A Past Genetic Bottleneck from Argentine Beans and a Selective Sweep Led to the Race Chile of the Common Bean (Phaseolus vulgaris L.).

The domestication process of the common bean gave rise to six different races which come from the two ancestral genetic pools, the Mesoamerican (Durango, Jalisco, and Mesoamerica races) and the Andean (New Granada, Peru, and Chile races). In this study, a collection of 281 common bean landraces from Chile was analyzed using a 12K-SNP microarray. Additionally, 401 accessions representing the rest of the five common bean races were analyzed. A total of 2543 SNPs allowed us to differentiate a genetic group of 165 accessions that corresponds to the race Chile, 90 of which were classified as pure accessions, such as the bean types 'Tórtola', 'Sapito', 'Coscorrón', and 'Frutilla'. Our genetic analysis indicates that the race Chile has a close relationship with accessions from Argentina, suggesting that nomadic ancestral peoples introduced the bean seed to Chile. Previous archaeological and genetic studies support this hypothesis. Additionally, the low genetic diversity (π = 0.053; uHe = 0.53) and the negative value of Tajima' D (D = -1.371) indicate that the race Chile suffered a bottleneck and a selective sweep after its introduction, supporting the hypothesis that a small group of Argentine bean genotypes led to the race Chile. A total of 235 genes were identified within haplotype blocks detected exclusively in the race Chile, most of them involved in signal transduction, supporting the hypothesis that intracellular signaling pathways play a fundamental role in the adaptation of organisms to changes in the environment. To date, our findings are the most complete investigation associated with the origin of the race Chile of common bean.

A Comprehensive Review on Chickpea (Cicer arietinum L.) Breeding for Abiotic Stress Tolerance and Climate Change Resilience.

Chickpea is one of the most important pulse crops worldwide, being an excellent source of protein. It is grown under rain-fed conditions averaging yields of 1 t/ha, far from its potential of 6 t/ha under optimum conditions. The combined effects of heat, cold, drought, and salinity affect species productivity. In this regard, several physiological, biochemical, and molecular mechanisms are reviewed to confer tolerance to abiotic stress. A large collection of nearly 100,000 chickpea accessions is the basis of breeding programs, and important advances have been achieved through conventional breeding, such as germplasm introduction, gene/allele introgression, and mutagenesis. In parallel, advances in molecular biology and high-throughput sequencing have allowed the development of specific molecular markers for the genus Cicer, facilitating marker-assisted selection for yield components and abiotic tolerance. Further, transcriptomics, proteomics, and metabolomics have permitted the identification of specific genes, proteins, and metabolites associated with tolerance to abiotic stress of chickpea. Furthermore, some promising results have been obtained in studies with transgenic plants and with the use of gene editing to obtain drought-tolerant chickpea. Finally, we propose some future lines of research that may be useful to obtain chickpea genotypes tolerant to abiotic stress in a scenario of climate change.

Soft tissue metric parameters, methods and aesthetic indices in implant dentistry: A critical review.

OBJECTIVES: The primary objective was to provide an overview of soft tissue metric parameters, methods, and aesthetic indices in implant dentistry. The secondary objective was to describe reliability and validity of aesthetic indices. MATERIALS AND METHODS: Two independent reviewers conducted an electronic literature search in Pubmed, Web of Science, Embase, and Cochrane databases up to October 2020 to identify studies on soft tissue metric parameters, methods, and aesthetic indices. Aesthetic indices were evaluated in terms of reliability and validity. Data extraction was performed by the same reviewers. RESULTS: Five metric parameters (papilla height, linear changes in soft tissue level, color assessment, soft tissue thickness, and profilometric soft tissue changes) registered by means of several methods (intra-oral registrations, radiographic assessments, digital analyses, and ultrasonic assessments), and 15 aesthetic indices (Papilla Index (PI), ad hoc questions scored with Visual Analogue Scales, Pink Esthetic Score (PES), Implant Crown Aesthetic Index (ICAI), Implant Aesthetic Score (IAS), Rompen Index, Subjective Esthetic Score, White Esthetic Score, Copenhagen Index, Complex Esthetic Index, Californian Dental Association Index (CDAI), Peri-Implant, and Crown Index, Functional Implant Prosthodontic Score, Implant Restoration Esthetic Index (IREI), and Mucosal Scarring Index (MSI)) could be identified. With respect to metric parameters and methods, intra-oral registrations were least accurate whereas profilometric soft tissue changes on the basis of digital surface models were most accurate. Six aesthetic indices showed good inter-rater reliability (PI, PES, ICAI, CDAI, IREI, and MSI). Good validity could only be shown for two indices (PES and CEI). Given this and on the basis of ease of use and ease of interpretation, PES qualified best for clinical research on single implants. None of the indices fulfilled the quality criteria for clinical research on multiple implants. CONCLUSION: Many soft tissue assessment methods with varying reliability and validity have been described and used, which hampers uniform reporting in implant dentistry. Clinical investigators are advised to measure linear and profilometric soft tissue changes using digital surface models, and to use a reliable and validated aesthetic index. Currently, PES qualifies best for aesthetic evaluation of single implants. An index is to be developed to assess the aesthetic outcome of rehabilitations on multiple implants.

