Niccolò Terzaroli

Allium ampeloprasum var. holmense (elephant garlic) is traditionally cultivated in “Val di Chiana”, an area between Umbria and Tuscany regions of Central Italy, under the name “Aglione della Valdichiana”. This product has recently increased in importance, becoming a key economic resource for local farmers. In 2019, phytosanitary problems of elephant garlic cloves ready for transplanting emerged in this cultivation area. Symptom/sign observations and fungal isolations were performed for cloves divided into four components (tunic, basal plate, reserve tissue and shoot) from six farms in the “Val di Chiana” area. Isolates obtained were identified, using partial β-tubulin (BenA) and calmodulin (CaM) or translation elongation factor 1α (tef1α) genes sequences, as belonging to Penicillium [P. allii (95%), P. citrinum (4%), P. brevicompactum (1%)] or Fusarium [F. oxysporum (81%), F. proliferatum (19%)]. Fusarium spp. were mainly associated with clove tunics and basal plates, while Penicillium spp. with basal plates, reserve tissues and shoots. Fungi often also developed from asymptomatic components, but a correlation was found between isolated pathogens and disease symptoms. Pathogenicity and virulence towards elephant garlic cloves were verified for a representative isolate of each identified species, and Penicillium allii was the most virulent. Strategies to control Fusarium and Penicillium spp. on cloves were assessed, including chemicals, a biocontrol agent, surface sterilization and heat treatment. Among these, treatments with Patriot Gold® (active ingredient [a.i.] Trichoderma asperellum TV1, approved in organic farming on crops similar to elephant garlic), or Signum® (a.i. boscalid + pyraclostrobin, approved for Integrated Pest Management systems on crops similar to elephant garlic), were effective in simultaneous reduction of Penicillium spp. and Fusarium spp. Transplanting of asymptomatic cloves combined with the use of the above treatments showed promising effects for pathogens control, and to assist elephant garlic crop establishment.

Main conclusion: Apomixis is a complex evolutionary trait with many possible origins. Here we discuss various clues and causes, ultimately proposing a model harmonizing the three working hypotheses on the topic. Asexual reproduction through seeds, i.e., apomixis, is the holy grail of plant biology. Its implementation in modern breeding could be a game-changer for agriculture. It has the potential to generate clonal crops and maintain valuable complex genotypes and their associated heterotic traits without inbreeding depression. The genetic basis and origins of apomixis are still unclear. There are three central hypothesis for the development of apomixis that could be: i) a deviation from the sexual developmental program caused by an asynchronous development, ii) environmentally triggered through epigenetic regulations (a polyphenism of sex), iii) relying on one or more genes/alleles. Because of the ever-increasing complexity of the topic, the path toward a detailed understanding of the mechanisms underlying apomixis remains unclear. Here, we discuss the most recent advances in the evolution perspective of this multifaceted trait. We incorporated our understanding of the effect of endogenous effectors, such as small RNAs, epigenetic regulation, hormonal pathways, protein turnover, and cell wall modification in response to an upside stress. This can be either endogenous (hybridization or polyploidization) or exogenous environmental stress, mainly due to oxidative stress and the corresponding ROS (Reacting Oxygen Species) effectors. Finally, we graphically represented this tangled web.

Key message: A new R-software procedure for fixed/random Diallel models was developed. We eased the diallel schemes approach by considering them as specific cases with different parameterisations of a general linear model. Diallel experiments are based on a set of possible crosses between some homozygous (inbred) lines. For these experiments, six main diallel models are available in literature, to quantify genetic effects, such as general combining ability (GCA), specific combining ability (SCA), reciprocal (maternal) effects and heterosis. Those models tend to be presented as separate entities, to be fitted by using specialised software. In this manuscript, we reinforce the idea that diallel models should be better regarded as specific cases (different parameterisations) of a general linear model and might be fitted with general purpose software facilities, as used for all other types of linear models. We start from the estimation of fixed genetical effects within the R environment and try to bridge the gap between diallel models, linear models and ordinary least squares estimation (OLS). First, we review the main diallel models in literature. Second, we build a set of tools to enable geneticists, plant/animal breeders and students to fit diallel models by using the most widely known R functions for OLS fitting, i.e. the ‘lm()’ function and related methods. Here, we give three examples to show how diallel models can be built by using the typical process of GLMs and fitted, inspected and processed as all other types of linear models in R. Finally, we give a fourth example to show how our tools can be also used to fit random/mixed effect diallel models in the Bayesian framework.

The production of seeds without sex is considered the holy grail of plant biology. The transfer of apomixis to various crop species has the potential to transform plant breeding, since it will allow new varieties to retain valuable traits thorough asexual reproduction. Therefore, a greater molecular understanding of apomixis is fundamental. In a previous work we identified a gene, namely APOSTART, that seemed to be involved in this asexual mode of reproduction, which is very common in Poa pratensis L., and here we present a detailed work aimed at clarifying its role in apomixis. In situ hybridization showed that PpAPOSTART is expressed in reproductive tissues from pre-meiosis to embryo development. Interestingly, it is expressed early in few nucellar cells of apomictic individuals possibly switching from a somatic to a reproductive cell as in aposporic apomixis. Moreover, out of 13 APOSTART members, we identified one, APOSTART_6, as specifically expressed in flower tissue. APOSTART_6 also exhibited delayed expression in apomictic genotypes when compared with sexual types. Most importantly, the SCAR (Sequence Characterized Amplified Region) derived from the APOSTART_6 sequence completely co-segregated with apomixis.

Heterosis is the superiority of an F₁ hybrid over its parents. Since this phenomenon is still unclear in melon, a half diallel experiment based on eight genetically distant breeding lines was conducted in six environments of Central Italy, assessing commercially important traits: yield, total soluble solids (TSS), and days to ripening (DTR). To estimate the additive (general combining ability; GCA) and the non-additive gene effects (specific combining ability; SCA), yield was analyzed by Griffing’s methods two and four, and the results were compared to the GGE (Genotype plus Genotype by Environment interaction) biplot methodology; TSS and earliness were evaluated only by Griffing’s method four. Overall, GCAs were significantly more relevant than SCAs for all examined traits. Least square means (LsM), mid-parent heterosis (MPH), best-parent heterosis (BPH), as well as Euclidean and Mahalanobis’ distances were calculated and compared with the genetic distance (GD). As a few correlations were found statistically significant (only for TSS), it was difficult to predict the value of a hybrid combination only by knowing the genetic distance of its parents. Despite this, heterosis was observed, indicating either the presence of epistatic effects (additive × additive interactions) and/or an underestimate of SCAs embedded within Griffing’s method. The significant Env × Entries source of variation suggests development of hybrids in specific environments. The results are discussed with a breeding perspective.

“Roveja di Civita di Cascia” is a landrace of Pisum sativum grown in marginal land habitats of the Apennines, Central Italy, and is one of the eleven herbaceous crop landraces listed in the Regional Register of local varieties. The objective of the present paper was to assess its genetic structure using 62 morphological traits and five microsatellites. As many as 55 traits showed significant differences with the control entries (P. sativum subsp. sativum var. arvense and var. sativum). We tested P. sativum Simple Sequence Repeats (SSRs) for their transferability to “Roveja”, and found that only 12 out of 35 performed well. Of these, we demonstrated that five were sufficient to assess the genetic structure of this landrace, characterized by several private alleles, differentiating it from Paladio and Bluemoon, which were used as controls. Phenotypic and genotypic data evidenced a genetic structure based on a blend of several pure-bred lines. The sustainability of on-farm landrace conservation is discussed.