Influence of Iron Chlorosis on Pigment and Protein Metabolism in Leaves of Nicotiana tabacum L

Experiments were conducted on Nicotiana tabacum, L. to study the relation in the grana among chlorophylls, carotenoids, and proteins. The effect of iron chlorosis on protein and pigment synthesis was studied at different stages of chlorosis using glycine-U-C¹⁴. Pigments were separated by thin layer chromatography.

Chlorophyll a, chlorophyll b, carotenoid, and protein contents of chloroplasts from chlorotic tissue were less than those of normal tissues. A 25% decrease in protein labeling and a 45% decrease in chlorophyll labeling was noted in deficient tissue compared to normal tissue even before chlorosis was perceptible. Both normal and iron deficient leaf discs which received iron in the incubation medium incorporated higher amounts of radioactive glycine into chlorophyll a and chlorophyll b at all stages of development than their respective counterparts not supplied with iron in the incubation medium. The presence of iron in the incubation medium reduced the amount of glycine incorporated into carotenes and xanthophylls, except where the tissue was severely chlorotic. This may be attributed to active competition for glycine between the iron-dependent- (chlorophyll) and iron-independent-(carotenoid) biosynthetic pathways. Incorporation of glycine into chloroplast pigments was lowest at severe chlorosis, probably due to a reduction in the overall enzyme activity.

     

Identification of a stress-induced protein in stem exudates of soybean seedlings root-infected with Fusarium solani f. sp. glycines

Sudden death syndrome of soybean (Glycine max) is caused by the soilborne fungus, Fusarium solani f. sp. glycines, that infects soybean roots. Besides root necrosis, symptoms include interveinal leaf chlorosis, necrosis and premature defoliation. It is proposed that a fungal toxin is produced in soybean roots and translocated to foliage. In this study, we isolated compounds from soybean stem exudates from plants that were either inoculated or not inoculated with F. solani f. sp. glycines. A protein with an estimated molecular mass of 17 kDa and designated as FISP 17 for F. solani f. sp. glycines-induced stress protein was identified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This protein occurred only in F. solani f. sp. glycines-infected soybean stem exudates. The N-terminal amino acid sequence of the purified protein had 100 % identity with a starvation-associated message 22 protein, and 80 and 78 % identity with purified bean pathogenesis-related proteins, PvPR1 and PvPR2, respectively. To determine if the protein was of plant or fungal origin, a synthetic peptide was designed based on the N-terminal sequence and used to raise a polyclonal antibody from rabbit. Western blot analysis showed that the antibody only reacted with a 17-kDa protein in F. solani f. sp. glycines-infected plant exudates, but no reaction occurred with healthy plant exudates or with culture filtrates of F. solani f. sp. glycines. This is the first report of the presence of a stress-induced protein in stem exudates of soybean seedlings root-infected with F. solani f. sp. glycines.     

Purification and characterization of potato ribonuclease P

Ribonuclease P (RNase P), a ribonucleo-protein endoribonuclease, responsible for 5′ maturation of precursor tDNA, is well characterized in bacteria, yeast and human, but not in plant. Attempt has been made to partially purify and characterize nuclear RNase P from potato. cDNAs encoding two putative protein subunits of potato ribonuclease P (RNase P), StPop5 and StRpp25, were picked from potato EST library based on homology with respective human RNase P protein subunits. Both the cDNAs, 435 bp long StPop5 and 765 bp long StRpp25, were RT-PCR amplified, cloned and sequenced. StPop5 exhibited 46 % nucleotide sequence similarity to the hPop5 sequence. The deduced amino acid sequence of StPop5 had 23 % identity and 35 % similarity to hPop5. Both hRpp25 and StRpp25 had 46 % nucleotide sequence homology, and 17 % identity and 27 % similarity at a length of 271 amino acids. The molecular masses of purified 6× His-tagged recombinant StPop5 and StRpp25 proteins were 18 kDa and 33 kDa respectively. Potato nuclear RNase P was partially purified from leaves employing DEAE-Sephacel anion-exchange chromatography, and from floral buds employing DEAE-Sephacel and HiTrap Q anion-exchange chromatography, ammonium sulfate precipitation and gel filtration chromatography. Immunoprecipitation with polyclonal antisera, raised against recombinant StPop5 and StRpp25, demonstrated association of these two proteins with floral bud RNase P activity but not with leaf RNase P activity.     

