Sally Hilton

Meeting the demand for crop production : the challenge of yield decline in crops grown in short rotations

There is a trend world‐wide to grow crops in short rotation or in monoculture, particularly in conventional agriculture. This practice is becoming more prevalent due to a range of factors including economic market trends, technological advances, government incentives, and retailer and consumer demands. Land‐use intensity will have to increase further in future in order to meet the demands of growing crops for both bioenergy and food production, and long rotations may not be considered viable or practical. However, evidence indicates that crops grown in short rotations or monoculture often suffer from yield decline compared to those grown in longer rotations or for the first time. Numerous factors have been hypothesised as contributing to yield decline, including biotic factors such as plant pathogens, deleterious rhizosphere microorganisms, mycorrhizas acting as pathogens, and allelopathy or autotoxicity of the crop, as well as abiotic factors such as land management practices and nutrient availability. In many cases, soil microorganisms have been implicated either directly or indirectly in yield decline. Although individual factors may be responsible for yield decline in some cases, it is more likely that combinations of factors interact to cause the problem. However, evidence confirming the precise role of these various factors is often lacking in field studies due to the complex nature of cropping systems and the numerous interactions that take place within them. Despite long‐term knowledge of the yield‐decline phenomenon, there are few tools to counteract it apart from reverting to longer crop rotations or break crops. Alternative cropping and management practices such as double‐cropping or inter‐cropping, tillage and organic amendments may prove valuable for combating some of the negative effects seen when crops are grown in short rotation. Plant breeding continues to be important, although this does require a specific breeding target to be identified. This review identifies gaps in our understanding of yield decline, particularly with respect to the complex interactions occurring between the different components of agro‐ecosystems, which may well influence food security in the 21st Century.

Impact of Shortened Crop Rotation of Oilseed Rape on Soil and Rhizosphere Microbial Diversity in Relation to Yield Decline

Oilseed rape (OSR) grown in monoculture shows a decline in yield relative to virgin OSR of up to 25%, but the mechanisms responsible are unknown. A long term field experiment of OSR grown in a range of rotations with wheat was used to determine whether shifts in fungal and bacterial populations of the rhizosphere and bulk soil were associated with the development of OSR yield decline. The communities of fungi and bacteria in the rhizosphere and bulk soil from the field experiment were profiled using terminal restriction fragment length polymorphism (TRFLP) and sequencing of cloned internal transcribed spacer regions and 16S rRNA genes, respectively. OSR cropping frequency had no effect on rhizosphere bacterial communities. However, the rhizosphere fungal communities from continuously grown OSR were significantly different to those from other rotations. This was due primarily to an increase in abundance of two fungi which showed 100% and 95% DNA identity to the plant pathogens Olpidium brassicae and Pyrenochaeta lycopersici, respectively. Real-time PCR confirmed that there was significantly more of these fungi in the continuously grown OSR than the other rotations. These two fungi were isolated from the field and used to inoculate OSR and Brassica oleracea grown under controlled conditions in a glasshouse to determine their effect on yield. At high doses, Olpidium brassicae reduced top growth and root biomass in seedlings and reduced branching and subsequent pod and seed production. Pyrenochaeta sp. formed lesions on the roots of seedlings, and at high doses delayed flowering and had a negative impact on seed quantity and quality.

