Walnut Orchards Research Articles

Development of a Multiplex PCR Assay to Detect Neofusicoccum parvum and Botryosphaeria dothidea in Walnut.

In walnut orchards, frequent symptoms of cankers and dieback (fruit blight, twig and branch cankers up to tree death) are caused by different agents, in particular by Botryosphaeriaceae, primarily Neofusicoccum parvum and Botryosphaeria dothidea. This study aimed at developing a sensitive, rapid, specific and internally controlled multiplex PCR assay for the detection of these species. The ability of the multiplex PCR, with an internal inhibition control (i.e. E. coli DNA), to specifically and successfully detect members of the two targeted species was validated using 11 different isolates for each fungal target (inclusivity) and 20 tree-associated fungal species different from B. dothidea or N. parvum (exclusivity). Applicability to plant materials was further investigated and showed an absence of amplification for asymptomatic husk or twig samples while the amplification profiles of symptomatic tissues ranged from no amplification to amplification of both species, in correlation with the observed fungal contamination level. In conclusion, we developed a rapid diagnostic tool for simultaneous detection of two major fungal pathogens of walnut. Although this protocol was tailored for walnut husks and twigs, applicability to any plant sample contaminated by these pathogens can be considered.

Aspects of introduction and prospects for the use of Juglans L. representatives in Central Russia

Background. Human health security is an important goal of contemporary society. The combination of beneficial properties in Juglans nuts makes them one of the most valuable plant genetic resources, capable of ensuring a wholesome supply of nutrients required by the human organism. The Persian walnut (Juglans regia L.) has the largest set of useful agronomic traits among the representatives of the genus, except for winter hardiness. Its cultivation in areas farther to the north from the conventional area of distribution for commercial walnut orchards would be possible if Juglans spp. with higher resistance to unfavorable climate conditions could be involved. Interspecies hybridization can be used to increase winter hardiness in walnut. The objective of this study was to analyze the gene pool of Juglans L. and screen its bioresource collection at the arboretum of the Russian Research Institute of Fruit Crop Breeding (VNIISPK) in the context of useful agronomic indicators promising for plant breeding and landscaping. Materials and methods. The study was carried out in the VNIISPK arboretum (Orel Province, 53°00’N, 36°00’E) where 7 Juglans spp. are maintained. Results and conclusion. The results of the screening highlighted J. mandshurica Maxim., J. cordiformis Maxim., and Vekhov’s hybrid (J. cinerea L. × J. mandshurica Maxim.) as the most resistant to adverse environmental effects. They are recommended for use in plant breeding and landscaping programs under the conditions of Central Russia. A seedling of cv. ‘Astakhovsky’ demonstrated the best agronomic indicators among the Persian walnut (J. regia) genotypes. It can be used by breeders to develop new cultivars with useful agronomic traits for the environmental conditions of the region of study.

Behavioral, Physiological, and Molecular Mechanisms Underlying the Adaptation of Helicoverpa armigera to the Fruits of a Marginal Host: Walnut (Juglans regia).

In northwest China, changes in cultivation patterns and the scarcity of preferred hosts have forced Helicoverpa armigera to feed on the marginal host walnut (Juglans regia). However, the mechanisms allowing this adaptation remain poorly understood. Here, we investigated the behavioral, physiological, and molecular mechanisms underlying the local adaptation of this pest to walnut fruits. The green husk and shell generally contained higher levels of phytochemicals than the kernel. Bioassays revealed that the phytochemical-rich green husk and shell were less preferred, reduced larval fitness and growth, and elevated the activity of detoxification enzymes compared to the nutrient-rich kernel, which were further supported by a larger number of upregulated detoxification genes in insects fed green husks or shells based on transcriptome sequencing. Together, these data suggest that P450 genes (LOC110371778) may be crucial to H. armigera adaptation to the phytochemicals of walnuts. Our findings provide significant insight into the adaptation of H. armigera to walnut, an alternative host of lower quality. Meanwhile, our study provides a theoretical basis for managing resistance to H. armigera larvae in walnut trees and is instrumental in developing comprehensive integrated pest management strategies for this pest in walnut orchards and other agricultural systems.

