Stomatal characteristics of Eucalyptus grandis clonal hybrids in response to water stress

This study describes the stomatal response occurring during water stress and subsequent recovery of three Eucalyptus grandis clonal hybrids. The aim was to investigate the degree to which stomatal conductance (g _s) and stomatal density differ between the clonal hybrids across seasons and in response to water stress. Plants from one E. grandis × E. camaldulensis (GC) and two E. grandis × E. urophylla (GU1 and GU2) clones were grown for 18 months in 80 l planting bags. Plants were subjected to three watering treatments: control (100% field capacity), chronic water stress (maintained at 15% of field capacity) and acute water stress (cyclic water stress, where water was withheld until leaf wilting point, and a subsequent period of recovery followed). Stomatal conductance was measured after 6, 12 and 18 months growth. At 12 months of age, the recovery of g _s 1, 2 and 7 d after rewatering (following acute water stress) was further investigated. The GC hybrid showed consistently higher g _s than the GU clones at each measurement period. Stomatal conductance was 24–66% higher during winter (after 12 months growth) than during summer. The recovery of stomatal conductance from acute water stress was more rapid in the GC clone than the GU clones. Chronic water stress was shown to decrease g _s in GU clones by up to 70%, but not in the GC clone. Water stress did not affect stomatal density or size. Remarkably, stomata were absent from the adaxial leaf surface of clone GU1 leaves, but not from the leaves of the other E. urophylla hybrid cross (GU2). Total biomass of the GC clone was significantly greater at 9 months growth, but after 18 months growth the GU1 clone had attained greater biomass accumulation (although not significantly). Measurement of g _s, transpiration, stomatal density and total biomass in the GU1 clone indicated stomatal sensitivity to water stress, a favourable trait during periods of drought. The differing growth strategies of the GU and GC clones could be partially explained by their differences in stomatal sensitivity in response to water stress.     

Testing of selected South African <Emphasis Type="Italic">Pinus</Emphasis> hybrids and families for tolerance to the pitch canker pathogen, <Emphasis Type="Italic">Fusarium circinatum</Emphasis>

Plantations of Pinus spp. constitute approximately 50% of the South African forestry industry. The first aim of this study was to develop a reliable inoculation technique to screen Pinus spp., for tolerance to infection by F. circinatum, which threatens pine forestry in South Africa. Inoculation of branches was compared with stem inoculations and we considered the number of branches or trees required to obtain statistically significant results. Furthermore, variation in the susceptibility of some Pinus families, clones and hybrids was considered. Results showed that branch inoculations were closely correlated with those from stem inoculations, and that it is important to consider branch and stem diameters when assessing susceptibility of trees. Subsequent trials using branch inoculations showed significant differences in F. circinatum tolerance amongst a range of pine species and hybrids of potential interest to forestry in South Africa. Significant differences in susceptibility were also found among clones of two P. radiata families. The most tolerant trees were P. elliottii × caribaea and P. patula × oocarpa hybrids, while the most susceptible species were P. patula, P. greggii and hybrids of these two. This is the first trial considering the susceptibility of Pinus hybrids, Pinus clones and some P. patula provenances, and the results indicate excellent potential for breeding for tolerance to pitch canker in South Africa. Application The accurate selection of disease tolerant planting stock for the South African forestry industry is crucially important for the continued sustainability of this important industry. The work described here provides valuable information on an artificial inoculation technique that will assist the industry in screening trees for tolerance to the pitch canker fungus, F. circinatum. It also provides some indication of the relative susceptibility of a number of Pinus spp., hybrids and families currently being evaluated in the country.     

Botryosphaeriaceae associated with Eucalyptus canker diseases in Colombia

The identities of Botryosphaeriaceae causing cankers on Eucalyptus in Colombia were investigated using morphological and DNA sequence comparisons. The pathogenicity of the species was also assessed on 42 Eucalyptus grandis clones planted at four different sites. Two species of the Botryosphaeriaceae were found to occur on E. grandis in Colombia. Neofusicoccum ribis was the more common species, and also the most pathogenic. Botryosphaeria dothidea was found only in one zone of Colombia, and was also less pathogenic than N. ribis. These two species could be distinguished easily based on DNA sequences of the ITS1/ITS2 rDNA region and EF1‐α, in addition to conidial sizes. Significant differences in resistance of clones to these pathogens were also evident from the various trials.     