Severe outbreak of Fusarium wilt on common beans (Phaseolus vulgaris) caused by Fusarium oxysporum in the Maule Region, central Chile.

Common bean (Phaseolus vulgaris L.) is an important grain legume cultivated worldwide as food for humans and livestock (Schwartz et al., 2005). Common beans in central Chile reach up to 3,893 ha from which 1,069 ha are located in the Maule region. Common bean is produced by small farmers who have limited access to fertilization, technical irrigation, and crop protection. In spring 2018, bean plants initially showed a slight yellowing and premature senescence 50 days after sowing (das) until showing wilting symptoms (70 -100 das) in Curepto fields (35 05'S; 72 01'W), Maule region. The basal part of affected plants displayed internal reddish-brown discoloration of the vascular tissues. Based on the plant external symptoms, we estimated an incidence between 15% and 45% in bean fields. Nine symptomatic plants were collected, and surface washed with sterile water and disinfested with 75% ethanol (v/v). Then small fragments (5-mm) from damage vascular tissue from each plant were cut and placed on Petri dishes containing PDA acidified with 0.5 ml/l of 92% lactic acid (APDA, 2%). The isolations were incubated for seven days at 25°C. Nine Fusarium-like isolates from single-spore on APDA (2%) became pale vinaceous, floccose with abundant aerial mycelium and dark vinaceous reverse colony, with a growing rate of 10.8 to 11.6 mm/d at 25°C (Lombard et al., 2019). Phialides were short, singular growing laterally on the mycelium. Macroconidia were hyaline, fusiform with basal foot cells shaped to pointed and apical cells tapered, 2-5 septate, and 28.6 to 47.6 (av. 38.1) µm long x 2.2 to 3.6 (av. 3.1) µm wide. Microconidia were hyaline, oval to ellipsoid, one-celled, and 4.5 to 10.9 (av. 6.1) µm long and 2.2 to 3.3 (av. 2.7) µm wide (n=50 spore). For molecular identification, three isolates (Curi-3.1, Be-8.1, and Be-11.3) were sequenced using PCR amplification of the partial sequences of beta-tubulin (BT) and translation elongation factor 1-α gene (TEF) (Lombard et al., 2019). NCBI BLAST analysis showed 99 to 100% similarity with sequences (TEF; BT) of strain CPC 25822 of Fusarium oxysporum. The maximum-likelihood phylogenetic analysis placed the Chilean isolates in the F. oxysporum complex clade. Chilean sequences were deposited into GenBank under accession numbers MW419125, MW419126, MW419127 (TEF) and MW419128, MW419129, MW419130 (BT). Pathogenicity tests (isolates Curi-3.1, Be-8.1, and Be-11.3) were conducted under greenhouse (15-28°C, 85%RH) on healthy bean plants (n=30) cv. Blanco Español INIA cultivated in pots (sand/peat moss/soil) at the University of Talca. Plants that are 30 days-old were inoculated using 200 µl of conidial suspension (106 conidia/ml) on wounded roots (crown). Control plants (n=10) were similarly inoculated with sterile distilled water. After 45 days, all inoculated plants with F. oxysporum isolates developed necrotic lesions on vascular tissue, and chlorosis, and wilting while control plants remained healthy. This experiment was conducted twice. The pathogen was reisolated (100%) from diseased plants and molecularly identified as F. oxysporum. To our knowledge, this is the report of a severe outbreak of F. oxysporum causing Fusarium yellows in P. vulgaris in the Maule region, Chile. Previously, F. oxysporum has been reported affecting tomato (Sepúlveda-Chavera et al., 2014) and blueberry in Chile (Moya-Elizondo et al., 2019).

