Survival, growth and gall formation by Phytolyma lata on Milicia excelsa established in mixed-species tropical plantations in Ghana

1 Milicia excelsa (Moraceae) is an important timber tree in much of Africa and when grown in monocultural plantations has been subject to nearly complete destruction by gall-forming psyllids in the genus Phytolyma. 2 We tested the impact on susceptibility to Phytolyma spp. of planting Milicia excelsa in various densities and species mixtures. Replicated plantings consisting of 11, 25, 50 and 100% Milicia in mixtures with Terminalia superba (Combretaceae) were established at a site surrounded by natural forest and in an agricultural area. 3 In addition at the agricultural site, 50–50 mixtures of Milicia excelsa with Albizia adianthifolia (Mimosaceae), Tectona grandis (Verbenaceae), Khaya ivorensis. (Meliaceae), Ceiba pentandra (Bombacaceae) and a mixture combining all species were planted. 4 Two months after planting, gall numbers were significantly lower in the 11% Milicia mixtures at the forest site. Gall numbers were also significantly lower at the forest site than at the agricultural site, for all densities of Milicia. At 6 months, dieback of Milicia resulting from gall formation had taken place equally on all seedlings, but survival of seedlings at the forest site was 40% higher than at the agricultural site.     

Changes in Phenolics, Polyphenol Oxidase and its Isoenzyme Patterns in Relation to Resistance in Taro against Phytophthora colocasiae

The effect of Phytophthora leaf blight disease, caused by Phytophthora colocasiae Raciborski, on the accumulation of phenolics and polyphenol oxidase (PPO) activity in ex vitro plants was studied in three resistant (DP‐25, Duradim and Jhankri) and one susceptible (N‐118) genotypes of taro [Colocasia esculenta (L). Schott]. The inoculation of taro leaves with P. colocasiae spores resulted in a quantitative change in both biochemical parameters and induction of PPO isoforms in resistant genotypes. The amount of phenolics was increased owing to blight by 68.02%, 58.87%, 52.67% and 11.50% in DP‐25, Duradim, Jhankri and N‐118, respectively. The per cent increase in PPO under stress over non‐stress condition was also highest in DP‐25 (49.14%) followed by Duradim (41.56%), Jhankri (40.55%) and N‐118 (17.08%). The resistant genotypes showed higher activity of PPO as compared with susceptible ones, which was reflected through its banding pattern in isoenzyme analysis, detecting four different isoforms. The intensity of the bands was higher in the resistant genotypes than in susceptible N‐118. The appearance of high intensity bands and/or reduction in the intensity of particular isoform(s) in the zymograms of all the three resistant taro genotypes studied, led to the apparent conclusion of linking PPO isoenzyme expression with blight resistance in taro. The blight incidence (per cent leaf infection and leaf area infection) was lower in the resistant genotypes than in susceptible, N‐118. The yield reduction owing to blight was below 20% in DP‐25, Jhankri and Duradim, while the same was more than 40% in N‐118. The phenolics and PPO activity was negatively correlated with disease incidence and yield reduction owing to blight. Based on the results of disease incidence, biochemical contents and yield, the pattern of stress tolerance was DP‐25 > Duradim > Jhankri > N‐118. The studied parameters, i.e. phenolics and PPO could be used as biochemical markers for leaf blight stress tolerance studies in taro.     

Biochemical markers as a useful tool for the early identification of Fusarium oxysporum f.sp. cubense,race 1 resistance banana clones

Abstract

Panama disease of banana (Musa spp) caused by the fungus Fusarium oxysporum f. sp. Cubense (FOC), is a serious constraint both to the commercial production of banana and cultivation for subsistence agriculture. Chemical control is not economically effective and is also hazardous to the environment and human health. Breeding for disease resistance is an alternative strategy, which leads to the development of resistance clones. Field evaluation is the most reliable method of screening for disease resistance, but it is demanding in terms of cost, manpower and space requirements. Another approach of screening hybrids at the sucker's stage (planting material) through biochemical markers has been found to be effective in early identification of resistant hybrids. The resistance mechanisms involving the role of phenol, PAL, oxidative enzymes like peroxidase (PO), polyphenol oxidase (PPO), superoxide dismutase (SOD), catalase and PR-proteins like chitinase, β-1-3 glucanase were studied and they showed relatively higher activity in resistant hybrids than susceptible hybrids. Isozyme analysis of peroxidase (PO) and polyphenol oxidase (PPO) was also carried out in cultivars and hybrids, which revealed the induction of specific isoforms in the resistant hybrids upon challenge inoculation. This could be a useful tool for early identification of F. oxysporum f. sp. cubense resistance banana clones.     