Quambalaria eucalypti found on Eucalyptus in Indonesia

The Eucalyptus plantation industry in Indonesia has expanded rapidly during the last few decades. During routine nursery disease surveys, symptoms of a leaf and shoot blight disease were detected on Eucalyptus mother plants. Isolates were obtained from symptomatic tissues and identified using DNA sequence analyses. Phylogenetic analyses showed that the isolates were those of Quambalaria eucalypti. Pathogenicity tests were conducted with isolates of Q. eucalypti on clones of E. pellita and E. grandis × E. pellita hybrids. These resulted in symptoms similar to those observed on naturally infected plants. Eucalyptus genotypes tested showed variation in their susceptibility, highlighting the potential to select and breed for resistance and thus to manage future outbreaks of the disease. This is the first report of the pathogen in Indonesia as well as in Southeast Asia.     

A serious new wilt disease of Eucalyptus caused by Ceratocystis fimbriata in Central Africa

In a recent survey of Eucalyptus clones in the Republic of Congo, Central Africa, a serious wilt and die-back disease of two different hybrid clones was observed. Affected trees ranged in age from approximately 6 months to 4 years. Isolations from symptomatic plant material consistently yielded a Ceratocystis species. On the basis of morphology and sequence data this fungus was identified as Ceratocystis fimbriata, a well-known wilt and canker pathogen of many economically important plants. The Eucalyptus isolates were compared with other Ceratocystis spp. based on sequence data generated from the ITS and 5.8S region of the rRNA operon. The results confirmed the identity of the Ceratocystis isolates from Eucalyptus as C. fimbriata and showed that they group with other C. fimbriata isolates from Brazil, South Africa and Europe. Inoculations on young Eucalyptus plants were conducted in the greenhouse and all three of the Congolese isolates tested, produced typical lesions in the bark and xylem. This study represents the first report of C. fimbriata as a pathogen of Eucalyptus in Africa. This is a serious new disease that will require considerable study in order to ensure that losses, caused by C. fimbriata, do not continue.     

Lesion size induced by Chrysoporthe fungal pathogens varies between Eucalyptus species and geographic locations in Zambia

Chrysoporthe canker disease is one of the serious threats facing planted Eucalyptus in southern Africa. In order to manage this disease, planting of less susceptible species of Eucalyptus has been promoted in many countries but it is not clear which species are less susceptible for Zambian environments. The purpose of this study was to test the response of different Eucalyptus species to Chrysoporthe fungal pathogens under field conditions to identify the least susceptible species to the disease. Two-year-old Eucalyptus trees (E. grandis, E. camaldulensis, E. cloeziana and E. tereticornis) growing in low- and high-rainfall locations were inoculated separately with three Chrysoporthe fungal species (C. austroafricana, C. syzygiicola and C. zambiensis) and the induced lesion size was measured six months after inoculation. Analysis of variance revealed that lesion size differed significantly between Eucalyptus species (F_3,948 = 1 978.8, p < 0.001). Lesion size on Eucalyptus species increased in the order E. camaldulensis < E. teriticornis < E. cloeziana < E. grandis. On average, trees growing in the higher-rainfall site produced significantly larger lesions than trees growing in the low-rainfall site (F_1,948 = 565, p < 0.001), suggesting that location influences disease severity. Smaller lesions on E. camaldulensis indicate lower susceptibility to Chrysoporthe canker disease, providing an opportunity to use this species in the selection and breeding for resistant Eucalyptus genotypes for plantation development in high disease-risk sites in Zambia.     

Susceptibility of Eucalyptus grandis to Cryphonectria cubensis

Artificial inoculation of rooted Eucalyptus grandis cuttings with Cryphonectria cubensis in the glasshouse showed that significant differences exist in the susceptibility of various Eucalyptus clones to the pathogen. A 61.3% difference in susceptibility was observed between the most resistant and susceptible group of clones. Results appeared, however, to have been influenced by genotype × environment interaction.     

Impact of Botryosphaeria,Diplodia and Neofusicoccum species on two Eucalyptus species and a hybrid: From pathogenicity to physiological performance

Species of Botryosphaeriaceae are associated with canker and dieback of Eucalyptus spp. worldwide, but little is known about their effect on the host physiology. The aim of this study was to evaluate the impact of Botryosphaeriaceae isolates from nine species in three genera (Botryosphaeria, Diplodia and Neofusicoccum), previously isolated from eucalypts, on three different Eucalyptus hosts (seedlings of E. nitens, cuttings of E. globulusand of E. globulus× E. cypellocarpa). An approach combining standard pathogenicity trials with evaluation of plant morpho‐physiological parameters was used. The size of the lesions produced revealed differences in fungal aggressiveness and host susceptibility. Isolates of Neofusicoccum kwambonambienseand Diplodia corticolawere the most aggressive, while Botryosphaeria dothidea and Diplodia seriataisolates were the least aggressive. In general, hybrid E. globulus× E. cypellocarpa plants developed smaller lesions, followed by E. nitens and E. globulus. Eucalyptus nitensplants showed minimal modifications on the morpho‐physiological profile when infected, although more severe symptoms and mortality were observed. This is probably due to a more variable genetic background of the plants. However, in general, no direct association between putative fungal aggressiveness and plant physiological disorders could be found. Results suggested that under optimal growth conditions plants manage to cope with pathogen attack and maintain their physiological performance.     

