Nutzung von In-vitro-Pflanzen der Kartoffel zur Prüfung der antiphytoviralen Wirkung von 2,4-Dioxohexahydro-1,3,5-triazin (DHT) in Kombination mit N-Cyanoguanidin

Suitability of in vitro potato plantlets for testing of antiphytoviral effect of a combination of 2,4-dioxo-hexahydro-1,3,5-triazine (DHT) and N-cyano guanidine The suitability of a semiquantitative determination of virus content for the detection of antiphytoviral effects of chemicals was demonstrated by ELISA directly applying at in vitro potato cultures systemically infected and chemotherapeutically treated. The combination of 2, 4-dioxohexahydro-1,3,5-triazine (DHT) and N-cyanoguanidine both applied at a concentration of 0.03% to the culture medium resulted in a high significant reduction of the relative concentration of the potato virus S in the, potato genotype ‘M-812820’ of 44–73 %. A phytotoxic influence of the substances was not observed. The combination of the two antiphytoviral substances had no effect against potato virus Y in explants of the variety ‘Ackersegen’. Differences in the susc, eptibility of various genotypes against biologically active substances were indicated.     

Verstärkung der antiphytoviralen Wirkung von 2,4-Dioxohexahydro-l,3,5-triazin durch Kombination mit Verbindungen mit Guanidinstruktur

Intensified antiphytoviral activity of 2,4-dioxohexahydro-l,3,5-triazine by combination with guanidines Combined application of the antiphytoviral compound 2,4-dioxohexahydro-l,3,5 triazine (DHT) and different guanidines (GDs) that were either unsubstituted or substituted only by low-molecular substituents reduced the concentration of potato virus χ (PVX) in leaves or Nicotiana tabacum L. cv. Samsun much more than application of either agent alone. In secondarily PVX-infected leaves, the activity of 2,4-dioxohexahydro- 1,3,5-triazine was increased by a larger number of GDs and to a greater degree than in inoculated ones. The activities of GDs against tobacco mosaic virus (TMV) in Nicotiana tabacum L. cv. Samsun as well as against brome grass mosaic virus (BRV) in Hordeum vulgare L. cv Vogelsanger Gold were only insignificantly increased by combination with DHT. On the other hand, in experiments with so-called identical potato eye cuttings, in which several eye cuttings were obtained from each potato tuber, one serving as a control and the others being treated with DHT, N-cyano guanidine or a combination of these substances, the number of cuttings with symptoms of potato leaf-roll virus (PLRV) could be much more greatly reduced by the combination than by the individual preparations. The number of cuttings with symptoms of potato virus Y and potato virus A was significantly reduced by treatment with the combined preparation, but not by treatment with DHT or N-cyano guanidine alone. Additional investigation with N-cyano-GD, and, beside this, with acetyl-GD and N N′ N triamino-GD indicated a close correlation between the diminution of numbers of potato eye cuttings with virus symptoms and the increase in tuber weight. The greater the reduction in the number of cuttings showing virus symptoms, the greater was the increase in tuber weight. These relationships were observed even in those cases where controls had been treated with ammonium nitrate solutions whose N contents equivalent to the N contents of the preparations. The observed effects of the preparations therefore are not attributable to N-fertilizing effects.     

Hemmung von Pflanzenviren durch Membranlipidanaloga unter besonderer Berücksichtigung von Alkanmonosulfonaten

Inhibition of plant viruses by membrane lipid analogs under special regard of alkane monosulfonates Under investigation was the inhibition of plant viruses by compounds, which could be comprehended as membrane lipid analogs because of some structural similarities to these lipids. Among these analogs, alkane monosulfonates (AMS, preparation “Emulgator E 30”) exhibit distinctly marked antiphytoviral activities, which are reported in detail. The mentioned AMS reduced the concentrations of PVX (potato virus X) as well as of TMV (tobacco mosaic virus) markedly in inoculated leaves of Nicotiana tabacum L. ‘Samsun’ and remarkably in secondarily infected ones. The formation of local lesions on leaves of hypersensitive hosts is influenced strongly by AMS. Likewise, the activity of the free virion is reduced in vitro. The mentioned effectsproved dependent on the length of the alkane chains and the concentration of AMS. Innertherapeutical effects of AMS have been pointed out. Several substances proved excellent coergists of AMS and vice versa, above all DHT (2,4-dioxohexahydro-1,3,5-triazine), several guanidines, a thiourea compound, a thiadiazole compound and ribavirin. In part full synergistic effects were obtained. AMS as well as combinations of AMS with some of the mentioned coergists also influence potato viroses. Using so-called identical potato eye cuttings, it was demonstrated that the mentioned treatments resulted in a significant and in part considerable reduction of the number of potato eye cutting plants with virus symptoms and in a partially high increase in the weight of tubers formed by the eye cutting plants. Likewise, the number of plants with virus symptoms was reduced and the tuber mass was increased by newly developed granulated AMS. A combined treatment with granulated AMS and granulated DHT resulted in a strong reduction of the number of potato eye cutting plants with virus symptoms, but the tuber mass was not augmented.     

