Mechanism of paraquat tolerance in perennial ryegrass

Abstract The mechanism of paraquat tolerance was investigated in lines of perennial ryegrass (Lolium perenne L.) which had been selected for resistance to the herbicide. Uptake, metabolism and translocation of paraquat were studied. Susceptible cultivars and a tolerant line were not found to differ in uptake of radioactive paraquat applied to the leaf surface or supplied to the cut ends of excised leaves. Distribution of herbicide within leaf tissue was similar in tolerant and susceptible plants and no metabolites of ¹⁴C-paraquat were detected in tolerant or susceptible plants treated with sub-lethal concentrations of the herbicide. Autoradiography and quantitative determinations showed much variation in translocation of ¹⁴C-paraquat out of treated leaves of intact plants, but the variation was not related to the degree of susceptibility to the herbicide. It is concluded that paraquat tolerance in perennial ryegrass is unlikely to depend upon reduced uptake, enhanced metabolism or altered translocation of the herbicide.     

Protection from Paraquat-Mediated Photo-Oxidative Stress by Glutathione in Poplar (Populus tremula×P. alba) Plants

Abstract: The sensitivity of hybrid poplar (Populus tremula × P. alba) to oxidative stress mediated by paraquat exposure was analysed with leaf discs from wild-type plants and plants expressing the bacterial cDNA of the enzymes of glutathione synthesis, namely gshI, encoding γ-glutamylcysteine synthetase (ECS), or gshII, encoding glutathione synthetase (GS), both in the cytosol. It was expected that leaf discs containing more than 2-fold elevated glutathione concentrations due to over-expression of ECS are less susceptible to paraquat exposure than wild-type plants and transformants over-expressing GS. However, neither over-expression of GS nor of ECS improved paraquat tolerance of the leaves. This result was surprising, because in wild-type plants reduced paraquat sensitivity of young compared with mature leaves coincided with ca. 30 % higher glutathione contents of the young leaves. Apparently, developmental changes in paraquat sensitivity of poplar leaves are controlled by factors different from glutathione contents. Feeding experiments with glutathione and its metabolic precursor γ-glutamylcysteine (EC) plus gly showed that glutathione can provide protection from paraquat-mediated photo-oxidative stress; but at least ca. 5-fold elevated glutathione levels are required for this effect in poplar leaves. Currently, such high glutathione levels have not been achieved by the application of plant molecular biology techniques. The significance of glutathione for the compensation of photo-oxidative stress is discussed.     

Integration of the cyanobacterial <Emphasis Type="Italic">DesA</Emphasis> gene for Δ12-acyl-lipid desaturase improves potato tolerance to paraquat-induced oxidative stress

The effect of potato plant (Solanum tuberosum L., cv. Desnitsa) transformation with the desA gene from Synechocystis sp. PCC 6803, encoding Δ12 acyl-lipid desaturase, on the development of plant tolerance to oxidative stress was studied. To initiate oxidative stress, plants were treated with 1 mM paraquat; this treatment enhanced oxidative processes in both wild-type and transformed potato plants via the activation of superoxide anion-radical generation. This resulted in the activated oxidation of membrane lipids and the formation of a great amount of fatty acids with coupled double bonds (conjugated dienes, CD), further breakdown of lipid molecules, and enhanced production of MDA in tissues of wild-type and transformed plants. The characteristics of oxidative stress, including lipid peroxidation, were less pronounced in transformants as compared with wild-type plants. After treatment with paraquat, activities of main antioxidant enzymes (superoxide dismutase, catalase, and peroxidase) were much higher in wild-type than in transformed plants. Thus, expression of inserted heterologous desA gene for Δ12 acyl-lipid desaturase in potato plants resulted in improved tolerance of transformants to oxidative stress due to the more efficient maintenance of stable cell membrane structure functioning, and this permits prevention of electron “jump” to oxygen and, as a result, of accelerated ROS generation. More developed and regularly arranged chloroplast membrane system in transformants may also favor their improved tolerance.     