Fine endophytes (Glomus tenue) are related to Mucoromycotina, not Glomeromycota

Fine endophytes (FE),Glomus tenue, are traditionally considered to be arbuscular mycorrhizal fungi (AMF) with distinctive microscopic morphology when stained. FE have fine hyphae (c. 1.5 lm diameter) which branch intra-cellularly in a distinctive fan-like pattern (Gianinazzi-Pearson et al., 1981; Abbott, 1982). The hyphae contain small swellings along their length, sometimes referred to as vesicle-like swellings (Hall, 1977). FE form arbuscules (or arbuscule-like structures) with fine elements in a tapered, conical shape (Greenall, 1963; Merryweather & Fitter, 1998). Spores of FE are very small (< 20 lm) compared to the majority of Glomeromycota, and colourless (Hall, 1977). Morphological variations indicate that FE may consist of multiple species (Thippayarugs et al., 1999), hence we use the term FE to indicate a species group. Within the kingdom Fungi, both morphological and genetic characteristics are used to determine taxonomic classification (St€ urmer, 2012). In 2001, all AMFwere placed within the phylum Glomeromycota (Sch€ ußler et al., 2001). In the listing of glomeromycotan species by Sch€ ußler &Walker (2010), some members of the genusGlomuswere not revised due to insufficient taxonomic knowledge, and this included FE. A key reason for classifying FE within the Glomeromycota was the presence of arbuscules, considered apomorphic for the phylum (Morton, 1990).However, the morphological features of root colonization by FE are distinct from other, coarse, AMF so their placement within the genus Glomus and the Glomeromycota was questioned (Hall, 1977; Sch€ ußler &Walker, 2010), and their status as mycorrhizal fungi is ambivalent. Accurate determination of FE usually requires magnification ≥9100, hence, where assessments of AMF colonization use lower magnifications theymay not be identified. Furthermore, FEmaybe undetected if samples are not processed within 2 d of harvesting (Orchard et al., 2016a). Nevertheless, FE are globally distributed and prolific within many ecosystems, examples include: pastures and native bushland of New Zealand (Crush, 1973) and Australia (Abbott&Robson, 1982;McGee, 1989),Venezuelan cloud forests (Rabatin et al., 1993), riverine and alpine regions of Europe (Read &Haselwandter, 1981; Turnau et al., 1999; Binet et al., 2011) and an old-field in the United States (Hilbig &Allen, 2015). However, the difficulty of isolating and, hence, genetically characterizing FE has hindered the determination of their phylogenetic placement.

The origins of replication of granuloviruses

The genomes of eight granuloviruses (GVs), have been analyzed for the presence of homologous regions (hrs) that may act as origins of replication. Thirteen 74–76-bp palindromes within 11 hrs have previously been identified in the Cydia pomonella GV (CpGV) genome and found to replicate in an infection-dependent DNA replication assay. We report a further palindrome within one of the hrs, which was found to replicate, bringing the total to 14 palindromes. We also report imperfect palindromes, with similar 13-bp end sequences to the CpGV palindromes, within the Adoxophyes orana GV, Cryptophlebia leucotreta GV (CrleGV), Choristoneura occidentalis GV and Phthorimaea operculella GV genomes. No hrs were detected in Agrotis segetum GV, and no additional hrs or palindromes, other than those published, were detected in the Plutella xylostella GV and Xestia c-nigrum GV genomes. Several putative hrs from the GVs were tested for replication in C. pomonella cells using a CpGV-dependent replication assay. Two CrleGV hrs were found to replicate at a low level.

A bacmid approach to the genetic manipulation of granuloviruses

A Cydia pomonella granulovirus (CpGV) bacmid has been constructed, which allows rapid and efficient production of recombinant baculoviruses in Escherichia coli. An 8.6kbp bacterial DNA cassette derived from the AcMNPV Bac-to-Bac system was ligated into a unique PacI restriction site within an intergenic region flanking the DNA ligase gene of the CpGV genome. The CpGV bacmids produced in E. coli were transfected into a CpGV-permissive C. pomonella cell line and the transfected cells fed to larvae to amplify the virus. The enhanced green fluorescent protein (EGFP) gene under the constitutive Drosophila heat-shock promoter was transposed into the mini-attTn7 transposition site, using a modified pFASTBAC donor plasmid, to generate a recombinant CpGV bacmid which caused infected larvae to glow under UV light. Targeted homologous recombination was also achieved in a recombinant proficient E. coli strain (BJ5183). A chloramphenicol acetyl transferase (CAT) gene replaced the cathepsin (v-cath) gene in the bacmid to produce a v-cath-deletion mutant. This is the first published report of a granulovirus bacmid, which will allow easy manipulation of the CpGV genome, enabling future studies on granulovirus genes and biology.

Resistance and resilience responses of a range of soil eukaryote and bacterial taxa to fungicide application

Highlights • We studied the resistance and resilience of soil microbial communities.• There was a significant concentration-dependent impact on dehydrogenase activity.• Significant impacts on nematode and fungal communities were also observed.

