Genetic variation in root development responses to salt stresses of quinoa

Soil salinity has become a serious environmental abiotic stress limiting crop productivity and quality. The root system is the first organ sensing the changes in salinity. Root development under elevated salinity is therefore an important indicator for saline tolerance in plants. Previous studies focused on varietal differences in morphological traits of quinoa under saline stresses; however, variation in root development responses to salinity remains largely unknown. To understand the genetic variation in root development responses to salt stress of quinoa, we conducted a preliminary screening for salinity response at two salinity levels of a diverse set of 52 quinoa genotypes and microsatellite markers were used to link molecular variation to that in root development responses to salt stresses of represented genotypes. The frequency distribution of saline tolerance index showed continuous variation in the quinoa collection. Cluster analysis of salinity responses divided the 52 quinoa genotypes into six major groups. Based on these results, six genotypes representative of groups I to VI including Black quinoa, 2-Want, Atlas, Riobamba, NL-6 and Sayaña, respectively, were selected to evaluate root development under four saline stress levels: 0, 100, 200 and 300 mM NaCl. Contrasts in root development responses to saline stress levels were observed in the six genotypes. At 100 mM NaCl, significant differences were not observed in root length development (RLD) and root surface development (RSAD) of most genotypes except Black quinoa; a significant reduction was observed in this genotype as compared to controls. At 200 mM NaCl, significant reduction was detected in RLD and RSAD in all genotypes showing this as the best concentration to discriminate among genotypes. The strongest inhibition of root development was found for all genotypes at 300 mM NaCl as compared to lower saline levels. Among genotypes, Atlas of group III shows as a saline-tolerant genotype confirming previous reports. Variation in root responses to salinity stresses is also discussed in relation to climate conditions of origins of the genotypes and reveal interesting guidelines for further studies exploring the mechanisms behind this aspect of saline adaptation.     

Cellulose content as a selection trait in breeding for kraft pulp yield in Eucalyptus urophylla

? The aim of this study was to investigate the effectiveness of using cellulose content, measured by the diglyme-HCl method, as a selection trait in breeding programs for kraft pulp yield in Eucalyptus urophylla.

? A total of 275 trees from sixty-two families were sampled from a thinned progeny trial of E. urophylla in northern Vietnam to evaluate cellulose content from breast-height increment cores. Among those, twenty unrelated trees were felled to evaluate cellulose content and pulp yield from breast-height disk samples.

? The regression of pulp yield of disk samples on cellulose content was strong either from disks (R ² = 0.83) or increment cores (R ² = 0.69). There was no significant difference in cellulose content between the provenances. The narrow-sense within-provenance heritability of cellulose content was 0.50 and the coefficient of additive genetic variation was 3.9%. Genetic correlations between cellulose content and growth (0.28–0.45) or wood basic density (?0.02) were not significantly different from zero.

? Breast-height increment core cellulose content measured by diglyme-HCl method is under strong genetic control and can be used to rank trees for pulp yield in E. urophylla plantations. Selection for increased cellulose content would have only minor effects on growth and wood basic density.

     

Agrobacterium-mediated transformation of the CrDAT gene and selection of transgenic periwinkle lines with a high vincristine accumulation

Catharanthus roseus contains vincristine and vinblastine, which are outstanding drugs for cancer. In the biosynthetic pathways of terpenoid indole alkaloids (TIAs) in C. roseus, deacetylvindoline 4-O-acetyltransferase (DAT) is a key enzyme that catalyses the last reaction of vindoline biosynthesis to form vinblastine and vincristine. In this study, the CrDAT transgene was transferred into the periwinkle by Agrobacterium-mediated transformation and generated transgenic periwinkle lines with an increase in vincristine accumulation. The C. roseus DAT gene was introduced into C. roseus plants and it was confirmed that CrDAT was successfully transferred into the genome of periwinkle plants and efficiently translated to synthesise recombinant DAT protein. Four transgenic periwinkle lines in T1 generation, T1-1, T1-3, T1-6, and T1-7, expressed recombinant DAT protein with the total protein content in the range of 2.86 μg.mg^?1 to 5.12 μg.mg^?1. Moreover, the vincristine contents of four transgenic lines increased by 1.63?2.48-fold compared to non-transgenic plants, ranging from 6.91 µg.g^?1 (fresh weight) to 10.53 µg.g^?1 (fresh weight). The T1-1 line had the highest vincristine content. Hence, the overexpression of the recombinant DAT protein can improve the vincristine accumulation of transgenic C. roseus plants.