Induced biotic responses to herbivory and associated cues in the Amazonian ant-plant Maieta poeppigii

Ants inhabiting ant‐plants can respond to cues of herbivory, such as the presence of herbivores, leaf damage, and plant sap, but experimental attempts to quantify the dynamic nature of biotic defenses have been restricted to a few associations between plants and ants. We studied the relationship between certain features of the ant‐shrub Maieta poeppigii Cogn. (Melastomataceae) and the presence or absence of ant patrolling on the leaf surface in plants occupied by the ant Pheidole minutula Mayr (Hymenoptera: Formicidae). We also carried out field experiments to examine ant behavior following plant damage, and the potential cues that induce ant recruitment. These experiments included clipping of the leaf apex, as well as the presentation of a potential herbivore (live termite worker) and a foliar extract from Maieta on treatment leaves. The presence of ants patrolling the leaves of M. poeppigii is influenced by the number of domatia on the plant. Ant patrolling on the leaves of M. poeppigii was constant throughout a 24 h cycle, but the mean number of patrolling ants decreased from young to mature leaves, and from leaves with domatia to those without domatia. There was an overall increase in the number of ants on experimental leaves following all treatments, compared to control leaves. Visual and chemical cues associated with herbivory are involved in the induction of ant recruitment in the Maieta–Pheidole system. The continuous patrolling behavior of ants, associated with their ability to respond rapidly to foliar damage, may result in the detection and repellence/capture of most insect herbivores before they can inflict significant damage to the leaves.     

Overexpression of bacterial catalase in tomato leaf chloroplasts enhances photo-oxidative stress tolerance

The Escherichia coli gene katE, which is driven by the promoter of the Rubisco small subunit gene of tomato, rbcS3C, was introduced into a tomato (Lycopersicon esculentum Mill.) by Agrobacterium tumefaciens‐mediated transformation. Catalase activity in progeny from transgenic plants was approximately three‐fold higher than that in wild‐type plants. Leaf discs from transgenic plants remained green at 24 h after treatment with 1 µm paraquat under moderate light intensity, whereas leaf discs from wild‐type plants showed severe bleaching after the same treatment. Moreover, ion leakage from transgenic leaf discs was significantly less than that from wild‐type leaf discs at 24 h after treatment with 1 µm paraquat and 10 mm H₂O₂, respectively, under moderate light intensity. To evaluate the efficiency of the E. coli catalase to protect the whole transgenic plant from the oxidative stress, transgenic and wild‐type plants were sprayed with 100 µm paraquat and exposed to high light illumination (800 µmol m^?2 s^?1). After 24 h, the leaves of the transgenic plants were less damaged than the leaves of the wild‐type plants. The catalase activity and the photosynthesis activity (indicated by the F_v/F_m ratio) were less affected by paraquat treatment in leaves of transgenic plants, whereas the activities of the chloroplastic ascorbate peroxidase isoenzymes and the ascorbate content decreased in both lines. In addition, the transgenic plants showed increased tolerance to the oxidative damage (decrease of the CO₂ fixation and photosystem II activity and increase of the lipid peroxidation) caused by drought stress or chilling stress (4 °C) under high light intensity (1000 µmol m^?2 s^?1). These results indicate that the expression of the catalase in chloroplasts has a positive effect on the protection of the transgenic plants from the photo‐oxidative stress invoked by paraquat treatment, drought stress and chilling stress.     

Tolerance to apical and foliar damage: costs and mechanisms in Raphanus raphanistrum

To study mechanisms underlying plant tolerance to herbivore damage, we used apical and foliar damage as experimental treatments to study whether there are similar tolerance mechanisms to different types of damage. We also studied whether tolerance to different types of damage are associated, and whether there is a cost involved in plant tolerance to different types of herbivore damage. Our greenhouse experiment involved 480 plants from 30 full-sib families of an annual weed Raphanus raphanistrum , wild radish, which were subjected to control and two different simulated herbivore damage treatments, apex removal and foliar damage of 30% of leaf area. Apical damage significantly decreased seed production, whereas foliar damage had no effect. There was a significant genetic variation for tolerance to foliar, but not apical damage. No costs were observed in terms of negative correlation between tolerance to either damage type and fitness of undamaged plants. Tolerances to apical and foliar damage were not significantly correlated with each other. We observed a larger number of significant associations between tolerance and reproductive traits than between tolerance and vegetative traits. Plant height and leaf size of damaged plants interacted in their association to tolerance to foliar damage. Inflorescence number and pollen quantity per flower of damaged plants were positively associated with tolerance to apical damage. In late-flowering genotypes, petal size of undamaged plants and pollen quantity of damaged plants were positively associated with tolerance to foliar damage. In summary, traits involved in floral display and male fitness were associated with plant tolerance to herbivore damage.     

Brassinolide alleviated the adverse effect of water deficits on photosynthesis and the antioxidant of soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.)