Histo-chemical changes induced by PGPR during induction of resistance in pearl millet against downy mildew disease

Plant growth-promoting rhizobacteria Bacillus pumilus strain INR-7 effectively induced downy mildew resistance in pearl millet. The histo-chemical analysis of B. pumilus INR-7 mediated systemic resistance showed that induced resistance is associated with the expression of hypersensitive response (HR), enhanced lignification, callose deposition, and hydrogen peroxide in addition to the increased expression of the defense enzymes β-1,3-glucanase, chitinase, phenylalanine ammonia lyase (PAL), peroxidase (POX), and polyphenol oxidase (PPO). There was rapid expression of HR in the resistant pearl millet as well as the susceptible seedlings induced by treatment with INR-7 after pathogen infection when compared to the susceptible seedlings, which expressed HR at later hours. Examination of inoculated pearl millet tissues by microscopy showed that lignin, callose, and hydrogen peroxide accumulated earlier and to higher levels in resistant and induced resistant seedlings. Accumulation of various defense enzymes was an immediate response to Sclerospora graminicola infection and preceded the development of induced resistance elicited by strain INR-7. Tissue print analysis showed that defense enzymes were found to be localized in the vascular bundles and revealed the visual difference in the expression pattern of β-1,3-glucanase, chitinase, PAL, POX, and PPO whose intensity varied among resistant, INR-7 treated, and susceptible pearl millet seedlings. This study clearly demonstrated that the differences between the responses, susceptible, INR-7 treated or resistant pearl millet seedlings recorded differences in the speed, intensity, and pattern of different histo-chemical responses to S. graminicola infection.     

Biochemical responses associated with resistance to bacterial stalk rot caused by Dickeya zeae in maize

Bacterial stalk rot (BSR) of maize caused by Dickeya zeae is an important disease in northwest region of India. In the current study, eighty maize lines were evaluated for resistance against this disease. Of these, 20 were moderately resistant, 25 were moderately susceptible and the rest were highly susceptible to BSR. Six lines from each set were randomly selected. Activities of three antioxidant enzymes, viz. phenylalanine ammonia lyase (PAL), peroxidase (POX) and polyphenol oxidase (PPO) were analysed from these three sets of maize lines representing different levels of resistance. A trend of elevated activity of PAL, POX and PPO was observed in all the three sets of maize lines. The results showed significantly more activity of these three enzymes in moderately resistant than highly susceptible maize lines. The activity of PAL and PPO peaked after 48 hr and of POX after 72 hr of challenge inoculation by D. zeae in all the maize lines. The activity of these enzymes further correlated negatively with disease development. Our results show that PAL, POX and PPO play an important role in contributing towards resistance in maize against BSR.     

Chitinase and peroxidase activities in sunflower hypocotyls: Effects of BTH and inoculation with <Emphasis Type="Italic">Plasmopara halstedii</Emphasis>

Systemic acquired resistance (SAR) can be induced in plants by incompatible pathogens, pathogen derived extracts, or certain chemicals as benzothiadiazole (BTH). The aim of this work was to compare changes in peroxidase and chitinase activities, enzymes considered as PR-proteins, caused by BTH and the pathogen Plasmopara halstedii. Hypocotyls from susceptible and resistant BTH-treated sunflower seedlings showed increased peroxidase and chitinase activities. Inoculation with P. halstedii increased chitinase and peroxidase activities in inoculated hypocotyls from susceptible but not from resistant sunflower seedlings.     

