Plant Breeding Approaches Research Articles

Engineering photosynthesis: progress and perspectives.

Photosynthesis is the basis of primary productivity on the planet. Crop breeding has sustained steady improvements in yield to keep pace with population growth increases. Yet these advances have not resulted from improving the photosynthetic process per se but rather of altering the way carbon is partitioned within the plant. Mounting evidence suggests that the rate at which crop yields can be boosted by traditional plant breeding approaches is wavering, and they may reach a “yield ceiling” in the foreseeable future. Further increases in yield will likely depend on the targeted manipulation of plant metabolism. Improving photosynthesis poses one such route, with simulations indicating it could have a significant transformative influence on enhancing crop productivity. Here, we summarize recent advances of alternative approaches for the manipulation and enhancement of photosynthesis and their possible application for crop improvement.

Climate Clever Clovers: New Paradigm to Reduce the Environmental Footprint of Ruminants by Breeding Low Methanogenic Forages Utilizing Haplotype Variation.

Mitigating methane production by ruminants is a significant challenge to global livestock production. This research offers a new paradigm to reduce methane emissions from ruminants by breeding climate-clever clovers. We demonstrate wide genetic diversity for the trait methanogenic potential in Australia’s key pasture legume, subterranean clover (Trifolium subterraneum L.). In a bi-parental population the broadsense heritability in methanogenic potential was moderate (H2 = 0.4) and allelic variation in a region of Chr 8 accounted for 7.8% of phenotypic variation. In a genome-wide association study we identified four loci controlling methanogenic potential assessed by an in vitro fermentation system. Significantly, the discovery of a single nucleotide polymorphism (SNP) on Chr 5 in a defined haplotype block with an upstream putative candidate gene from a plant peroxidase-like superfamily (TSub_g18548) and a downstream lectin receptor protein kinase (TSub_g18549) provides valuable candidates for an assay for this complex trait. In this way haplotype variation can be tracked to breed pastures with reduced methanogenic potential. Of the quantitative trait loci candidates, the DNA-damage-repair/toleration DRT100-like protein (TSub_g26967), linked to avoid the severity of DNA damage induced by secondary metabolites, is considered central to enteric methane production, as are disease resistance (TSub_g26971, TSub_g26972, and TSub_g18549) and ribonuclease proteins (TSub_g26974, TSub_g26975). These proteins are good pointers to elucidate the genetic basis of in vitro microbial fermentability and enteric methanogenic potential in subterranean clover. The genes identified allow the design of a suite of markers for marker-assisted selection to reduce rumen methane emission in selected pasture legumes. We demonstrate the feasibility of a plant breeding approach without compromising animal productivity to mitigate enteric methane emissions, which is one of the most significant challenges to global livestock production.

Development of transgenic Brassica juncea lines for reduced seed sinapine content by perturbing phenylpropanoid pathway genes.

Sinapine is a major anti-nutritive compound that accumulates in the seeds of Brassica species. When ingested, sinapine imparts gritty flavuor in meat and milk of animals and fishy odor to eggs of brown egg layers, thereby compromising the potential use of the valuable protein rich seed meal. Sinapine content in Brassica juncea germplasm ranges from 6.7 to 15.1 mg/g of dry seed weight (DSW) which is significantly higher than the prescribed permissible level of 3.0 mg/g of DSW. Due to limited natural genetic variability, conventional plant breeding approach for reducing the sinapine content has largely been unsuccessful. Hence, transgenic approach for gene silencing was adopted by targeting two genes—SGT and SCT, encoding enzymes UDP- glucose: sinapate glucosyltransferase and sinapoylglucose: choline sinapoyltransferase, respectively, involved in the final two steps of sinapine biosynthetic pathway. These two genes were isolated from B. juncea and eight silencing constructs were developed using three different RNA silencing approaches viz. antisense RNA, RNAi and artificial microRNA. Transgenics in B. juncea were developed following Agrobacterium-mediated transformation. From a total of 1232 independent T0 transgenic events obtained using eight silencing constructs, 25 homozygous lines showing single gene inheritance were identified in the T2 generation. Reduction of seed sinapine content in these lines ranged from 15.8% to 67.2%; the line with maximum reduction had sinapine content of 3.79 mg/g of DSW. The study also revealed that RNAi method was more efficient than the other two methods used in this study.