Leveraging Digital Technologies for Carbon Footprint Tracking in Perennial Cultivations: A Case Study of Walnut Orchard Establishment in Central Greece

The present paper aims to quantify the carbon emissions associated with the establishment of 15 walnut orchards (“Juglans californica”) in the greater area of Magnisia, Greece, with the use of a carbon footprint tool interconnected to a Farm Management Information System. The data collection spanned the first five years following the planting of the trees, providing a comprehensive view of the emissions during this critical establishment phase. Over the five-year period examined (February 2019–December 2023), the results revealed net carbon emissions amounting to 13.71 tn CO2 eq ha−1, with the calculated emissions showing an increasing trend from the first year through the fifth year. Scope 1 (7.38 tn CO2 eq ha−1) and Scope 2 (3.71 tn CO2 eq ha−1) emissions emerged as the most significant, while irrigation (drip irrigation) and fertilizing practices were identified as the highest contributors to emissions. This study highlights the significance of using integrated digital tools for monitoring the performance of cultivations rather than standalone tools that are currently widely available. Integrated tools that incorporate various applications simplify data collection, encourage accurate record-keeping, and facilitate certification processes. By automating data entry and calculations, these tools reduce human error during agricultural carbon management and save time; thus, the integration of digital monitoring tools is vital in improving data accuracy, streamlining certification processes, and promoting eco-friendly practices, crucial for the evolving carbon market.

Genetic diversity and population structure of Botryosphaeria dothidea and Neofusicoccum parvum on English walnut (Juglans regia L.) in France

Botryosphaeriaceae species are the major causal agents of walnut dieback worldwide, along with Diaporthe species. Botryosphaeria dothidea and Neofusicoccum parvum are the only two Botryosphaeriaceae species associated with this recently emergent disease in France, and little is known about their diversity, structure, origin and dispersion in French walnut orchards. A total of 381 isolates of both species were genetically typed using a sequence-based microsatellite genotyping (SSR-seq) method. This analysis revealed a low genetic diversity and a high clonality of these populations, in agreement with their clonal mode of reproduction. The genetic similarity among populations, regardless of the tissue type and the presence of symptoms, supports the hypothesis that these pathogens can move between fruits and twigs and display latent pathogen lifestyles. Contrasting genetic patterns between N. parvum populations from Californian and Spanish walnut orchards and the French ones suggested no conclusive evidence for pathogen transmission from infected materials. The high genetic similarity with French vineyards populations suggested instead putative transmission between these hosts, which was also observed with B. dothidea populations. Overall, this study provides critical insight into the epidemiology of two important pathogens involved in the emerging dieback of French walnut orchards, including their distribution, potential to mate, putative origin and disease pathways.

Soil Fertility Dynamics and Identify the Most Limiting Nutrients Affecting Walnut Productivity in Nepal

This study aimed to assess the soil nutrient status in walnut orchards of different ages in Jajarkot district, Nepal, to understand soil fertility dynamics and identify the most limiting nutrients affecting walnut productivity. The research employed a randomized complete block design (RCBD) with three treatments representing different age groups of walnut orchards (1-5 years, 6-10 years, and 11-15 years), each replicated seven times across various municipalities. The study was conducted in Jajarkot district, Karnali Province, Nepal, encompassing municipalities including Nalgad, Junichadey, and Barekot, from March to April 2023. Soil samples were collected from multiple depths (1, 2, and 3 feet) in each orchard and analyzed for pH, soil organic matter (SOM), total nitrogen, available phosphorus, and available potassium. Data analysis included descriptive statistics, one-way analysis of variance (ANOVA), and correlation analyses to explore relationships between soil parameters and orchard age. The study showed that significant variations were observed among different age groups of walnut orchards for soil pH, SOM, nitrogen, phosphorus, and potassium levels. Soil pH decreased with orchard age, while SOM, nitrogen, and phosphorus tended to increase with orchard age. Phosphorus was identified as the most limiting nutrient across all sampled soils, followed by nitrogen and potassium. Moreover, strong correlations were found between orchard age and soil N (r = 0.894, p < 0.01) and P (r = 0.776, p < 0.01), underscoring age-dependent nutrient dynamics. The study highlights the critical role of orchard age in shaping soil nutrient dynamics in walnut orchards. Older orchards exhibited higher levels of SOM, nitrogen, and phosphorus, indicating the accumulation of organic matter and nutrients over time. Phosphorus emerged as the primary limiting nutrient, essential for root growth, flowering, and fruiting in walnut trees. These findings underscore the importance of targeted fertilization strategies to optimize soil fertility and sustain long-term walnut productivity in the region.