Effect of DHT (2,4-Dioxohexahydro-1,3,5-triazine) as Well as a Combination of DHT, Alkane Monosulfonate and N-Phenyl-N'-p-carboxyphenyl Thiourea on Tomato Mosaic Virus in Infected Glasshouse Tomato Plants

In two consecutive trials, three treatments of tomato plants of the variety Rivermoon with the antiphytoviral substance DHT (2,4-dioxohexahydro-1,3,5-triazine, 0.15%) 2 d a.i. and 2 and 7 d p.i. reduced the concentration of ToMV (tomato mosaic virus) by 62.3 and 60.5%. The average tomato yields increased by 40 and 26 % compared with the virus diseased, untreated controls, but the yields of the healthy controls were not achieved. A combined treatment of DHT, alkane monosulphonate and N-phenyl-N'-p-carboxyphenyl thiourea only resulted in a minor reduction in the ToMV titre and only a slight increase in yield, compared with the DHT-solo treatment.     

The Mode of Inhibition of TMV- and PVX-Induced RNA-Dependent RNA Polymerases by some Antiphytoviral Drugs

Six nucleobase and nucleoside analogues (6-aminouracil [6-AU], 6-aminothymin [6-AT], 6-fluorothymin [6-FT], 2,4-dioxohexahydro-1,3,5-triazine [DHT], 9-(2,3-dihydroxypropyl) adenine [DHPA] and 1-β-D-ribofuranosyl-1,2,4-triazol-3-carboxamid [ribavirin]) with potential antiphytoviral activity were tested in an in vitro replication system for their effect on tobacco mosaic virus (TMV)-and potato virus X(PVX)-induced RNA dependent RNA polymerase. 6-AU, 6-AT, 6-FT, and DHPA inhibited both polymerases which may be a target of these compounds. A Putative noncompetitive mode of inhibition is discussed. Whereas ribavirin did not affect the in vitr RNA synthesis in the established systems, the uracil-derivative DHT inhibited only the PVX-specific enzyme.     

Chemotherapeutical Elimination of Potato Virus X from Potato Stem Cuttings

Potato virus X was completely eliminated from all infected potato stem cuttings grown in nutrient media containing 0.02 or 0.03 % 2,4-dioxohexahydro-1,3,5-triazine (DHT). When DHT was added to the media in concentrations of 0.01 and 0.005% the efficiency by which the virus was eliminated differed between the varieties tested. The method is less time-consuming than the generally used meristem (axillary-) tip culture in combination with chemo- or thermotherapy.     

Beziehungen zwischen dem Wirkungsgrad antiphytoviraler Substanzen und dem Niveau der quantitativen Sortenresistenz sowie der Virulenz von Virusisolaten an Virus/Wirt-Systemen der Kartoffel

Relationship between the efficiency of antiphytoviral substances, the degree of quantitative resistance of cultivars, and the virulence of virus isolates on virus/host-systems of potato. In vitro systematically infected with potato virus S, potato virus M or potato virus X stem cuttings of different potato varieties were treated with 2,4-dioxohexahydro-1,3,5-triazine and Ribavirin. The reduction of virus replication by chemotherapy differed between varieties depending on their level of quantitative resistance and the virulence of virus isolates. An increasing resistance level of cultivars and a decreasing virulence of the virus isolates resulted in a relative enhancement of the inhibitory of the antiphytoviral substances.     

