Yves Desjardins

A polyphenol-rich cranberry extract protects from diet-induced obesity, insulin resistance and intestinal inflammation in association with increased Akkermansia spp. population in the gut microbiota of mice

Objective The increasing prevalence of obesity and type 2 diabetes (T2D) demonstrates the failure of conventional treatments to curb these diseases. The gut microbiota has been put forward as a key player in the pathophysiology of diet-induced T2D. Importantly, cranberry (Vaccinium macrocarpon Aiton) is associated with a number of beneficial health effects. We aimed to investigate the metabolic impact of a cranberry extract (CE) on high fat/high sucrose (HFHS)-fed mice and to determine whether its consequent antidiabetic effects are related to modulations in the gut microbiota. Design C57BL/6J mice were fed either a chow or a HFHS diet. HFHS-fed mice were gavaged daily either with vehicle (water) or CE (200 mg/kg) for 8 weeks. The composition of the gut microbiota was assessed by analysing 16S rRNA gene sequences with 454 pyrosequencing. Results CE treatment was found to reduce HFHS-induced weight gain and visceral obesity. CE treatment also decreased liver weight and triglyceride accumulation in association with blunted hepatic oxidative stress and inflammation. CE administration improved insulin sensitivity, as revealed by improved insulin tolerance, lower homeostasis model assessment of insulin resistance and decreased glucose-induced hyperinsulinaemia during an oral glucose tolerance test. CE treatment was found to lower intestinal triglyceride content and to alleviate intestinal inflammation and oxidative stress. Interestingly, CE treatment markedly increased the proportion of the mucin-degrading bacterium Akkermansia in our metagenomic samples. Conclusions CE exerts beneficial metabolic effects through improving HFHS diet-induced features of the metabolic syndrome, which is associated with a proportional increase in Akkermansia spp. population.

Pineapple (Ananas comosus L. Merr) micropropagation in temporary immersion systems

Abstract A procedure for the mass propagation of pineapple plants (Ananas comosus L. Merr) using a temporary immersion technique is described. This procedure involved three distinct phases in the automated temporary immersion system: shooting, bud differentiation and elongation. To establish this protocol, we used in vitro shoots obtained from established liquid culture as starting materials. Three culture methods (solid, liquid and temporary immersion) were compared. Temporary immersion increased the multiplication rate and fresh and dry weight after 42 days. Conventional micropropagation (liquid medium) and temporary immersion were compared in combination with paclobutrazol. Paclobutrazol promoted the formation of compact bud clusters with limited leaf development. The highest multiplication rate (106) was found when ex-plants were cultured in shooting medium (MS+2.1 mg/l BA+0.3 mg/l NAA) supplemented with 1 mg/l PB for 7 weeks. A 10-l temporary immersion bioreactor was used to test two approaches during elongation stage: reduction of the shoot-formation period or decrease of the initial number of explants. The highest number of competent and uniform plants (191.8 plant/l) was achieved when shoots were cultured for 4 weeks in shooting medium supplemented with PB.

Effects of sucrose on photosynthesis and phosphoenolpyruvate carboxylase activity of in vitro cultured strawberry plantlets

Photosynthesis and phosphoenolpyruvate carboxylase activity were investigated in 5, 10 and 28 day-old micropropagated strawberry plantlets (Fragaria x ananassa Duch. cv Kent) rooted in vitro with different levels of sucrose (0, 1, 3 and 5%) on cellulose plugs (Sorbarods). The photosynthetic capability was influenced by the level of sucrose in the culture medium with the largest rates of photosynthesis corresponding to the cultures with 0 and 1% sucrose. The apparent quantum yield and the ratio of variable fluorescence to maximum fluorescence were also reduced in plantlets cultured with 3 or 5% sucrose as compared to those with 0 or 1%. Phosphoenolpyruvate carboxylase activity was largest 5 and 10 days after the onset of culture and decreased in the absence of sucrose in the culture medium. At 28 days after the onset of culture, the activity of this carboxylating enzyme was lower than at the beginning of culture and independent of the concentration of sucrose in the culture medium. Phosphoenolpyruvate carboxylase appears to be an important carboxylating enzyme in micropropagated plantlets.