Harnessing microbiome‐based biotechnologies for sustainable mitigation of nitrous oxide emissions

SummaryAchieving the Sustainable Development Goal of climate change mitigation within this century will require adoption of new innovative technologies to control emissions of nitrous oxide (N2O), an important greenhouse gas leading to global warming. This is particularly important in the face of growing fertilizer consumption and continuous land degradation. Currently used tools to mitigate N2O emissions are based on agrochemical inputs and agronomic practices. Emerging technologies include plant breeding approaches to manipulate microbiome activities in agro‐ecosystems, and microbial biotechnology approaches for in situ microbiome manipulation and engineering via use of biochemical, cellular and genome‐editing methods. This article assessed the likely contribution of microbial biotechnology to the mitigation of N2O emissions and discussed how to facilitate the development of environmental‐friendly microbiome‐based biotechnology for sustainable climate change mitigation.

Variation in Nutritional Composition of Australian Mungbean Varieties

Australian mungbean production is primarily focused in Central and Southern Queensland and Northern NSW, with around 95% of the total mungbean crop exported as a commodity to overseas markets. Significant improvement in pulse crops such as mungbean have primarily been achieved from plant breeding approaches focused on increasing yield and disease resistance. Whilst this remains crucial for the ongoing success and production of the crop, further improvements could be achieved through an increased understanding of nutritional variation, varietal performance and the impact of environmental and agronomic factors on overall nutritional quality. In this survey, the primary objective was to evaluate and compare the nutritional composition of three commercial Australian mungbean varieties (Crystal, Satin II and Celera II-AU), grown in different regions in Queensland (Warra and Hermitage sites) and New South Wales (Liverpool Plains and Northern NSW sites). Mungbean varieties were evaluated in terms of visual seed appearance, measuring seed colour and size, prior to comprehensive nutritional evaluation of milled mungbean flours in order to determine the extent of variation between varieties and regions. Moisture, protein, ash, fat, dietary fibre (total, insoluble and soluble fibre), starch and amylose composition, starch pasting properties (RVA profile) and amino acid compositions were evaluated and compared. This survey may potentially lead to a larger scale evaluation in the future, broadening the scope to include other significant Australian pulse crops such as faba bean and chickpea. Ultimately, the information gathered from this survey may assist plant breeders, producers and processors in regard to improving, growing, processing and value-adding Australian mungbean for both domestic and export markets.

Periclinal chimera technique: new plant breeding approach.

Plant interspecific periclinal chimeras are a mosaic formed by tissues from two species. They are manipulated here as an efficient plant breeding tool for cassava root yields. In this study, plants synthesized from two chimeras, designated as chimera 2 and chimera 4, were characterized morphologically and cytologically to unravel the origin of their tissue layers (L2 and L3). Root yield of the two chimeras was also evaluated. Chimera 2 that was developed from graft union between Manihot fortalezensis (F) as scion and M. esculenta (E) as rootstock and the same in chimera 4 was developed from grafting triploid cassava cultivar (2n = 54) (C) as scion and M. pohlii (P) (2n = 36) as rootstock. A new method of inducing interspecific chimeras without using hormones was also tested in this study. Five combinations between four cassava cultivars on one side and M. fortalezensis and an interspecific hybrid (M. glaziovii x M. esculenta) on the other side were experimented to determine compatibility between the parents. Wild species always gave L2 and L3, independent of being used as rootstock or scion. L3 is responsible for producing pericycle. Thus, its performance was different in each chimera due to specific epigenetic interaction. Of 48 grafts, it was obtained one chimera giving a percentage of 2.1% that is little lower than using hormones but much efficient to use. Chimera induction efficiency in this investigation was the same when using hormones. Thus, our new, less labor, and more cost-effective technique is as much efficient as hormones and is much potential to employ as an effective plant breeding method boosting cassava root yield.