Brassinolide (BR) is a relatively new plant growth regulator. To test whether BR could be used to increase tolerance to water deficits in soybean, the effects of BR application on photosynthesis, assimilate distribution, antioxidant enzymes and seed yield were studied. BR at 0.1 mg l⁻¹ was foliar applied at the beginning of bloom. Two levels of soil moisture (80% field capacity for well-watered control and 35% for drought-stressed treatment) were applied at pod initiation. BR treatment increased biomass accumulation and seed yield for both treatments. Drought stress inhibited translocation of assimilated ¹⁴C from the labeled leaf, but BR increased the translocation for both treatments. Drought stress depressed chlorophyll content and assimilation rate (A), while chlorophyll content and A of BR-treated plants were greater than that of drought-stressed plants. BR treatment increased maximum quantum yield of PS II, the activity of ribulose-1,5-bisphosphate carboxylase, and the leaf water potential of drought-stressed plants. Treatment with BR also increased the concentration of soluble sugars and proline, and the activities of peroxidase and superoxide dismutase of soybean leaves when drought-stressed. However, it decreased the malondialdehyde concentration and electrical conductivity of leaves under drought stress. This study show that BR can be used as a plant growth regulator to enhance drought tolerance and minimize the yield loss of soybean caused by water deficits.     

Oxidative stress tolerance and longevity in Arabidopsis: the late‐flowering mutant gigantea is tolerant to paraquat

Recent genetic analyses of longevity in animals have revealed that long-lived strains are more tolerant to environmental stresses. To investigate whether extended longevity in Arabidopsis also correlates with an increase in stress tolerance, the response was tested of 11 late-flowering mutants to the superoxide radical-generating herbicide paraquat. A tight correlation between flowering time and paraquat tolerance was found when plants were exposed to low doses of herbicide. Furthermore, the mutant gigantea (gi-3) with the longest delay in flowering time had a high tolerance level to paraquat-induced oxidative stress. All the tested gi alleles had an increased tolerance to paraquat toxicity compared to wild-type, although the actual levels of tolerance differed. In addition, the gi-3 mutant was more tolerant to hydrogen peroxide. These results suggest that the link between longevity and oxidative stress resistance in plants is similar to that found in animals, implying that this phenomenon may be general for all aerobic organisms.     

Effect of foliar‐applied potassium silicate on coffee leaf infection by Hemileia vastatrix

Coffee leaf rust, caused by Hemileia vastatrix, is the most devastating disease of coffee. Since limited information is available in the literature on silicon (Si) affecting plant diseases in coffee, this study was designed to investigate foliar application of potassium silicate (PS), a source of soluble (Si), on infection process of coffee leaf rust at the microscopic level. The foliar Si concentration for plants sprayed with water and PS has no significant difference (0.24 and 0.30 dag kg^?1, respectively). X‐ray microanalysis indicated that the deposition of Si on the leaves of the plants that were sprayed with PS was greater in comparison to the leaf samples from the plants sprayed with water. Rust severity on leaves of plants sprayed with water or sprayed with PS reached 44% and 32%, respectively, at 36 days after inoculation (dai). Plates of polymerised PS were observed on the leaf surfaces of the plants sprayed with the product, in contrast to its absence on the leaf surfaces of plants sprayed with water. At 36 dai, a greater number of uredia were observed on the leaf surfaces of plants sprayed with water in comparison to the leaf surfaces of plants sprayed with PS. On fractured leaf tissues that were sprayed with PS, less fungal colonisation was observed in comparison to the leaves of plants sprayed with water. In conclusion, the results of this study suggest that the effect of foliar‐applied Si on the control of the coffee leaf rust development may be attributed to the physical role of the polymerised PS, its osmotic effect against urediniospores germination, or both.     

Effect of 2-aminoethanol pretreatment on the antioxidant enzyme activity in <Emphasis Type="Italic">Zea mays</Emphasis> under oxidative stress

Exposure of plants to stress may result in liberation of 2-aminoethanol as an inducer of alarm reaction that activates cellular resistance and tolerance mechanisms. In the present study, the effect of exogenous 2-aminoethanol on stimulation of tolerance mechanisms was investigated in maize (Zea mays L.) plants treated with the herbicide paraquat. Maize shoots were pretreated with 2-aminoethanol. Two days later the shoots were treated with paraquat. Pretreatment with 2-aminoethanol increased catalase and guaiacol peroxidase activities and cysteine content and decreased MDA and hydrogen peroxide content in maize plants treated with paraquat. Based on the results of this study, it can be suggested that strengthening of the antioxidant mechanisms by plant pretreatment with amino alcohol could be helpful in the development of a broad tolerance to adverse stress conditions.     