Apple Peel Polyphenols and Their Beneficial Actions on Oxidative Stress and Inflammation

Since gastrointestinal mucosa is constantly exposed to reactive oxygen species from various sources, the presence of antioxidants may contribute to the body’s natural defenses against inflammatory diseases. Hypothesis To define the polyphenols extracted from dried apple peels (DAPP) and determine their antioxidant and anti-inflammatory potential in the intestine. Caco-2/15 cells were used to study the role of DAPP preventive actions against oxidative stress (OxS) and inflammation induced by iron-ascorbate (Fe/Asc) and lipopolysaccharide (LPS), respectively. Results The combination of HPLC with fluorescence detection, HPLC-ESI-MS TOF and UPLC-ESI-MS/MS QQQ allowed us to characterize the phenolic compounds present in the DAPP (phenolic acids, flavonol glycosides, flavan-3-ols, procyanidins). The addition of Fe/Asc to Caco-2/15 cells induced OxS as demonstrated by the rise in malondialdehyde, depletion of n-3 polyunsaturated fatty acids, and alterations in the activity of endogenous antioxidants (SOD, GPx, G-Red). However, preincubation with DAPP prevented Fe/Asc-mediated lipid peroxidation and counteracted LPS-mediated inflammation as evidenced by the down-regulation of cytokines (TNF-α and IL-6), and prostaglandin E2. The mechanisms of action triggered by DAPP induced also a down-regulation of cyclooxygenase-2 and nuclear factor-κB, respectively. These actions were accompanied by the induction of Nrf2 (orchestrating cellular antioxidant defenses and maintaining redox homeostasis), and PGC-1α (the “master controller” of mitochondrial biogenesis). Conclusion Our findings provide evidence of the capacity of DAPP to reduce OxS and inflammation, two pivotal processes involved in inflammatory bowel diseases.

PHYSIOLOGY OF EFFECTS OF TEMPORARY IMMERSION BIOREACTORS ON MICROPROPAGATED PINEAPPLE PLANTLETS

SummaryTemporary immersion bioreactors are an efficient tool for plant mass propagation because they increase multiplication rate and plant quality. Little knowledge is available on the ecosystem and physiological behavior of plantlets when using this new culture technique. In order to evaluate the effects of the conditions on physiological change of pineapple plantlets, a factorial experiment was conducted, where axillary clusters were cultured under two levels of photosynthetic photon flux (PPF): 30 μmol m−2s−1 (low) and 225 μmol m−2s−1 (high), using two culture methods (conventional micropropagation in liquid medium and a temporary immersion bioreactor) during the elongation phase. CO2 concentration in the headspace volume container was measured during a whole cycle of temporary immersion (3h). At the time before the next immersion period, the levels of CO2 increased significantly to 14171 μmol mol−1 at high PPF. The maximal photosynthetic rate as well as the maximum quantum yield of photosystem II were low for plantlets cultivated in the femporary immersion bioreactor at high PPF. However, these plantlets showed large increases in sugar and nitrogen uptake and also increases in dry weight and foliar area. These results indicate that shoot growth did not totally depend on the photosynthesis process. In vitro pineapple plantlets appeared to use more nutrients in the culture medium than those from photosynthesis. In summary, temporary immersion bioreactor-derived plantlets showed remarkable nutrient uptake, indicating a higher photo-mixotrophic metabolism.

A tripartite culture system for endomycorrhizal inoculation of micropropagated strawberry plantlets in vitro

The objective of the current investigation was to develop a reliable method to obtain vesicular arbuscular mycorrhizae (VAM) in micropropagated plantlets and to determine their influence on growth. An in vitro system for culturing the VA mycorrhizal fungus Glomus intraradices with Ri T-DNA-transformed carrot roots or nontransformed tomato roots was used in this study as a potential active source of inoculum for the colonization of micropropagated plantlets. After root induction, micropropagated plantlets grown on cellulose plugs (sorbarod) were placed in contact with the primary mycorrhizae in growth chambers enriched with 5000 ppm CO2 and fed with a minimal medium. After 20 days of tripartite culture, all plantlets placed in contact with the primary symbiosis were colonized by the VAM fungus. As inoculum source, 30-day-old VA mycorrhizal transformed carrot roots had a substantially higher infection potential than 5-, 10-or 20-day-old VAM. Colonized plantlets had more extensive root systems and better shoot growth than control plants. The VAM symbiosis reduced the plantlet osmotic potential. This response may be a useful pre-adaptation for plantlets during transfer to the acclimatization stage.

