Effect of sugarcane cropping systems on herbicide losses in surface runoff

Herbicide runoff from cropping fields has been identified as a threat to the Great Barrier Reef ecosystem. A field investigation was carried out to monitor the changes in runoff water quality resulting from four different sugarcane cropping systems that included different herbicides and contrasting tillage and trash management practices. These include (i) Conventional - Tillage (beds and inter-rows) with residual herbicides used; (ii) Improved - only the beds were tilled (zonal) with reduced residual herbicides used; (iii) Aspirational - minimum tillage (one pass of a single tine ripper before planting) with trash mulch, no residual herbicides and a legume intercrop after cane establishment; and (iv) New Farming System (NFS) - minimum tillage as in Aspirational practice with a grain legume rotation and a combination of residual and knockdown herbicides. Results suggest soil and trash management had a larger effect on the herbicide losses in runoff than the physico-chemical properties of herbicides. Improved practices with 30% lower atrazine application rates than used in conventional systems produced reduced runoff volumes by 40% and atrazine loss by 62%. There were a 2-fold variation in atrazine and >10-fold variation in metribuzin loads in runoff water between reduced tillage systems differing in soil disturbance and surface residue cover from the previous rotation crops, despite the same herbicide application rates. The elevated risk of offsite losses from herbicides was illustrated by the high concentrations of diuron (14mugL-1) recorded in runoff that occurred >2.5months after herbicide application in a 1st ratoon crop. A cropping system employing less persistent non-selective herbicides and an inter-row soybean mulch resulted in no residual herbicide contamination in runoff water, but recorded 12.3% lower yield compared to Conventional practice. These findings reveal a trade-off between achieving good water quality with minimal herbicide contamination and maintaining farm profitability with good weed control.

Composite Medicago truncatula plants harbouring Agrobacterium rhizogenes-transformed roots reveal normal mycorrhization by Glomus intraradices

Composite plants consisting of a wild-type shoot and a transgenic root are frequently used for functional genomics in legume research. Although transformation of roots using Agrobacterium rhizogenes leads to morphologically normal roots, the question arises as to whether such roots interact with arbuscular mycorrhizal (AM) fungi in the same way as wild-type roots. To address this question, roots transformed with a vector containing the fluorescence marker DsRed were used to analyse AM in terms of mycorrhization rate, morphology of fungal and plant subcellular structures, as well as transcript and secondary metabolite accumulations. Mycorrhization rate, appearance, and developmental stages of arbuscules were identical in both types of roots. Using Mt16kOLI1Plus microarrays, transcript profiling of mycorrhizal roots showed that 222 and 73 genes exhibited at least a 2-fold induction and less than half of the expression, respectively, most of them described as AM regulated in the same direction in wild-type roots. To verify this, typical AM marker genes were analysed by quantitative reverse transcription-PCR and revealed equal transcript accumulation in transgenic and wild-type roots. Regarding secondary metabolites, several isoflavonoids and apocarotenoids, all known to accumulate in mycorrhizal wild-type roots, have been found to be up-regulated in mycorrhizal in comparison with non-mycorrhizal transgenic roots. This set of data revealed a substantial similarity in mycorrhization of transgenic and wild-type roots of Medicago truncatula, validating the use of composite plants for studying AM-related effects.

In vitro REGENERATION OF PIGEON PEA USING LEAF EXPLANTS

Pigeon pea ( Cajanus cajan (L.) Millsp.) is a drought tolerant pulse legume, mainly grown for grain in the semi-arid tropics, particularly in Africa. Pigeon pea production in countries like Kenya is faced with a number of challenges, particularly lack of high quality seeds. The objective of this study was to develop an in vitro regeneration system for pigeon pea varieties grown in Kenya, that is amenable to genetic transformation. In vitro regeneration of pigeon pea varieties, KAT 60/8 and ICEAP 00557, commonly grown in Kenya was achieved using leaf explants from in vitro grown seedlings, through callus initiation, followed by shoot and root induction. For callus initiation, MS media supplemented with 0.5-4 mg l-1 2, 4-D and TDZ separately were tested, and IBA at 0.1, 0.5 and 1 mg l-1 was tested for rooting of shoots. Embryogenic calli was obtained on MS containing 2, 4- D; whereas TDZ induced non-embryogenic callus alone or with shoots directly on explants. Indirect shoot regeneration frequency of 6.7 % was achieved using 1 mg l-1 2, 4-D-induced embryogenic callus obtained using KAT 60/8 explants. Whereas direct shoot regeneration frequencies of 20 and 16.7% were achieved using ICEAP 00557 and KAT 60/8 explants, using 0.5 mg l-1 and 2 mg l-1 TDZ, respectively. Optimum rooting was achieved using 0.5 mg l-1 IBA; and up to 92% rooted shoots were successfully established in soil after acclimatisation. Genotype and hormone concentrations had a significant (P<0.05) influence on callus, shoot and root induction. The protocol developed can be optimised for mass production and genetic transformation of KAT 60/8 variety.