Antiphytovirale Verbindungen mit nichtzyklischer Azin Struktur

Antiphytoviral compounds with noncyclic azine structutre The antiphytoviral activities of variously substituted compounds with noncylic azine structures have been studied f a total of 90 tested compunds 42 had the effect of more or less strong inhibiting the concentration of potato virus X (PVX) in inoculated and / or secondarily infected leaves of Nicotiana tabacum L. cv Samsun (Table.1) An effect on the virion of PVX in vitro couldn't be observed (Table.2) Thus the substances may interact with the virus replication. Some of them also reduced the number of local lesions caused by tabacco mosaic virus on leaves of Nicotiana glutinosa L. (Table-3) Several Compounds proved exellent synergists of 2,4- dioxohexahydro- 1,3,5- triazine (DHT = 5 azadihydrouracil; Table 4) Pyridine- 3- aldehyde S ethyl isothiosemicarbazone and the Cu complex of 1- ethyl-isatine S- ethyl isothisosemicarbazone when used in combination with 2,4 dioxohexahydro- 1,3,5, triazine, greatly increased the mass of potato tubers produced from plantlets grown up from patato eye cutting as compared with the indentical control Simultaneously the mentioned substances reduced the number of symptom bearing eye cutting plants Quinoline 2- aldehyde- N- oxide S- allyl-isothiosemicarbazone had the effect of very greatly reducing the number of symptom-bearing plants without substantially influencing the mass of tubers (Table 5). Thus some compounds with noncyclic azine structure especially when used in combinitation with DHT, may be of high interest for practical application Comparing the structures of compounds with noncyclic azine structure active against plant or human viruses it should be noted that antiphytoviral compounds active in these two different virus host systems often are closely related structurally.     

Obtaining a High Percentage of Explants with Negative Serological Reactions against viruses by Combining Potato Meristem Culture with Antiphytoviral Chemotherapy

A high percentage of plantlets, in which PVX, PVM, and PVS were no longer detectable by precipitin tests was obtained from 100% virus-infected stocks of the potato varieties Rosa and Priobskij rannija by combining meristem (tip) culture with antiphytoviral chemotherapy. When 2,4 - dioxohexahydro 1,3,5-triazinc (DHT) was added to the nutrient media, no virus was detectable serologically in 72% of the corresponding explants of the variety Rosa and in 66% of that of the variety Priobskij rannija, respectively. Contrary to this, all untreated control explants proved virusinfected. Treatment with 2-thiouracil resulted in 46.6%, with cyanoguanidine in 28.6% of plantlets, in which no virus was detectable serologically.     

Effect of mixed viral infections (potato virus Y-potato leafroll virus) on biology and preference of vectors Myzus persicae and Macrosiphum euphorbiae (Hemiptera: Aphididae)

Mixed viral infections of heterologous viruses such as Potato virus Y (family Potyviridae, genus Potyvirus, PVY) and Potato leafroll virus (family Luteoviridae, genus Polerovirus, PLRV) are a regular occurrence in Idaho's potato, Solanum tuberosum (L.), cropping systems. An increased number of plant samples from Idaho's potato fields over the past 2 yr has serologically tested positive for both PVY and PLRV via double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and exhibited more severe symptoms than singly-infected plants (PVY or PLRV). Several studies have extensively examined the mixed infection phenomenon but to the best of our knowledge, none have examined the effect of such infections on vector biology and preference. Laboratory studies were conducted to examine the effect of mixed viral (PVY-PLRV) infection on the fecundity and preference of two of the most efficient PVY and PLRV vectors, the green peach aphid, Myzus persicae (Sulzer), and the potato aphid, Macrosiphum euphorbiae (Thomas) (Hemiptera: Aphididae). M. persicae and M. euphorbiae adults were clip-caged (one adult per cage) to leaflets of PVY, PLRV, PVY-PLRV-infected, and noninfected potato plants. The number of nymphs produced in all four treatments was recorded after 96 h. M. persicae and M. euphorbiae fecundity was significantly higher on mixed infected plants than on singly infected plants or noninfected plants. Preference of alatae and apterae of M. persicae and M. euphorbiae was determined with the use of settling bioassays. Both alatae and apterae of M. persicae and M. euphorbiae preferentially settled on PVY-PLRV-infected plants than on singly infected plants (PVY or PLRV) or noninfected plants.     