Abbreviation: CrDAT - Catharanthus roseus Deacetylvindoline-4-O-Acetyl Transferase; D4H - Deacetoxyvindoline 4-hydroxylase; ELISA - Enzyme-Linked Immunosorbent Assay Monoterpene indole alkaloid; T0, T1 - Generations of transgenic plants; TIAs - Terpenoid indole alkaloids; WT- The wild-type tobacco plants (non transgenic plant); 35S - Cauliflower mosaic virus 35S promoter     

Regional Sediment Deficits in the Dutch Lowlands: Implications for Long-Term Land-Use Options (8 pp)

Background, Aim and Scope   Coastal and river plains are the surfaces of depositional systems, to which sediment input is a parameter of key-importance. Their habitation and economic development usually requires protection with dikes, quays, etc., which are effective in retaining floods but have the side effect of retarding sedimentation in their hinterlands. The flood-protected Dutch lowlands (so-called dike-ring areas) have been sediment-starved for up to about a millennium. In addition to this, peat decomposition and soil compaction, brought about by land drainage, have caused significant land subsidence. Sediment deficiency, defined as the combined effect of sediment-starvation and drainage-induced volume losses, has already been substantial in this area, and it is expected to become urgent in view of the forecasted effects of climate change (sea-level rise, intensified precipitation and run-off). We therefore explore this deficiency, compare it with natural (Holocene) and current human sediment inputs, and discuss it in terms of long-term land-use options. Materials and Methods: We use available 3D geological models to define natural sediment inputs to our study area. Recent progress in large-scale modelling of peat oxidation and compaction enables us to address volume loss associated with these processes. Human sediment inputs are based on published minerals statistics. All results are given as first-order approximations. Results: The current sediment deficit in the diked lowlands of the Netherlands is estimated at 136 ± 67 million m3/a. About 85% of this volume is the hypothetical amount of sediment required to keep up with sea-level rise, and 15% is the effect of land drainage (peat decomposition and compaction). The average Holocene sediment input to our study area (based on a total of 145 km3) is ~14 million m3/a, and the maximum (millennium-averaged) input ~26 million m3/a. Historical sediment deficiency has resulted in an unused sediment accommodation space of about 13.3 km3. Net human input of sediment material currently amounts to ~23 million m3/a. Discussion: As sedimentary processes in the Dutch lowlands have been retarded, the depositional system's natural resilience to sea-level rise is low, and all that is left to cope is human countermeasure. Preserving some sort of status quo with water management solutions may reach its limits in the foreseeable future. The most viable long-term solutions therefore seem a combination of allowing for more water in open country (anything from flood-buffer zones to open water) and raising lands that are to be built up (enabling their lasting protection). As to the latter, doubling or tripling the use of filling sand in a planned and sustained effort may resolve up to one half of the Dutch sediment deficiency problems in about a century. Conclusions: Conclusions, Recommendations and Perspectives. We conclude that sediment deficiency – past, present and future – challenges the sustainable habitation of the Dutch lowlands. In order to explore possible solutions, we recommend the development of long-term scenarios for the changing lowland physiography, that include the effects of Global Change, compensation measures, costs and benefits, and the implications for long-term land-use options. Recommendations and Perspectives: -     

Variable-timing,fixed-rate application of cattle biogas effluent to rice using a leaf color chart: microcosm experiments in Vietnam

ABSTRACT

Livestock production plays a leading role in agricultural land-use change. Producing biogas from livestock waste and subsequently using the biogas effluent as fertilizer for crops is a promising option to solve environmental problems resulting from expanding livestock production. However, it is difficult to promptly and accurately measure the nitrogen (N) concentration of effluent for farmers in developing countries, making precise N management difficult. The objectives of the current study were (1) to evaluate the feasibility of variable-timing, fixed-rate application of cattle biogas effluent using a leaf color chart (LCC) for rice (Oryza sativa L.) and (2) to determine the optimum LCC threshold for grain yield. We conducted two microcosm experiments in the Mekong Delta of Vietnam in 2018 using eight treatments of N-fertilizer application. In the Zero treatment, we applied no N. In the Estd treatment, we split-applied N as effluent (E) at fixed rate and timing as the standard method. In E2.75, E3.00, E3.25, E3.50, and E3.75, we applied effluent whenever the LCC value went below 2.75, 3.00, 3.25, 3.50, and 3.75, respectively. In U3.25, we applied N as urea (U) whenever the LCC value fell below 3.25. The total effluent-N application rate ranged from 90 to 210 kg N ha^?1 season^?1. Rice growth was normal but there was a substantial yield gap between the two microcosm experiments due to the seasonal difference in solar radiation. Rice yield tended to increase with increasing LCC threshold. There was a positive linear relationship between LCC and chlorophyll content (SPAD) values (R ² = 0.73–0.79). Grain yield was well explained (R ² = 0.70–0.89) by the seasonal mean LCC or SPAD value. Plant total N uptake increased with increasing LCC threshold, but the three calculated indices of N use efficiency (NUE) – apparent N recovery, agronomic NUE, and internal NUE – were not always improved with a higher LCC threshold. Our results showed that the tested variable-timing, fixed-rate strategy for the application of cattle biogas effluent was feasible and the optimum LCC threshold for grain production was 3.75 under the current microcosm conditions.     

