Camelina sativa (L. Crantz) Fresh Forage Productive Performance and Quality at Different Vegetative Stages: Effects of Dietary Supplementation in Ionica Goats on Milk Quality

The research meant to study the productive performances of Camelina sativa and the effects of feeding Camelina fresh forage harvested during five phenological stages (I: main stem elongation; II: maximum stem elongation: III: inflorescence appearance; IV: flowering; V: fruit set visible) on the yield, chemical composition and fatty acid profile of milk from autochthonous Ionica goats. Goats were randomly assigned to two groups (n = 15) that received a traditional forage mixture (Control) or Camelina forage harvested at different stages (CAM). The field experiment was conducted in two years; no significant differences between years were recorded for any of the Camelina production traits. The total biomass increased (p < 0.05) from phase I (1.4 t/ha) to phase V (5.2 t/ha). The distribution of stem, leaves and pod also changed during growth, showing a significant increase of stem from 40.8 to 45.6% and of pod from 0 to 19.4%, whereas leaves decreased from 59.2 to 35.1%. The milk yield and chemical composition were unaffected by the diet, while supplementation with Camelina forage increased milk CLA content (on average 1.14 vs. 0.78%). A markedly higher concentration of PUFAs was found in milk from goats fed Camelina harvested during the last three phenological stages. The index of thrombogenicity of milk from the CAM fed goats was significantly lower compared to the control group. In conclusion, Camelina sativa is a multi-purpose crop that may be successfully cultivated in Southern Italy regions and used as fresh forage for goat feeding. Milk obtained from Camelina fed goats showed satisfactory chemical and fatty acid composition, with potential benefits for human health.

What's past is past, mostly: Brassicaceae host plants mask the feedback from the previous year's soil history on bacterial communities, except when the Brassicaceae hosts experience drought

Previous soil history and the current plant hosts are two plant-soil feedbacks that operate at different time-scales to influence the structure soil bacterial communities. In this study, we used a MiSeq metabarcoding strategy to describe the impact of five Brassicaceae host plant species, and three different soil histories, on the structure of their bacterial root and rhizosphere communities at full flower. We found that the Brassicaceae host plants were consistently significant in structuring the bacterial communities. Four host plants (Sinapis alba, Brassica napus, B. juncea, B. carinata) formed nearly the same bacterial communities, regardless of soil history. Camelina sativa host plants structured phylogenetically distinct bacterial communities compared to the other hosts, particularly in their roots. Soil history established the previous year was only a significant factor for bacterial community structure when the feedback of the Brassicaceae host plants was weakened, potentially due to limited soil moisture during a dry year. Understanding how plant-soil feedbacks operate at different time-scales and are involved in how microbial communities are structured is a pre-requisite for employing microbiome technologies in improving agricultural systems.

First Report of Powdery Mildew Caused by Erysiphe cruciferarum on Camelina sativa in Montana

Camelina sativa (L.) Crantz, also known as false flax, is an annual flowering plant in the family Brassicaceae and originated in Europe and Asia. In recent years, it is cultivated as an important biofuel crop in Europe, Canada, and the northwest of the United States. In June of 2021, severe powdery mildew was observed on C. sativa ‘Suneson’ plants under greenhouse conditions (temperature 18.3°C/22.2°C, night/day) in Bozeman, Montana (45°40'N, 111°2'W). The disease incidence was 80.67% (150 pots, one plant per pot). White ectophytic powdery mildew including mycelia and conidia were observed on the upper leaves, usually developed from bottom tissues to top parts, also present on stems and siliques. Mycelia on leaves were amphigenous and in patches, often spreading to become effused. These typical symptoms were similar to a previous report of powdery mildew on Broccoli raab (Koike and Saenz 1997). Appressoria are lobed, and foot cells are cylindrical with size 18 to 26 × 7 to 10 μm. Conidia are cylindrical and produced singly, with a size of 35 to 50 × 12 to 21 μm and a length : width ratio greater than two (Koike and Saenz 1997). No chasmothecia were observed under the greenhouse conditions. The symptoms and fungal microscopic characters are typical of Pseudoidium anamorph of Erysiphe (Braun 1995). The specific measurements and characteristics are consistent with previous records of Erysiphe cruciferarum Opiz ex L. Junell (Braun and Cook 2012; Vellios et al. 2017). To identify the pathogen, the partial internal transcribed spacer (ITS) region of rDNA of sample CPD-1 was amplified using primers ITS1 and ITS4 (White et al. 1990). The amplicons were sequenced, and the resulting 559-bp sequence was deposited in GenBank (CPD-1, Accession number: OK160719). A GenBank BLAST search of the ITS sequences showed an exact match (100% query cover, E-value 0, and 100% identity 559/559 bp) with those of E. cruciferarum on hosts Brassica sp. (KY660929.1), B. juncea from Vietnam (KM260718.1) and China (KT957424.1). A phylogenetic tree was generated with the CPD-1 ITS sequence with several of ITS sequences of close species with different hosts obtained from the GenBank. Isolate CPD-1 was grouped with pathogens from Brassica hosts rather than the holotype strain (KU672364.1) from papaveraceous hosts. To fulfill Koch's postulates, pathogenicity was confirmed through inoculation by dusting conidia onto leaves of seven healthy, potted, 14-day-old C. sativa seedlings (cv. Suneson). Seven non-inoculated plants served as a control treatment. The plants were incubated in a greenhouse with a temperature of 18°C (night) to 22°C (day). The inoculated plants developed similar symptoms after 7 days, whereas the control plants remained symptomless. The fungus on the inoculated plants was morphologically identical to that was originally observed on the diseased plants. Though many Brassica spp. have been known to be infected by E. cruciferarum throughout the world, powdery mildew of C. sativa cultivar Crantz in natural conditions by E. cruciferarum has been reported only in the province of Domokos in Central Greece (Vellios et al. 2017). To our knowledge, this is the first report of powdery mildew caused by E. cruciferarum on C. sativa in Montana. Though the powdery mildew on C. sativa was observed in the greenhouse conditions in this work, it poses a potential threat to the production of this biofuel crop in the northwest of the United States.