Identification of structural and chemical traits in Juglans regia associated with host plant preference of Batocera horsfieldi

1. Common walnut Juglans regia L. suffered devastating damage from the white‐striped longhorn beetle Batocera horsfieldi (Hope) in southwest China, which resulted in a dramatic decrease of local walnut production. However, the control of this wood‐boring pest with traditional methods is difficult as a result of its cryptic feeding habit.
2. We proposed that developing host‐plant resistance is a more effective way of controlling this insect with long‐term benefits. One way of improving host‐plant resistance is to identify plant traits related to host‐plant preference of targeted insects.
3. In the present study, we investigated the differences in feeding and oviposition preferences of B. horsfieldi on four J. regia cultivars in the field and tested whether such preferences are associated with structural and chemical traits in plant branches.
4. The results obtained showed that the host‐plant resistance of J. regia cultivars to B. horsfieldi was related to increased wood density, higher amounts of cellulose and lignin, and higher contents of tannins, total flavonoids and total phenolics, whereas the susceptible cultivars were characterized by thicker bark, higher contents of water, sugar and soluble protein.
5. The identified traits involved in host‐plant preference can subsequently contribute to the selection and breeding of resistant J. regia cultivars to B. horsfieldi.

     

Inducible Proteins in Citrus Rootstocks with Different Tolerance Towards the Root Rot Pathogen Phytophthora palmivora

Activities of defence‐related proteins (β‐1,3‐glucanases, chitinases and peroxidases) and concentrations of total soluble phenolics were measured in roots and leaves of non‐infected and infected plants to investigate the response of different citrus rootstock genotypes to the root rot pathogen Phytophthora palmivora Butler. Infection with the pathogen increased concentrations of total proteins, total phenolics and β‐1,3‐glucanase activity in roots of all genotypes, and increases were associated with the extent of root mass reductions and thus susceptibility of the plants. Root chitinase and root peroxidase levels were slightly reduced or unaltered upon infection. β‐1,3‐Glucanase activity was also elevated in leaves of infected plants, but increases did not differ between tolerant and susceptible rootstocks. Effects of root infection on leaves were typically the reverse of effects on roots for chitinase‐ and peroxidase levels and more pronounced in susceptible rootstock genotypes. Although differences in enzyme expression were observed between susceptible and tolerant citrus seedlings, effects were usually associated with disease progression, and not with resistance to P. palmivora. It is suggested that increased activities of the proteins and soluble phenolics studied are not implicated in the primary defence to Phytophthora root diseases, but may contribute to the inhibition of the pathogen during infection in tolerant citrus.     

Comparative analysis of activities of vital defence enzymes during induction of resistance in pearl millet against downy mildew

Pearl millet [Pennisetum glaucum (L.) R. Br.] has the seventh largest annual production in the world giving it significant economic importance. Although generally well adapted to the growing conditions in arid and semi-arid regions, major constraints to yields are susceptibility to downy mildew disease caused by the oomycete Sclerospora graminicola (Sacc.) Schroet. Induction of resistance against downy mildew disease of pearl millet has been well established using various biotic and abiotic inducers. The present study demonstrated the comparative analysis of the involvement of the important defence enzymes like β-1,3-Glucanase, chitinase, phenylalanine ammonia-lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO) and lipoxygenase (LOX) during induced systemic resistance (ISR) mediated by inducers like Benzo(1,2,3)-thiadiazole-7-carbothionic acid-S-methyl ester (BTH), Beta amino butyric acid (BABA), Chitosan and Cerebroside against pearl millet downy mildew disease. Native-PAGE showed six POX isozymes in all categories of uninoculated pearl millet seedlings and maximum intensity of bands was noticed in resistant seedlings. After inoculation in Cerebroside-treated seedlings, there were seven isoforms, POX-4 was not present in any other seedlings. Native-PAGE analysis showed the presence of five PPO isozymes in all categories of uninoculated pearl millet seedlings and after inoculation seven isoforms of PPO-7 were noticed, and the intensity of banding was more in resistant and Cerebroside-treated seedlings. The isoforms PPO-3 were present as an extra band after inoculation in all seedlings. Isoform PPO-7, though found in all seedlings, was very prominent in Chitosan- and Cerebroside-treated seedlings. β-1,3-Glucanase Native-PAGE analysis showed the presence of only one isozyme in all categories of uninoculated/inoculated pearl millet seedlings. Glu-1 isozyme was very prominent in all seedlings including resistant and susceptible seedlings. Among the induced resistant seedlings, highest intensity was observed in Cerebroside-treated seedlings. Native-PAGE analysis showed the presence of three LOX isozymes in all categories of uninoculated pearl millet seedlings, and the intensity of banding pattern was very low in BTH-treated seedlings. LOX-1 and LOX-2 were very prominent in resistant, Chitosan- and Cerebroside-treated seedlings. Upon inoculation, one extra band, LOX-3, was exclusively noticed in Cerebroside-treated seedlings. In inoculated seedlings, LOX-1, LOX-2 and LOX-4 were very prominent in Chitosan Cerebroside-treated seedlings compared to other seedlings.     

