Green manuring is a conservation agricultural practice that improves soil quality and crop yield. However, increasing the active nitrogen (N) and carbon (C) pools during green manure (GM) amendment may accelerate soil N transformation and stimulate N loss. Previous studies have reported the effects of cover crop incorporation on N2O emission; however, the driving mechanisms and other N losses remain unclear. Therefore, we conducted a comprehensive meta-analysis of 109 published articles (517 paired observations) to clarify the effects of GM amendment on soil reactive N (Nr) losses (N2O emissions, NH3 volatilization, and N leaching and runoff), N pools, and N cycling functional gene abundance. The results showed that green manuring increased soil microbial biomass N (MBN) and NO3--N concentrations and stimulated N2O emission but significantly lowered N leaching and yield-scaled NH3 volatilization. Practices of green manuring made a dominant contribution to the variation in N2O emissions and NH3 volatilization after GM application. Furthermore, applying legume-based GM, using N derived from GM (GMN) as an additional input, and short-term GM amendment each stimulated N2O emissions. In contrast, adopting non-legume GM, using GMN to partially substitute mineral N, and applying GM to the soil surface or paddy field mitigated NH3 loss during GM amendment. Additionally, the variation in NH3 volatilization was positively related to soil pH and N application rate (NAR) but had a negative relationship with mean annual precipitation (MAP). This study highlighted the marked effects of green manuring on soil N retention and loss. Agricultural operations that adopt GM amendment should select suitable GM species and optimize mineral N inputs to minimize N loss.

Malnutrition is a global concern in older adults, as it negatively affects morbidity and mortality. While higher animal protein intake may help prevent and treat malnutrition, it might also increase the risk of chronic diseases and death. Conversely, vegetable protein intake might have a lower anabolic effect and not be as effective to improve nutritional status. We studied whether animal and vegetable protein intake are associated with changes in nutritional status in older adults.
We used pooled data from two Spanish cohorts: the Seniors-ENRICA 1 and Seniors-ENRICA 2.
2,965 community-dwelling adults aged 62-92 years.
Protein intake was estimated at baseline via an electronic, validated diet history. Nutritional status was assessed at baseline and after 2.6 years with the GLIM (Global Leadership Initiative on Malnutrition) phenotypic criteria: weight loss, low body mass index, and reduced muscle mass. The odds of improvements in nutritional status were assessed with logistic regression models, extensively adjusted for potential confounders.
Higher animal and vegetable protein intake were associated with improvements in nutritional status [odds ratios (95% confidence intervals) per 0.25 g/kg/day were 1.15 (1.00, 1.32) and 1.77 (1.35, 2.32), respectively]. Cereal protein intake drove most of the latter association [2.07 (1.44, 2.98)]. Replacing 0.25 g/kg/day of total animal protein, meat, or fish protein (but not dairy or egg protein) with vegetable protein was associated with improvements in nutritional status [1.54 (1.13, 2.09), 1.70 (1.20, 2.41), and 1.77 (1.18, 2.64), respectively].
Higher animal and, especially, vegetable protein intake were associated with improvements in nutritional status in older adults. Replacing total animal protein, meat, or fish protein with vegetable protein may help improve malnutrition.

Mutualism is thought to be more prevalent in the tropics than temperate zones and may therefore play an important role in generating and maintaining high species richness found at lower latitudes. However, results on the impact of mutualism on latitudinal diversity gradients are mixed, and few empirical studies sample both temperate and tropical regions. We investigated whether a latitudinal diversity gradient exists in the symbiotic microbial community associated with the legume Chamaecrista nictitans. We sampled bacteria DNA from nodules and the surrounding soil of plant roots across a latitudinal gradient (38.64-8.68 °N). Using 16S rRNA sequence data, we identified many non-rhizobial species within C. nictitans nodules that cannot form nodules or fix nitrogen. Species richness increased towards lower latitudes in the non-rhizobial portion of the nodule community but not in the rhizobial community. The microbe community in the soil did not effectively predict the non-rhizobia community inside nodules, indicating that host selection is important for structuring non-rhizobia communities in nodules. We next factorially manipulated the presence of three non-rhizobia strains in greenhouse experiments and found that co-inoculations of non-rhizobia strains with rhizobia had a marginal effect on nodule number and no effect on plant growth. Our results suggest that these non-rhizobia bacteria are likely commensals-species that benefit from associating with a host but are neutral for host fitness. Overall, our study suggests that temperate C. nictitans plants are more selective in their associations with the non-rhizobia community, potentially due to differences in soil nitrogen across latitude.