表达PVY和PLRV双价外壳蛋白基因马铃薯的抗病性研究

表达马铃薯Y病毒(PVY)和马铃薯卷叶病毒(PLRV)双价外壳蛋白基因的马铃薯(Solanum tubero-sum L.)栽培品种“Favorita”和“虎头”,经摩擦接种PVY和用桃蚜接种PLRV后,观察症状并用ELISA测定病毒滴度。结果表明,两个品种转双价CP基因的各株系,接种病毒后表现无症状或症状轻微,其中PVY和PLRV平均滴度均较不转基因对照植株低。不同品种对PVY和PLRV的抗性比较表明,转双价CP基因的“Favorita”对PVY抗性较明显,而转双价CP基因的“虎头”则对PLRV抗性较对PVY抗性明显。不同转基因株系抗病毒水平不同。“Favorita”9个转双价CP基因株系中有6个株系PVY滴度较未转基因对照降低52.5％～90.0％,而“虎头”7个转双价CP基因株系中有4个株系PLRV含量较对照降低53.0％～98.0％。在抗性株系中还出现一些抗1种病毒或抗2种病毒的抗性较强的单株。     

双价外壳蛋白基因植物表达载体的构建及马铃薯转基因植株的鉴定

在克隆了马铃薯Ｘ病毒（ＰＶＸ）、马铃薯Ｙ 病毒（ＰＶＹ）和马铃薯卷叶病毒（ＰＬＲＶ）的外壳蛋白基因的基础上,构建同时包含ＰＶＸ和ＰＶＹ 与ＰＶＹ 和ＰＬＲＶ 两个外壳蛋白基因植物表达框架的表达载体,通过农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ）介导转化烟草（Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ ）和生产上常用的几个马铃薯（Ｓｏｌａｎｕｍ ｔｕｂｅｒｏｓｕｍ ）优良品种：“Ｆａｖｏｒｉｔａ”、“虎头”、“克４”。经ＰＣＲ检测证明外源基因已整合到植物的染色体上,得到批量转基因植株。在转ＰＶＸ＋ＰＶＹ 外壳蛋白基因的烟草上接种ＰＶＸ （５ μｇ／ｍ Ｌ）、ＰＶＹ（２０ μｇ／ｍ Ｌ）病毒,得到有一定抗性的植株     

Construction of Plant Expression Vectors and Identification of Transgenic Potato Plants

Potato virus X (PVX), potato virus Y (PVY) and potato leaf roll virus (PLRV) infection in potato may result in the loss of centrification of seed potatoes and affect the quality and yield of potatoes in agricultural production. The authors cloned coat protein (cp) genes of PVX, PVY and PLRV and constructed two kinds of plant expression vector which contain PVX and PVY or PVY and PLRV cp genes. Three major commercial cultivars of potato and one cultivar of tobacco were transformed via Agrobacterium tumefaciens mediated procedure. Transgenic plants were confirmed by PCR analysis. Transgenic tobacco plants containing both PVX and PVY cp genes were significantly resistant to PVX and PVY infection via mechanical inoculation.     

Accumulation of potato virus Y is enhanced in Solatium brevidens also infected with tobacco mosaic virus or potato spindle tuber viroid

The accumulation of potato virus Y?(PVY?) and potato leaf roll virus (PLRV) was studied in plants of Solanum brevidens co-infected with each of six viruses or a viroid. Virus could not be detected by ELISA in plants of S. brevidens infected solely with PVY. However, accumulation of PVY was increased c. 1000-fold in plants doubly infected with tobacco mosaic virus or potato spindle tuber viroid (PSTVd). PVY titres in doubly infected plants of S. brevidens were between 1% and 0.1% of those found in the PVY-susceptible interspecific Solanum hybrid DTO-33. Double infections of 5. brevidens by PVY and alfalfa mosaic virus or potato viruses M, S, T or X did not significantly enhance PVY accumulation. Accumulation of PLRV was not enhanced in plants co-infected with any of the six viruses or PSTVd.     