Role of Pseudomonas fluorescens and INA against Black Rot of Cabbage

Cabbage (Brassica oleracea var. capitata) is an important vegetable crop among crucifers. It is affected by a bacterial disease known as black rot. Black rot is caused by Xanthomonas campestris pv. campestris a disease of worldwide importance. The present study highlights the effect of biotic inducer—Pseudomonas fluorescens—and an abiotic inducer—2,6‐dichloro‐isonicotinic acid—in combating black rot, followed by their effect on the seed treatment and disease incidence, role of antioxidant enzymes followed by validation of the defence‐related genes by quantitative real‐time PCR. The resistant (Pusa mukta) and the highly susceptible (NBH boss) cabbage cultivars were analysed for defence‐related enzymes such as peroxidase and superoxide dismutase. An increase in total peroxidase and superoxide dismutase activity was observed upon inoculation with X. campestris pv. campestris. The activity was greater in resistant cultivar when compared to susceptible ones. Both enzyme activity assays and qPCR analyses for the expression of the defence genes in susceptible and resistant cultivars demonstrated that the peroxidase gene was up‐regulated in resistant cultivar compared to susceptible cultivar. The present study proved that P. fluorescens‐induced resistance against X. campestris pv. campestris in cabbage seedlings is more efficient as compared to the use of INA—abiotic inducer.     

Induction of resistance against downy mildew on sunflower by rhizobacteria

Abstract

Induction of resistance to downy mildew caused by Plasmopara halstedii in sunflower was studied after treatment with PGPR (plant growth promoting rhizobacteria) strain INR7 (Bacillus spp). Treatment of sunflower seeds with 1×10⁸cfu/ml of PGPR strain INR7 resulted in decreased disease severity and offered 51 and 54% protection under green house and field conditions, respectively. The induction of resistance to P. halstedii by PGPR strain INR7 was accompanied by the accumulation of various host defense-related enzymes in susceptible sunflower seedlings. Enhanced activation of catalase (CAT), phenylalanine ammonia-lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO) and chitinase (CHI) was evident at 6, 9, 12, 12 and 12h post inoculation, respectively, in sunflower seedlings raised from seeds treated with PGPR strain INR7. This enhanced and early activation of defense-related responses in the susceptible cultivar after treatment with PGPR strain INR7 was comparable to that in the resistant cultivar. The results indicate that PGPR strain INR7 induced resistance against P. halstedii in sunflower is mediated through enhanced expression of defense mechanism.     

Reactive Oxygen Species are not Increased in Resistant Oat Genotypes Challenged by Crown Rust Isolates

Crown rust (Puccinia coronata Corda f.sp. avenae) can devastate oats (Avena sativa). Oxidative stress is part of the resistance mechanism in several pathosystems, but in the oat–crown rust system, it is unclear, especially regarding partial resistance. We evaluated the effects of P. coronata on oxidative stress in oat cultivars: URS 21 (partially resistant), Leggett (race‐specific resistant), URS22 and Clintland 64 (susceptibles). Seedlings and plants were inoculated with P. coronata uredospores. Cultivars were assessed for antioxidant enzyme activity and the reactive oxygen species (ROS) hydrogen peroxide and superoxide. Due to the importance of the partial resistance of URS21, this cultivar and URS 22 were also appraised for total phenolics and the relative expression of oxidative stress genes. Postinoculation, Leggett and URS 21 showed no increased peroxide levels. The susceptible cultivars increased ROS and ascorbate peroxidase activity. Clintland 64 increased also catalase activity, whereas URS 22 increased glutathione reductase and the expression of genes encoding antioxidant enzymes. URS 21 showed almost no antioxidant enzyme induction. Shortly after inoculation, URS 21 showed increased expression of genes encoding lipoxygenase and peroxidase. Cultivars URS 21 and Leggett accumulated cell wall fluorescent compounds, phenolics being detected in the former. Oxidative stress appears not to cause the hypersensitive response in this pathosystem, but late ROS accumulation did occur in the susceptible cultivars. Cultivar URS 21 may, differently from other known mechanism to date, reduce ROS accumulation by increasing the level of phenolics, resulting in later pathogen and cell death, showing non‐specific resistance to races of the pathogen also at seedling stage.     