Chemical control of flowering time.

Flowering at the right time is of great importance; it secures seed production and therefore species survival and crop yield. In addition to the genetic network controlling flowering time, there are a number of much less studied metabolites and exogenously applied chemicals that may influence the transition to flowering as well as flower opening. Increased emphasis on research within this area has the potential to counteract the negative effects of global warming on flowering time, especially in perennial crop plants. Perennial crops have a requirement for winter chill, but winters become increasingly warm in temperate regions. This has dramatic effects on crop yield. Different strategies are therefore being developed to engineer flowering time to match local growing conditions. The majority of these efforts are within plant breeding, which benefits from a substantial amount of knowledge on the genetic aspects of flowering time regulation in annuals, but less so in perennials. An alternative to plant breeding approaches is to engineer flowering time chemically via the external application of flower-inducing compounds. This review discusses a variety of exogenously applied compounds used in fruit farming to date, as well as endogenous growth substances and metabolites that can influence flowering time of annuals and perennials.

Microbiome Selection Could Spur Next-Generation Plant Breeding Strategies.

“No plant is an island too…”Plants, though sessile, have developed a unique strategy to counter biotic and abiotic stresses by symbiotically co-evolving with microorganisms and tapping into their genome for this purpose. Soil is the bank of microbial diversity from which a plant selectively sources its microbiome to suit its needs. Besides soil, seeds, which carry the genetic blueprint of plants during trans-generational propagation, are home to diverse microbiota that acts as the principal source of microbial inoculum in crop cultivation. Overall, a plant is ensconced both on the outside and inside with a diverse assemblage of microbiota. Together, the plant genome and the genes of the microbiota that the plant harbors in different plant tissues, i.e., the ‘plant microbiome,’ form the holobiome which is now considered as unit of selection: ‘the holobiont.’ The ‘plant microbiome’ not only helps plants to remain fit but also offers critical genetic variability, hitherto, not employed in the breeding strategy by plant breeders, who traditionally have exploited the genetic variability of the host for developing high yielding or disease tolerant or drought resistant varieties. This fresh knowledge of the microbiome, particularly of the rhizosphere, offering genetic variability to plants, opens up new horizons for breeding that could usher in cultivation of next-generation crops depending less on inorganic inputs, resistant to insect pest and diseases and resilient to climatic perturbations. We surmise, from ever increasing evidences, that plants and their microbial symbionts need to be co-propagated as life-long partners in future strategies for plant breeding. In this perspective, we propose bottom–up approach to co-propagate the co-evolved, the plant along with the target microbiome, through – (i) reciprocal soil transplantation method, or (ii) artificial ecosystem selection method of synthetic microbiome inocula, or (iii) by exploration of microRNA transfer method – for realizing this next-generation plant breeding approach. Our aim, thus, is to bring closer the information accrued through the advanced nucleotide sequencing and bioinformatics in conjunction with conventional culture-dependent isolation method for practical application in plant breeding and overall agriculture.

Soybean spermidine concentration: Genetic and environmental variation of a potential ‘anti-aging’ constituent

Soybean seed is one of the richest food sources of spermidine and other polyamines. Recent findings from human and animal models have confirmed spermidine as a potential anti-aging substance acting through the initiation of autophagy pathways as well as through antioxidant and anti-inflammatory properties. As this might be of relevance for selecting soybeans for soy food production, the present research addresses the natural variation of spermidine concentration of soybean to determine the influences of genotype and environmental factors on spermidine and other polyamines, and to study possible relationships between spermidine and major seed quality traits. Sixteen early maturity soybean genotypes were grown near Vienna, Austria for three seasons, and harvest samples were subject to ultra-high performance liquid chromatography (UHPLC) for determining concentrations of polyamines and free amino acids. Based on individual samples, spermidine concentration ranged between 167 and 291mgkg−1 dry seed, and both genotype and growing season significantly affected spermidine level. Spermidine concentration was closely correlated to putrescine but was not related to seed protein content or other major seed constituents determined by NIRS analysis. These results demonstrate the feasibility of plant breeding approaches to modify the spermidine level of soybean which might support the future development of functional soy foods.