Study on Soil Fertility Characteristics of Walnut Orchards with Different Parent Materials and Soil Types in Gyaca County, Tibet

Walnut trees on the Tibetan Plateau have sustainable and nutritious characteristics. They grow naturally without the use of any chemical fertilizers and pesticides. Therefore, the soil fertility of walnut orchards is a key factor influencing walnut yield and quality. This experiment measured and analyzed the basic soil nutrient content, microbial biomass carbon (MBC), nitrogen (MBN), phosphorus (MBP), and leaf nutrient indicators in five types of representative walnut orchard soil parent materials in Gyaca County to clarify the fertility characteristics of different soil types and their correlations with walnut nutrients, providing a basis for the sustainable development of the plateau walnut industry. The results showed that there were significant differences in soil organic matter (SOM), total nitrogen (TN), available nutrient contents, and microbial activity among soil types with different parent materials. The comprehensive representation of available nutrient levels in the soils is as follows: gray-cinnamon soil and calcareous alluvial soil > mountain meadow soil > plateau prairie soil and brush prairie soil. The average MBC, MBN, and MBP contents in the five soils were 368.8 mg/kg, 28.4 mg/kg, and 23.6 mg/kg, respectively, and the content in the topsoil (0–20 cm) was higher than that of the subsoil (20–40 cm). The total nitrogen (PN), total phosphorus (PP), and total potassium (PK) contents in walnut leaves were found to be closely related to the soil physicochemical properties and microbial activity, among which five indicators, including soil TN, available nitrogen (AN), SOM, MBC, and slowly available potassium (SAK), had the greatest impact on leaf nutrient levels. These findings are valuable for the future management and development of walnut orchards. Therefore, for different soil parent materials and soil types, improving the soil fertility and actively applying nitrogen-rich organic fertilizers should be prioritized to improve the yield and quality of Tibetan Plateau walnuts.

Linking soil organic carbon characteristics, nutrient stoichiometry, and microbial community to eco-enzymatic stoichiometry within aggregates in different aged walnut plantations

Research on the variations of microbial attributes, C and nutrient properties, eco-enzymatic activities and their stoichiometry in different aged walnut orchards is essential for the sustainable development of walnut gardens. Here, four walnut orchards of various ages (0-, 7-, 14-, and 21-years) were selected in Hebei province, China, to evaluate the temporal changes in the above-mentioned indices within aggregates based on thermal gravimetric analysis, phospholipid fatty acid analysis and fluorometric assays. Results revealed that as the walnut plantation ages or aggregate sizes increased, the quantity and thermal stability of organic C exhibited increasing and decreasing trends, respectively. Long-term walnut plantation could increase C- and P- acquiring enzyme activities, and decrease N-acquiring enzyme activities in larger aggregates. Eco-enzymatic stoichiometry analyses demonstrated that the microbial C and P co-limitation increased with aggregate sizes or walnut plantation ages, although long-term walnut planting (14- and 21-years) and larger aggregates (>0.25 mm) provided more and easily available C resources for microbes. The aggravated C limitation (or P limitation) could be ascribed to the increased the ratio between microbial biomass C and organic carbon content (or the increased fungi/bacteria and soil N/P ratios) in the elder walnut plantations or larger aggregates. Overall, the study's results can provide several valuable insights (e.g., the old orchards can appropriately apply more P fertilizer) into the sustainable development of walnut gardens from the perspective of microbial nutrient demand.