Loci underlying resistance to manganese toxicity mapped in a soybean recombinant inbred line population of ‘Essex2019; x ‘Forrest’

Resistance to manganese toxicity is associated with some soybean (Glycine max) cultivars grown on acidic soils or in hydroponics. Previously random amplified polymorphic DNA (RAPD) markers had seemed to identify 4 quantitative trait loci (QTL), regions that might underlie resistance to manganese toxicity in a recombinant inbred line (RIL) population derived from ‘Essex’ x ‘Forrest’. Our objective was to identify microsatellite markers linked to these, or additional, QTL for resistance to manganese toxicity in a separate assay. Two hundred and forty microsatellite markers and 100 RILs were used to construct a map. The response of five plants per genotype to manganese was measured by leaf chlorosis (scored from 0–5) and root necrosis (scored from 0–5) from 7–28 days after treatment with 125 μM of manganese in hydroponics. The experiment was repeated. ANOVA and MapMaker/QTL were used to identify regions underlying the responses. Three genomic regions on different linkage groups were found to contain QTL for resistance to necrosis during manganese toxicity. The regions located on linkage groups C2 (BARC__S att291),I(BARC__S att239)andG(OP__O EO2)wereeachsignificantlyassociated(P<0.005, R ²=20%) with root necrosis at 7 days after treatment. The regions all derived the beneficial allele from Essex. One of the previously identified RAPD associated root necrosis QTL was identified in this new study. However, no QTL for leaf chlorosis were detected (P<0.005) and none of the RAPD identified leaf chlorosis QTL could be identified. We conclude that root and leaf resistance to manganese toxicity are environmentally sensitive quantitative traits determined by separate loci of different number and magnitude of effect.     

Analysis of nutrient deficiencies affecting in vitro growth and development of <Emphasis Type="Italic">Eucalyptus dunnii</Emphasis> Maiden

Although basal medium optimization is a key factor in the success of tissue culture, its mineral composition is frequently disregarded when optimizing in vitro propagation protocols. A previous work on Eucalyptus dunnii micropropagation suggests that excessive callus formation and leaf chlorosis are related to specific nutritional conditions of the basal media. Recently, a novel basal medium based on the mineral nutrient analysis of E. dunnii young stump shoots was developed and successfully tested in plant regeneration and micropropagation of E. dunnii, avoiding all these issues. Considering this basal medium as an ideal growth condition, a mild deprivation of each macro and micronutrient and of the total organic fraction was imposed to E. dunnii in vitro cultures for 30 d. As a result, K, Mg, Mn, Cl, Zn, Mo, Ni or Co deprivation quantitatively affected growth and development of axillary shoots. Moreover, leaf chlorosis and the development of organogenic callus under Fe deficiency, and leaf drop along with shoot tip necrosis under N deficiency were observed. These symptoms suggest that nutrient content in E. dunnii tissues needs to be above 420.3 mg kg^?1 for Fe and 27.7 g kg^?1 for N to avoid the symptoms of leaf chlorosis and shoot tip necrosis. Additionally, the main role of Mn in quantitative responses and the antagonism between ions, especially for Mg/K and Mg/Zn, were denoted by the multivariate analysis. Overall, these results make a relevant contribution to the optimization of in vitro propagation of E. dunnii and other hard-to-propagate related species.     

Activity of ascorbate-dependent H2O2-scavenging enzymes and leaf chlorosis are enhanced in magnesium- and potassium-deficient leaves, but not in phosphorus-deficient leaves