Bis-thiourea Metal Complexes with Enhanced Antiphytoviral Activity

The activity of bis-thiourea, diallyl-bis-thiourea and in part of diphenyl-bis-thiourea against potato virus X (PVX) can be augmented considerably by complexation of the mentioned compounds with Cu^±±, Ni^±±, and Co^±±. Contrary to this, complexation with Fe^±± and Cd^±± does not augment or reduce the antiphytoviral activity. Most antiphytoviral active are the diallyl-bis-thiourea complexes with Cu^±±, Ni^±± and Co^±±. The antiphytoviral activity of the Cu^±± and Cd^±± complexes with bisthiourea can be additionally augmented by combined treatment with small amounts of 2,4-dioxohexahydro-l,3,5-triazine (DHT).     

Characterization of the expression and inheritance of potato leafroll virus (PLRV) and potato virus Y (PVY) resistance in three generations of germplasm derived from Solanum etuberosum

Potato virus Y (PVY) and potato leafroll virus (PLRV) are two of the most important viral pathogens of potato. Infection of potato by these viruses results in losses of yield and quality in commercial production and in the rejection of seed in certification programs. Host plant resistance to these two viruses was identified in the backcross progeny of a Solanum etuberosum Lindl. somatic hybrid. Multiple years of field evaluations with high-virus inoculum and aphid populations have shown the PVY and PLRV resistances of S. etuberosum to be stably expressed in two generations of progeny. However, while PLRV resistance was transmitted and expressed in the third generation of backcrossing to cultivated potato (Solanum tuberosum L. subsp. tuberosum), PVY resistance was lost. PLRV resistance appears to be monogenic based on the inheritance of resistance in a BC₃ population. Data from a previous evaluation of the BC₂ progeny used in this study provides evidence that PLRV resistance was partly conferred by reduced PLRV accumulation in foliage. The field and grafting data presented in this study suggests that resistance to the systemic spread of PLRV from infected foliage to tubers also contributes to the observed resistance from S. etuberosum. The PLRV resistance contributed by S. etuberosum is stably transmitted and expressed through sexual generations and therefore would be useful to potato breeders for the development of PLRV resistant potato cultivars.     

Volatiles from potato plants infected with potato leafroll virus attract and arrest the virus vector, Myzus persicae (Homoptera: Aphididae)

The influence of viral disease symptoms on the behaviour of virus vectors has implications for disease epidemiology. Here we show that previously reported preferential colonization of potatoes infected by potato leafroll virus (genus Polerovirus) (luteovirus) (PLRV) by alatae of Myzus persicae, the principal aphid vector of PLRV, is influenced by volatile emissions from PLRV-infected plants. First, in our bioassays both differential immigration and emigration were involved in preferential colonization by aphids of PLRV-infected plants. Second, M. persicae apterae aggregated preferentially, on screening above leaflets of PLRV-infected potatoes as compared with leaflets from uninfected plants, or from plants infected with potato virus X (PVX) or potato virus Y (PVY). Third, the aphids aggregated preferentially on screening over leaflet models treated with volatiles collected from PLRV-infected plants as compared with those collected from uninfected plants. The specific cues eliciting the aphid responses were not determined, but differences between headspace volatiles of infected and uninfected plants suggest possible ones.     

Microbial populations,fungal species diversity and plant pathogen levels in field plots of potato plants expressing theBacillus thuringiensis var.tenebrionis endotoxin

The environmental release of genetically engineered (transgenic) plants may be accompanied by ecological effects including changes in the plant-associated microflora. A field release of transgnic potato plants that produce the insecticidal endotoxin ofBacillus thuringiensis var.tenebrionis (Btt) was monitored for changes in total bacterial and fungal populations, fungal species diversity and abundance, and plant pathogen levels. The microflora on three phenological stages of leaves (green, yellow and brown) were compared over the growing season (sample days 0, 21, 42, 63 and 98) for transgenic potato plants, commercial Russet Burbank potato plants treated with systemic insecticide (Di-Syston) and commercial Russet Burbank potato plants treated with microbialBtt (M-Trak). In addition, plant and soil assays were performed to assess disease incidence ofFusarium spp.,Pythium spp.,Verticillium dahliae, potato leaf roll virus (PLRV) and potato virus Y (PVY). Few significant differences in phylloplane microflora among the plant types were observed and none of the differences were persisent. Total bacterial populations on brown leaves on sample day 21 and on green leaves on sample day 42 were significantly higher on the transgenic potato plants. Total fungal populations on gree leaves on sample day 63 were significantly different among the three plant types; lowest levels were on the commerical potato plants treated with systemic insecticide and highest levels were on the commercial potato plants treated with microbialBtt. Differences in fungal species assemblages and diversity were correlated with sampling dates, but relatively consistent among treatments.Alternaria alternata, a common saprophyte on leaves and in soil and leaf litter, was the most commonly isolated fungus species for all the plant treatments. Rhizosphere populations of the soilborne pathogensPythium spp.,Fusarium spp. andV. dahliae did not differ between the transgenic potato plants and the commercial potato plants treated with systemic insecticide. The incidence of tuber infection at the end of the growing season by the plant pathogenV. dahliae was highest for the transgenic potato plants but this difference was related to longer viability of the transgenic potato plants. This difference in longevity between the transgenic potato plants and the commercial + systemic insecticide potato plants also made comparison of the incidence of PVY and PLRV problematic. Our results indicate that under field conditions the microflora of transgenicBtt-producing potato plants differed minimally from that of chemically and microbially treated commerical potato plants.     

