Dataset Information

Direct cryopreservation of winter-acclimated buds of Dracocephalum austriacum (Lamiaceae) from field material.

ABSTRACT: This study develops protocols for the micropropagation and cryopreservation of Dracocephalum austriacum (Lamiaceae). It is a perennial herbaceous plant that overwinters with ground-level sprouts and is classified as critically endangered in Europe. In vitro cultures were initiated from seeds on growth-regulator-free Murashige & Skoog (MS) medium after nicking the seed coat. Propagation via shoot culture was achieved on ½ MS medium with 1 µM benzyl adenine (BAP). Rooting on various indole-3-butyric acid (IBA)-media was not reliable, but the rooting success was 80% after 10 weeks on medium with 1 µM BAP. Two starting materials underwent cryopreservation: (1) shoot tips from cold-acclimated in vitro plantlets and (2) axillary buds from winter shoots from field plants. For the cryopreservation of in vitro shoots, plant vitrification solution (PVS)3 and incubation over ice yielded the best results (~?34% regeneration success). However, regeneration using winter acclimated buds were 100, 76 and 30% for collections in December, February and March, respectively, using the same protocol. Moreover, the ploidy levels of cryopreserved plantlets were estimated using flow cytometry. The use of winter-acclimated field material of temperate herbaceous plants or subshrubs has high potential as explant source for cryopreservation and calls for exploring this technique for other species.

SUBMITTER: Rasl T

PROVIDER: S-EPMC7307384 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Direct cryopreservation of winter-acclimated buds of <i>Dracocephalum austriacum</i> (Lamiaceae) from field material.

Rasl Thomas T Schalk Mona M Temsch Eva E Kodym Andrea A

Plant cell, tissue and organ culture 20200519 1

This study develops protocols for the micropropagation and cryopreservation of <i>Dracocephalum austriacum</i> (Lamiaceae). It is a perennial herbaceous plant that overwinters with ground-level sprouts and is classified as critically endangered in Europe. In vitro cultures were initiated from seeds on growth-regulator-free Murashige & Skoog (MS) medium after nicking the seed coat. Propagation via shoot culture was achieved on ½ MS medium with 1 µM benzyl adenine (BAP). Rooting on various indole- ...[more]

PMID: 32587434

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Project description:This study compared the photosynthetic performance and the global gene expression of the winter hardy wheat Triticum aestivum cv Norstar grown under non-acclimated (NA) or cold-acclimated (CA) condition at either ambient CO2 or elevated CO2 (EC). CA Norstar maintained comparable light saturated and CO2 saturated rates of photosynthesis but lower quantum requirements for photosystem II and non photochemical quenching relative to NA plants even at EC. Neither NA nor CA plants were sensitive to feedback inhibition of photosynthesis at EC. Global gene expression using microarray combined with bioinformatics analysis revealed that genes affected by EC were 3 times higher in NA (1022 genes) compared to CA (372 genes) Norstar. The most striking effect was the down-regulation of genes involved in the plant defense responses in NA Norstar. In contrast, cold acclimation reversed this down regulation due to the cold induction of genes involved in plant pathogenesis resistance, and cellular and chloroplast protection. These results suggest that EC have less impact on plant performance and productivity in cold adapted winter hardy plants in the northern climates compared to warmer environments. Selection for cereal cultivars with constitutively higher expression of biotic stress defense genes may be necessary under EC during the warm growth period and in warmer climates. Twelve replicate pots with 3 plants per pot were grown at either NA or CA conditions at either ambient or EC. To minimize any possible chamber effects, the 12 replicate pots at each growth condition were distributed between two growth chambers. A total of 3 biological replicates for each condition were used for microarray analyses. Each biological replicate sample was obtained by pooling three whole fully expanded 3rd leaves from different plants harvested randomly. The different biological samples of Norstar winter wheat leaves were ground in dry ice to fine powder and total RNA was extracted with trizol (Invitrogen, Burlington, ON, CA). Total RNA was cleaned using RNeasy plant mini kit (Qiagen) and integrity was determined on agarose gel and on a bioanalyser (Agilent 2100). Synthesized cDNAs were transcribed to cRNAs with the 3M-bM-^@M-^YIVT labelling kit (Santa Clara, CA, USA) and hybridized to the Affymetrix wheat genome array (Santa Clara, Ca, USA) at the McGill University and GM-CM-)nome QuM-CM-)bec Innovation Centre (Montreal, Qc, CA). The experimental design consisted of three biological replicates for each of the four growth conditions, 1: Non-acclimated (NA) at ambient CO2 (AC) (NAAC); 2: Cold-acclimated (CA) at ambient CO2 (AC) (CAAC); 3: Non-acclimated (NA) at elevated CO2 (EC) (NAEC); 4: Cold-acclimated (CA) at elevated CO2 (EC) (CAEC). Thus, a total of 3 biological samples from each of the 4 growth conditions described above were used for hybridizations.

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Dataset Information

Direct cryopreservation of winter-acclimated buds of Dracocephalum austriacum (Lamiaceae) from field material.

Publications

Direct cryopreservation of winter-acclimated buds of <i>Dracocephalum austriacum</i> (Lamiaceae) from field material.

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