The effect of phosphorus (P), potassium (K), and magnesium (Mg)deficiency on the development of leaf symptoms (chlorosis andnecrosis) and activities of ascorbate-dependent H₂O₂ scavengingenzymes (ascorbate peroxidase, monodehydroascorbate reductase,dehydroascorbate reductase, and glutathione reductase) was studiedin bean (Phaseolus vulgans) plants over a 12 d period of growthin nutrient solution. With increasing plant age Mg- and K-deficientleaves developed severe interveinal chlorosis and, accordingly,chlorophyll concentrations were reduced. However, in P-deficientleaves neither chlorosis nor necrosis appeared; the leaves remaineddark green and even at an advanced stage of P deficiency, chlorophyllconcentrations were still higher than those of control plants.In K- and, particularly, Mg-deficient leaves with an increasein severity of leaf chlorosis, activity of ascorbate-dependentH₂O₂- scavenging enzymes was progressively increased. In contrast,in P-deficient leaves, as in leaves of the control plants, activityof H₂O₂-scavenging enzymes remained at a low level over the12 d period. Accordingly, compared with P-deficient and controlplants, Mg- and K-deficient leaves with elevated anti-oxidativepotential showed much higher resistance to chlorophyll destructionby the herbicide paraquat. Elevated levels of H₂O₂-scavengingenging enzymes in Mg- and K-deficient leaves indicate a higherproduction of H₂O₂ and related toxic O₂ species. It Is suggestedthat in Mg- and K-deficient leaves, utilization of photoreductantsin CO₂ fixation is restricted because of impaired export andthus accumulation of photosynthates. This disturbance mightlead to enhanced photoreduction of molecular O₂ to toxic O₂species causing chlorophyll destruction (chlorosis), a processwhich is not important in P-deficient leaves where export ofsucrose is not affected. Key words: Bean, hydrogen peroxide detoxification, leaf chlorosis, magnesium nutrition, oxygen activation, phosphorus nutrition, potassium nutrition     

Inhibition of Phytophthora species by secondary metabolites produced by the dark septate endophyte Phialocephala europaea

Dark septate fungal root endophytes of the Phialocephala fortinii s.l.–Acephala applanata species complex (PAC) are widely distributed throughout the temperate and subtropical regions of the Northern Hemisphere. Previous studies have shown that some PAC members are pathogenic, others suppress oomycete root pathogens and some have no obvious effect on their Norway spruce (Picea abies) host. The activity of 85 PAC isolates against Phytophthora citricola s.l. was investigated by co-culture on plates. We identified a strain of Phialocephala europaea that significantly reduced the growth of P. citricola in vitro. Characterization of its extracellular metabolites resulted in the identification of four major compounds, sclerin, sclerolide, sclerotinin A, and sclerotinin B. These compounds are known for their positive as well as negative effects on plant growth. We found that sclerin and sclerotinin inhibited the growth of P. citricola in vitro at 150 μg ml^?1 (～1 mM). This is the first report of their production by Phialocephala and of activity of these compounds against an oomycete. Therefore, our data suggest that some PAC might reduce disease resulting from P. citricola by the production of antibiotics and plant growth promoting metabolites.     

Diatrypaceae species overlap between vineyards and natural ecosystems in South Africa

Diatrypaceae species collected from 33 woody hosts occurring near vineyards were characterised. Plants showed symptoms of dieback, necrosis and cankers. Phylogenetic analyses based on ITS-rDNA and the β-tubulin gene separated the 265 isolates obtained into 14 species. The five most frequently encountered species, Cryptovalsa ampelina, Eutypa cremea, Eutypa lata, Eutypella citricola and Eutypella microtheca, were also found on grapevine in a previous study in South Africa. Several plant species, therefore, serve as hosts for these pathogens. First reports in South Africa include Cryptosphaeria ligniota, Eutypella leprosa and Eutypella australiensis. Host ranges of Cryptosphaeria multicontinentalis, Eu. australiensis and Eu. microtheca were also expanded. Diatrypaceae species caused brown discolouration when inoculated onto wounded grapevine tissues, which suggests that cross-infections are possible between grapevine and other woody hosts. Campelina., Eu. microtheca and Eu. citricola were the most virulent species and their virulence was comparable to that of Eu. lata, a known pathogen causing Eutypa dieback.     

Effects of ozone and Phytophthora citricola on non-structural carbohydrates of European beech (Fagus sylvatica) saplings

A lysimeter study was performed to monitor long term effects of chronic ozone enrichment on saplings of European beech (Fagus sylvatica L). After 3 years of ozone exposure a root infection with Phytophthora citricola Swada was established in the fourth year to study the interaction between elevated ozone and the root infection on the carbon budget of beech saplings. By using quantitative PCR no differences in root infection with P. citricola were observed between the ozone treatments. In contrast to the first 3 years of ozone exposure, sucrose and starch concentrations in leaves were diminished in ozone treated plants in the fourth year. The root infection reduced sucrose concentrations in leaves. Starch reserves of the heterotrophic biomass were not affected by any treatments. Thus 4 years of ozone exposure and 1 year of P. citricola root infection had only limited effect on carbohydrate metabolism in beech saplings.     