Verticillium wilt is a major threat to many crops, among them alfalfa (Medicago sativa). The model plant Medicago truncatula, a close relative of alfalfa was used to study the genetic control of resistance towards a new Verticillium alfalfae isolate. The accidental introduction of pathogen strains through global trade is a threat to crop production and such new strains might also be better adapted to global warming. Isolates of V. alfalfae were obtained from alfalfa fields in Iran and characterized. The Iranian isolate AF1 was used in a genome-wide association study (GWAS) involving 242 accessions from the Mediterranean region. Root inoculations were performed with conidia at 25°C and symptoms were scored regularly. Maximum Symptom Score and Area under Disease Progess Curve were computed as phenotypic traits to be used in GWAS and for comparison to a previous study with French isolate V31.2 at 20°C. This comparison showed high correlation with a shift to higher susceptibility, and similar geographical distribution of resistant and susceptible accessions to AF1 at 25°C, with resistant accessions mainly in the western part. GWAS revealed 30 significant SNPs linked to resistance towards isolate AF1. None of them were common to the previous study with isolate V31.2 at 20°C. To confirm these loci, the expression of nine underlying genes was studied. All genes were induced in roots following inoculation, in susceptible and resistant plants. However, in resistant plants induction was higher and lasted longer. Taken together, the use of a new pathogen strain and a shift in temperature revealed a completely different genetic control compared to a previous study that demonstrated the existence of two major QTLs. These results can be useful for Medicago breeding programs to obtain varieties better adapted to future conditions.

Snack alternatives based on common beans (Phaseolus vulgaris L.) have been developed to promote pulse consumption. The purpose of this study was to evaluate the chemical composition, sensory acceptance and the effect of common bean baked snack (CBBS) consumption on blood lipid levels in participants with overweight and altered blood lipid levels. A sensory evaluation by 80 untrained judges was carried out using a hedonic scale. A randomized crossover 2 × 2 trial was performed, where 20 participants with overweight and one blood lipid alteration consumed 32 g of CBBS or did not consume it (control) for four weeks. Blood samples were taken to quantify the triglycerides, total cholesterol, LDL-c, HDL-c, ApoB-100, glucose and insulin. Furthermore, anthropometric, dietary and physical activity parameters were recorded. The overall acceptance of CBBS was similar compared to popcorn (p > 0.05). The consumption of CBBS reduced the apolipoprotein B-100 levels (p = 0.008). This reduction could be associated with the additional dietary fiber consumption during the CBBS period (p = 0.04). Although it did not improve any other blood lipid or glucose parameters (p > 0.05), it did not affect them either, which means that the CBBS could be consumed without compromising cardiovascular health.

