Effective approaches to study the plant-root knot nematode interaction.

With the growing global demands on sustainable food production, one of the biggest challenges to agriculture is associated with crop losses due to parasitic nematodes. While chemical pesticides have been quite successful in crop protection and mitigation of damage from parasites, their potential harm to humans and environment, as well as the emergence of nematode resistance, have necessitated the development of viable alternatives to chemical pesticides. One of the most promising and targeted approaches to biocontrol of parasitic nematodes in crops is that of RNA interference (RNAi). In this study we explore the possibility of using biostimulants obtained from metabolites of soil streptomycetes to protect wheat (Triticum aestivum L.) against the cereal cyst nematode Heterodera avenae by means of inducing RNAi in wheat plants. Theoretical models of uptake of organic compounds by plants, and within-plant RNAi dynamics, have provided us with useful insights regarding the choice of routes for delivery of RNAi-inducing biostimulants into plants. We then conducted in planta experiments with several streptomycete-derived biostimulants, which have demonstrated the efficiency of these biostimulants at improving plant growth and development, as well as in providing resistance against the cereal cyst nematode. Using dot blot hybridization we demonstrate that biostimulants trigger a significant increase of the production in plant cells of si/miRNA complementary with plant and nematode mRNA. Wheat germ cell-free experiments show that these si/miRNAs are indeed very effective at silencing the translation of nematode mRNA having complementary sequences, thus reducing the level of nematode infestation and improving plant resistance to nematodes. Thus, we conclude that natural biostimulants produced from metabolites of soil streptomycetes provide an effective tool for biocontrol of wheat nematode.

/pdf/rnai-based-biocontrol-of-wheat-nematodes-using-natural-poly-1ryj3rzeby.pdf

RNAi-Based Biocontrol of Wheat Nematodes Using Natural Poly-Component Biostimulants

The genetic diversity of bananas (Musa spp.), one of the most economically important crops in Tanzania, is underestimated with scarce reports available. In addition, cultivation of banana is severely constrained by plant parasitic nematodes (PPN). We assessed genetic diversity and population structure of 159 banana varieties from four agro-ecological zones (Northern, Southern highlands, Lake and Zanzibar islands) of Tanzania using 20 SSR markers analysed with UPGMA method. We also, assessed the PPN occurrence through isolation and counting of PPN from each banana genotype where leaf samples were collected. SSR primer pairs were polymorphic; and generated 63 distinct reproducible bands. The polymorphic information content values of each SSR marker ranged from 0.50 to 0.75 indicating high level of genetic diversity among banana varieties. The 159 banana varieties were grouped into two clusters: A and B, based on UPGMA cluster and population structure analysis. A total of 128 independent and 31 overlapping genotypes were identified. Higher kinship among genotypes was observed in cluster A compared with cluster B. Some of the clusters in A and B genotypes showed high genetic distance. The most prevalent and abundant nematode species was Pratylenchus goodeyi with a mean count of 63%, followed by Radopholus similis (31%) and P. coffeae (6%). The results from this study provides a foundation for understanding genetic variability of bananas existing in Tanzania and PPN occurrence that will be valuable information for breeding disease and pest-resistant bananas with high yield traits.

/pdf/plant-parasitic-nematodes-occurrence-and-genetic-diversity-4dv0ognban.pdf

Plant parasitic nematodes occurrence and genetic diversity of banana cultivars grown in Tanzania

The need for biofortification of staple crops has become the concern of Modern Nutritional Genomics in producing staple crops with the ability of accumulating high micronutrients. Beta-carotene (Provitamin A) biosynthesis pathway is a stepwise process which is coded by two main genes; phytoene synthase (PSY) and lycopene beta-cyclase (LYB) . These two important genes LYB 7 and PSY 1 1 were amplified, sequenced and analyzed within various diploid and triploid plantain landraces to identify SNPs present and to study their potentials for mapping the quantitative trait loci in those genes brought about by climatic variation within plantain landraces over time and also reveal their phylogenetic relationship. A total of 7 SNPs were found in the two genes from 68 genotypes used for PSY 11 and 35 plantain genotypes for LYB 7 gene. A frequency of 1 SNP per 80bp was detected in LYB 7 and I SNP per 32bp in PSY 11 gene of 160 bp sequences each. The allelic diversity was 0.0286 in LYB 7 and 0.0147 to 0.0294 in PSY 11. The phylogenetic relationship revealed that the genotypes for LYB 7 gene follow two distinct lines of evolution whereas in PSY 11 gene, the genotypes are similar with Zue and Sel advancing a bit. From the research, the high frequency of SNPs detected showed that it is sufficient in mapping the quantitative trait loci associated with high beta-carotene synthesis and could be incorporated in plantain breeding program.