Contrasting ozone × pathogen interaction as mediated through competition between juvenile European beech (Fagus sylvatica) and Norway spruce (Picea abies)

Based on the growth-differentiation balance theory (GDB) and the influence of tropospheric ozone (O₃) on plants, we hypothesized that pre-conditioning with elevated O₃ reduces adverse effects of the root rot pathogen Phytophthora citricola Sawada. To this end a 2-year phytotron study with juvenile European beech (Fagus sylvatica L.) and (Picea abies [L.] Karst.) grown in mixture was performed. The hypothesis was tested on phenological, leaf and root morphological as well as physiological aspects of plant performance. Contrasting with spruce, elevated O₃ limited leaf and root biomass development, photosynthetic performance and N uptake of beech. The growth limitation by O₃ conveyed increased resistance in beech against the pathogen. Conversely, spruce displayed enhanced susceptibility in the combined O₃/P. citricola treatment. The hypothesis was supported in the case of beech rather than spruce. Nevertheless, conclusions support GDB regarding the trade-off between growth and stress defense, although compliance appears to be species-specific.     

Some Biochemical Evidence on the Selective Insecticide Toxicity between the Two Aphids,Aphis citricola and Myzus malisuctus (Homoptera: Phididae), and Their Predator,Harmonia axyridis (Coleoptera: Coccinellidae)

This experiment was carried out to compare the differences in biochemical enzyme activity on the selective insecticide toxicity between the two species of aphid, Aphis citricola van der Goot and Myzus malisuctus Matsumura, and their predator, Harmonia axyridis Pallas. Esterase activities between the two species of aphids and between the two stages of H. axyridis were significant different. Glutathione S-traasferase (GST) activity toward 1-chloro-2, 4-dinitrobenzene (CDNB) was much higher than 1, 2-dichloro-4-nirobenzene (DCNB) in all species tested. No DCNB conjugation was detected in A. citricola and M. malisuctus. The predator, H. axyridis, had much higher GST activity than the preys, A. citricola and M. malisuctus. GST activity toward CDNB in H. axyridis adult was highest, even 6.2-fold higher activity than H. axyridis larva. M. malisuctus had much higher GST activity than A. citricola. The degree of acetylcholinesterase (AChE) inhibition by phosphamidon among all three species tested was significantly varied. The concentration of phosphamidon required for 50% AChE inhibition was lowest in H. axyridis larva, while highest in M. malisuctus. There fore, elevated GST activity and target-site insensitivity may be largely associated with the differential susceptibility between larva and adult of H. axyridis. However, differential susceptibility between A. citricola and M. malisuctus may be due to other various biochemical mechanisms responsible for the multiple selective toxicity, including elevated GST activity and target-site insensitivity.     

Influence of Conformation of M. tuberculosis RNase P Protein Subunit on Its Function

RNase P is an essential enzyme that processes 5'' end leader sequence of pre-tRNA to generate mature tRNA. The bacterial RNase Ps contain a RNA subunit and one protein subunit, where the RNA subunit contains the catalytic activity. The protein subunit which lacks any catalytic activity, relaxes the ionic requirements for holoenzyme reaction and is indispensable for pre-tRNA cleavage in vivo. In the current study, we reconstituted the M. tuberculosis RNase P holoenzyme in vitro. We prepared the RNase P protein through two different strategies that differ in the conditions under which the recombinant M. tuberculosis protein, expressed in E. coli was purified. The mycobacterial RNase P protein which was purified under native conditions subsequent to isolation from inclusion bodies and in vitro renaturation, was capable of cleaving pre-tRNA specifically without the requirement of RNase P RNA. However, the preparation that was purified under denaturing conditions and refolded subsequently lacked any inherent pre-tRNA processing activity and cleaved the substrate only as a component of the holoenzyme with the RNA subunit. We found that the two RNase P protein preparations attained alternative conformations and differed with respect to their stability as well.     