MultispeQ Beta: a tool for large-scale plant phenotyping connected to the open PhotosynQ network

Large-scale high-throughput plant phenotyping (sometimes called phenomics) is becoming increasingly important in plant biology and agriculture and is essential to cutting-edge plant breeding and management approaches needed to meet the food and fuel needs for the next century. Currently, the application of these approaches is severely limited by the availability of appropriate instrumentation and by the ability to communicate experimental protocols, results and analyses. To address these issues, we have developed a low-cost, yet sophisticated open-source scientific instrument designed to enable communities of researchers, plant breeders, educators, farmers and citizen scientists to collect high-quality field data on a large scale. The MultispeQ provides measurements in the field or laboratory of both, environmental conditions (light intensity and quality, temperature, humidity, CO2 levels, time and location) and useful plant phenotypes, including photosynthetic parameters—photosystem II quantum yield (ΦII), non-photochemical exciton quenching (NPQ), photosystem II photoinhibition, light-driven proton translocation and thylakoid proton motive force, regulation of the chloroplast ATP synthase and potentially many others—and leaf chlorophyll and other pigments. Plant phenotype data are transmitted from the MultispeQ to mobile devices, laptops or desktop computers together with key metadata that gets saved to the PhotosynQ platform (https://photosynq.org) and provides a suite of web-based tools for sharing, visualization, filtering, dissemination and analyses. We present validation experiments, comparing MultispeQ results with established platforms, and show that it can be usefully deployed in both laboratory and field settings. We present evidence that MultispeQ can be used by communities of researchers to rapidly measure, store and analyse multiple environmental and plant properties, allowing for deeper understanding of the complex interactions between plants and their environment.

Identification, Characterization, and Expression Analysis of Cell Wall Related Genes in Sorghum bicolor (L.) Moench, a Food, Fodder, and Biofuel Crop.

Biomass based alternative fuels offer a solution to the world's ever-increasing energy demand. With the ability to produce high biomass in marginal lands with low inputs, sorghum has a great potential to meet second-generation biofuel needs. Despite the sorghum crop importance in biofuel and fodder industry, there is no comprehensive information available on the cell wall related genes and gene families (biosynthetic and modification). It is important to identify the cell wall related genes to understand the cell wall biosynthetic process as well as to facilitate biomass manipulation. Genome-wide analysis using gene family specific Hidden Markov Model of conserved domains identified 520 genes distributed among 20 gene families related to biosynthesis/modification of various cell wall polymers such as cellulose, hemicellulose, pectin, and lignin. Chromosomal localization analysis of these genes revealed that about 65% of cell wall related genes were confined to four chromosomes (Chr. 1–4). Further, 56 tandem duplication events involving 169 genes were identified in these gene families which could be associated with expansion of genes within families in sorghum. Additionally, we also identified 137 Simple Sequence Repeats related to 112 genes and target sites for 10 miRNAs in some important families such as cellulose synthase, cellulose synthase-like, and laccases, etc. To gain further insight into potential functional roles, expression analysis of these gene families was performed using publically available data sets in various tissues and under abiotic stress conditions. Expression analysis showed tissue specificity as well as differential expression under abiotic stress conditions. Overall, our study provides a comprehensive information on cell wall related genes families in sorghum which offers a valuable resource to develop strategies for altering biomass composition by plant breeding and genetic engineering approaches.

ICQG 5 on Madison's Isthmus links genomics and statistics of complex traits.

ICQG 5 on Madison's Isthmus links genomics and statistics of complex traits.