Efeito inseticida de vicilinas isoladas de sementes de erythrina velutina em condições de semi-campo para moscas das frutas (ceratitis capitata)

Chitin-binding vicilins from legume seeds (Erythrina velutina. Canavalia ensiformes and Phaseolus vulgares) were isolated by ammonium sulfate followed by affinity chromatography on a chitin column. Effect of these vicilins on female adults of Ceratitis capitata was examined by bioassay and in a semi-field assay model. Mechanism of action of the vicilins was determined by in vivo digestibility and chitin affinity. Among the tested vicilins, E. velutina when added to diet caused strong effect on mortality at 10% dose. This insecticidal property was tested in a semi-field assay which showed the same effect observed in laboratory conditions, where doses of 10% and 15% were lethal to female adults of C. capitata. These deleterious effects were not only associated to the binding to chitin structures present in peritrophic membrane, but principally to its low digestibility in the C. capitata digestive tract. This fact was confirmed because chiting binding proteins as WGA and the other tested vicilins were not toxic to female adults of C. capitata due susceptibility of these proteins to digestive enzymes of the insects. By other side EvV was more resistant to digestive enzymes, causing deleterious effects on female adults of C. capitata. These results showed that EvV may be part of the pest management programs or an alternative in plant improvement program in the population control of this fruticulture pest

SSR genetic diversity assessment of popular pigeonpea varieties in Malawi reveals unique fingerprints

Background: Pigeonpea ( Cajanus cajan L. Millsp.) is a drought tolerant legume of the Fabaceae family and the only cultivated species in the genus Cajanus. It is mainly cultivated in the semi-arid tropics of Asia and Oceania, Africa and America. In Malawi, it is grown as a source of food and income and for soil improvement in intercropping systems. However, varietal contamination due to natural outcrossing causes significant quality reduction and yield losses. In this study, 48 polymorphic SSR markers were used to assess the diversity among all pigeonpea varieties cultivated in Malawi to determine if a genetic fingerprint could be identified to distinguish the popular varieties. Results: A total of 212 alleles were observed with an average of 5.58 alleles per marker and a maximum of 14 alleles produced by CCttc019 (Marker 40). Polymorphic information content (PIC), ranged from 0.03 to 0.89 with an average of 0.30. A neighbor-joining tree produced 4 clusters. The most commonly cultivated varieties, which include released varieties and cultivated land races, were well-spread across all the clusters observed, indicating that they generally represented the genetic diversity available in Malawi, although substantial variation was evident that can still be exploited through further breeding. Conclusion: Screening of the allelic data associated with the five most popular cultivated varieties, revealed 6 markers – CCB1, CCB7, Ccac035, CCttc003, Ccac026 and CCttc019 – which displayed unique allelic profiles for each of the five varieties. This genetic fingerprint can potentially be applied for seed certification to confirm the genetic purity of seeds that are delivered to Malawi farmers.

An investigation into the properties of non-digestible carbohydrates that selectively promote colonic propionate production