Evaluation of fungicides for management of Botryosphaeriaceae associated with dieback in Australian walnut orchards

Dieback of fruiting spurs, stems and branches of walnut trees (Juglans regia L.), caused by Botryosphaeriaceae, is widespread in walnut orchards in Australia. Five species of Botryosphaeriaceae (Diplodia seriata, Dothiorella omnivora, Neofusicoccum macroclavatum, N. parvum, and Spencermartinsia viticola) were recovered from the Australian walnut orchards in a previous study, with D. seriata and N. parvum being the most prevalent. The present study evaluated inhibitory effects of ten fungicides on mycelium growth of those five species and on conidium germination of D. seriata and N. parvum. It investigated the preventative and curative efficacy of selected fungicides on disease incidence in glasshouse and field trials. In vitro experiments showed that nine of the fungicides reduced mycelium growth, and all ten inhibited conidium germination, but to varying extents. Tebuconazole, prochloraz manganese chloride, fluazinam, fludioxonil and pyraclostrobin were the most effective for inhibiting mycelium growth (EC50 < 0.14 µg a.i. mL-1), whereas pyraclostrobin, fluxapyroxad, fluopyram, penthiopyrad and tebuconazole were the most effective for inhibiting conidium germination (EC50 < 2.2 µg a.i. mL-1). In planta experiments with five fungicides confirmed that preventative treatments had greater efficacy than curative treatments. A field trial with four commercial fungicide formulations demonstrated that tebuconazole and tebuconazole + fluopyram provided protection of walnut trees for the longest period. The field trial also confirmed the efficacy of pyraclostrobin and the inhibitory effect of fluazinam. This study is the first in Australia to evaluate fungicides in different classes and with different modes of action for efficacy against Botryosphaeriaceae recovered from walnut orchards in Australia, and provides a wider selection of active ingredients for a fungicide rotation programme than that which is currently available to the Australian walnut industry.

Phytophthora and Phytopythium species associated with walnut dieback in plantations of northern Patagonia, Argentina

AbstractThe walnut industry in Argentina has grown in recent years, especially in northern Patagonia. However, this expansion has also brought new challenges, including the emergence of diseases threatening the health and productivity of walnut plantations. One of the most important diseases affecting walnut crops worldwide is ‘ink disease’ or ‘walnut dieback'. The aim of this study was to characterize ink disease, identify the aetiological agents, and estimate its incidence in walnut orchards of northern Patagonia, Argentina. Symptomatic trees were observed in the seven surveyed walnut orchards, showing a range of symptoms, including chlorosis and wilting, stem cankers and desiccation. In vivo assay and molecular analyses revealed that not only Phytophthora (P.) but also Phytopythium (Ph.) species are the causal agents of the disease, with P. drechsleri, Ph. mercuriale and Ph. vexans being the most virulent over the commonly used rootstock Juglans hindsii. Notably, Ph. vexans was the most frequently isolated pathogen, suggesting its prevalence and potential impact on the regional walnut industry. Overall, this study provides critical insights into the prevalence and impact of Phytophthora and Phytopythium species on walnut health in northern Patagonia. The presence of multiple pathogenic oomycete species raises concerns about the health of walnut orchards, as well as the potential risk of pathogen transmission to other fruit crops in the region. The findings will help growers to develop better cultural practices and implement measures to guarantee the sustainability of the walnut industry in Patagonia.

Morphological and pomological assessments of seedling-originated walnut (Juglans regia L.) trees to select the promising late-leafing genotypes.