Carbon nutrition in vitro — regulation and manipulation of carbon assimilation in micropropagated systems

Micropropagation is one of the first biotechnologies to be extensively used by industry. This technique allows efficient and rapid multiplication of many fruit, vegetable and ornamental species and is now widely used in the trade. However, despite its extraordinary potential, it is still confronted with many problems. The stimulation of natural propagation capacity, which is desirable in terms of production efficiency, induces several anatomical, morphological and physiological changes that affect the growth of plantlets very seriously upon transfer to ex vitro conditions. A complete understanding of the conditions leading to the appearance of an aberrant morphology and physiology in vitro must be achieved in order to optimize growth of the plantlets and their survival upon transfer to acclimatization. This knowledge is necessary to increase the profitability of tissue culture laboratories and to extend the technique to industrial, agronomic and forestry crops.

Synthesis of active oryzacystatin I in transgenic potato plants

SummaryTransformation of potato (Solanum tuberosum L.) with cysteine proteinase inhibitor (PI) genes represents a potential way of controlling the major insect pest Colorado potato beetle (CPB; Leptinotarsa decemlineata Say). The present study describes the Agrobacterium-mediated transformation of potato (cv. Kennebec) with an oryzacystatin I (OCI) cDNA clone linked to a CaMV 35S promoter. The transgenic plants accumulated active OCI in potato leaves, as demonstrated by the papain-inhibitory activity of transgenic plant leaf extracts. In addition to their anti-papain activity, the extracts also caused a partial but significant inhibition of CPB digestive proteinases, similar to that observed with pure inhibitors. Recombinant OCI did not alter the activity of the major potato leaf endogenous proteinases, which seemed to be of the serine-type. Therefore we suggest that the OCI cDNA can be used for the production of CPB-resistant transgenic potato plants without interfering with endogenous proteinases of these plants.

Photosynthesis and transpiration of in vitro cultured asparagus plantlets

Abstract Nodal sections of asparagus ( Asparagus officinalis L.) were cultured and rooted on a modified Murashige and Skoog (MS) medium for 10 weeks. These plants were then acclimatized for 5 weeks. Photosynthesis and transpiration of in vitro plantlets, acclimatized plantlets and control seedlings were measured with an open gas exchange system. Rates of photosynthesis of in vitro-cultured plantlets were as high as those of seedlings grown in a greenhouse, while their rates of transpiration were much higher. Photosynthetic capacity of in vitro-cultured asparagus plantlets was sufficiently high to support autotrophic growth during the period of acclimatization. Photosynthesis of acclimatized plantlets was always considerably lower than that of in vitro plantlets except at high concentrations of CO 2 . Evaportranspiration of acclimatized plantlets was comparable to that of seedlings. These results suggest that high water loss incurred by in vitro shoots imposes severe limitations on newly formed shoots in acclimatization thus reducing whole plant photosynthesis. Protecting in vitro plantlets from water stress is the most important factor to consider in order to ensure their survival during the acclimatization period.

Exogenous sucrose can decrease in vitro photosynthesis but improve field survival and growth of coconut (cocos nucifera l.) in vitro plantlets

SummaryCoconut (Cocos nucifera L.) plantlets grown in vitro often grow slowly when transferred to the field possibly, due to a limited photosynthetic capacity of in vitro-cultured plantlets, apparently caused by the sucrose added to growth medium causing negative feedback for photosynthesis. In this paper, we tested the hypothesis that high exogenous sucrose will decrease ribulose 1,5-bisphosphate carboxylase (Rubisco) activity and photosynthesis resulting in limited ex vitro growth. Plantlets grown with high exogenous sucrose (90 gl−1) had reduced photosynthetic activity that resulted in a poor photosynthetic response to high levels of light and CO2. These plantlets also had low amounts of Rubisco protein, low Rubisco activity, and reduced growth despite showing high survival when transferred to the field. Decreasing the medium’s sucrose concentration from 90 to 22.5 gl−1 or 0 gl−1 resulted in increased photosynthetic response to light and CO2 along with increased Rubisco and phosphoenolpyruvate carboxylase (PEPC) activities and proteins. However, plantlets grown in vitro without exogenous sucrose died when transferred ex vitro, whereas those grown with intermediate exogenous sucrose showed intermediate photosynthetic response, high survival, fast growth, and ex vitro photosynthesis. Thus, exogenous sucrose at moderate concentration decreased photosynthesis but increased survival, suggesting that both in vitro photosynthesis and exogenous sucrose reserves contribute to field establisment and growth of coconut plantlets cultured in vitro.