The effect of the pine woolly aphid (Pineus pini) on survival,growth and natural selection in Scots pine (Pinus sylvestris) in Iceland

1. Scots pine Pinus sylvestris was originally introduced to Iceland in the beginning of the 20th Century. Extensive plantings started in the late 1940s and, in total, 2–3 million Scots pine seedlings were planted, mainly originating from two counties in northern Norway. Part of this plant material was imported as seedlings.
2. Pine woolly aphid Pineus pini was introduced to Iceland before 1940, most likely on imported seedlings in 1937.
3. High mortality of Scots pine, concurrent with high infestation of the pine woolly aphid, was observed in Iceland during the late 1950s and 1960s and planting was discontinued.
4. Provenance trials with Scots pine were established in Iceland in 2004–2006. They consisted of 15 provenances from Norway, four from Finland, four from Scotland, one from Russia, one from the Austrian Alps and three first generation Scots pine provenances from Iceland, collected from survivors of the epidemic in the 1950s and 1960s. In total, there were 28 provenances.
5. The Icelandic provenances had significantly lower P. pini infestation than all the provenances of non‐Icelandic origin, which indicates that natural selection in Scots pine in Iceland has occurred in favour of individuals less susceptible to P. pini.

     

Peroxidase: a marker for Ascochyta blight resistance in chickpea

On the basis of incidence of appearance of Ascochyta blight symptoms after artificial inoculation of 25-day-old chickpea seedlings with 10 different pathotypes of Ascochyta rabiei, GL94011, PBG5 and C214 have been classified as resistant, moderately resistant and susceptible, respectively, to Ascochyta blight. In none of the pathotypes, peroxidase (PO) activity could be detected in culture medium and mycelium. Healthy tissues of GL94011 have almost three times the PO activity in comparison with that of C214. Resistant and moderately resistant genotypes showed 30–60% upregulation of PO activity against infection by A. rabiei whereas it was only 3–6% in susceptible genotype C214. These results indicate the possibility of using PO as a marker of Ascochyta blight resistance.     

Pollen genotype selection for a simply inherited qualitative factor determining resistance to chlorsulfuron in maize

Pollen genotype selection for genes expressed in both the haploid and diploid phases of the plant life cycle can lead to correlated responses detectable in the sporophyte. A pollen selection was conducted in two genetic backgrounds of maize (Zea mays L.) for chlorsulfuron resistance, conferred by the XA17 allele. Plants of two backcross (BC) lines segregating 1 (heterozygote, resistant) : 1 (homozygote, susceptible) for chlorsulfuron resistance were used as pollen donor. Selection treatment was applied during microspore development, and tassels were cut about 10 days before anthesis and placed into a liquid medium with or without 40 mg l^–1 of chlorsulfuron. Pollen was used to fertilize an unrelated susceptible genotype (tester). The resulting testcrosses (TC) were evaluated in the greenhouse by spraying seedlings with chlorsulfuron at 23 g ha^–1. Non-selected TC progenies derived from heterozygous BC plants showed a proportion of resistant and susceptible plants close to the expected 1:1 ratio, while non-selected TC progenies derived from homozygous BC plants showed susceptible plants only. Selected TC progenies derived from heterozygous BC plants showed a frequency of resistant plants ranging from 89% to 100%. BC plants homozygous for the susceptible allele, subjected to selection treatment, gave poor seed set, and no resistant plants were found in their TC progenies. Resistant TC plants obtained through pollen genotype selection were selfed, and the proportion of resistant plants was close to 3:1 in all selfed families, in accordance with the hypothesis that all of them inherited the XA17 allele through selection. In this study, pollen genotype selection was extremely effective, and its effect persisted in the second sporophytic generation. Received: 19 November 1999 / Accepted: 30 April 2000     

Potential damage by Acromyrmex ant species in pine plantations in southern Brazil