Endophytic bacteria in Miscanthus seed: implications for germination, vertical inheritance of endophytes, plant evolution and breeding

AbstractWith growing interest in the role of microbiomes, and symbionts in particular, the aim of this study was to determine the diversity of the bacterial endophyte population within Miscanthus and to ascertain the extent of vertical transmission via the seed. A great diversity of endophytic bacteria was found in all parts of the mature plant (rhizome, root, stem and leaf), and in seedlings grown from sterilized seed grown in sterile conditions. A total of three phyla and five families of bacteria were identified as cultures compared to 19 phyla and 85 families using 16S rDNA amplification and sequencing. Not all cultured bacteria could be identified by 16S rDNA, implying that the true diversity is even greater. More bacterial diversity was identified in sterile‐grown seedlings than in all parts of the mature plant combined, 17 and 13 phyla, respectively, with 11 in common. Five phyla were present in all plant samples examined. Vertical transmission via the seed may therefore be a major source of endophytes in Miscanthus, presumably supplemented by ingress of soil bacteria as the plant grows. Bacteria identified from the mature plant were predominantly similar to known bacterial sequences in GenBank, but a small number from the stem and many from the seed were novel, potentially adapted to an in planta life cycle. Endophytic bacteria were found to form spores and other dense structures, and this provides a mechanism for long‐term survival and seed transmission. The staining of germinating seeds identified bacteria at the root tip of the emerging radicle. We propose that seed transmission of bacterial endophytes requires adaptation of both plant and microbes, plays a role in germination and has evolutionary significance and implications for future plant breeding approaches, in Miscanthus and more widely.

De novo construction of a "Gene-space" for diploid plant genome rich in repetitive sequences by an iterative Process of Extraction and Assembly of NGS reads (iPEA protocol) with limited computing resources.

BackgroundThe continuing increase in size and quality of the “short reads” raw data is a significant help for the quality of the assembly obtained through various bioinformatics tools. However, building a reference genome sequence for most plant species remains a significant challenge due to the large number of repeated sequences which are problematic for a whole-genome quality de novo assembly. Furthermore, for most SNP identification approaches in plant genetics and breeding, only the “Gene-space” regions including the promoter, exon and intron sequences are considered.ResultsWe developed the iPea protocol to produce a de novo Gene-space assembly by reconstructing, in an iterative way, the non-coding sequence flanking the Unigene cDNA sequence through addition of next-generation DNA-seq data. The approach was elaborated with the large diploid genome of pea (Pisumsativum L.), rich in repetitive sequences. The final Gene-space assembly included 35,400 contigs (97 Mb), covering 88 % of the 40,227 contigs (53.1 Mb) of the PsCam_low-copy Unigen set. Its accuracy was validated by the results of the built GenoPea 13.2 K SNP Array.ConclusionThe iPEA protocol allows the reconstruction of a Gene-space based from RNA-Seq and DNA-seq data with limited computing resources.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-016-1903-z) contains supplementary material, which is available to authorized users.

Enhancing onion breeding using molecular tools

AbstractBulb onion (Allium cepaL.) is an ancient crop that is thought to have originated in Central Asia and has been cultivated for over 5000 years. Classical genetic and plant breeding approaches have been used to improve onion yield, quality, and resistance against biotic and abiotic stresses. However, its biennial life cycle, cross‐pollinated nature and high inbreeding depression have proved challenging for the characterization and breeding of improved traits. New technologies, notably next‐generation sequencing, are providing researchers with the genomic resources and approaches to overcome these challenges. Using these genomic technologies, molecular markers are being rapidly developed and utilized for germplasm analysis and mapping in onion. These new tools and knowledge are allowing the integration of molecular and conventional breeding to speed up onion improvement programmes. In this review, we outline recent progress in onion genomics and molecular genetics and prospects for enhancing onion yield and quality in the future.