First report of bacterial wilt caused by Ralstonia solanacearum on eucalypts in South Africa

Ralstonia solanacearum, the causal agent of bacterial wilt, has one of the widest host ranges of all phytopathogenic bacteria. This pathogen was first reported on Eucalyptus spp. in the late 1980s in Brazil. Since then, there have been reports of its occurrence on this host in Australia, China and Venezuela. Early in 1997, an 18‐month‐old clonally propagated Eucalyptus grandis × Eucalyptus camaldulensis (GC) hybrid in Zululand, KwaZulu/Natal, showed signs of wilting. The vascular tissue of infected trees was dicoloured and bacterial exudation was produced from cut surfaces. The bacterium was consistently isolated from diseased tissue, purified and identified as R. solanacearum biovar 3 race 1, using the BioLog bacterial identification system. Inoculation trials were conducted on three E. grandis × E. camaldulensis clones (GC515, GC550 and GC505). Clone GC550 displayed wilting after 3 days and all cuttings subsequently died. Clones GC515 and GC505 appeared to be less susceptible with cuttings not showing signs of disease until 7 days after inoculation. After 14 days, 90 and 80%, respectively, of cuttings of these two clones had died. This is the first report of bacterial wilt on Eucalyptus in South Africa.     

Occurrence and pathogenicity of Neofusicoccum parvum and N. mangiferae on ornamental Tibouchina species

Species in the Botryosphaeriaceae are important canker pathogens of woody plants, including Eucalyptus spp. The recent discovery of the Eucalyptus pathogen, Chrysoporthe austroafricana, on ornamental Tibouchina trees raised the question as to whether Tibouchina spp. might be alternative hosts for other Eucalyptus pathogens. Therefore, the aim of this study was to consider whether species of the Botryosphaeriaceae occurring on Eucalyptus spp. might also occur on ornamental Tibouchina spp. Isolations were made from Tibouchina trees in South Africa, New Zealand and Australia. Isolates were identified using morphological characteristics and DNA‐based techniques. Neofusicoccum parvum and N. mangiferae were identified from the samples. Pathogenicity trials on T. urvilleana showed that both species are pathogenic to this host.     

Cryphonectria canker on Tibouchina in Colombia

Cryphonectria canker, caused by Cryphonectria cubensis, has limited the development of new Eucalyptus plantations in tropical and subtropical regions. The pathogen is commonly found on Eucalyptus, but its occurrence on other hosts in the Myrtaceae has also been documented. In this study C. cubensis is reported as the causal agent of a serious canker disease on Tibouchina spp. (Melastomataceae) in Colombia. We used morphological studies, pathogenicity tests on Eucalyptus and Tibouchina, and a phylogenetic study using partial ribosomal DNA sequence data. This is the first record of C. cubensis on a host outside the Myrtaceae.     

Diseases on Eucalyptus species in Zimbabwean plantations and woodlots

Zimbabwe embarked on planting Eucalyptus species in the early 1900s. Based on a robust breeding programme, it has become a major source of seed for other countries in and outside Africa. Tree health surveys conducted on Eucalyptus in some east and southern African countries over the past two decades have revealed several important fungal diseases that were previously not known in the region, but little is known regarding these problems in Zimbabwe. The aim of this study was to identify important Eucalyptus diseases across Zimbabwe's agroclimatic regions. Morphological characteristics and DNA sequence data were used to identify pathogens collected to species level. Widespread stem canker diseases, caused by species belonging to the Botryosphaeriaceae and Teratosphaeriaceae, and leaf spot diseases caused by fungi in the Capnodiales, were identified. Armillaria root and stem rot was restricted to a single site in the Eastern Highlands. Fungi that could cause canker or blue stain of timber were isolated from recently harvested stumps and included species of Ceratocystis and Ophiostoma. This study is the first to identify Eucalyptus pathogens to species level in Zimbabwe and we report for the first time the presence of the stem canker pathogen T. gauchensis in southern Africa. The results will provide a foundation for the formulation of future disease management strategies in the country.     