LeNRT1.1 Improves Nitrate Uptake in Grafted Tomato Plants under High Nitrogen Demand.

Grafting has become a common practice among tomato growers to obtain vigorous plants. These plants present a substantial increase in nitrogen (N) uptake from the root zone. However, the mechanisms involved in this higher uptake capacity have not been investigated. To elucidate whether the increase in N uptake in grafted tomato plants under high N demand conditions is related to the functioning of low- (high capacity) or high-affinity (low capacity) root plasma membrane transporters, a series of experiments were conducted. Plants grafted onto a vigorous rootstock, as well as ungrafted and homograft plants, were exposed to two radiation levels (400 and 800 µmol m-2 s-1). We assessed root plasma membrane nitrate transporters (LeNRT1.1, LeNRT1.2, LeNRT2.1, LeNRT2.2 and LeNRT2.3) expression, Michaelis‒Menten kinetics parameters (Vmax and Km), root and leaf nitrate reductase activity, and root respiration rates. The majority of nitrate uptake is mediated by LeNRT1.1 and LeNRT1.2 in grafted and ungrafted plants. Under high N demand conditions, vigorous rootstocks show similar levels of expression for LeNRT1.1 and LeNRT1.2, whereas ungrafted plants present a higher expression of LeNRT1.2. No differences in the uptake capacity (evaluated as Vmax), root respiration rates, or root nitrate assimilation capacity were found among treatments.

Long-term non-invasive and continuous measurements of legume nodule activity.

Symbiotic nitrogen fixation is a process of considerable economic, ecological and scientific interest. The central enzyme nitrogenase reduces H(+) alongside N2 , and the evolving H2 allows a continuous and non-invasive in vivo measurement of nitrogenase activity. The objective of this study was to show that an elaborated set-up providing such measurements for periods as long as several weeks will produce specific insight into the nodule activity's dependence on environmental conditions and genotype features. A system was developed that allows the air-proof separation of a root/nodule and a shoot compartment. H2 evolution in the root/nodule compartment can be monitored continuously. Nutrient solution composition, temperature, CO2 concentration and humidity around the shoots can concomitantly be maintained and manipulated. Medicago truncatula plants showed vigorous growth in the system when relying on nitrogen fixation. The set-up was able to provide specific insights into nitrogen fixation. For example, nodule activity depended on the temperature in their surroundings, but not on temperature or light around shoots. Increased temperature around the nodules was able to induce higher nodule activity in darkness versus light around shoots for a period of as long as 8 h. Conditions that affected the N demand of the shoots (ammonium application, Mg or P depletion, super numeric nodules) induced consistent and complex daily rhythms in nodule activity. It was shown that long-term continuous measurements of nodule activity could be useful for revealing special features in mutants and could be of importance when synchronizing nodule harvests for complex analysis of their metabolic status.

An RNA sequencing transcriptome analysis reveals novel insights into molecular aspects of the nitrate impact on the nodule activity of Medicago truncatula.

The mechanism through which nitrate reduces the activity of legume nodules is controversial. The objective of the study was to follow Medicago truncatula nodule activity after nitrate provision continuously and to identify molecular mechanisms, which down-regulate the activity of the nodules. Nodule H2 evolution started to decline after about 4 h of nitrate application. At that point in time, a strong shift in nodule gene expression (RNA sequencing) had occurred (1,120 differentially expressed genes). The most pronounced effect was the down-regulation of 127 genes for nodule-specific cysteine-rich peptides. Various other nodulins were also strongly down-regulated, in particular all the genes for leghemoglobins. In addition, shifts in the expression of genes involved in cellular iron allocation and mitochondrial ATP synthesis were observed. Furthermore, the expression of numerous genes for the formation of proteins and glycoproteins with no obvious function in nodules (e.g. germins, patatin, and thaumatin) was strongly increased. This occurred in conjunction with an up-regulation of genes for proteinase inhibitors, in particular those containing the Kunitz domain. The additionally formed proteins might possibly be involved in reducing nodule oxygen permeability. Between 4 and 28 h of nitrate exposure, a further reduction in nodule activity occurred, and the number of differentially expressed genes almost tripled. In particular, there was a differential expression of genes connected with emerging senescence. It is concluded that nitrate exerts rapid and manifold effects on nitrogenase activity. A certain degree of nitrate tolerance might be achieved when the down-regulatory effect on late nodulins can be alleviated.