Genetic analysis of iron chlorosis tolerance in Prunus rootstocks

The high economic losses caused by the occurrence of iron chlorosis in Prunus orchards in the Mediterranean area justifies the implementation of breeding programs to generate high-performance rootstocks for different edaphoclimatic area conditions. For that reason, the genetic control of iron chlorosis tolerance was studied in an F₁ population derived from a three-way interspecific cross between a Myrobalan plum (P 2175) and an almond?×?peach hybrid (Felinem). Several phenotypic measurements were assessed to guarantee an accurate data set for genetic analysis. SPAD (Soil and Plant Analyzer Development) values, chlorophyll concentration, and visual diagnostic symptoms were highly correlated with leaf chlorosis in trees. SPAD value was the most reliable measure, since it was an objective, unbiased, and non-destructive method. Two significant quantitative trait loci (QTLs) involved in SPAD and chlorophyll concentration were identified for Felinem in linkage groups 4 and 6. Both QTLs were detected in four of the six consecutive years of the experiment. For P 2175, two of the three putative QTLs identified, pspad4.1 and chl4.1, were placed in linkage group 4. These QTLs were related to the SPAD values and chlorophyll concentration, respectively, and co-localized with QTLs detected in the Felinem map affecting the same traits. Candidate gene PFIT, related to iron metabolism, was localized within the confidence interval of the QTL in linkage group 4. This research suggests an association of this chromosome region with tolerance to iron chlorosis in Prunus, and it provides a first approach to localize candidate genes involved in tolerance to this abiotic stress.     

Lack of Types 1 and 2A Protein Serine(P)/Threonine(P) Phosphatase Activities in Chloroplasts

Protein phosphatase activity in crude leaf extracts and in purified intact chloroplasts of wheat (Triticum aestivum) and pea (Pisum sativum) was analyzed using exogenously supplied phosphoproteins or endogenous thylakoid proteins. Leaf extracts contain readily detectable amounts of protein phosphatase activity measured with either phosphohistone or phosphorylase a, substrates of mammalian protein phosphatases. No significant chloroplast protein phosphatase activity was detected using these exogenous phosphoproteins. The dephosphorylation of endogenous thylakoid light-harvesting chlorophyll a/b binding proteins in situ was inhibited by fluoride, but not by microcystin-LR or okadaic acid, diagnostic inhibitors of mammalian types 1 and 2A protein phosphatases. Additionally, no evidence for a pea chloroplast alkaline phosphatase activity was found using β-glycerolphosphate or 4-methylum-belliferyl phosphate as substrates. From these results, we conclude that phosphohistone and phosphorylase a are not useful substrates for chloroplast thylakoid protein phosphatase activity and that the chloroplast enzymes may not fit into one of the canonical classifications currently used for protein phosphatases.     

Defence reactions in the apoplastic proteome of oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis> var. <Emphasis Type="Italic">napus</Emphasis>) attenuate <Emphasis Type="Italic">Verticillium longisporum</Emphasis>growth but not disease symptoms

Background

Verticillium longisporum is one of the most important pathogens of Brassicaceae that remains strictly in the xylem during most stages of its development. It has been suggested that disease symptoms are associated with clogging of xylem vessels. The aim of our study was to investigate extracellular defence reactions induced by V. longisporum in the xylem sap and leaf apoplast of Brassica napus var. napus in relation to the development of disease symptoms, photosynthesis and nutrient status.

Results

V. longisporum (strain VL43) did not overcome the hypocotyl barrier until 3 weeks after infection although the plants showed massive stunting of the stem and mild leaf chlorosis. During this initial infection phase photosynthetic carbon assimilation, transpiration rate and nutrient elements in leaves were not affected in VL43-infected compared to non-infected plants. Proteome analysis of the leaf apoplast revealed 170 spots after 2-D-protein separation, of which 12 were significantly enhanced in response to VL43-infection. LS-MS/MS analysis and data base searches revealed matches of VL43-responsive proteins to an endochitinase, a peroxidase, a PR-4 protein and a β-1,3-glucanase. In xylem sap three up-regulated proteins were found of which two were identified as PR-4 and β-1,3-glucanase. Xylem sap of infected plants inhibited the growth of V. longisporum.