Wound response of Eucalyptus clones after inoculation with Cryphonectria cubensis

Twenty-five different Eucalyptus grandis clones were artificially wounded and inoculated with a virulent isolate of Cryphonectria cubensis. The capacity of wounds to close through callus production was correlated with the relative susceptibility of these clones to infection by C. cubensis. Clones with the greatest capacity to close wounds were those that were also most tolerant to C. cubensis infection. Those with a lower capacity to close wounds were most susceptible to Cryphonectria canker.     

管芯片技术通量检测杨树溃疡病病菌的应用

正基因芯片技术因能对生物样本进行快速、高通量的定性和定量分析,而在植物病害诊断中也得到普遍的应用[1]。树木溃疡病是指引起木本植物树皮上出现溃疡腐烂等症状的病害[2]。引起杨树溃疡病的真菌种类较多,加之其无性型和有性型多样,所以,关于杨树溃疡病病原菌的报道多有不同。另外,受地理位置、寄主植物关系及分类体系的限制,迫切需要发展不依赖于常规分离培养的病原鉴定新技术。     

Evaluation of fungicides for the control of Botryosphaeria corticola on cork oak in Catalonia (NE Spain)

Botryosphaeria canker of cork oak (Quercus suber L.), caused by Botryosphaeria corticola, is a well-known factor contributing to the decline of this forest species and is a serious disease in the main cork-producing countries of the Western Mediterranean basin. In this work, fungicides with the potential for controlling the disease were selected using a combination of in vitro tests and field trials. Fourteen fungicides in different chemical families were tested in vitro to evaluate their effect on inhibition of mycelial growth. The most effective fungicides were benomyl, carbendazim, cyprodinil + fludioxonil, thiabendazol and thiophanate-methyl; these compounds were then further tested under field conditions. In the field trials, carbendazim and thiophanate-methyl were the most effective fungicides, as shown by efficacy indices, in reducing debarked trunk surface affected by cankers (76% and 50–65%, respectively). These results suggest that the occurrence of Botryosphaeria canker on cork oak can be reduced effectively with the application of selected fungicides after cork removal from the trees.     

Screening wild cherry (Prunus avium) for resistance to bacterial canker by laboratory and field tests

Currently, bacterial canker caused by Pseudomonas syringae is a major cause of dieback and tree death in wild cherry (Prunus avium) plantations. The evaluation of breeding collections is needed to produce less susceptible clones or cultivars. Resistance tests were performed using excised shoots (1 and 2 years old) from 79 clones in the laboratory. A subset of 10 clones was also tested in the field. The clones were inoculated with four to seven isolates of a set of 15 isolates of P. s. pv. morsprunorum, P. s. pv. syringae, P. s. pv. persicae, P. syringae pv. avii and P. fluorescens. In the laboratory tests, older and larger shoots were more susceptible. In the field test, size and age of the shoots were not related to girdling by the bacterial canker. Two‐year‐old shoots were best for clonal discrimination. Correlations between 1 and 2‐year‐old shoots were significant but not high. The isolates varied a lot between experiments, but as the clone × isolate interactions were always low, breeding could thus be facilitated. The ranking of clones was conserved quite well between two laboratory tests, but not between two others. Good agreement was found for the best clones in the laboratory tests and in the field test. However, the two worst clones in the latter were among the best in one laboratory test. At least two independent tests in the laboratory are needed to evaluate resistance/susceptibility of clones. Broad sense heritability for resistance varied from 0.27 to 0.51. Although moderate, such heritability clearly encourages a breeding approach to reduce the problem of bacterial canker.     

Pitch canker ratings of longleaf pine clones correlate with Fusarium circinatum infestation of seeds and seedling mortality in containers

The incidence and severity of pitch canker was rated among 16 clones of mature longleaf pine (Pinus palustris) in a seed orchard and cones were collected from five of those clones across the range of pitch canker disease ratings. Seed were extracted and commercially processed by clone and the percentages contaminated with Fusarium circinatum determined. Seed from each clone were sown either without treatment or with one of three fungicide treatments in a soil‐less mix at a commercial container nursery to evaluate the effects of F. circinatum (syn. F. subglutinans) on seed and seedling survival. The percent of seed with F. circinatum correlated with the pitch canker ratings in the orchard for the year of collection (r = 0.88, p = 0.05) and, when sown without fungicide treatment, with the number of seedlings produced (r = ?0.94, p = 0.01) and with seedling mortality after germination (r = 0.92, p = 0.02). The same orchard clones were more symptomatic of pitch canker through three annual surveys. Fusarium circinatum was isolated from a higher percent of seed from more symptomatic clones and a lower percent of their seed produced plantable seedlings. We propose that removing such clones from seed collections should reduce seedling mortality at the nursery.     