RNA-seq transcriptome profiling reveals that Medicago truncatula nodules acclimate N2 fixation before emerging P deficiency reaches the nodules.

Legume nodules are plant tissues with an exceptionally high concentration of phosphorus (P), which, when there is scarcity of P, is preferentially maintained there rather than being allocated to other plant organs. The hypothesis of this study was that nodules are affected before the P concentration in the organ declines during whole-plant P depletion. Nitrogen (N2) fixation and P concentration in various organs were monitored during a whole-plant P-depletion process in Medicago truncatula. Nodule gene expression was profiled through RNA-seq at day 5 of P depletion. Until that point in time P concentration in leaves reached a lower threshold but was maintained in nodules. N2-fixation activity per plant diverged from that of fully nourished plants beginning at day 5 of the P-depletion process, primarily because fewer nodules were being formed, while the activity of the existing nodules was maintained for as long as two weeks into P depletion. RNA-seq revealed nodule acclimation on a molecular level with a total of 1140 differentially expressed genes. Numerous genes for P remobilization from organic structures were increasingly expressed. Various genes involved in nodule malate formation were upregulated, while genes involved in fermentation were downregulated. The fact that nodule formation was strongly repressed with the onset of P deficiency is reflected in the differential expression of various genes involved in nodulation. It is concluded that plants follow a strategy to maintain N2 fixation and viable leaf tissue as long as possible during whole-plant P depletion to maintain their ability to react to emerging new P sources (e.g. through active P acquisition by roots).

The activity of nodules of the supernodulating mutant Mtsunn is not limited by photosynthesis under optimal growth conditions.

Legumes match the nodule number to the N demand of the plant. When a mutation in the regulatory mechanism deprives the plant of that ability, an excessive number of nodules are formed. These mutants show low productivity in the fields, mainly due to the high carbon burden caused through the necessity to supply numerous nodules. The objective of this study was to clarify whether through optimal conditions for growth and CO2 assimilation a higher nodule activity of a supernodulating mutant of Medicago truncatula (M. truncatula) can be induced. Several experimental approaches reveal that under the conditions of our experiments, the nitrogen fixation of the supernodulating mutant, designated as sunn (super numeric nodules), was not limited by photosynthesis. Higher specific nitrogen fixation activity could not be induced through short- or long-term increases in CO2 assimilation around shoots. Furthermore, a whole plant P depletion induced a decline in nitrogen fixation, however this decline did not occur significantly earlier in sunn plants, nor was it more intense compared to the wild-type. However, a distinctly different pattern of nitrogen fixation during the day/night cycles of the experiment indicates that the control of N2 fixing activity of the large number of nodules is an additional problem for the productivity of supernodulating mutants.

N2 Fixation, N Transfer, and Land Equivalent Ratio (LER) in Grain Legume-Wheat Intercropping: Impact of N Supply and Plant Density.