Incomplete reprogramming of cell-specific epigenetic marks during asexual reproduction leads to heritable phenotypic variation in plants

Plants differ from animals in their capability to easily regenerate fertile adult individuals from terminally differentiated cells [1]. This unique developmental plasticity is commonly observed in nature where many species can reproduce asexually through the ectopic initiation of organogenic or embryogenic developmental programs [2, 3]. However, it is not currently known if this developmental reprogramming is coupled to a global epigenomic resetting, or what impact it has on the phenotype of the clonal progeny. Here we show that plants asexually propagated via induction of a zygotic developmental program do not fully reset cell-specific epigenetic imprints. These imprints are instead inherited even over multiple rounds of sexual reproduction, becoming fixed in hybrids and resulting in heritable molecular and physiological phenotypes that depend on the founder cell used. Our results demonstrate how novel phenotypic variation in plants can be unlocked through the incomplete reprogramming of cell-specific epigenetic marks during asexual propagation.

Partial maintenance of organ-specific epigenetic marks during plant asexual reproduction leads to heritable phenotypic variation

Significance While clonally propagated individuals should share identical genomes, there is often substantial phenotypic variation among them. Both genetic and epigenetic modifications induced during regeneration have been associated with this phenomenon. Here we investigated the fate of the epigenome after asexual propagation by generating clonal individuals from differentiated somatic cells through the manipulation of a zygotic transcription factor. We found that phenotypic novelty in clonal progeny was linked to epigenetic imprints that reflect the organ used for regeneration. Some of these organ-specific imprints can be maintained during the cloning process and subsequent rounds of meiosis. Our findings are fundamental for understanding the significance of epigenetic variability arising from asexual reproduction and have significant implications for future biotechnological applications. Plants differ from animals in their capability to easily regenerate fertile adult individuals from terminally differentiated cells. This unique developmental plasticity is commonly observed in nature, where many species can reproduce asexually through the ectopic initiation of organogenic or embryogenic developmental programs. While organ-specific epigenetic marks are not passed on during sexual reproduction, the fate of epigenetic marks during asexual reproduction and the implications for clonal progeny remain unclear. Here we report that organ-specific epigenetic imprints in Arabidopsis thaliana can be partially maintained during asexual propagation from somatic cells in which a zygotic program is artificially induced. The altered marks are inherited even over multiple rounds of sexual reproduction, becoming fixed in hybrids and resulting in heritable molecular and physiological phenotypes that depend on the identity of the founder tissue. Consequently, clonal plants display distinct interactions with beneficial and pathogenic microorganisms. Our results demonstrate how novel phenotypic variation in plants can be unlocked through altered inheritance of epigenetic marks upon asexual propagation.

Plant rhizosphere selection of plasmodiophorid lineages from bulk soil : the importance of “hidden” diversity

Microbial communities closely associated with the rhizosphere can have strong positive and negative impacts on plant health and growth. We used a group-specific amplicon approach to investigate local scale drivers in the diversity and distribution of plasmodiophorids in rhizosphere/root and bulk soil samples from oilseed rape (OSR) and wheat agri-systems. Plasmodiophorids are plant- and stramenopile-associated protists including well known plant pathogens as well as symptomless endobiotic species. We detected 28 plasmodiophorid lineages (OTUs), many of them novel, and showed that plasmodiophorid communities were highly dissimilar and significantly divergent between wheat and OSR rhizospheres and between rhizosphere and bulk soil samples. Bulk soil communities were not significantly different between OSR and wheat systems. Wheat and OSR rhizospheres selected for different plasmodiophorid lineages. An OTU corresponding to Spongospora nasturtii was positively selected in the OSR rhizosphere, as were two genetically distinct OTUs. Two novel lineages related to Sorosphaerula veronicae were significantly associated with wheat rhizosphere samples, indicating unknown plant-protist relationships. We show that group-targeted eDNA approaches to microbial symbiont-host ecology reveal significant novel diversity and enable inference of differential activity and potential interactions between sequence types, as well as their presence.