Engineering a feedback inhibition-insensitive plant dihydrodipicolinate synthase to increase lysine content in Camelina sativa seeds

Camelina sativa (camelina) is emerging as an alternative oilseed crop due to its short growing cycle, low input requirements, adaptability to less favorable growing environments and a seed oil profile suitable for biofuel and industrial applications. Camelina meal and oil are also registered for use in animal and fish feeds; however, like meals derived from most cereals and oilseeds, it is deficient in certain essential amino acids, such as lysine. In higher plants, the reaction catalyzed by dihydrodipicolinate synthase (DHDPS) is the first committed step in the biosynthesis of lysine and is subject to regulation by lysine through feedback inhibition. Here, we report enhancement of lysine content in C. sativa seed via expression of a feedback inhibition-insensitive form of DHDPS from Corynebacterium glutamicums (CgDHDPS). Two genes encoding C. sativa DHDPS were identified and the endogenous enzyme is partially insensitive to lysine inhibition. Site-directed mutagenesis was used to examine the impact of alterations, alone and in combination, present in lysine-desensitized DHDPS isoforms from Arabidopsis thaliana DHDPS (W53R), Nicotiana tabacum (N80I) and Zea mays (E84K) on C. sativa DHDPS lysine sensitivity. When introduced alone, each of the alterations decreased sensitivity to lysine; however, enzyme specific activity was also affected. There was evidence of molecular or structural interplay between residues within the C. sativa DHDPS allosteric site as coupling of the W53R mutation with the N80V mutation decreased lysine sensitivity of the latter, but not to the level with the W53R mutation alone. Furthermore, the activity and lysine sensitivity of the triple mutant (W53R/N80V/E84T) was similar to the W53R mutation alone or the C. glutamicum DHDPS. The most active and most lysine-insensitive C. sativa DHDPS variant (W53R) was not inhibited by free lysine up to 1 mM, comparable to the C. glutamicums enzyme. Seed lysine content increased 13.6 -22.6% in CgDHDPS transgenic lines and 7.6–13.2% in the mCsDHDPS lines. The high lysine-accumulating lines from this work may be used to produce superior quality animal feed with improved essential amino acid profile.

Green Chemistry Production of Codlemone, the Sex Pheromone of the Codling Moth (Cydia pomonella), by Metabolic Engineering of the Oilseed Crop Camelina (Camelina sativa)

Synthetic pheromones have been used for pest control over several decades. The conventional synthesis of di-unsaturated pheromone compounds is usually complex and costly. Camelina (Camelina sativa) has emerged as an ideal, non-food biotech oilseed platform for production of oils with modified fatty acid compositions. We used Camelina as a plant factory to produce mono- and di-unsaturated C12 chain length moth sex pheromone precursors, (E)-9-dodecenoic acid and (E,E)-8,10-dodecadienoic acid, by introducing a fatty acyl-ACP thioesterase FatB gene UcTE from California bay laurel (Umbellularia californica) and a bifunctional ∆9 desaturase gene Cpo_CPRQ from the codling moth, Cydia pomonella. Different transgene combinations were investigated for increasing pheromone precursor yield. The most productive Camelina line was engineered with a vector that contained one copy of UcTE and the viral suppressor protein encoding P19 transgenes and three copies of Cpo_CPRQ transgene. The T2 generation of this line produced 9.4% of (E)-9-dodecenoic acid and 5.5% of (E,E)-8,10-dodecadienoic acid of the total fatty acids, and seeds were selected to advance top-performing lines to homozygosity. In the T4 generation, production levels of (E)-9-dodecenoic acid and (E,E)-8,10-dodecadienoic acid remained stable. The diene acid together with other seed fatty acids were converted into corresponding alcohols, and the bioactivity of the plant-derived codlemone was confirmed by GC-EAD and a flight tunnel assay. Trapping in orchards and home gardens confirmed significant and specific attraction of C. pomonella males to the plant-derived codlemone.