Intercropping legumes with cereals can lead to increased overall yield and optimize the utilization of resources such as water and nutrients, thus enhancing agricultural efficiency. Legumes possess the unique ability to acquire nitrogen (N) through both N2 fixation and from the available N in the soil. However, soil N can diminish the N2 fixation capacity of legumes. It is postulated that in intercropping, legumes uptake N mainly through N2 fixation, leaving more soil N available for cereals. The latter, in turn, has larger root systems, allowing it to explore greater soil volume and absorb more N, mitigating its adverse effects on N2 fixation in legumes. The goal of this study was to evaluate how the supply of N affects the intercropping of faba beans (Vicia faba L.) and peas (Pisum sativum L.) with wheat under varying plant densities and N levels. We measured photosynthetic traits, biomass production, the proportion of N derived from air (%Ndfa) in the shoot of the legumes, the N transferred to the wheat, and the land equivalent ratio (LER). The results revealed a positive correlation between soil N levels and the CO2 assimilation rate (An), chlorophyll content, and N balance index (NBI) in wheat. However, no significant effect was observed in legumes as soil N levels increased. Transpiration (E) increased in wheat intercropped with legumes, while stomatal conductance (gs) increased with N addition in all crops. Water use efficiency (WUE) decreased in faba beans intercropped with wheat as N increased, but it showed no significant change in wheat or peas. The shoot dry matter of wheat increased with the addition of N; however, the two legume species showed no significant changes. N addition reduced the %Ndfa of both legume species, especially in monoculture, with peas being more sensitive than faba beans. The intercropping of wheat alleviated N2 fixation inhibition, especially at high wheat density and increased N transfer to wheat, particularly with peas. The LER was higher in the intercropping treatments, especially under limited N conditions. It is concluded that in the intercropping of wheat with legumes, the N2 fixation inhibition caused by soil N is effectively reduced, as well as there being a significant N transfer from the legume to the wheat, with both process contributing to increase LER.

Nutritional Characterization of Chilean Landraces of Common Bean.

Common bean (Phaseolus vulgaris L.) is the primary grain legume cultivated worldwide for direct human consumption due to the high nutritional value of its seeds and pods. The high protein content of common beans highlights it as the most promising source of plant-based protein for the food industry. Additionally, landraces of common bean have great variability in nutritional traits, which is necessary to increase the nutritional quality of elite varieties. Therefore, the main objective of this study was to nutritionally characterize 23 Chilean landraces and 5 commercial varieties of common bean to identify genotypes with high nutritional value that are promising for the food industry and for genetic improvement programs. The landrace Phv23 ('Palo') was the most outstanding with high concentrations of minerals such as P (7.53 g/kg), K (19.8 g/kg), Mg (2.43 g/kg), Zn (52.67 mg/kg), and Cu (13.67 mg/kg); essential amino acids (364.8 mg/g protein); and total proteins (30.35 g/100 g seed). Additionally, the landraces Phv9 ('Cimarrón'), Phv17 ('Juanita'), Phv3 ('Araucano'), Phv8 ('Cabrita/Señorita'), and Phv4 ('Arroz') had a high protein content. The landrace Phv24 ('Peumo') stood out for its phenolic compounds (TPC = 218.1 mg GA/100 g seed) and antioxidant activity (ORAC = 22,167.9 µmol eq trolox/100 g extract), but it has moderate to low mineral and protein concentrations. In general, the concentration of nutritional compounds in some Chilean landraces was significantly different from the commercial varieties, highlighting their high nutritional value and their potential use for the food industry and for genetic improvement purposes.

Assessing Grain Quality Changes in White and Black Rice under Water Deficit.

Rice is an essential diet component for a significant portion of the population worldwide. Due to the high water demand associated with rice production, improving water use efficiency and grain quality is critical to increasing the sustainability of the crop. This species includes rice varieties with diverse pigmentation patterns. Grain quality, including industrial, nutritional, and functional quality traits, of two black rice genotypes and a commercial white rice cultivar were evaluated in different locations and under different water regimes. Flooding produced higher grain weight compared to alternate wetting and drying irrigation. A high correlation was found between grain color, total phenolic content (TPC), and antioxidant activity. The black rice genotypes showed higher TPC levels and antioxidant capacity, mainly due to higher levels of cyanidin 3-O-glucoside. The phenolic profile varied between whole and polished grains, while mineral composition was influenced by location and irrigation regime. In turn, the environment influenced grain quality in terms of industrial and nutritional characteristics, with significant differences in quality between whole and polished grains. This study provides valuable information on the genotype-environment relationship in rice and its effect on grain quality, which could contribute to selecting genotypes for an appropriate environment.

Meta-QTL Analysis for Yield Components in Common Bean (Phaseolus vulgaris L.).