Cowpea (Vigna unguiculata [L.] Walp., diploid, 2n = 22) is a major crop used as a protein source for human consumption as well as a quality feed for livestock. It is drought and heat tolerant and has been bred to develop varieties that are resilient to changing climates. Plant adaptation to new climates and their yield are strongly affected by flowering time. Therefore, understanding the genetic basis of flowering time is critical to advance cowpea breeding. The aim of this study was to perform genome-wide association studies (GWAS) to identify marker trait associations for flowering time in cowpea using single nucleotide polymorphism (SNP) markers. A total of 368 accessions from a cowpea mini-core collection were evaluated in Ft. Collins, CO in 2019 and 2020, and 292 accessions were evaluated in Citra, FL in 2018. These accessions were genotyped using the Cowpea iSelect Consortium Array that contained 51,128 SNPs. GWAS revealed seven reliable SNPs for flowering time that explained 8-12% of the phenotypic variance. Candidate genes including FT, GI, CRY2, LSH3, UGT87A2, LIF2, and HTA9 that are associated with flowering time were identified for the significant SNP markers. Further efforts to validate these loci will help to understand their role in flowering time in cowpea, and it could facilitate the transfer of some of this knowledge to other closely related legume species.

The global rise in obesity and type 2 diabetes has generated significant interest in regulating the glycaemic impact of staple foods. Wheat breads (white or wholemeal) are popular staples, but have a high-glycaemic index, due to the highly digestible wheat starch. Reducing the glycaemic potency of white bread is challenging because the bread-making conditions are mostly conducive to starch gelatinisation. Cellular legume powders are a new source of type 1 resistant starch, where the starch is encapsulated by dietary fibre in the form of intact plant cell walls. The starch in these cell powders is less susceptible to gelatinisation and digestion than starch in conventional legume flours. However, legume cell resilience to baking conditions and the effects of this ingredient on glycaemic responses and product quality are unknown. Here we show that the integrity of cell wall fibre in chickpea powder was preserved on baking and this led to a ~40% reduction in in vivo glycaemic responses (iAUC120) to white bread rolls (~50 g available carbohydrate and 12 g wheat protein per serving) when 30% or 60% (w/w) of the wheat flour was replaced with intact cell powder. Significant reductions in glycaemic responses were achieved without adverse effects on bread texture, appearance or palatability. Starch digestibility analysis and microscopy confirmed the importance of cell integrity in attenuating glycaemic responses. Alternative processing methods that preserve cell integrity are a new, promising way to provide healthier low glycaemic staple foods; we anticipate that this will improve dietary options for diabetes care.

Given the high prevalence of cardiovascular disease (CVD) among older adults, especially in men compared to women, the aim of this study was to examine nut and legume consumption in relation to Framingham risk score (FRS) and cardiovascular risk factors in older adult men.
In this cross-sectional study, we recruited 267 men aged 60-74 years old. Diet was measured using a validated and reliable food frequency questionnaire. We used FRS to predict a 10-year chance of developing CVD among participants.
Nut consumption was inversely associated with serum levels of low-density lipoprotein cholesterol (LDL-C) (Odds ratio (OR): 0.36, 95% confidence interval (CI): 0.17, 0.59; P < 0.001) and directly associated with high-density lipoprotein cholesterol (HDL-C) level (OR: 2.24, 95% CI: 1.18, 3.89; P = 0.001). In addition, higher intake of legumes was associated with lower serum levels of LDL-C (OR: 0.28, 95% CI: 0.15, 0.49; P < 0.001) and higher serum levels of HDL-C (OR: 2.14, 95% CI: 1.20, 3.72; P = 0.005). However, no associations were observed between nuts and legumes on FRS and other CVD risk factors.
Overall, nut and legume consumption was associated with higher serum levels of HDL-C and lower serum levels of LDL-C. Future studies are required to elucidate the association between nut and legume consumption and CVD risk factors to characterize gender differences.