紫茎泽兰叶片水浸液对蚕豆的化感效应

外来入侵植物可以通过淋溶、自然挥发、根系分泌和植株凋落物分解等途径向周围环境释放化感物质,抑制伴生植物的生长、发育。该研究以不同浓度紫茎泽兰(Eupatorium adenophorum)叶片水浸液处理蚕豆(Vicia faba)种子,研究紫茎泽兰叶片水浸液对蚕豆根尖细胞微核、染色体畸变、细胞凋亡、蚕豆幼苗叶片叶绿素和N含量、光合生理特性、生物量的影响。结果表明:(1)紫茎泽兰叶片水浸液处理显著抑制蚕豆根尖的伸长和细胞的有丝分裂,并诱导蚕豆根尖细胞染色体畸变和细胞微核的产生,有丝分裂指数随着叶片水浸液浓度增加而减小,根尖细胞微核率随叶片水浸液浓度增加而增大,高浓度叶片水浸液处理对蚕豆根尖细胞的凋亡及坏死有明显影响。(2)紫茎泽兰叶片水浸液处理引起蚕豆幼苗叶片的叶绿素和N含量显著降低,并导致蚕豆幼苗最大净光合速率和生物量的显著下降。总之,紫茎泽兰叶片水浸液可能引起蚕豆根尖的氧化损伤和抑制根尖的伸长,且叶片水浸液的抑制作用呈现一定的剂量效应。紫茎泽兰叶片水浸液对蚕豆根尖的损伤和抑制作用可能影响了植株对氮素的吸收,进而对蚕豆幼苗光合生理表现以及生物量积累产生显著负面效应。     

The influence of form of deposit on the phytotoxicity of MCPA, paraquat and glyphosate applied as individual drops

MCPA, paraquat and glyphosate were applied as individual drops (200–400 μm) to pot-grown plants of radish (Raphanus sativus) or wild oat (Avena fatua), using concentrations appropriate to very low volume applications of these herbicides. For a given dose per plant, herbicide activity was unaffected by concentration of MCPA or paraquat but was enhanced as the concentration of glyphosate was increased. The activity of all three herbicides on both species was affected by variation of the site of application but not by drop size. On radish the greatest activity resulted when paraquat was applied to the cotyledons, glyphosate to the interveinal areas of true leaves and MCPA to the veins of true leaves. This is discussed in relation to herbicide mobility and local toxicity following applications at high concentration.     

The mechanism of resistance to paraquat is strongly temperature dependent in resistant Hordeum leporinum Link and H. glaucum Steud.

Paraquat-resistant biotypes of the closely-related weed species Hordeum leporinum Link and H. glaucum Steud. are highly resistant to paraquat when grown during the normal winter growing season. However, when grown and treated with paraquat in summer, these biotypes are markedly less resistant to paraquat. This reduced resistance to paraquat in summer is primarily a result of increased temperature following herbicide treatment. The mechanism governing this decrease in resistance at high temperature was examined in H. leporinum. No differences were observed between susceptible and resistant biotypes in the interaction of paraquat with isolated thylakoids when assayed at 15, 25, or 35 °C. About 98 and 65% of applied paraquat was absorbed through the leaf cuticle of both biotypes at 15 and 30 °C, respectively. Following application to leaves, more herbicide was translocated in a basipetal direction in the susceptible biotype compared to the resistant biotype at 15 °C. However, at 30 °C more paraquat was translocated in a basipetal direction in the resistant biotype. Photosynthetic activity of young leaf tissue from within the leaf sheath which had not been directly exposed to paraquat was measured 24 h after treatment of plants with para. quat. This activity was inhibited in the susceptible biotype when plants were maintained at either 15 °C or 30 °C after treatment. In contrast, photosynthetic activity of such tissue of the resistant biotype was not inhibited when plants were maintained at 15 °C after treatment, but was inhibited at 30 °C. The mechanism of resistance in this biotype of H. leporinum correlates with decreased translocation of paraquat and decreased penetration to the active site. This mechanism is temperature sensitive and breaks down at higher temperatures.We are grateful to Zeneca Agrochemicals, Jealotts Hill, Berkshire, UK who provided [¹⁴C]paraquat. E.P. was supported through a Ph.D. scholarship from the Australian International Development Assistance Bureau and C.P. was the recipient of an Australian Research Council Postdoctoral Fellowship.     

