Effects of green-manure and tillage management on soil microbial community composition, nutrients and tree growth in a walnut orchard

This study characterized the effect of green manures (February orchid, hairy vetch, rattail fescue and a no-green-manure control) and the termination method (flail or disk) on nutrient contents, enzyme activities, microbial biomass, microbial community structure of rhizosphere soil and vegetative growth of walnut tree. All three selected green manures significantly enhanced the water content, organic C, total N and available P. The rattail fescue significantly decreased the mineral N. Total organic C, total N, mineral N and available P were significantly greater under flail than under disk. Hairy vetch and February orchid significantly improved levels of soil β-glucosidase, N-acetyl-glucosaminidase and acid phosphatase activity, whereas rattail fescue improved only β-glucosidase activity. All of the green manures significantly decreased phenoloxidase activity. β-glucosidase, N-acetyl-glucosaminidase and acid phosphatase activities were significantly greater under flail relative to disk. The termination method had no significant effect on phenoloxidase activity. The different types of green manures and termination methods significantly altered the soil microbial biomass and microbial community structure. The green-manure treatments were characterized by a significantly greater abundance of Gram-positive (Gram +) bacteria, total bacteria and saprophytic fungi compared to the control. Hairy vetch significantly decreased the abundance of arbuscular mycorrhizal fungi (AMF) while February orchid and rattail fescue increased their abundance compared to the no-green-manure treatment. The abundance rates of Gram+ bacteria, actinomycetes, saprophytic fungi and AMF were significantly greater in soils under flail than under disk. In terms of vegetative growth of walnut tree, hairy vetch showed the greatest positive effects. The growth of walnut tree was significantly greater under flail relative to disk. Our results indicate that green-manure application benefits the rhizosphere soil micro-ecology, rhizosphere soil nutrient contents and tree growth. Overall, the hairy vetch and flail combined treatment is recommended for walnut orchards in northern China.

Comparative Genomics of Xanthomonaseuroxanthea and Xanthomonas arboricola pv. juglandis Strains Isolated from a Single Walnut Host Tree

The recent report of distinct Xanthomonas lineages of Xanthomonas arboricola pv. juglandis and Xanthomonas euroxanthea within the same walnut tree revealed that this consortium of walnut-associated Xanthomonas includes both pathogenic and nonpathogenic strains. As the implications of this co-colonization are still poorly understood, in order to unveil niche-specific adaptations, the genomes of three X. euroxanthea strains (CPBF 367, CPBF 424T, and CPBF 426) and of an X. arboricola pv. juglandis strain (CPBF 427) isolated from a single walnut tree in Loures (Portugal) were sequenced with two different technologies, Illumina and Nanopore, to provide consistent single scaffold chromosomal sequences. General genomic features showed that CPBF 427 has a genome similar to other X. arboricola pv. juglandis strains, regarding its size, number, and content of CDSs, while X. euroxanthea strains show a reduction regarding these features comparatively to X. arboricola pv. juglandis strains. Whole genome comparisons revealed remarkable genomic differences between X. arboricola pv. juglandis and X. euroxanthea strains, which translates into different pathogenicity and virulence features, namely regarding type 3 secretion system and its effectors and other secretory systems, chemotaxis-related proteins, and extracellular enzymes. Altogether, the distinct genomic repertoire of X. euroxanthea may be particularly useful to address pathogenicity emergence and evolution in walnut-associated Xanthomonas.

Complete Genome Sequence Obtained by Nanopore and Illumina Hybrid Assembly of Xanthomonas arboricola pv. juglandis CPBF 427, Isolated from Buds of a Walnut Tree

ABSTRACT We report the genome sequence of Xanthomonas arboricola pv. juglandis strain CPBF 427, which was isolated from early-season buds of a diseased walnut tree, suggesting overwinter potential. This study provides a consistent genomic reference for this pathovar and may contribute to addressing the overwinter survival of these walnut pathogens.

Flooding by California Rivers Results in Walnut Scion Infections by Species of Phytophthora

Many walnut orchards were inundated by flooding from the Feather and Stanislaus Rivers in winter and spring 2017 and developed bleeding cankers in trunk, root, and crown tissues exposed to the water. Orchard surveys and diagnostic isolations associated Phytophthora pini, P. chlamydospora, and P. gonapodyides with the cankers in 2017. Pathogenicity of P. pini was confirmed in seedlings and excised shoots of Juglans regia, but the other species caused negligible amounts of disease. Feather River and associated flood waters were assayed using culture-independent sequencing of rRNA gene amplicons and pear baiting methods; 14 species of Phytophthora were detected, including P. chlamydospora and P. gonapodyides, but not P. pini. Severe and prolonged walnut orchard flooding from rivers, such as occurred in 2017, places diverse mixtures of Phytophthora species from multiple sources into close, infective proximity with susceptible walnut tree scions. Systemic chemical or genetic protection strategies may be valuable for orchards subject to such flooding.

Complete Genome Sequences of Walnut-Associated Xanthomonas euroxanthea Strains CPBF 367 and CPBF 426 Obtained by Illumina/Nanopore Hybrid Assembly

ABSTRACT We present the complete genome sequences of two Xanthomonas euroxanthea strains isolated from buds of a walnut tree. The whole-genome sequences of strains CPBF 367 and CPBF 426 consist of two circular chromosomes of 4,923,218 bp and 4,883,254 bp and two putative plasmids of 45,241 bp and 17,394 bp, respectively. These data may contribute to the understanding of Xanthomonas species-specific adaptations to walnut.

Forty years of study on interactions between walnut tree and arbuscular mycorrhizal fungi. A review

Walnut trees are among the most important hardwood species in the northern hemisphere, ecologically and economically. They are mainly cultivated for timber and nut production but are also attractive ornamental trees in parks. Establishing walnut orchards is difficult because seedlings have a coarse root architecture and few of them survive to transplanting. Planting success is mainly determined by the root system morphology and the nutrient status of the seedlings, so that rhizosphere conditions are critical for plant performance. Walnut trees can associate with soil-borne arbuscular mycorrhizal fungi, which are obligate biotrophs. In this association, plant-produced carbon compounds are traded against fungus-acquired soil mineral nutrients. The beneficial effect of arbuscular mycorrhizal symbiosis on hardwood seedling quality and field performance has long been known, but an integrated view is lacking about the effects of arbuscular mycorrhizas on walnut cropping. Therefore, we surveyed the literature published over the last 40 years to provide up-to-date knowledge on the relationships between arbuscular mycorrhizas and walnut trees. Our review outlines the major following points: (1) the arbuscular-mycorrhiza-mediated nutrient uptake capacity of walnut trees is associated with first- to third-order roots, and fibrous tip-ended roots are dependent on arbuscular mycorrhizal fungi, whereas pioneer roots are not; (2) early inoculation with arbuscular mycorrhizal fungi improves the survival and seedling performance attributes of transplanted walnut trees: biotization enhances walnut transplant success by increasing the number of lateral roots and plant P uptake, but these benefits are fungus- and host-dependent; (3) in the context of walnut agroforestry, deeply rooted walnut trees play a role as reservoirs of arbuscular mycorrhizal fungal propagules for the surrounding vegetation, but tree shade and soluble phosphate availability decrease walnut mycorrhizal dependency; and (4) the arbuscular mycorrhizal mycelium mediates the transport of juglone and thus plays a role in walnut tree allelopathy.