The potential of hyperspectral images and partial least square regression for predicting total carbon,total nitrogen and their isotope composition in forest litterfall samples

Purpose

The main objective of this study was to examine the potential of using hyperspectral image analysis for prediction of total carbon (TC), total nitrogen (TN) and their isotope composition (δ¹³C and δ¹⁵N) in forest leaf litterfall samples.

Materials and methods

Hyperspectral images were captured from ground litterfall samples of a natural forest in the spectral range of 400–1700 nm. A partial least-square regression model (PLSR) was used to correlate the relative reflectance spectra with TC, TN, δ¹³C and δ¹⁵N in the litterfall samples. The most important wavelengths were selected using β coefficient, and the final models were developed using the most important wavelengths. The models were, then, tested using an external validation set.

Results and discussion

The results showed that the data of TC and δ¹³C could not be fitted to the PLSR model, possibly due to small variations observed in the TC and δ¹³C data. The model, however, was fitted well to TN and δ¹⁵N. The cross-validation R² _cv of the models for TN and δ¹⁵N were 0.74 and 0.67 with the RMSE_cv of 0.53% and 1.07‰, respectively. The external validation R² _ex of the prediction was 0.64 and 0.67, and the RMSE_ex was 0.53% and 1.19 ‰, for TN and δ¹⁵N, respectively. The ratio of performance to deviation (RPD) of the predictions was 1.48 and 1.53, respectively, for TN and δ¹⁵N, showing that the models were reliable for the prediction of TN and δ¹⁵N in new forest leaf litterfall samples.

Conclusions

The PLSR model was not successful in predicting TC and δ¹³C in forest leaf litterfall samples using hyperspectral data. The predictions of TN and δ¹⁵N values in the external litterfall samples were reliable, and PLSR can be used for future prediction.

     

A food-grade system for inducible gene expression in Lactobacillus plantarum using an alanine racemase-encoding selection marker

Food-grade gene expression systems for lactic acid bacteria are useful for applications in the food industry. We describe a new food-grade host/vector system for Lactobacillus plantarum based on pSIP expression vectors and the use of the homologous alanine racemase gene (alr) as selection marker. A new series of expression vectors were constructed by exchanging the erythromycin resistance gene (erm) in pSIP vectors by the L. plantarum WCFS1 alr gene. The vectors were applied for the overexpression of β-galactosidase genes from L. reuteri L103 and L. plantarum WCFS1 in an alr deletion mutant of L. plantarum WCFS1. The expression levels obtained in this way, i.e. without the use of antibiotics, were comparable to the levels obtained with the conventional system based on selection for erythromycin resistance. The new system is suitable for the production of ingredients and additives for the food industry.     

Bordetella bronchiseptica aroA mutant as a live vaccine vehicle for heterologous porcine circovirus type 2 major capsid protein expression

Porcine circovirus type 2 (PCV2) infections cause important respiratory diseases in the pig industry and are associated with many bacterial, mycoplasmal, and viral complications. In this study, a heterologous PCV2 major capsid protein (MCP) was expressed in the Bordetella bronchiseptica aroA mutant strain (BBS-MCP) and used as a live vaccine vehicle. Mice and pigs were immunized with live BBS-MCP via the intranasal route. The antibodies against MCP were induced successfully in the serum as determined by ELISA. In the PCV2 challenge experiment, viral DNA was removed successfully from the lymph nodes of pigs vaccinated with live BBS-MCP. Overall, BBS-MCP is believed to be a good candidate for the development of a live-attenuated vaccine against PCV2 infections.