Proline protects <Emphasis Type="Italic">Atropa belladonna</Emphasis> plants against nickel salt toxicity

Atropa belladonna L. plants were grown in water culture for 8 weeks before the nutrient medium was supplemented with NiCl₂ to final concentrations of 0 (control treatment), 50, 100, 150, 200, 250, and 300 μM. After 4 days of plant growing in the presence of nickel chloride, the content of water, proline, Ni, Fe, free polyamines, as well as lipid peroxidation rates were measured. The addition of 100–150 μM Ni to the medium significantly reduced the fresh weight increments and water content in comparison with these parameters for untreated plants; 200 μM Ni caused serious, although nonlethal damage to the plants, whereas 250 and 300 μM Ni proved to be lethal. In the aboveground organs, the major part of Ni was accumulated in the apical leaves. When the plants were treated with 200 μM Ni, the Ni content in apical leaves was 220 μg/g dry wt, while Ni content in roots reached 1500 μg/g dry wt. The treatment of plants with proline in the presence of 200 μM Ni inhibited Ni accumulation in tissues. The proline-treated plants exhibited elevated iron content in leaves and especially in roots and were characterized by comparatively low rates of lipid peroxidation and by sustained leaf water status. When 200 μM Ni was applied, the content of free putrescine decreased, while the contents of spermine and spermidine in leaves increased appreciably with respect to the control values. The toxic effect of nickel was accompanied not only by an enhanced accumulation of high- molecular-weight polyamines but also by their oxidative degradation, which was evident from the 14-fold increase in the content of 1,3-diaminopropane. The protective effect of exogenous proline in the presence of high nickel concentrations was manifested in lowered lipid peroxidation rates, alleviation of iron deficiency, and in retarded oxidative degradation of polyamines.     

Sustained accumulation of methyl salicylate alters antioxidant protection and reduces tolerance of holm oak to heat stress

Methyl salicylate (MeSA) is thought to have a major role in biotic and abiotic stresses by acting as a signal to trigger the oxidative burst, which is needed to activate gene expression in plant stress responses. To assess the potential effects of sustained foliar accumulation of MeSA on plant stress tolerance, the extent of photo- and antioxidant protection, lipid peroxidation and visual leaf area damage were evaluated in MeSA-treated ( c. 60 nl l⁻¹ in air) and control holm oak ( Quercus ilex L.) plants exposed to heat stress. Control plants showed an increase in foliar MeSA levels up to 1.8 nmol [gDW]⁻¹ as temperature increased and they displayed tolerance to temperatures as high as 45°C, which might be attributed, at least in part, to enhanced xanthophyll de-epoxidation and increases in ascorbate and α-tocopherol. MeSA-treated plants showed a sustained foliar accumulation of this compound, with values ranging from 10 to 23 nmol [gDW]⁻¹ throughout the experiment. These plants showed lower ascorbate and tocopherol levels and higher oxidative damage at 50°C than controls, as indicated by enhanced malondialdehyde levels and leaf area damage and lower maximum efficiency of PSII photochemistry ( F _v/ F _m ratio). These results demonstrate that a sustained foliar accumulation of MeSA is detrimental to plant function and that it can reduce thermotolerance in holm oak by altering antioxidant defences.     

Uniconazole-induced tolerance of soybean to water deficit stress in relation to changes in photosynthesis, hormones and antioxidant system

This study investigated whether uniconazole confers drought tolerance to soybean and if such tolerance is correlated with changes in photosynthesis, hormones and antioxidant system of leaves. Soybean plants were foliar treated with uniconazole at 50 mg L-1 at the beginning of bloom and then exposed to water deficit stress at pod initiation for 7 d. Uniconazole promoted biomass accumulation and seed yield under both water conditions. Plants treated with uniconazole showed higher leaf water potential only in water-stressed condition. Water stress decreased the chlorophyll content and photosynthetic rate, but those of uniconazole-treated plants were higher than the stressed control. Uniconazole increased the maximum quantum yield of photosystemand ribulose-1,5-bisphosphate carboxylase/oxygenase activity of water-stressed plants. Water stress decreased partitioning of assimilated 14C from labeled leaf to the other parts of the plant. In contrast, uniconazole enhanced translocation of assimilated 14C from labeled leaves to the other parts, except stems, regardless of water treatment. Uniconazole-treated plants contained less GA3, GA4 and ABA under well-watered condition than untreated plants, while the IAA and zeatin levels were increased substantially under both water conditions, and ABA concentration was also increased under water stressed condition. Under water-stressed conditions, uniconazole increased the content of proline and soluble sugars, and the activities of superoxide dismutase and peroxidase in soybean leaves but not the malondialdehyde content or electrical conductivity. These results suggest that uniconazole-induced tolerance to water deficit stress in soybean was related to the changes of photosynthesis, hormones and antioxidant system of leaves.     