1. Acromyrmex species primarily attack Pinus taeda plantations during the first months after planting, with more intense damage in the first 30 days.
2. We evaluated potential damage by Acromyrmex crassispinus and Acromyrmex subterraneus in 30-day-old P. taeda plantations in southern Brazil by assessing the number of attacked plants and the distance reached by each colony, along with the losses in plant development resulting from different levels of defoliation.
3. Both species were significant pests in newly established pine plantations. A single colony can attack a substantial number of seedlings (up to 453 seedlings). Colonies with larger nests attacked more plants and ranged farther; A. crassispinus reached 59 m and A. subterraneus 87 m.
4. After 10 years, no significant losses in plant development were seen in seedlings defoliated less than 50% at 30 days after planting. But when defoliation of the young plants attained 75%, 100%, and 100%, including the cut of the apical meristem, volume losses reached 32%, 37%, and 43%, respectively.
5. Leaf-cutting ant controls should be carried out in plantations of P. taeda within 30 days of planting to avoid attacks. When seedlings are defoliated more than 75%, they should be replanted to avoid future losses in final wood volume.

     

Wyerone Acid Phytoalexin Synthesis and Peroxidase Activity as Markers for Resistance of Broad Beans to Chocolate Spot Disease

Infection of broad bean (Vicia faba L.) leaves with Botrytis fabae (Sard.) resulted in wyerone acid phytoalexin accumulation and increase in peroxidase activities in resistant and susceptible broad bean cultivars. Rapid wyerone acid accumulation was observed in leaves of resistant cultivars that reached levels greater than twofold of the susceptible cultivars. Following infection of broad bean cultivars, either resistant or susceptible to B. fabae, wyerone acid synthesis in resistant cultivars peaked at 5 days then declined, whereas in susceptible cultivars synthesis gradually decreased. While the ethanol‐extract of wyerone acid significantly reduced B. fabae spore germination with increased concentrations, the phytoalexin had no effect on in vitro B. fabae germ‐tube growth. Peroxidase activity in infected leaves of resistant was 10 times higher than the susceptible cultivars and eight times higher in uninfected leaves of the resistant than the susceptible cultivars. Peroxidase activity in infected leaves was greater than twofold in resistant cultivars and less than twofold in susceptible cultivars, when compared with uninfected leaves. The ratio of peroxidase activity in infected to uninfected leaves increased over time in susceptible cultivars but remained unchanged in resistant cultivars. Peroxidase activity in uninfected and infected leaves and wyerone acid biosynthesis in infected broad bean plants were successfully used to ascertain the resistance and susceptibility of four broad bean cultivars to B. fabae. Using wyerone acid synthesis and peroxidase activity as preliminary markers for resistance of broad bean to chocolate spot disease, caused by B. fabae, is suggested in this study.     

Enhancement of Defence Responses against Bayoud Disease by Treatment of Date Palm Seedlings with an Hypoaggressive Fusarium oxysporum Isolate

Pretreatment of date palm seedlings with an hypoaggressive Fusarium isolate (AHD) protected them partially from further infection by Fusarium oxysporum f.sp. albedinis (Foa), the Bayoud disease pathogen. No mortality occurred during 2–3 months of incubation in plants pretreated with AHD, as opposed to aggressive isolate (ZAG) inoculated controls where up to 100% mortality was observed 15–30 days after inoculation. Such protection involved biochemical interactions between the host plant and AHD since no direct competition or antagonism was revealed between AHD and ZAG. The examination of the accumulation of phenolics and peroxidase activity, two parameters previously reported to be involved in date palm resistance to Foa, indicated that the response to AHD was correlated with the ability of pretreated date palm tissues to establish a faster defence response in the roots of both susceptible and resistant cultivars. Plants pretreated with AHD accumulated higher amounts of phenolics, mainly non‐constitutive hydroxycinnamic acid derivatives, which play a crucial role in date palm defence against Foa, as previously described by our group. These compounds were accumulated along with the constitutive caffeoylshikimic acids. A faster induction of peroxidase activity in response to Foa was also recorded in plants pretreated with AHD. Given the multi‐component nature of these induced responses, AHD could be part of integrated disease management strategies for a sustainable control of the palm tree Bayoud disease.