In many parts of the world, including Iran, walnut (Juglans regia L.) production is limited by late-spring frosts. Therefore, the use of late-leafing walnuts in areas with late-spring frost is the most important method to improve yield. In the present study, the phenotypic diversity of 141 seedling genotypes of walnut available in the Senejan area, Arak region, Markazi province, Iran was studied based on morphological traits to obtain superior late-leafing genotypes in the cropping seasons of 2022 and 2023. Based on the results of the analysis of variance, the studied genotypes showed a significant variation in terms of most of the studied morphological and pomological traits. Therefore, it is possible to choose genotypes for different values ​​of a trait. Kernel weight showed positive and significant correlations with leaf length (r = 0.32), leaf width (r = 0.33), petiole length (r = 0.26), terminal leaflet length (r = 0.34), terminal leaflet width (r = 0.21), nut length (r = 0.48), nut width (r = 0.73), nut weight (r = 0.83), kernel length (r = 0.64), and kernel width (r = 0.89). The 46 out of 141 studied genotypes were late-leafing and were analyzed separately. Among late-leafing genotypes, the length of the nut was in the range of 29.33-48.50mm, the width of the nut was in the range of 27.51-39.89mm, and nut weight was in the range of 8.18-16.06g. The thickness of shell was in the range of 1.11-2.60mm. Also, kernel length ranged from 21.97-34.84mm, kernel width ranged from 21.10-31.09mm, and kernel weight ranged from 3.10-7.97g. Based on important and commercial traits in walnut breeding programs, such as nut weight, kernel weight, kernel percentage, kernel color, and ease of kernel removal from nuts, 15 genotypes, including no.92, 91, 31, 38, 33, 18, 93, 3, 58, 108, 16, 70, 15, 82, and 32 were superior and could be used in walnut breeding programs in line with the introduction of new cultivars and the revival of traditional walnut orchards to commercialize them.

An exploratory analysis of the structure of tetranychid and phytoseiid assemblages in walnut orchards in California.

Spider mites were considered secondary pests of walnut production in California, under the control of phytoseiid predators. Due to increased importance as walnut pests in recent decades there is renewed interest in the structure and function of the associated phytoseiid assemblage. In this study we report the results from a 3 year survey of the tetranychid and phytoseiid assemblages in walnut orchards in the Central Valley of California. The survey was conducted to determine the range and dominance of web-spinning Tetranychus species present, to investigate the species richness and dominance of the phytoseiid species present, and to explore whether patterns of variation in the relative abundance of phytoseiid species could be explained by one or more explanatory variables. Tetranychus urticae was the dominant spider mite in all growing regions and years with T. pacificus and T. turkestani also present in orchards in the southern San Joaquin Valley. Phytoseiid species richness declined with latitude among the three walnut growing regions and of the 13 species found Amblyseius similoides, Euseius stipulatus, Galendromus occidentalis and Typhlodromus caudiglans were the most abundant and widespread species present. Mean proportional abundance significantly increased from early (mid May-July) to late (August-mid October) season and from southern to northern growing regions for Type II and IV predators, but significantly decreased from early to late season and from southern to northern growing regions for Type III predators. The mean proportional abundance of Type II predators, particularly G. occidentalis, significantly increased and that of Type III predators significantly decreased with mean Tetranychus density in individual orchards. The current survey provides a more in-depth analysis of mite assemblages in walnut orchards in California and can be used to better inform adaptive management strategies for integrated mite management in the future.

Analysis of the Impact of Agricultural Production Type and Traffic on Nitrogen Oxide Emissions

The protection of air from pollution is one of the basic principles of sustainable development, and this article analyzes the joint impact of various types of agricultural production and traffic on nitrogen oxide (NOx) emissions. The goal of this research is the development of a new methodology for assessing the combined impact of different types of agricultural production, traffic load, and traffic flow structure on the emission of nitrogen oxides. For the analysis of the joint impact of traffic and agricultural production, a new impact assessment methodology was developed, which is based on two selected impact prediction models. The following two models were used to predict the impact: the BioGrace model and the microsimulation model VISSIM. The developed methodology was applied to a case study in real field conditions. The results showed that the difference in the emission of NOx gases, depending on the type of agricultural production, indicates a reduction in NOx emissions in the walnut–winter-wheat system by 30%, compared to the production of winter wheat in an open field. The statistical analysis showed that there is no statistically significant difference between NOx emissions for different vehicle arrival scenarios for the same traffic load. The results of the joint impact show that the lowest NOx emission was found in walnut orchards and counted traffic. In this example, the difference between the one-hour emission load of three different agricultural types of production and traffic is not large; however, in the cumulative load, it can lead to significant differences. Emphasizing the importance of adopting new agricultural practices and the impact of traffic and the share of freight and heavy goods vehicles in agricultural production, we position them as a proactive and sustainable response to the critical ecological and agricultural challenges highlighted in European and Croatian policy.