Conclusion

V. longisporum infection did not result in drought stress or nutrient limitations. Stunting and mild chlorosis were, therefore, not consequences of insufficient water and nutrient supply due to VL43-caused xylem obstruction. A distinct array of extracellular PR-proteins was activated that might have limited Verticillium spreading above the hypocotyl. In silico analysis suggested that ethylene was involved in up-regulating VL43-responsive proteins.

     

Effects of Tabtoxinine-beta-Lactam on Nitrogen Metabolism in Avena sativa L. Roots

The effects of tabtoxinine-β-lactam (T-β-L) on nitrate uptake and glutamine synthetase (GS) and nitrate reductase (NR) activities in roots of Avena sativa seedlings were determined. Seven-day-old oat seedlings placed in a 10 mm KNO₃ and 0.5 mm T-β-L solution for 24 hours took up T-β-L and lost approximately 90% of their root GS activity. [³H]-T-β-L taken up by roots of seven-day-old oat seedlings was associated with GS immunoprecipitated from the extract of these roots. Total nitrate uptake and in vivo NR activity were decreased approximately 50% in the T-β-L treated roots. However, T-β-L uptake did not affect the induction phases of nitrate uptake or reduction, nor did it inhibit in vitro NR activity. Thus, the decrease in nitrate uptake and reduction is a secondary effect of T-β-L action. Roots of seven-day-old oat seedlings were inoculated with Pseudomonas syringae pv tabaci (Tox+) and the pathogen population in the rhizosphere was estimated by dilution plate count; 6 × 10¹³ bacteria were recovered after 3 days, as compared to the original inoculation with 7 × 10⁹ bacteria, indicating a significant growth of the pathogen in the rhizosphere. The bacteria recovered from the rhizosphere caused chlorosis in tobacco leaves and produced T-β-L in culture; 1 × 10¹⁴ bacteria were recovered from roots of seedlings inoculated with P. syringae pv tabaci (Tox−) using the same inoculation and assay procedure as for the pv tabaci (Tox+). Extracts of surface-sterilized roots previously inoculated with P. syringae pv tabaci (Tox+) did not produce viable bacterial cultures when plated out on a complete medium. Oat seedlings growing in sand culture and inoculated with P. syringae pv tabaci (Tox+) had developed chlorosis, and root GS activity had declined to less than 10% of controls after 3 days. Conversely, seedlings inoculated with P. syringae pv tabaci (Tox−) never developed chlorosis and maintained normal levels of GS activity. All oat plants inoculated with P. syringae pv tabaci (Tox+) died within 7 days after inoculation as compared to the plants inoculated with P. syringae pv tabaci (Tox−) which grew to maturity.     

Reduced Photosystem II Activity and Accumulation of Viral Coat Protein in Chloroplasts of Leaves Infected with Tobacco Mosaic Virus

We previously reported (A Reinero, RN Beachy 1986 Plant Mol Biol 6:291-301) that coat protein (CP) of tobacco mosaic virus (TMV) accumulates in chloroplasts of systemically infected leaves. To determine the significance of such interaction we examined electron transport rates in chloroplasts containing different levels of TMV-CP. Tobacco (Nicotiana tabacum L.) plants were infected with either a TMV strain inducing chlorosis or with a strain inducing mild symptoms, and both the accumulation pattern of TMV-CP inside chloroplasts as well as the rates of photosynthetic electron transport were followed. The CP of the TMV strain inducing chlorosis was detected inside chloroplasts 3 days after infection, and thereafter accumulated at a rapid rate, first in the stroma and then in the thylakoid membranes. On the other hand, the CP of the TMV strain that caused only mild symptoms accumulated in chloroplasts to lower levels and little CP was associated with the thylakoids. In vivo and in vitro measurements of electron transport revealed that photosystem II activity was inhibited in plants infected with the aggressive TMV strain while no reduction was observed in plants infected with the mild strain. The capacity of chloroplasts to synthesize proteins was equivalent in organelles isolated from healthy and virus-infected leaves. The possibility that a large accumulation of TMV-CP inside chloroplasts may affect photosynthesis in virus-infected plants by inhibiting photosystem II activity is discussed.     

