Abstract Modern plant breeding is still a time-consuming and costly process, even with the most advanced technologies such as gene editing. Hence, there is an urgent need to develop alternative means for plant trait manipulation and plant protection. RNA interference (RNAi) is a conserved cellular mechanism mediated by naturally occurring double-stranded RNA (dsRNA) and small RNAs (sRNAs) that can target mRNAs for destruction or transcript reduction. Here, we review the potential of technology based on RNAi, called spray-induced gene silencing (SIGS), as an alternative or adjunct to breeding for manipulation of endogenous gene expression in plants or pathogen control. SIGS based on exogenous application of RNA molecules in plants may be especially useful in reducing pest or pathogen impacts, thereby ameliorating biotic stresses and increasing the agronomic performance of crops.

Microbiota in the gut play essential roles in human health. Prebiotics are non-digestible complex carbohydrates that are fermented in the colon, yielding energy and short chain fatty acids, and selectively promote the growth of Bifidobacteria and Lactobacillae in the gastro-intestinal tract. Fructans and inulin are the best-characterized plant prebiotics. Many vegetable, root and tuber crops as well as some fruit crops are the best-known sources of prebiotic carbohydrates, while the prebiotic-rich grain crops include barley, chickpea, lentil, lupin, and wheat. Some prebiotic-rich crop germplasm have been reported in barley, chickpea, lentil, wheat, yacon, and Jerusalem artichoke. A few major quantitative trait loci and gene-based markers associated with high fructan are known in wheat. More targeted search in genebanks using reduced subsets (representing diversity in germplasm) is needed to identify accessions with prebiotic carbohydrates. Transgenic maize, potato and sugarcane with high fructan, with no adverse effects on plant development, have been bred, which suggests that it is feasible to introduce fructan biosynthesis pathways in crops to produce health-imparting prebiotics. Developing prebiotic-rich and super nutritious crops will alleviate the widespread malnutrition and promote human health. A paradigm shift in breeding program is needed to achieve this goal and to ensure that newly-bred crop cultivars are nutritious, safe and health promoting.

Background: The genetic diversity of maize in Peru includes several landraces (within race clusters) and modern open pollinated and hybrid cultivars that are grown by farmers across various regions, thereby making this country a secondary center of diversity for this crop. A main topic of controversy in recent years refers to the unintended presence of transgenic events in locally grown cultivars at main centers of crop diversity. Peru does not yet have biosafety regulations to control or permit the growing of genetically modified crops. Hence, the aim of this research was to undertake a survey in the valley of Barranca, where there were recent claims of authorized transgenic maize grown in farmers fields as well as in samples taken from feed storage and grain or seed trade centers. Results: A total of 162 maize samples (134 from fields, 15 from local markets, eight from the collecting centers of poultry companies, from the local trading center and four samples from seed markets) were included for a qualitative detection by the polymerase chain reaction (PCR) of Cauliflower Mosaic Virus (CaMV) 35S promoter (P35S) and nopaline synthase terminator (Tnos) sequences, as well as for six transgenic events, namely BT11, NK603, T25, 176, TC1507 and MON810. The 134 maize samples from farmers fields were negative for Cry1Ab delta-endotoxin insecticidal protein and enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) using lateral flow strips. The PCR analysis did not detect any of the six transgenic events in samples from farmers fields, local markets, seed trading shops and the local collecting center. There were four transgenic events (T25, NK603, MON810 and TC1507) in grain samples from the barns of poultry companies. Conclusions: This research could not detect, at the 95% probability level, transgenes in farmers' fields in the valley of Barranca. The four transgenic events in grain samples from barns of poultry companies were not surprising because Peru imports maize, mainly for animal feed, from Argentina and the United States that are known for growing transgenic maize.

The use of transgenic crops is steadily increasing around the world, led by soybean (based on total area) and maize (in terms of total number of countries). Transgenic maize is grown in at least 17 countries across four continents: Africa, America, Asia and Europe. The comprehensive global spread of transgenic maize has significant implications for organizations involved in germplasm conservation and genetic enhancement; particularly as some countries require a GMO-free declaration when receiving shipments of maize germplasm. This article describes the protocol used by the International Maize and Wheat Improvement Center (CIMMYT) for monitoring unintentional transgene flow in maize genebank and breeding plots. The protocol is based on polymerase chain reaction (PCR) markers for detecting specific recombinant DNA sequences in bulked samples collected from sentinel plots. To date, no unintentional transgene flow has been detected in CIMMYT fields of maize genebank accessions or breeding materials.

Simple sequence repeats (SSR) or micro-satellites are becoming standard DNA markers for plant genome analysis and are being used as markers in marker assisted breeding. De novo generation of micro-satellite markers through laboratory-based screening of SSR-enriched genomic libraries is highly time consuming and expensive. An alternative is to screen the public databases of related model species where abundant sequence data is already available. All the genomic sequences of Medicago from the public domain database were searched and analysed of di, tri, and tetra nucleotide repeats. Of the total of about 156,000 sequences which were searched, 7325 sequences were found to contain repeat motif and may yield SSR which will yield product sizes of around 200 bp. Of these the most abundantly found repeats were the tri-nucleotide (5210) group. Except for a very small proportion (436), these link to the gene annotation database at TIGR (<A HREF="http://www.tigr.org/">http://www.tigr.org</A>). To facilitate further exploration of this resource, a dynamic database with options to search and link to other resources is available at (<A HREF="http://www.icrisat.org/text/research/grep/homepage/genomics/medssrs1.asp">http://www.icrisat.org/text/research/grep/homepage/genomics/medssrs1.asp</A>) and on CDs from <A HREF="mailto:V.Mahalakshmi@cgiar.org">V.Mahalakshmi@cgiar.org</A>.