Factors affecting the detection of potato leafroll virus in potato foliage by enzyme-linked immunosorbent assay

Using antiserum globulins that reacted only weakly with plant materials, potato leafroll virus (PLRV) at 10 ng/ml was detected consistently by enzyme-linked immunosorbent assay (ELISA). The reaction with PLRV particles was slightly impaired in potato leaf extracts that were diluted less than 10^-1 but not at greater dilutions. Antiserum globulins that reacted more strongly with plant materials could be used satisfactorily for coating microtitre plates but were unsuitable for conjugating with enzyme. The detection end-point of PLRV, in leaf sap of potato cv. Cara plants grown from infected tubers in the glasshouse, was about 10^-2 and the virus was reliably detected in extracts of composite samples of one infected and 15 virus-free leaves. PLRV concentration was much less in extracts of roots or stolons than in leaf extracts. The virus was detected in infected leaves of all 27 cultivars tested. PLRV was readily detectable 2 wk before symptoms of secondary infection developed in field-grown plants of cv. Cara and Maris Piper and remained so for at least 5 wk. Its concentration was slightly greater in old than in young leaves and was similar to that in glasshouse-grown plants. In field-grown plants of cv. Maris Piper with primary infection, PLRV was detected in tip leaves 21–42 days after lower leaves were inoculated by aphids; in some shoots it later reached a concentration, in tip leaves, similar to that in leaves with secondary infection. Symptoms of primary infection developed in the young leaves of some infected shoots but were inconspicuous and were not observed until at least a week after PLRV was detected by ELISA.     

Expression of a luteoviral movement protein in transgenic plants leads to carbohydrate accumulation and reduced photosynthetic capacity in source leaves

Elucidating the role of viral genes in transgenic plants revealed that the movement protein (MP) from tobacco mosaic virus is responsible for altered carbohydrate allocation in tobacco and potato plants. To study whether this is a general feature of viral MPs, the movement protein MP17 of potato leafroll virus (PLRV), a phloem-restricted luteovirus, was constitutively expressed in tobacco plants. Transgenic lines were strongly reduced in height and developed bleached and sometimes even necrotic areas on their source leaves. Levels of soluble sugars and starch were significantly increased in source leaves. Yet, in leaf laminae the hexose—phosphate content was unaltered and ATP reduced to only a small extent, indicating that these leaves were able to maintain homeostatic conditions by compartmentalization of soluble sugars, probably in the vacuole. On the contrary, midribs contained lower levels of soluble sugars, ATP, hexose—phosphates and UDP-glucose supporting the concept of limited uptake and catabolism of sucrose in the phloem. The accumulation of carbohydrates led to a decreased photosynthetic capacity and carboxylation efficiency of ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) probably owing to decreased expression of photosynthetic proteins. In parallel, levels of pathogenesis-related proteins were elevated which may be the reason for the obtained limited resistance against the unrelated potato virus Y (PVY)^N in the transgenic tobacco plants. Ultrathin sections of affected leaves harvested from 2-week-old plants revealed plasmodesmal alterations in the phloem tissue while plasmodesmata between mesophyll cells were indistinguishable from wild-type. These data favour the phloem tissue to be the primary site of PLRV MP17 action in altering carbohydrate metabolism.