Short chain fatty acids (SCFA), including propionate, are produced by the bacterial fermentation of carbohydrates in the colon. Propionate has many potential roles in health, including inhibiting cholesterol synthesis, de novo lipogenesis and increasing satiety. The profile of SCFA produced is determined by both the substrate available and the bacteria present and may be influenced by environmental conditions within the lumen of the colon. Whilst it may be beneficial to increase colonic propionate production, dietary strategies to achieve this are unproven. Adding propionate to food leads to poorer organoleptic properties, and oral propionate is absorbed in the small intestine. The optimum way to selectively increase colonic propionate would be to select fermentable carbohydrates that selectively promote propionate production. To date, few studies have undertaken a systematic assessment of the factors leading to increased colonic propionate production making the selection of propiogenic carbohydrates challenging. The aim of this thesis was to identify the best carbohydrates for selectively increasing propionate production, and to explore the factors which control propionate production. This work started with a systematic review of the literature for evidence of candidate carbohydrates, which led to a screen of ‘propiogenic’ substrates using in vitro batch fermentations and mechanistic analysis of the impact of pH, bond linkage and orientation using a range of sugars, polysaccharides and fibre sources. A new unit for SCFA production was developed to allow comparison of results from in vitro studies encompassing a range different methodologies found in the literature. The systematic review found that rhamnose yielded the highest rate and proportion of propionate production whereas, for polysaccharides, β-glucan ranked highest for rate and guar gum ranked highest for molar production, but this was not replicated across all studies. Thus, no single NDC was established as highly propiogenic. Some substrates appeared more propiogenic than others and when these were screened in vitro. Laminarin, and other β-glucans ranked highest for propionate production. Legume fibre and mycoprotein fibre were also propiogenic. A full complement of glucose disaccharides were tested to examine the role glycosidic bond orientation and position on propionate production. Of the glucose disaccharides tested, β(1-4) bonding was associated with increased proportion of propionate and α(1-1) and β(1-4) increased the rate and proportion of butyrate production. In conclusion, it appears that for fibre to affect satiety, high intakes of fibre are needed, and which a major mechanism is thought to occur via propionate. Within this thesis it was identified that rather than selecting specific fibres, increasing overall intakes of highly fermentable carbohydrates is as effective at increasing propionate production. Selecting carbohydrates with beta-bonding, particularly laminarin and other β(1-4) fermentable carbohydrates leads to marginal increases in propionate production. Compared with targeted delivery of propionate to the colon, fermentable carbohydrates examined in this thesis have lesser and variable effects on propionate production. A more complete understanding of the impact of bond configurations in polysaccharides, rather than disaccharides, may help selection or design of dietary carbohydrates which selectively promote colonic propionate production substrates for inclusion in functional foods. Overall this study has concluded that few substrates are selectively propiogenic and the evidence suggests that similar changes in propionate production may be achieved by modest changes in dietary fibre intake

NITROUS OXIDE EMISSIONS IN COVER CROP-BASED CORN PRODUCTION SYSTEMS

Nitrous oxide (N2O) is a potent greenhouse gas; the majority of N2O emissions are the result of agricultural management, particularly the application of N fertilizers to soils. The relationship of N2O emissions to varying sources of N (manures, mineral fertilizers, and cover crops) has not been well-evaluated. Here we discussed a novel methodology for estimating precipitation-induced pulses of N2O using flux measurements; results indicated that short-term intensive time-series sampling methods can adequately describe the magnitude of these pulses. We also evaluated the annual N2O emissions from corn-cover crop (Zea mays; cereal rye [Secale cereale], hairy vetch [Vicia villosa], or biculture) production systems when fertilized with multiple rates of subsurface banded poultry litter, as compared with tillage incorporation or mineral fertilizer. N2O emissions increased exponentially with total N rate; tillage decreased emissions following cover crops with legume components, while the effect of mineral fertilizer was mixed across cover crops.

Vicilinas de sementes de leguminosas selvagens:purificação, caracterização, efeito de deletério e mecanismo de ação para bruquídeos e para fungos fitopatogênicos

Globulins fractions of legume seeds of Crotalaria pallida, Erytrina veluntina and Enterolobium contortisiliquum were isolated and submitted to assays against serine, cysteine and aspartic proteinases, as also amylase present in midgut of C. maculatus and Z. subfasciatus. Hemagglutination assays indicated presence of a lectin in E. veluntina globulin fractions. This lectin had affinity to human erythrocytes type A, B and O. Vicilins were purified by chromatography on Sephacryl S-300 followed of a chromatography on Sephacryl S-200, which was calibrated using protein markers. Vicilins from C. pallida (CpV) and E. veluntina (EvV) seeds had a molecular mass of 124.6 kDa and E. contortisiliquum a molecular mass of 151kDa. Eletrophoresis in presence of SDS showed that CpV was constituted by four subunities with apparent molecular mass of 66, 63, 57 and 45 kDa, EvV with three subunities with apparent molecular mass of 45kDa and EcV four subunities, two with 37.1 kDa and two with 25.8 kDa. Non denaturantig eletrophoresis displayed single bands with high homogeneity, where CpV had lower acidic behavior. All vicilins are glycoproteins with carbohydrate contents at 1 to1.5%. Bioassays were done to detect deleterious effects of vicilins against C. maculatus and Z. subfasciatus larvae. CpV, EvV and EcV exhibited a WD50 of 0.28, 0.19 and 1.03%; LD50 0.2, 0.26, and 1.11% respectively to C. maculatus. The dose responses of CpV, EvV and EcV to Z. subfasciatus were: WD50 of 0.12, 0.14, 0.65% and LD50 of 0.09, 0.1, and 0.43% respectively. The mechanism of action of these proteins to bruchids should be based on their properties of bind to chitin present in mid gut of larvae associated with the low digestibility of vicilin. In assays against phytopatogenous fungus, only EcV was capable of inhibit F. solani growth at concentrations of 10 and 20 µg and its action mechanism should be also based in the affinity of EcV to chitin present in the fungi wall