Development and application of marker-assisted reverse breeding using hybrid maize germplasm

Humankind has been through different periods of agricultural improvement aiming at enhancing our food supply and the performance of food crops. In recent years, whole genome sequencing and deep understanding of genetic and epigenetic mechanisms have facilitated new plant breeding approaches to meet the challenge of growing population, dwindling resources, and changing climate. Here we proposed a simple and fast molecular breeding method, marker-assisted reverse breeding (MARB), which will revert any maize hybrid into inbred lines with any level of required similarity to its original parent lines. Since all the pericarp DNA of a hybrid is from the maternal parent, whereas one half of the embryo DNA is from the maternal parent and the other half from the paternal parent, so we firstly extract DNA from seed embryo and pericarp of a selected elite hybrid separately and then we derived the genotypes of the two parents with high-density single nucleotide polymorphism (SNP) chips. The following marker-assisted selection was performed based on an Illumina low-density SNP chip designed with 192 SNPs polymorphic between the two parental genotypes, which were uniformly distributed on 10 maize chromosomes. This method has the advantages of fast speed, fixed heterotic mode, and quick recovery of beneficial parental genotypes compared to traditional pedigree breeding using elite hybrids. Meanwhile, MARB has the advantage of not requiring sophisticated transformation and double haploid (DH) technologies over RNA interference (RNAi)-mediated reverse breeding. In addition, MARB can also be used with feed corn harvested from big farms, which is often similar to F2 populations, and the relevant transgenes in the population can be eliminated by marker-assisted selection. As a result, the whole global commercial maize hybrids can be utilized as germplasm for breeding with MARB technology. Starting with an F2 population derived from an elite hybrid, our experiment indicates that with three cycles of marker-assisted selection, selected lines could recover over 80% of the parental genotypes and associated beneficial genes in a fixed heterotic mode. The success application of MARB in maize suggests that this technology is applicable to any hybrid crop to breed new inbreds with improved hybrid performance but the same heterotic mode. As chip technology becomes cheap, it would be expected that polymorphism screening and following marker-assisted selection could be done with one all-purpose high density chip. Several issues associated with MARB were discussed, including its rationale, efficiency and advantages, along with food/feed and environmental safety issues and applications of MARB in variety protection and marker-assisted plant breeding.

Binucleation to breed new plant species adaptable to their environments

Classical plant breeding approaches may fall short to breed new plant species of high environmental and ecological interests. Biotechnological and genetic manipulations, on the other hand, may hold more effective capabilities to circumvent the limitations of sexual incompatibility and conventional breeding programs. Given that plant cells encompass multiple copies of organellar genomes (mitochondrial and plastidial genomes), an important question could be raised about whether an artificial attempt to duplicate the nuclear genome might also be conceivable through a binucleation approach (generating plant cells with 2 nuclei from 2 different plant species) for potential production of new polyploidies that would characterize new plant species. Since the complexities of plant genomes are the result of multiple genome duplications, an artificial binucleation approach would thus be of some interest to eventually varying plant genomes and producing new polyploidy from related or distal plant species. Here, I discuss the potentiality of such an approach to engineer binucleated plant cells as a germ of new plant species to fulfill some environmental applications such as increasing the biodiversity and breeding new species adaptable to harsh environmental stresses and increasing green surfaces to reduce atmospheric pollutions in arid lands with poor vegetation.