Manganese Toxicity in Sugarcane Plantlets Grown on Acidic Soils of Southern China

Ratoon sugarcane plantlets in southern China have suffered a serious chlorosis problem in recent years. To reveal the causes of chlorosis, plant nutrition in chlorotic sugarcane plantlets and the role of manganese (Mn) in this condition were investigated. The study results showed that the pH of soils growing chlorotic plantlets ranged from 3.74 to 4.84. The symptoms of chlorosis were similar to those of iron (Fe) deficiency while the chlorotic and non-chlorotic plantlets contained similar amount of Fe. Chlorotic plantlets had 6.4-times more Mn in their leaf tissues compared to the control plants. There was a significantly positive correlation between Mn concentration in the leaves and the exchangeable Mn concentration in the soils. Moreover, leaf Mn concentration was related to both seasonal changes in leaf chlorophyll concentration and to the occurrence of chlorosis. Basal stalks of mature sugarcanes contained up to 564.36 mg·kg^-1 DW Mn. Excess Mn in the parent stalks resulted in a depress of chlorophyll concentration in the leaves of sugarcanes as indicated by lower chlorophyll concentration in the leaves of plantlets emerged from basal stalks. Ratoon sugarcane plantlets were susceptible to chlorosis due to high Mn accumulation in their leaves (456.90–1626.95 mg·kg^-1 DW), while in planted canes chlorosis did not occur because of low Mn accumulation (94.64–313.41mg·kg^-1 DW). On the other hand, active Fe content in chlorotic plantlets (3.39 mg kg^-1 FW) was only equivalent to 28.2% of the concentration found in the control. These results indicate that chlorosis in ratoon sugarcane plantlets results from excessive Mn accumulated in parent stalks of planted cane sugarcanes grown on excessive Mn acidic soils, while active Fe deficiency in plantlets may play a secondary role in the chlorosis.     

Analysis of Putative Apoplastic Effectors from the Nematode,Globodera rostochiensis,and Identification of an Expansin-Like Protein That Can Induce and Suppress Host Defenses

The potato cyst nematode, Globodera rostochiensis, is an important pest of potato. Like other pathogens, plant parasitic nematodes are presumed to employ effector proteins, secreted into the apoplast as well as the host cytoplasm, to alter plant cellular functions and successfully infect their hosts. We have generated a library of ORFs encoding putative G. rostochiensis putative apoplastic effectors in vectors for expression in planta. These clones were assessed for morphological and developmental effects on plants as well as their ability to induce or suppress plant defenses. Several CLAVATA3/ESR-like proteins induced developmental phenotypes, whereas predicted cell wall-modifying proteins induced necrosis and chlorosis, consistent with roles in cell fate alteration and tissue invasion, respectively. When directed to the apoplast with a signal peptide, two effectors, an ubiquitin extension protein (GrUBCEP12) and an expansin-like protein (GrEXPB2), suppressed defense responses including NB-LRR signaling induced in the cytoplasm. GrEXPB2 also elicited defense response in species- and sequence-specific manner. Our results are consistent with the scenario whereby potato cyst nematodes secrete effectors that modulate host cell fate and metabolism as well as modifying host cell walls. Furthermore, we show a novel role for an apoplastic expansin-like protein in suppressing intra-cellular defense responses.     

Comparison of soil tests and leaf analysis as methods of diagnosing Zn deficiency in British Columbia apple orchards

Soil Zn extracted by 0.25M MgCl₂ or DTPA as an index of Zn availability was compared to Zn or P:Zn concentration in leaves for 40 commercial apple orchards in southern British Columbia. Sampled trees included a wide range of ages and cultivar/rootstock combinations. Leaves were sampled from the midportion of current season's shoots at four times; May–June 1984, July 1984, May 1985 and July 1985. At the same time the severity of the four Zn deficiency symptoms, rosetting, blind bud, little leaf and chlorosis was assessed. Little relationship was found between soil and leaf Zn measures, except in July 1984, when both terminal leaf Zn and P:Zn concentrations varied directly with 0.25M MgCl₂ extractable soil Zn. Soil Zn extracted by 0.25M MgCl₂ was more closely related to severity of deficiency symptoms on the trees at all four samples dates than was DTPA-soil Zn or leaf Zn concentration. Blindbud was the most useful indicator of deficiency in samples collected in May whereas chlorosis was the most useful one in July.