The Effect of Applying Mineral Fertilizers Through Drip Irrigation and Furrow Method on the Movement of Mobile Nutrients in the Soil and the Growth, Development, and Accumulation of Nutrients in the Walnut Leaves

A task was set to reveal the effect of water-soluble fertilizers of new composition and quality on the content of available nutrients at different depths of the soil, depending on the movement of the irrigation water, irrigation time, as well as its effect on the growth, yielding capacity and the quality of fruit in commercial size walnut orchards. The research has shown that the movement of nutrients in the soil is completely dependent on the movement of water when applying water-soluble complex fertilizers with drip irrigation. Therefore, the amount of the water should be adjusted so that the nutrients are available to the plant’s root system at all depths, improving the growth, and increasing the content of the basic nutrients in the leaves.Fertilizers with less water-solubility are mainly preserved at a 0 cm - 30 cm depth. When the root system grows and spreads to the inter-row spaces, it is appropriate to periodically fertilize also the inter-row spacing by the furrow method.

Economic and Profitability Analysis of Walnut Production in Kashmir Valley, India

The Jammu and Kashmir union territory is the largest producer of walnuts in India, and this crop provides an important source of livelihood for many farmers. This study aims to measure the economic efficiency and profitability of walnut orchards and explore constraints in cultivation in the Kashmir Valley. It relies on a crosssectional database collected from 240 walnut growers in the study area during the 2018/19 production period. Results reveal that walnut cultivation is highly laborintensive as it incurs 80 percent of total production costs. The cost-benefit ratio of 1:5.35 per hectare indicates better economic prospects for the walnut industry in Kashmir Valley. The factors affecting productivity include farmyard manure, labor, chemical fertilizers, plant density, women participation, and information. The regression coefficients of production analysis, marginal value product, and marginal factor cost ratio indicate that there is ample scope for the expansion of walnut cultivation in the research area. However, walnut growers are confronted by several problems that tend to be location specific. The study calls for policy intervention concerning improved access to extension services, credit, and farmer training programs to boost walnut production in the study region.

Cost–Benefit Evaluation of Walnut and Jujube Orchards under Fruit Tree–Crop Intercropping Conditions in Southern Xinjiang

Fruit planting plays an essential role in achieving zero hunger, ensuring food security, and increasing the income of local people. As one of China’s principal fruit-planting areas, southern Xinjiang possesses more than 80% of the total fruit-planting areas of Xinjiang. However, the spatial distribution, extent, and types of fruit trees remain unknown, generating uncertainty in calculating the economic benefits of orchards in this area. In this paper, we used walnut and jujube orchards under fruit tree–crop intercropping conditions in the Hotan Oasis in southern Xinjiang as the research object. Walnut and jujube orchards were precisely characterized using object-oriented and decision tree classification methods. Then, the economic benefits for farmers were estimated based on field surveys and statistical data. From 2003 to 2020, the area of jujube orchards rapidly increased from 1.91 × 103 ha to 33.59 × 103 ha, while that of walnut orchards steadily increased from 18.05 × 103 ha to 40.24 × 103 ha. The expansion areas of walnut orchards were mainly transformed from other orchards in the oasis, while the expansion areas of jujube orchards mainly originated from newly cultivated land in the desert. The increases in areas and yields largely offset the increase in planting costs and the decrease in purchase prices, resulting in an increasing trend in the total net income of the orchards. From 2003 to 2020, the total net income of walnut orchards increased by 68.96%, and the total net income of jujube orchards significantly increased by 23.37 times. However, the output/input ratios of walnut orchards under intercropping conditions were approximately two times higher than those of jujube orchards under monocropping conditions. The increase in investment slightly offset the decreases in yields and purchase prices, resulting in increases of 5.24% and 18.64% in the output/input ratios of walnuts and jujubes, respectively, in orchards exhibiting standardized cultivation. In the future, it is necessary to significantly expand the area of orchards involving standardized cultivation and improve the yield and quality of fruits, thereby increasing the yields and income levels of local farmers.