Common bean is one of the most important legumes produced and consumed worldwide because it is a highly valuable food for the human diet. However, its production is mainly carried out by small farmers, who obtain average grain yields below the potential yield of the species. In this sense, numerous mapping studies have been conducted to identify quantitative trait loci (QTL) associated with yield components in common bean. Meta-QTL (MQTL) analysis is a useful approach to combine data sets and for creating consensus positions for the QTL detected in independent studies. Consequently, the objective of this study was to perform a MQTL analysis to identify the most reliable and stable genomic regions associated with yield-related traits of common bean. A total of 667 QTL associated with yield-related traits reported in 21 different studies were collected. A total of 42 MQTL associated with yield-related traits were identified, in which the average confidence interval (CI) of the MQTL was 3.41 times lower than the CIs of the original QTL. Most of the MQTL (28) identified in this study contain QTL associated with yield and phenological traits; therefore, these MQTL can be useful in common bean breeding programs. Finally, a total of 18 candidate genes were identified and associated with grain yield within these MQTL, with functions related to ubiquitin ligase complex, response to auxin, and translation elongation factor activity.

Plant-growth-promoting bacteria from rhizosphere of Chilean common bean ecotype (Phaseolus vulgaris L.) supporting seed germination and growth against salinity stress.

Salinity abiotic stress is increasing day by day due to continuous global warming and climate change. This is also becoming one of the major causes behind the reduction in crop production. Plant-bacteria interaction plays an essential role in improving crop yield without using any chemical fertilizers. The present study aims to characterize the interaction between plant-growth-promoting bacteria (PGPB) and their role in mitigating salinity stress for local variety crops. Therefore, in this work, two PGPB, namely, Bacillus proteolyticus Cyn1 and Bacillus safensis Cyn2, were isolated from rhizospheric soil of the Chilean common bean ecotype "Sapito" (Phaseolus vulgaris L.), and their PGPB traits were analyzed. Cyn1 can produce NH3 and HCN and also secrete siderophores, whereas Cyn2 produced NH3 and siderophores but responded negatively to HCN production. Both the isolated bacteria have shown a positive result for ACC deaminase production, phosphate solubilization, and catalase enzyme secretion. Under all three tested abiotic stresses, i.e., temperature, water, and salinity, both the bacteria and their consortium have demonstrated positive responses. Cyn1 under temperature stress and water stress can produce a biofilm network to combat the stress. While under salinity stress, both the PGPB isolates indicated the production of stress components and cytoplasmic inclusion bodies. Based on the response, among all other abiotic stresses, salinity stress was chosen for further plant-bacteria interaction study and growth. Visible root colonization of the bacteria has been observed in comparison to the control. The germination index was 100% for all experimental setups of seed bacterization, both under control conditions and salinity stress. Both bacteria responded with good PGP traits that helped in the growth of healthy plants after the bacterial treatment in final pot experiments. Additionally, the consortium and the plants treated with Cyn1 have demonstrated high production of photosynthetic pigments in both experimental setups. Both B. proteolyticus Cyn1 and B. safensis Cyn2 have shown promising PGP characters and efficient response against toxicity related to salinity. Hence, both of these bacteria and consortium can be used for improved agricultural production of Chilean native common beans in the near future.

Improving bitter pit prediction by the use of X-ray fluorescence (XRF): A new approach by multivariate classification.