http://www.openscienceonline.com/author/download?paperId=62&stateId=8000&fileType=3

Single nucleotide polymorphism (SNP) markers discovery within Musa spp (plantain landraces, AAB genome) for use in beta carotene (Provitamin A) trait mapping

Plant pathogenic fungi may cause crop losses that affect the world economy. Although one of the most effective ways to combat plant pathogens is a chemical control, alternative methods have become necessity as a result of environmental pollution and residue problems caused by pesticides used in agriculture. The mechanism of RNA interference (RNAi) has been developed to completely prevent or decrease the production of protein which is an expression of a specific gene. Due to the degeneracy of mRNA chain which is complementary of double-stranded RNA (dsRNA) entered into cells is prevented the production of protein.  RNA silencing is very important for many organisms and microorganisms. This natural phenomenon can be exploited to control agronomically relevant plant diseases, based on the demonstration that in vitro feeding of dsRNA can signal Post transcriptional gene silencing (one of the RNA silencing methods) of target genes in various plant pests and pathogens, such as insects, nematodes and fungi.  In other words, as well as determining a function of specific gene and developing of new plant various, RNA silencing was also begun to use for developing resistant plant varieties against biotic and abiotic factors by the suppression of gene expression. This biotechnological method, termed host-induced gene silencing (HIGS), has emerged as a promising alternative in plant protection because it combines high selectivity for the target organism with minimal side effects, as compared with chemical treatments. In recent years, the significant developments related to the use of HIGS in management against plant pathogenic fungi ( Puccinia striiformis f.sp. tritici , Blumeria graminis, Fusarium verticillioides etc.) was obtained. In this review, it is mentioned from the mechanism of HIGS and studies related to the use against plant pathogenic fungi.

/pdf/a-new-approach-in-management-against-plant-fungal-disease-z21n1bgt06.pdf

A New Approach in Management Against Plant Fungal Disease: Host Induced Gene Silencing

High gibberellic acid content in plantain helps to increase its shelf life while parthenocarpy is a desirable agronomic trait in this crop. Gene fragments coding for gibberellic acid (JID 775) and parthenocarpy (DELA 381), were amplified, sequenced and analysed to detect SNPs present and to study their potentials for mapping the quantitative trait loci associated with these traits in plantain landraces. Gene sequences were analysed to elucidate phylogenetic relationships among the studied genotypes. A total of 49 SNPS and 7 Indels were found in the two genes in plantain. Out of the 31 genotypes comprising 27 triploids and 4 diploids analysed for JID 775, a frequency of 1SNP per 23bp and 1 Indel per 128bp was detected. Nineteen genotypes, including 13 triploids and 6 diploids, were analysed for DELA 381 gene, and a frequency of 1 SNP per 21bp and 1 Indel per 142bp was observed. The allelic diversity ranged from 0.0526 to 0.3684 in DELA 381 and 0.0323 to 0.6129 in JID 775. The phylogenetic tree for DELA 381 showed that diploid plantains had evolved in a separate line from the triploids. In JID 775, a diploid plantain (KM5) has advanced to have a similar gene sequence as the triploids. The high frequency of SNPs detected showed that it is sufficient in mapping the quantitative trait loci associated with the two agronomic traits studied in diploid and triploid plantains and could be incorporated in plantain breeding.

Single Nucleotide Polymorphism Markers Discovery in Musa Spp (Plantain Landraces, AAB Genome) and its Potentials for Use in Gibberellic Acid and Parthenocarpy Trait Mapping

Gene Silencing Technologies in Creating Resistance to Plant Diseases

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Single Nucleotide Polymorphism Markers Discovery in Musa Spp (Plantain Landraces, AAB Genome) and its Potentials for Use in Gibberellic Acid and Parthenocarpy Trait Mapping

Gene Silencing Technologies in Creating Resistance to Plant Diseases