Pathogenicity and progression of Botryosphaeriaceae associated with dieback in walnut orchards in Australia

AbstractDieback of the fruiting spurs of walnut (Juglans regia) results in yield losses in walnut orchards in Australia. Botryosphaeriaceae spp. have been implicated as the causative agents, with Diplodia seriata and Neofusicoccum parvum reported as the most prevalent species. Pathogenicity assays demonstrated inter- and intra-species variation with N. parvum more virulent than D. seriata. Both species were re-isolated beyond the dieback lesion. At 12 months post-inoculation on one-year-old trunks, N. parvum and D. seriata were re-isolated at 31.5 and 9 mm respectively beyond lesion edges. The lesion length as well as pathogen progression beyond the lesion increased over time. The pathogen progressed faster acropetally than basipetally. Following wound healing at the site of inoculation, and in the absence of any external lesion, both pathogens continued to move through the xylem, causing internal discolouration of the vascular tissues. Both mycelia and conidia were pathogenic, with mycelia showing higher infection rates. There was a strong linear relationship between the dieback length caused by the two types of inocula. As little as two conidia were able to infect wounded tissues. Although wounds were susceptible to infection for more than 4 months post-wounding for N. parvum and 1 month for D. seriata, the highest disease incidence occurred in the first week following wounding. The implications for the walnut industry for appropriate control strategies are discussed.

Influence of climate factors on population density and damage of the leopard moth, Zeuzera pyrina L., in walnut orchards, Iran.

The effect of climate factors (temperature, humidity, precipitation, and frost days) on the population changes, damage, and infestation area of the leopard moth, Zeuzera pyrina L., was studied during 2006-2018 in four parts of Iran including Saman, Arak, Najaf-abad, and Baft. For trend analysis, the Mann-Kendall test was run on time series data of both climate and pest population. According to the results, the annual mean (Kendall's statistics, T = 0.64 and 0.48), annual minimum (T = 0.60 and 0.42), and January mean (T = 0.64 and 0.61, respectively) temperatures showed increasing trends in Saman and Najaf-abad. Moreover, the annual mean minimum and January temperatures (T = 0.41 and 0.45, respectively) in Arak and the annual mean maximum temperature (T = 0.79) in Baft showed increasing trends. The number of frost days/year (Kendall's statistics, T = -0.63, -0.53, -0.32 and -0.37) and annual mean relative humidity (T = -0.43, -0.63, -0.64 and -0.42, respectively) showed decreasing trends in Saman, Arak, Baft, and Najaf-abad stations. Trend analysis indicated significant increases in the mean number of moths caught (T = 0.59, 0.76 and 0.90), the percentage of infested branches/tree (T = 0.66, 0.58, and 0.90), the number of active holes/tree (T = 0.79, 0.55, and 0.68) and the infested areas (T = 0.99, 0.73, and 0.98, respectively) in Saman, Arak and Najaf-abad stations. According to stepwise regression, the mean temperatures of January, autumn, and winter were the most effective variables for increasing Z. pyrina damage and population, while relative humidity and the number of frost days played the major role in reducing it.

Seasonal population dynamics of Aceria erineus (Nalepa) (Acari: Eriophyidae) on Walnut trees in Kashmir

Population fluctuation of Aceria erineus in relation to abiotic factors on different walnut orchards of Kashmir was roving surveyed in 27 agricultural sites in nine districts of Kashmir during May 2018 to April 2019. In each district, three sites were selected randomly and the incidence of the mite was recorded at fortnightly intervals throughout the crop growth period. Field surveys indicated the presence of the pest in the entire surveyed region in varying intensity with maximum incidence recorded in Kupwara (21.77±1.39 mites per leaf), Baramulla (19.27±1.09 per leaf), Shopian (18.4±2.47 per leaf), Budgam (18.33±1.24 per leaf) and Pulwama (18.24±1.75 per leaf) respectively. Significant positive correlations of mite populations with minimum temperature were found in Shopian and all the districts of north Kashmir. Significant negative correlation was also found between mite population and rainfall except Anantnag district where it was non-significant. Maximum and minimum temperatures as well as sunshine hours had non-significant positive correlation, whereas relative humidity had non-significant negative correlation with the mite population.