Bitter pit (BP) is one of the most relevant post-harvest disorders for apple industry worldwide, which is often related to calcium (Ca) deficiency at the calyx end of the fruit. Its occurrence takes place along with an imbalance with other minerals, such as potassium (K). Although the K/Ca ratio is considered a valuable indicator of BP, a high variability in the levels of these elements occurs within the fruit, between fruits of the same plant, and between plants and orchards. Prediction systems based on the content of elements in fruit have a high variability because they are determined in samples composed of various fruits. With X-ray fluorescence (XRF) spectrometry, it is possible to characterize non-destructively the signal intensity for several mineral elements at a given position in individual fruit and thus, the complete signal of the mineral composition can be used to perform a predictive model to determine the incidence of bitter pit. Therefore, it was hypothesized that using a multivariate modeling approach, other elements beyond the K and Ca could be found that could improve the current clutter prediction capability. Two studies were carried out: on the first one an experiment was conducted to determine the K/Ca and the whole spectrum using XRF of a balanced sample of affected and non-affected 'Granny Smith' apples. On the second study apples of three cultivars ('Granny Smith', 'Brookfield' and 'Fuji'), were harvested from two commercial orchards to evaluate the use of XRF to predict BP. With data from the first study a multivariate classification system was trained (balanced database of healthy and BP fruit, consisting in 176 from each group) and then the model was applied on the second study to fruit from two orchards with a history of BP. Results show that when dimensionality reduction was performed on the XRF spectra (1.5 - 8 KeV) of 'Granny Smith' apples, comparing fruit with and without BP, along with K and Ca, four other elements (i.e., Cl, Si, P, and S) were found to be deterministic. However, the PCA revealed that the classification between samples (BP vs. non-BP fruit) was not possible by univariate analysis (individual elements or the K/Ca ratio).Therefore, a multivariate classification approach was applied, and the classification measures (sensitivity, specificity, and balanced precision) of the PLS-DA models for all cultivars evaluated ('Granny Smith', 'Fuji' and 'Brookfield') on the full training samples and with both validation procedures (Venetian and Monte Carlo), ranged from 0.76 to 0.92. The results of this work indicate that using this technology at the individual fruit level is essential to understand the factors that determine this disorder and can improve BP prediction of intact fruit.

Morphological and Molecular Characterization of Selected Chilean Runner Bean (Phaseolus coccineus L.) Genotypes Shows Moderate Agronomic and Genetic Variability.

The runner bean is the third most economically important Phaseolus species, which is cultivated on small-scale agriculture for the production of immature pods or to obtain dry seeds. However, despite the economic importance and agronomic potential of this species, the runner bean has been little studied from the genetic standpoint. Therefore, the main objective of this study was to characterize ten selected lines of runner bean obtained from Central (Santiago) and Southern (Valdivia and Villarica) Chile based on morphological and agronomic traits. In addition, the genetic variability of these lines was determined using 12 Inter-Simple Sequence Repeat (ISSR) markers to evaluate the potential of this germplasm for breeding and commercial purposes. As a result, the lines from Central Chile were characterized, and had a higher number of pods per plant compared to the Southern lines, although the size and weight of their seeds were lower. Moreover, a low level of genetic diversity (He = 0.251) was encountered in this population. Finally, this is one of the first studies that generate relevant and novel information on the morphological, agronomic and genetic characterization of the P. coccineus germplasm present in Chile.

A randomized controlled study comparing guided bone regeneration with connective tissue graft to reestablish buccal convexity at implant sites: A 1-year volumetric analysis.

OBJECTIVES: To volumetrically compare guided bone regeneration (GBR) with connective tissue graft (CTG) to reestablish convexity at the buccal aspect of single implants. MATERIALS AND METHODS: Patients with a single tooth gap in the anterior maxilla and horizontal alveolar defect were enrolled in a single-blind randomized clinical trial (RCT). All sites had a buccopalatal bone dimension of at least 6 mm, received a single implant, and were randomly allocated to the control (GBR) or test group (CTG) to reestablish buccal soft tissue convexity. Patients received a provisional crown at 3 months and a permanent crown at 6 months. Primary outcomes were volumetric increase (mm3 ) and linear increase (mm) in buccal soft tissue profile (BSP) within a well-defined area of interest at fixed time points. Alveolar process deficiency was a secondary outcome. RESULTS: Twenty-one patients were included per group (control: 11 females, mean age 51; test: 9 females, mean age 48). After 1 year, GBR resulted in a significant volumetric increase of 20.74 mm3 (P < .001) corresponding to linear increase in BSP of 1.30 mm (P < .001). For CTG, this was 15.86 (P < .001) and 1.19 mm (P < .001), respectively. The changes over time in volume (P = .173) and BSP (P = .241) were not significantly different between the groups. Twenty-nine percentage and 26% of the final volumetric increase was the result of installing and altering prosthetic components in the control and test groups, respectively. Alveolar process deficiency significantly reduced from pre-op to 1 year following GBR (P < .001) and CTG (P < .001). The difference between the groups was not significant (P = .342). However, 58% of the patients treated with GBR and 38% treated with CTG failed to show perfect soft tissue convexity at the buccal aspect. CONCLUSION: GBR as well as CTG are effective in reducing horizontal alveolar defects for aesthetic purposes. However, in about half of the cases, either strategy failed to optimally reestablish buccal convexity.