Vicilina de sementes da leguminosa selvagem Anadenanthera macrocarpa: purificação, caracterização, efeito deletério e mecanismo de ação para Callosobruchus maculatus

Grains and legume seeds are foods that form the basis of the diets of many cultures around the world, winch contritbute to the daily nutrient requirements of humans. Vicilins (7S globulin) are storage proteins found in legume seeds, and may have an additional function constitutive defense of the embryo against pests and pathogens. In this work the vicilin from Anadenanthera macrocarpa - AmV (red-angico), was purified and partially characterized, its effect on development and larval survival and adult emergence of Callosobruchus maculatus was evaluated by determination of LD50, WD50 and ED50 in system bioassay. Purification of vicilin was initiated by the chitin affinity chromatography and then gel filtration (Superdex 75 Tricorn 10x300 mm) FPLC system followed by reverse phase chromatography (C8 phenomenex) on HPLC system. Bioassays WD50 and LD50 for larvae were 0.32% and 0.33% (w:w) respectively, since the ED50 for adults was 0.096%. The probable mechanism of action was evaluated by testing digestibility of AmV in vitro, and observed for the involvement of two fragments vicilins immunoreactive against polyclonal Anti-vicilin from Erythrina velutina (Anti-EvV) about of 22 and 13 kDa chitin binding. The AmV in its native form has been recognized by the anti-EvV, indicating that there is a conserved region in the vicilin and is probably corresponding to the chitin binding domains. These results point to a new vicilin chitin binding that can subsequently be used as a possible biopesticide protein source, in order to control insect pest C. maculatus and confirm literature findings that demonstrate vicilin in the presence of different kinds of ligands to conserved regions chitin not yet characterized

A pigeonpea gene confers resistance to Asian soybean rust in soybean.

Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, is one of the most economically important crop diseases, but is only treatable with fungicides, which are becoming less effective owing to the emergence of fungicide resistance. There are no commercial soybean cultivars with durable resistance to P. pachyrhizi, and although soybean resistance loci have been mapped, no resistance genes have been cloned. We report the cloning of a P. pachyrhizi resistance gene CcRpp1 (Cajanus cajan Resistance against Phakopsora pachyrhizi 1) from pigeonpea (Cajanus cajan) and show that CcRpp1 confers full resistance to P. pachyrhizi in soybean. Our findings show that legume species related to soybean such as pigeonpea, cowpea, common bean and others could provide a valuable and diverse pool of resistance traits for crop improvement.

Aluminium tolerance in bean traditional cultivars from Madeira

Common bean (Phaseolus vulgaris L.) is the most important legume crop in the world, providing low-cost, high quality protein, minerals and dietary fiber for human nutrition. The crop was originated from diversity centers in America and exhibits adaptation abilities to different environmental conditions, including soil with low pH. Acid soils occupy 30% of the agro ecosystem areas in the world. In Madeira, acid Andosols and unsatured Cambisols are the dominant groups of soils. Generally, under acidic and infertile conditions, besides of H+ toxicity, soluble aluminium (Al) is the most important abiotic factor limiting plant development and crop productivity. In the field, the hidden roots are also affected and the reduction of root growth under Al stress can be clearly observed in early stages. Seedlings of fifty bean accessions from the Archipelago of Madeira were tested under controlled conditions in the presence of 50 mM Al at pH 4.4. In general, the tested germplasm appeared to be sensitive or very sensitive to Al toxicity. However, fifteen traditional cultivars clearly exhibited elevated Al-tolerance, with an average root relative elongation (RRE) exceeding 50%, while top six accessions surpassed the 60% RRE mark. The Madeira bean germplasm is a valuable resource for sustainable crop production in acid soils and it could be used as parental lines in breeding programs aimed for Al tolerance in common beans.