First Report of Neofusicoccum australe Causing Branch Dieback of English Walnut cv. Chandler in Central Chile.

The English walnut (Juglans regia L.) is the second most important fruit crop of importance in Chile, with 43,700 hectares mainly in the Central Valley (www.odepa.cl, 2022). For several seasons symptoms of a branch dieback have been observed in walnut orchards with 3 to 50% of trees incidence levels. During the 2020 winter season (July) a total of 150 symptomatic spurs of 15 trees were sampled from an 8-year-old walnut cv. Chandler orchard located in Buin (33°42' S, 70° 42' W). The collected spurs showed external and internal brown necroses, starting from the tip with well-defined margins. The symptomatic tissue was cut in to small pieces (5 x 4 x 2 mm), surface disinfected by dipping in a 10% solution made from a commercial bleach solution (4,9% NaOHCl), rinsed twice in sterile water and plated on APDA (PDA Difco laboratories acidified with lactic acid (2,5 ml of 25% (vol/vol) per liter of medium). After five days at 20 °C in darkness, fast-growing, white-grey turning to black colonies were obtained, tentatively classified as a member of the Botryosphaeraceae family and two single-spore isolates (SS1, SS2) were selected for identification. Colony mycelia were first white and turned to light grey, dark grey or black, with tufts of mouse gray aerial mycelia. The pycnidia and conidia production was induced by inoculating autoclaved pine needles placed on APDA an incubation for 25 to 30 days at 20 °C in darkness. Black pycnidia solitary and globose were obtained producing hyaline, aseptate, fusiform to obovoid conidia with truncated ends with dimensions of (22.6-) 19.1 ± 1.4 (-13.3) x (6.7-) 5.5 ± 0.5 (-3.7) µm and 3.5 length/width ratio (n=100). Both isolates were identified using dichotomous keys confirming the description of Crous et al, 2006 as Neofusicoccum australe. The identification was molecularly confirmed by amplifying the nuclear ribosomal gene 5,8S (ITS1-5.8S-ITS2) using the ITS1/ITS4 primers, a partial region of β-tubulin gene (Bt2a/Bt2b), and the translation elongation factor 1-α gene (TEF1) with TEF1-728F/TEF1-986R primers. The BLASTn search revealed 100% of identity for ITS and TEF according to sequences of N. australe reference strains MT587467.1 and MK759852.1, respectively; and over 99% for β-tubulin compared to N. australe strain KX464929.1. The DNA sequences were submitted to the GenBank (ITS, OP142414, OP142416; BT, OP209981, OP209978; and TEF OP209979, OP209980) for SS1 and SS2 isolates, respectively, and deposited in the fungal collection of CChRGM - INIA, Chillán, Chile (RGM 3409 and 3410). Pathogenicity of both isolates was tested in 8-year-old asymtomatic English walnut cv. Chandler in the field during 2020 spring season, by cutting transversally 15 twigs of different tress and inoculating with a 5 day-old PDA plug. An equal number of wounded twigs were inoculated with a sterile PDA plug and served as control. After six months, all inoculated twigs developed the same necrotic lesions observed in field of 2.0 to 10.1 cm (SS1) and 1.9 to 10.8 cm (SS2) in length while control twigs showed only a scar without any dieback tissues. The inoculated pathogens of N. australe were recovered from the diseased tissues, thus fulfilling Koch's postulates. A similar dieback of walnut was reported in Chile, which caused Diplodia mutila (Díaz et al, 2018), and N. parvum (Luna et al, 2022) while N. australe has been reported in other hosts (Auger et al, 2013, Besoain et al, 2013). To the best of our knowledge, this is the first report of N. australe associated with walnut branch dieback in Chile.