Ammonium acts systemically while nitrate exerts an additional local effect on Medicago truncatula nodules.

Symbiotic nitrogen fixation (SNF) has a high energetic cost for legume plants; legumes thus reduce SNF when soil N is available. The present study aimed to increase our understanding regarding the impacts of the two principal forms of available N in soils (ammonium and nitrate) on SNF. We continuously measured the SNF of Medicago truncatula under controlled conditions. This permitted nodule sampling for comparative transcriptome profiling at points connected to the nodules' reaction following ammonium or nitrate applications. The N component of both ions systemically induced a rhythmic pattern of SNF, while the activity in control plants remained constant. This rhythmic activity reduced the per-day SNF. The nitrate ion had additional local effects; the more pronounced were a strong downregulation of leghaemoglobin, nodule cysteine-rich (NCR) peptides and nodule-enhanced nicotianamine synthase (neNAS). The neNAS has proven to be of importance for nodule functioning. Although other physiological impacts of nitrate on nodules were observed (e.g. nitrosylation of leghaemoglobin), the main effect was a rapid ion-specific and organ-specific change in gene expression levels. Contrastingly, during the first hours after ammonium applications, the transcriptome remained virtually unaffected. Therefore, nitrate-induced genes could be key for increasing the nitrate tolerance of SNF.

Regulation of Symbiotic Nitrogen Fixation in Legume Root Nodules.

In most legume nodules, the di-nitrogen (N2)-fixing rhizobia are present as organelle-like structures inside their root host cells. Many processes operate and interact within the symbiotic relationship between plants and nodules, including nitrogen (N)/carbon (C) metabolisms, oxygen flow through nodules, oxidative stress, and phosphorous (P) levels. These processes, which influence the regulation of N2 fixation and are finely tuned on a whole-plant basis, are extensively reviewed in this paper. The carbonic anhydrase (CA)-phosphoenolpyruvate carboxylase (PEPC)-malate dehydrogenase (MDH) is a key pathway inside nodules involved in this regulation, and malate seems to play a crucial role in many aspects of symbiotic N2 fixation control. How legumes specifically sense N-status and how this stimulates all of the regulatory factors are key issues for understanding N2 fixation regulation on a whole-plant basis. This must be thoroughly studied in the future since there is no unifying theory that explains all of the aspects involved in regulating N2 fixation rates to date. Finally, high-throughput functional genomics and molecular tools (i.e., miRNAs) are currently very valuable for the identification of many regulatory elements that are good candidates for accurately dissecting the particular N2 fixation control mechanisms associated with physiological responses to abiotic stresses. In combination with existing information, utilizing these abundant genetic molecular tools will enable us to identify the specific mechanisms underlying the regulation of N2 fixation.

Nitrate application or P deficiency induce a decline in Medicago truncatula N2-fixation by similar changes in the nodule transcriptome.

Nitrogen fixation of Medicago truncatula is regulated by the nitrogen status of leaves through inducing a repeatedly occurring 24-h nodule activity rhythm that reduces per day nitrogen fixation. The hypotheses of the present study were that (1) long-term moderate whole-plant P deficiency in Medicago truncatula induces an according daily rhythm in nitrogenase activity comparable to that induced by nitrate application and (2), the changes in the nodule transcriptome that go along with a strong nitrogenase activity decline during the afternoon would be similar under P deficiency or after nitrate supply. The nodules of plants in a low P treatment developed a rhythmic pattern of activity that resembled the pattern following nitrate application. A comprehensive, RNAseq-based comparative transcriptome profiling of nodules during a repeated part of the rhythm revealed similarities between P deficiency versus nitrate supply. Under both treatments, the formation of nitrogenase was targeted by a reduction in the expression of genes for nodule-specific cysteine-rich peptides (NCR), and possibly also by a disturbance of the inner cell iron allocation. A strong reduction in the expression of leghemoglobin is likely to have restricted the supply of oxygen for respiration.