Sequencing and analysing of the Arabidopsis thaliana genome, the first plant kingdom genome to be unraveled, will always remain a scientific landmark. International initiatives to sequence rice, the most important cereal in Asia, are underway. However as functional information piles up in Arabidopsis and rice, researchers working in other crops will benefit from this new knowledge and apply it to their studied plants or crop species. The increasing role of public databases of model organisms and bio-informatics in data mining, presents a new opportunity as well as a challenge to researchers to develop more focused molecular tools for gene discovery and deployment. The work presented in here describes how such an approach has benefited sorghum, a rainfed semi-arid troprical cereal.

Integrated pest management has historically placed great hopes on host plant resistance. However, conventional host-plant resistance to insects involves quantitative traits at several loci. As a result, the progress has been slow and difficult to achieve. With the advent of genetic transformation techniques, it has become possible to clone and insert genes into the crop plants to confer resistance to insect pests. Resistance to insects has been demonstrated in transgenic plants expressing genes for delta-endotoxins from Bacillus thuringiensis (Bt), protease inhibitors, enzymes and plant lectins. Most of the plant derived genes produce chronic rather than toxic effects and some insect pests are not sensitive to some of these factors. The potential of plant derived genes can be realised by deploying them in combination with host plant resistance and exotic genes. Genes conferring resistance to insects have been inserted into crop plants such as maize, cotton, potato, tobacco, potatoes, rice, broccoli, lettuce, walnuts, apples, alfalfa and soybean. Genetically transformed crops with Bt genes have been deployed for cultivation in USA, China and Australia. However, very little has been done to use this technology for improving crop production in the harsh environments of the tropics, where the need for increasing food production is most urgent. International agricultural research centres, advanced research institutes and the seed sector should make an effort to use these new tools for increasing food production in poorer regions of the world. There is an urgent need to develop a scientifically sound strategy to deploy exotic and plant derived genes for minimising the extent of losses caused by insect pests. Equally important is the need for following the biosafety regulations, more responsible public debate, social attitude and better presentation of the benefits for a rational deployment of the genetically transformed plants. <A NAME="Article"></A>

The triploid plantain landrace Obino l’Ewai (Musa spp., AAB genome) has been crossed with a wild diploid banana (M. acuminata subsp. burmannica var. ‘Calcutta 4’, AA genome) to generate full-sib diploid and tetraploid hybrids combining good agronomic performance and disease resistance. Microsatellite marker analysis of the parental genotypes confirmed the highly heterozygous nature of both parental genotypes. Comparative analysis of 2x and 4x full-sib hybrids with their parental genotypes indicated that tetraploid hybrids are generally more closely related to Obino l’Ewai than their diploid full-sibs. Based on VNTR analysis it is possible to identify those hybrids, which may be most useful in subsequent breeding of secondary triploid hybrids. There was a significant (P<0.05) negative association between the VNTR-based genetic similarity of hybrids to Obino l’Ewai and a phenotypic distance index based on eight agronomic descriptors. However, there was no association between the molecular genetic similarity of hybrids to Calcutta 4 and the respective phenotypic distance index. Many microsatellite markers generated an unexpectedly high number of amplification products from AA and AAB genotypes plus their progeny which may suggest the presence of a high frequency of loci duplication in both A and B genomes, in addition to the detection of heterozygous and/or homoeologous loci.

This article reviews some of the highlights of modern plant biotechnology and discusses the potential applications of biotechnology in the betterment of farming systems in the next millennium. Plant biotechnology will facilitate the farming of crops with multiple durable resistance to pests and diseases, particularly in the absence of pesticides. Likewise, transgenes or marker-assisted selection may assist in the development of high yielding crops, which will be needed to feed the world and save land for the conservation of plant biodiversity in natural habitats. Hence, crops should be engineered to meet the demands and needs of consumers. The genetic base of crop production can be preserved and widen by an integration of biotechnology tools in conventional breeding. Similarly targeting specific genotypes to particular cropping systems may be facilitated by understanding specific gene-by-environment interaction(s) with the aid of molecular research. High quality crops with improved nutritional and health characteristics as well as other aspects of added-value may be obtained through multidisciplinary co-operation among plant breeders, biotechnologists, and other plant scientists. Co-ordinated efforts between consumers, policy makers, farmers and researchers will be required to convert the various aspects of a crop ideotype into components of new and improved farming systems of the next millennium.

Bananas and plantains (Musa spp.) are the most important tropical fruit crops. They form an integral component of the farming systems in the humid agroecological zones of the tropics. A broad array of applied cell and molecular techniques are increasingly being used worldwide to facilitate and enhance the handling and improvement of plantain and banana germplasm. Tissue culture is used for germplasm exchange, conservation and rapid multiplication, while in vitro seed germination (based on embryo culture or rescue) plays a critical role in generating hybrid plants. DNA marker systems have been developed in Musa to assist germplasm management, selection within the breeding pool or gene introgression from wild species, and for disease diagnosis. Likewise, genetic transformation using the particle gun method or through Agrobacterium co-cultivation shows potential for the genetic betterment of the crop. This article discusses the applications of biotechnology for the genetic enhancement of banana and plantain. It highlights current advances by research teams across the world and reviews progress in molecular breeding of Musa by the International Institute of Tropical Agriculture and its collaborators