Breaking through the feed barrier: options for improving forage genetics

Pasture based on perennial ryegrass (Lolium perenne L.) and white clover (Trifolium repens L.) is the foundation for production and profit in the Australasian pastoral sectors. The improvement of these species offers direct opportunities to enhance sector performance, provided there is good alignment with industry priorities as quantified by means such as the forage value index. However, the rate of forage genetic improvement must increase to sustain industry competitiveness. New forage technologies and breeding strategies that can complement and enhance traditional approaches are required to achieve this. We highlight current and future research in plant breeding, including genomic and gene technology approaches to improve rate of genetic gain. Genomic diversity is the basis of breeding and improvement. Recent advances in the range and focus of introgression from wild Trifolium species have created additional specific options to improve production and resource-use-efficiency traits. Symbiont genetic resources, especially advances in grass fungal endophytes, make a critical contribution to forage, supporting pastoral productivity, with benefits to both pastures and animals in some dairy regions. Genomic selection, now widely used in animal breeding, offers an opportunity to lift the rate of genetic gain in forages as well. Accuracy and relevance of trait data are paramount, it is essential that genomic breeding approaches be linked with robust field evaluation strategies including advanced phenotyping technologies. This requires excellent data management and integration with decision-support systems to deliver improved effectiveness from forage breeding. Novel traits being developed through genetic modification include increased energy content and potential increased biomass in ryegrass, and expression of condensed tannins in forage legumes. These examples from the wider set of research emphasise forage adaptation, yield and energy content, while covering the spectrum from exotic germplasm and symbionts through to advanced breeding strategies and gene technologies. To ensure that these opportunities are realised on farm, continuity of industry-relevant delivery of forage-improvement research is essential, as is sustained research input from the supporting pasture and plant sciences.

Modernization in plant breeding approaches for improvingbiotic stress resistance in crop plants

Biotic stresses are a potential threat to global food security. The origin of new pathogens and insect races due to climatic andgenetic factors is a major challenge for plant breeders in breeding biotic stress resistant crops. Yield losses due to biotic stresses haveresulted in 800 million people underfed in the world. Reduced yield due to biotic stresses and increasing food demand put internationalfood security at risk as 70% more food will be required in 2050. This review describes and compares the conventional and moleculargenetics methods being used for breeding biotic stress resistant crops. In the past, classical breeding approaches like introduction,hybridization, composite crossing, multiline, and backcross breeding were utilized for this purpose. However, these methods wereslow, expensive, and hectic for developing resistance in crops. Furthermore, breakdown of resistance due to fast evolving pathogenscould not be coped with using these time consuming methods. Therefore, molecular genetics approaches like mutation, marker assistedselection (MAS), genomics, recombinant DNA technology, targeted induced local lesions in genome (TILLING), and virus inducedgene silencing (VIGS) were adapted by breeders to develop effective resistance in crop plants in a shorter time. TILLING, being anontransgenic method, is expected to become the most powerful tool for this purpose.

Impacts of Agricultural Nitrogen on the Environment and Strategies to Reduce these Impacts

Nitrogen is a primary limitation to plant productivity, a major energy and economic input in agroecosystems, and a primary source of environmental pollution via gaseous and leaching losses. The success of the Green Revolution was largely due to breeding of superior plant varieties and the massive use of fertilizers, especially synthetic N-fertilizers, to fulfil the higher yield potential of the new varieties. Today, agriculture uses over 100 million metric tons of fertilizer-N each year, which has doubled the flux of N through the terrestrial N-cycle. Production of fertilizer-N, via the Haber-Bosch process, consumes an enormous amount of fossil fuel, and produces a matching amount of CO2. Likewise, distribution and mechanical application of fertilizer to fields consume fossil fuels and add to atmospheric CO2. About a half of the fertilizer-N applied to fields is captured and used by the target crop, while the rest is lost to the environment. A small percentage is converted to the potent greenhouse gas, N2O, which contributes substantially to the total greenhouse gas emissions from agriculture. Current high-input agricultural systems are not sustainable, in part because they rely on finite fossil fuels and in part because of their negative effects on environmental systems.Several approaches are being taken to reduce the use and environmental impacts of fertilizer-N, while maintaining or even increasing plant productivity, including greater use of symbiotic nitrogen fixation in legumes, use and improvement of associative nitrogen fixation in non-legumes, and the development of synthetic nitrogen-fixing systems for key crop species. Plant breeding approaches to improve nitrogen use efficiency in crop species and management strategies to reduce nitrogen losses from fields are also underway and have met with some success. Basic research is still required to fully understand nitrogen dynamics in the complex and varied agroecosystems, which are affected by interactions between plants, soil, microbes, climate and other environmental variables.Understanding nitrogen dynamics and transformations in agroecosystems will help us to manage better the nitrogen resources in these systems, which will increase agricultural sustainability and reduce its impact on the environment and climate.