Comparison of terpenes in extracts from the resin and the bark of the resinous stem canker of Chamaecyparis obtusa and Thujopsis dolabrata var. hondae

A monoterpene and 15 diterpenes were isolated from the ethyl acetate extracts of the bark-glued resin from the resinous stem canker ofThujopsis dolabrata var.hondae Makino. A monoterpene (nezukone20) and 4 diterpenes (acetyl torulosol5, acetyl isocupressic acid8, acetyl abietinol11, and 7-methoxytotarol18) were characteristic constituents of the ethyl acetate extracts but were absent in then-hexane extracts from the resinous stem canker ofT. dolabrata var.hondae. These terpenes were first isolated fromT. dolabrata var.hondae andT. dolabrata. The available literature suggests that diterpene18 is a new compound. The resinous stem canker ofChamaecyparis obtusa Endlicher contained larger amounts of manool1,trans-communic acid6, and ferruginol12 and smaller amounts of isocupressic acid7 and abietinol10 than the resinous stem canker ofT. dolabrata var.hondae. The concentration of18 was less than 2% in the extracts, and the resinous stem canker ofC. obtusa lacked this compound. The resinous stem cankers ofC. obtusa andT. dolabrata var.hondae provided extracts 15.6 and 4.96 times, respectively, heavier than the healthy ones. Large differences in the ratios and compositions of terpenes were also observed between the resinous stem canker and the healthy trees. Terpenes isolated from the extracts contained many kinds of diterpene, especially the labdane-type diterpenes, in these diseased trees. These results suggest that the presence of labdane-type diterpenes is closely associated with the resinous stem canker or the causal fungi of this disease.Part of this report was presented at the 50th annual meeting of the Japan Wood Research Society, Kyoto, April 2000     

Future outlook for Pinus patula in South Africa in the presence of the pitch canker fungus (Fusarium circinatum)

Approximately 50% of the area planted to softwood trees in South Africa has been established with Pinus patula, making it the most important pine species in the country. More effort has gone into developing this species for improved growth, tree form and wood properties than with any other species. This substantial investment has been threatened in the last 10 years by the pitch canker fungus, Fusarium circinatum. The fungus infects and contaminates nursery plants and, once transferred to the field, causes severe mortality of young trees in the first year after establishment. Although nurserymen have some control of the disease, it is recognised that the best long-term solution to mitigate damage because of F. circinatum infection is to identify tolerant species, clones and hybrids for deployment in plantations in the future. Research has shown that alternative species such as P. tecunumanii, P. maximinoi and P. elliottii are suitable for warm sites. Pine hybrids, particularly between P. patula and the high-elevation sources of P. tecunumanii, appear to be a suitable replacement on subtemperate and temperate sites. Although these alternative species and hybrids are more sensitive to subfreezing temperatures than P. patula, their planting range can be increased by including cold tolerance as a selection criterion. Future breeding efforts will most certainly focus on improving the tolerance of pure P. patula to F. circinatum, which can be achieved by identifying specific family crosses and tolerant clones. The commercial deployment of disease-tolerant control-pollinated P. patula and hybrid families will most likely be established as rooted cuttings, which requires more advanced propagation technology. In the long term, new seed orchards comprised of P. patula clones tolerant to F. circinatum could be used to produce seed for seedling production.     

Effect of environment on the response of Eucalyptus clones to inoculation with Cryphonectria cubensis

The ascomycete Cryphonectria cubensis causes severe losses in Eucalyptus plantations in South Africa and selection programmes for disease tolerance are necessary. The aim of this study was to use two C. cubensis inoculation trials, planted at different locations to assess the disease susceptibility of the clones and the effect of the environment on disease development. These two trials consisted each of 21 Eucalyptus clones (E. grandis, E. grandis × camaldulensis and E. grandis × urophylla). All trees were inoculated with a single virulent strain of C. cubensis and lesion widths measured 6 and 12 months after inoculation. Clones differed significantly in their tolerance to C. cubensis. Further, disease severity differed depending on the geographical location of the trial. A significant clone × locality (genotype × environment) interaction was observed. Therefore, screening for disease resistance should take place only in areas where clones will be commercially grown.