulation of the VaXTH2 gene is not affected by mannitol. Furthermore, fusicoccin, which promotes acidification and growth, up-regulates VaXTH1 expression, but not VaXTH2 expression. Thus, the two XTH genes are committed to different steps of the cell wall dynamics in the same cell type at different stages of phloem fiber development, and are regulated by IAA in different ways.

Fabaceae↗

Diurnal regulation of water transport and aquaporin gene expression in maize roots: contribution of PIP2 proteins.

In maize (Zea mays) roots, xylem water transfer supported by root pressure occurs during the day and is less important at night. Diurnal modifications of osmotic pressure gradient between medium and xylem could not explain the oscillation of water flux in young maize roots during the day-night cycle. We observed a high turgor pressure of root cortical cells associated with a high flux. In maize roots, ZmPIP transcripts oscillate during the day-night cycle exhibiting some characteristics of genes regulated by a circadian mechanism. The PIP protein level profile is different from the mRNA pattern. Moreover, ZmPIP1 and ZmPIP2 protein levels are differentially regulated during the light and dark period and in response to continuous darkness suggesting different roles for both classes of PIP. Finally, our results suggest that aquaporins from ZmPIP2 subgroup may contribute to root water transfer by cellular pathway that occurs during the light and the dark period of the day-night cycle.

Amino Acid Sequence↗

Clarification of cinnamoyl co-enzyme A reductase catalysis in monolignol biosynthesis of Aspen.

Cinnamoyl co-enzyme A reductase (CCR), one of the key enzymes involved in the biosynthesis of monolignols, has been thought to catalyze the conversion of several cinnamoyl-CoA esters to their respective cinnamaldehydes. However, it is unclear which cinnamoyl-CoA ester is metabolized for monolignol biosynthesis. A xylem-specific CCR cDNA was cloned from aspen (Populus tremuloides) developing xylem tissue. The recombinant CCR protein was produced through an Escherichia coli expression system and purified to electrophoretic homogeneity. The biochemical properties of CCR were characterized through direct structural corroboration and quantitative analysis of the reaction products using a liquid chromatography-mass spectrometry system. The enzyme kinetics demonstrated that CCR selectively catalyzed the reduction of feruloyl-CoA from a mixture of five cinnamoyl CoA esters. Furthermore, feruloyl-CoA showed a strong competitive inhibition of the CCR catalysis of other cinnamoyl CoA esters. Importantly, when CCR was coupled with caffeoyl-CoA O-methyltransferase (CCoAOMT) to catalyze the substrate caffeoyl-CoA ester, coniferaldehyde was formed, suggesting that CCoAOMT and CCR are neighboring enzymes. However, the in vitro results also revealed that the reactions mediated by these two neighboring enzymes require different pH environments, indicating that compartmentalization is probably needed for CCR and CCoAOMT to function properly in vivo. Eight CCR homologous genes were identified in the P. trichocarpa genome and their expression profiling suggests that they may function differentially.

Acyl Coenzyme A↗

A novel function of abscisic acid in the regulation of rice (Oryza sativa L.) root growth and development.

Plant roots retain developmental plasticity and respond to environmental stresses or exogenous plant growth regulators by undergoing profound morphological and physiological alteration. In this study, we investigated the effects of exogenous ABA on root growth and development in Taichung native 1 (TN1) rice. Exogenous application of 10 microM ABA leads to swelling, root hair formation and initiation of lateral root primodia in the tips of young, seminal rice roots. Cortex cells increased in size and were irregularly shaped. ABA treatment significantly increased 2, 3, 5-triphenyl tetrazolium chloride (TTC) reductase ability in the root tips and the exudation rate of xylem sap. In addition, the K(+) ion content in xylem sap increased nearly 2-fold, but not that of Ca(2+) or Mg(2+). Analysis of proteins expressed in the root tips identified several ABA-induced or -repressed proteins, including actin depolymerization factor (ADF), late embryo abundant protein (LEA), putative steroid membrane-binding protein, ferredoxin thionine reductase and calcium-binding protein. The effects of ABA on root morphogenesis change were Ca(2+) dependent and required the participation of calmodulin and de novo protein synthesis. A model is presented that illustrates how ABA acts through a potential cellular and signal transduction mechanism to induce morphological and physiological changes in rice roots.

Abscisic Acid↗

Calcium regulation of sodium hypersensitivities of sos3 and athkt1 mutants.

T-DNA disruption mutations in the AtHKT1 gene have previously been shown to suppress the salt sensitivity of the sos3 mutant. However, both sos3 and athkt1 single mutants show sodium (Na+) hypersensitivity. In the present study we further analyzed the underlying mechanisms for these non-additive and counteracting Na+ sensitivities by characterizing athkt1-1 sos3 and athkt1-2 sos3 double mutant plants. Unexpectedly, mature double mutant plants grown in soil clearly showed an increased Na+ hypersensitivity compared with wild-type plants when plants were subjected to salinity stress. The salt sensitive phenotype of athkt1 sos3 double mutant plants was similar to that of athkt1 plants, which showed chlorosis in leaves and stems. The Na+ content in xylem sap samples of soil-grown athkt1 sos3 double and athkt1 single mutant plants showed dramatic Na+ overaccumulation in response to salinity stress. Salinity stress analyses using basic minimal nutrient medium and Murashige-Skoog (MS) medium revealed that athkt1 sos3 double mutant plants show a more athkt1 single mutant-like phenotype in the presence of 3 mM external Ca2+, but show a more sos3 single mutant-like phenotype in the presence of 1 mM external Ca2+. Taken together multiple analyses demonstrate that the external Ca2+ concentration strongly impacts the Na+ stress response of athkt1 sos3 double mutants. Furthermore, the presented findings show that SOS3 and AtHKT1 are physiologically distinct major determinants of salinity resistance such that sos3 more strongly causes Na+ overaccumulation in roots, whereas athkt1 causes an increase in Na+ levels in the xylem sap and shoots and a concomitant Na+ reduction in roots.

Adaptation, Physiological↗

The expression of different superoxide dismutase forms is cell-type dependent in olive (Olea europaea L.) leaves.

Superoxide dismutase (SOD) is a key antioxidant enzyme present in prokaryotic and eukaryotic cells as a first line of defense against the accumulation of superoxide radicals. In olive leaves, the SOD enzymatic system was characterized and was found to be comprised of three isozymes, an Mn-SOD, an Fe-SOD and a CuZn-SOD. Transcript expression analysis of whole leaves showed that the three isozymes represented 82, 17 and 0.8% of the total SOD expressed, respectively. Using the combination of laser capture microdissection (LCM) and real-time quantitative reverse transcription-PCR (RT-PCR), the expression of these SOD isozymes was studied in different cell types of olive leaves, including spongy mesophyll, palisade mesophyll, xylem and phloem. In spongy mesophyll cells, the isozyme proportion was similar to that in whole leaves, but in the other cells the proportion of expressed SOD isozymes was different. In palisade mesophyll cells, Fe-SOD was the most abundant, followed by Mn-SOD and CuZn-SOD, but in phloem cells Mn-SOD was the most prominent isozyme, and Fe-SOD was present in trace amounts. In xylem cells, only the Mn-SOD was detected. On the other hand, the highest accumulation of superoxide radicals was localized in vascular tissue which was the tissue with the lowest level of SOD transcripts. These data show that in olive leaves, each SOD isozyme has a different gene expression depending on the cell type of the leaf.

DNA, Plant↗