Stratospheric Ozone Depletion: High Arctic Tundra Plant Growth on Svalbard is not Affected by Enhanced UV-B after 7 years of UV-B Supplementation in the Field

The response of tundra plants to enhanced UV-B radiation simulating 15 and 30% ozone depletion was studied at two high arctic sites (Isdammen and Adventdalen, 78° N, Svalbard).The set-up of the UV-B supplementation systems is described, consisting of large and small UV lamp arrays, installed in 1996 and 2002. After 7 years of exposure to enhanced UV-B radiation, plant cover, density, morphological (leaf fresh and dry weight, leaf thickness, leaf area, reproductive and ecophysiological parameters leaf UV-B absorbance, leaf phenolic content, leaf water content) were not affected by enhanced UV-B radiation. DNA damage in the leaves was not increased with enhanced UV-B in Salix polaris and Cassiope tetragona. DNA damage in Salix polaris leaves was higher than in leaves of C. tetragona. The length of male gametophyte moss plants of Polytrichum hyperboreum was reduced with elevated UV-B as well as the number of Pedicularis hirsuta plants per plot, but the inflorescence length of Bistorta vivipara was not significantly affected. We discuss the possible causes of tolerance of tundra plants to UV-B (absence of response to enhanced UV-B) in terms of methodology (supplementation versus exclusion), ecophysiological adaptations to UV-B and the biogeographical history of polar plants     

Chemical aspects of host‐acceptance behaviour in the bird cherry‐oat aphid Rhopalosiphum padi: host‐acceptance signals used by different morphs with the same genotype

Host acceptance by gynoparae and winged virginoparae of the bird cherry‐oat aphid Rhopalosiphum padi (L.) is investigated utilizing leaves and aqueous extracts of the primary and secondary hosts, as well as nonhost plants. Gynoparae are specialized to reproduce on bird cherry Prunus padus L., whereas virginoparae reproduce and feed on various grasses. Host acceptance is assessed using levels of reproduction and survival for adults, as well as survival for nymphs. Little is known of host acceptance by nymphs. The data show that gynoparae and winged virginoparae produce nymphs almost exclusively on their host plants, bird cherry and barley leaves, respectively, over 72 h. When tested with aqueous plant extracts, however, gynoparae produce nymphs almost exclusively on bird cherry extract and progeny numbers are found to be similar to those on plant leaves. Few nymphs are produced on artificial diet. By contrast, winged virginoparae produce nymphs on aqueous extracts of barley, bird cherry and bean, as well as on artificial diet. The numbers of nymphs deposited by gynoparae are similar on aqueous extracts of bird cherry leaves collected at different times during the growing season. When extracts from leaves of various Prunus species are tested, only leaves of P. padus and Prunus virginiana stimulate parturition. Oviparae, the sexual female nymphs of gynoparae, survive well for 96 h on both bird cherry and barley leaves but not on bean seedlings, whereas nymphs of winged virginoparae survive well only on barley leaves. They do not survive for 96 h on any plant‐leaf extracts, although they do survive on artificial diet.     

Morphological changes and increase of resistance to oxidative stress by overexpression of the <Emphasis Type="Italic">LebZIP2</Emphasis> gene in <Emphasis Type="Italic">Nicotiana benthamiana</Emphasis>

The tomato bZIP2-encoding gene was inserted into the Nicotiana benthamiana genome using Agrobacterium-mediated transformation to characterize resistance to oxidative stress and two herbicides, glyphosate and paraquat. We produced transgenic tobacco plants using the LebZIP2 gene, which were then utilized to examine salt stress and herbicide resistance through oxidative mechanisms. Transgenic LebZIP2-overexpressing plants were examined using specific primers for selection marker genes (PCR using genomic DNA) and target genes (RT-PCR). Based on microscopic examination, we observed an increase in leaf thickness and cell number in transgenic plants. The electrolyte leakage of leaves suggested that LebZIP2-overexpressing lines were weak tolerant to NaCl stress and resistant to methyl viologen. During our analysis, transgenic lines were exposed to different herbicides. Transgenic plants showed an increased tolerance based on visual injury, as well as an increased biomass. Based on these results, the LebZIP2 gene may be involved in oxidative stress tolerance and cell development in plants.