BACKGROUND: The increasing importance of plant-based proteins in the food sector makes a reliable compositional analysis of plant-based high-protein ingredients a necessity. Specifically, the quantification of short-chain carbohydrates is relevant for multiple areas, including food product development, food labelling and fundamental food chemistry and food technology research. Commonly used extraction procedures for subsequent high-performance liquid chromatographic separation and quantification of short-chain carbohydrates have been discussed controversially regarding a range of complications that can potentially lead to inaccurate sugar determination. The present study compares the sugar levels in wheat flour and wholemeal wheat flour determined with different aqueous and ethanolic extraction procedures. These procedures included measures to prevent enzyme activity and microbial growth, which represent two of the most relevant challenges in sugar extraction from food samples.
RESULTS: Differences in sugar levels (sum of sucrose/maltose, glucose and fructose) as high as 1.8% dry matter (wheat flour) were observed between the employed extraction procedures. Ethanolic extraction (80% ethanol in ultrapure water) with the use of the antimicrobial agent sodium azide but without Carrez clarification was identified as most promising for sugar determination in plant-based high-protein ingredients.
CONCLUSIONS: A screening of high-protein ingredients derived from cereals (wheat gluten), pseudocereals (quinoa, amaranth, buckwheat) and legumes (soy, pea, lupin, lentil, carob, chickpea, faba bean) concerning their levels of sucrose, maltose, glucose and fructose confirmed the applicability of the chosen extraction procedure. © 2021 Society of Chemical Industry.

Although fruits and vegetable consumption has been shown to be associated with lower risks of mortality, cancers, and cardiovascular disease (CVD), there are limited data from China on the shape of the association. This study aimed to quantify the relationship between levels of fruit, vegetable, and legume consumption with the risk of major CVD, CVD mortality, cancer incidence, cancer mortality, and all-cause mortality.
In the baseline survey, participants attended 1 of 115 (45 urban and 70 rural) communities from 12 provinces to complete a standardized questionnaire, and undergo a physical examination between 2005 and 2009, and were followed up till 2017 (for the current analysis). Diet was assessed through in-person interviews by using validated food-frequency questionnaires. The clinical outcomes were adjudicated centrally by trained physicians using standardized definitions. Cox frailty models were used to explore the associations between fruit, vegetable, and legume consumption with the risk of all-cause, CVD, and cancer mortality.
A total of 41 243 participants were eligible for inclusion in the analyses. The average combined average daily intake of fruit, vegetable, and legume was 2.97 [standard deviation (SD) 1.22] servings per day. During a median follow-up of 8.9 years [interquartile range (IQR) 6.7-9.9 years], we recorded 1893 major CVDs, 794 cancer events, and 1324 deaths, with 411 CVD deaths and 429 cancer deaths. In the models adjusted for age, sex, and center (random effect), a higher total intake of fruit, vegetable, and legume was inversely associated with CVD mortality, cancer incidence, cancer mortality, and all-cause mortality. After adjusting for additional covariates, the associations were evidently attenuated and only the association with all-cause mortality (hazard ratio [HR] trend 0.92, 95% CI 0.86-0.98, p trend = 0.021) remained significant, with a non-significant trend for major CVD (HR trend 1.02, 95% CI 0.97-1.08, p trend = 0.449), CVD mortality (HR trend 0.94, 95% CI 0.84-1.06, p trend = 0.301), cancer incidence (HR trend 0.97, 95% CI 0.89-1.06, p trend = 0.540), or cancer mortality (HR trend 0.92, 95% CI 0.82-1.04, p trend = 0.174). Compared with the reference group, the risk of all-cause mortality was the lowest for four to five servings of total daily intake of fruit, vegetable, and legume (HR 0.73, 95% CI 0.55-0.97), and did not show a further decrease for the higher intake group. Separately, fruit intake was associated with a lower risk of all-cause mortality (HR trend 0.92, 95% CI 0.86-0.99, p trend = 0.020) and legume intake was associated with a lower risk of major CVD (HR trend 0.95, 95% CI 0.90-0.99, p trend = 0.028) and all-cause mortality (HR trend 0.94, 95% CI 0.89-0.99, p trend = 0.020) in the fully adjusted models.
This prospective study suggests that Chinese people with daily consumption of four to five servings (equivalent to 500-625 g/day) of fruit, vegetable, and legume demonstrated the lowest mortality, which conveys an encouraging message to the public that lifestyle modification to increase fruit, vegetable, and legume intakes may have greater beneficial effects on reducing all-cause mortality.