Aluminium tolerance in bean traditional cultivars from Madeira

Common bean (Phaseolus vulgaris L.) is the most important legume crop in the world, providing low-cost, high quality protein, minerals and dietary fiber for human nutrition. The crop was originated from diversity centers in America and exhibits adaptation abilities to different environmental conditions, including soil with low pH. Acid soils occupy 30% of the agro ecosystem areas in the world. In Madeira, acid Andosols and unsatured Cambisols are the dominant groups of soils. Generally, under acidic and infertile conditions, besides of H+ toxicity, soluble aluminium (Al) is the most important abiotic factor limiting plant development and crop productivity. In the field, the hidden roots are also affected and the reduction of root growth under Al stress can be clearly observed in early stages. Seedlings of fifty bean accessions from the Archipelago of Madeira were tested under controlled conditions in the presence of 50 mM Al at pH 4.4. In general, the tested germplasm appeared to be sensitive or very sensitive to Al toxicity. However, fifteen traditional cultivars clearly exhibited elevated Al-tolerance, with an average root relative elongation (RRE) exceeding 50%, while top six accessions surpassed the 60% RRE mark. The Madeira bean germplasm is a valuable resource for sustainable crop production in acid soils and it could be used as parental lines in breeding programs aimed for Al tolerance in common beans.

Diet composition of a declining steppe bird the Little Bustard (Tetrax tetrax) in relation to farming practices

Foraging strategies and diet selection play an essential role in individual survival and reproductive success. The study of feeding ecology becomes crucial when it concerns endangered species such as the Little Bustard (Tetrax tetrax), whose populations are suffering strong declines as a consequence of agricultural intensification. Despite the fact that several populations are overwintering in areas affected by agricultural transformation, nothing is known about how feeding behavior responds to these changes. We studied for the first time the winter diet composition of the Little Bustard in Spain and compared it between areas with two different farming systems: dry and irrigated farmland. Diet was studied through the micro-histological analysis of 357 droppings collected in 16 locations across the wintering range of the Little Bustard in Spain. Up to 62 plant species were identified. Most consumed species were cultivated legumes (46.7%) and dicotyledon weeds (45.6%), while monocotyledons were scarcely consumed (7.7%). Diet composition differed significantly between dry and irrigated farmland areas. In irrigated areas, diet was mainly composed of legumes, in particular alfalfa (Medicago sativa). In contrast, in dry farmland areas diet was more diverse, composed mainly of weeds (Compositae, Papaveraceae, and Cruciferae) and also cultivated legumes, particularly vetch (Vicia sativa). These results suggest that legume crops could be an effective measure to improve habitat quality in areas with scarce food resources. However, in the case of irrigated areas, the strong reliance on alfalfa could make the Little Bustard more vulnerable to changes in land use. This study is the first step to understand the winter trophic requirements of the endangered Little Bustard, but further research is necessary to understand the food requirements of this species during the entire annual cycle.

Heat shock response in bacteria with large genomes: lessons from rhizobia

Rhizobia are important soil bacteria due to their ability to establish nitrogen-fixing symbioses with legume plants. In this dual lifestyle, as free-living bacteria or as plant symbiont, rhizobia are often exposed to different environmental stresses. The present chapter overviews the current knowledge on the heat shock response of rhizobia, highlighting how these large genome bacteria respond to heat from a transcriptional point of view. Response to heat shock in rhizobia involves genome wide changes in the transcriptome that may affect more than 30% of the genome and involve all replicons. In addition to the expected upregulation of genes already known to be involved in stress response (dnaK, groEL, ibpA, clpB), the reports on the heat shock response in rhizobia also showed particular aspects of stress response in these resourceful bacteria. The transcriptional response to heat in rhizobia includes the overexpression of a large number of genes involved in transcription and carbohydrate transport and metabolism. Additional studies are needed in order to better understand the transcriptional regulation of stress response in bacteria with large genomes.

Stylosanthes guianensis cv. BRS Bela managed as organic fodder bank for supplementation of lactation crossbred cows in the Cerrado.

2016