Efficient DNA isolation from moroccan arar tree [Tetraclinis articulata (Vahl) Masters] leaves and optimization of the rapd-pcr molecular technique

Efficient DNA isolation from Moroccan Arar tree [Tetraclinis articulata (Vahl) Masters] leaves and optimization of the RAPD-PCR molecular technique. Molecular genetic analysis of Arar tree [Tetraclinis articulata (Vahl) Masters] is often limited by the availability of fresh tissue and an efficient and reliable protocol for high quality genomic DNA extraction. In this study, two DNA extraction protocols were specifically developed for extracting high quality genomic DNA from Arar tree leaves: modified QIAgen DNA Kit and protocol developed by Ouenzar et al. (1998). DNA yield and purity were monitored by gel electrophoresis and by determining absorbance at UV (A260/A280 and A260/A230). Both ratios were between 1.7 and 2.0, indicating that the presence of contaminating metabolites was minimal. The DNA yield obtained ranged between 20 to 40 μg/g of plant materiel. The Ouenzar and collaborators protocol gave higher yield but was more time consuming compared to QIAgen Kit. However, both techniques gave DNA of good quality that is amenable to RAPD-PCR reactions. Additionally, restriction digestion and PCR analyses of the obtained DNA showed its compatibility with downstream applications. Randomly Amplified Polymorphic DNA profiling from the isolated DNA was optimized to produce scorable and clear amplicons. The presented protocols allow easy and high quality DNA isolation for genetic diversity studies within Arar tree

The 8D05 Parasitism Gene of Meloidogyne incognita Is Required for Successful Infection of Host Roots

Parasitism genes encode effector proteins that are secreted through the stylet of root-knot nematodes to dramatically modify selected plant cells into giant-cells for feeding. The Mi8D05 parasitism gene previously identified was confirmed to encode a novel protein of 382 amino acids that had only one database homolog identified on contig 2374 within the Meloidogyne apla genome. Mi8D05 expression peaked in M. incognita parasitic second-stage juveniles within host roots and its encoded protein was limited to the subventral esophageal gland cells that produce proteins secreted from the stylet. Constitutive expression of Mi8D05 in transformed Arabidopsis thaliana plants induced accelerated shoot growth and early flowering but had no visible effects on root growth. Independent lines of transgenic Arabidopsis that expressed a double-stranded RNA complementary to Mi8D05 in host-derived RNA interference (RNAi) tests had up to 90% reduction in infection by M. incognita compared with wild-type control plants, suggesting that Mi8D05 plays a critical role in parasitism by the root-knot nematode. Yeast two-hybrid experiments confirmed the specific interaction of the Mi8D05 protein with plant aquaporin tonoplast intrinsic protein 2 (TIP2) and provided evidence that the Mi8D05 effector may help regulate solute and water transport within giant-cells to promote the parasitic interaction

The interaction of the novel 30C02 cyst nematode effector protein with a plant β-1,3-endoglucanase may suppress host defence to promote parasitism

Phytoparasitic nematodes secrete an array of effector proteins to modify selected recipient plant cells into elaborate and essential feeding sites. The biological function of the novel 30C02 effector protein of the soybean cyst nematode, Heterodera glycines, was studied using Arabidopsis thaliana as host and the beet cyst nematode, Heterodera schachtii, which contains a homologue of the 30C02 gene. Expression of Hg30C02 in Arabidopsis did not affect plant growth and development but increased plant susceptibility to infection by H. schachtii. The 30C02 protein interacted with a specific (AT4G16260) host plant β-1,3-endoglucanase in both yeast and plant cells, possibly to interfere with its role as a plant pathogenesis-related protein. Interestingly, the peak expression of 30C02 in the nematode and peak expression of At4g16260 in plant roots coincided at around 3–5 d after root infection by the nematode, after which the relative expression of At4g16260 declined significantly. An Arabidopsis At4g16260 T-DNA mutant showed increased susceptibility to cyst nematode infection, and plants that overexpressed At4g16260 were reduced in nematode susceptibility, suggesting a potential role of host β-1,3-endoglucanase in the defence response against H. schachtii infection. Arabidopsis plants that expressed dsRNA and its processed small interfering RNA complementary to the Hg30C02 sequence were not phenotypically different from non-transformed plants, but they exhibited a strong RNA interference-mediated resistance to infection by H. schachtii. The collective results suggest that, as with other pathogens, active suppression of host defence is a critical component for successful parasitism by nematodes and a vulnerable target to disrupt the parasitic cycle

Plant-driven selection of microbes in the rhizosphere and plant-microbe feedbacks

  L’effet des plantes sur les microorganismes du sol et les rétroactions entre les microbes et les plantes sont devenus le focus de beaucoup de recherche. Les dernières avancées dans le domaine des interactions plantes-microbes montrent que les plantes sont capables de façonner le microbiome de leur rhizosphère via des mécanismes divers. Dans cette revue, nous rassemblons des résultats de multiples études sur le rôle des plantes dans l’altération de la structure et des fonctions des microorganismes au niveau de la rhizosphère. En outre, nous discutons les rôles de différentes substances phytochimiques dans la médiation de ces effets. Finalement, nous soulignons que l’enrichissement sélectif de microorganismes spécifiques a aussi bien des rétroactions négatives, avec l’accumulation de pathogènes dans la rhizosphère, que positives, suite au recrutement de microflore bénéfique. Une meilleure compréhension des mécanismes qui régissent la sélection de microorganismes ayant des rétroactions positives ouvrira de nouvelles possibilités pour l’amélioration de la production agricole. Mots-clés: Plantes, microorganismes, rhizosphère, rétroactions.Plant impacts on soil microbial communities and plant-microbe feedbacks have become the focus of much research. Recent advances in plant-microbe interactions investigations show that plants are able to shape their rhizosphere microbiome through diverse mechanisms. In this review, we gather findings from across multiple studies on the role of plants in altering the structure and functions of microbes in the rhizosphere. In addition, we discuss the roles of diverse phytochemicals in mediating these effects. Finally, we highlight that selective enrichment of specific microorganisms in the rhizosphere has either negative feedbacks, with pathogen accumulation in the rhizosphere; or, perhaps most importantly, positive feedbacks as a result of the recruitment of a beneficial microflora. Insights into the mechanisms that underpin plant selection of microbial communities with positive feedbacks will provide new opportunities to increase crop production. Keywords: Plants, microbes, rhizosphere, feedbacks

Plant-driven selection of microbes in the rhizosphere and plant-microbe feedbacks

  L’effet des plantes sur les microorganismes du sol et les rétroactions entre les microbes et les plantes sont devenus le focus de beaucoup de recherche. Les dernières avancées dans le domaine des interactions plantes-microbes montrent que les plantes sont capables de façonner le microbiome de leur rhizosphère via des mécanismes divers. Dans cette revue, nous rassemblons des résultats de multiples études sur le rôle des plantes dans l’altération de la structure et des fonctions des microorganismes au niveau de la rhizosphère. En outre, nous discutons les rôles de différentes substances phytochimiques dans la médiation de ces effets. Finalement, nous soulignons que l’enrichissement sélectif de microorganismes spécifiques a aussi bien des rétroactions négatives, avec l’accumulation de pathogènes dans la rhizosphère, que positives, suite au recrutement de microflore bénéfique. Une meilleure compréhension des mécanismes qui régissent la sélection de microorganismes ayant des rétroactions positives ouvrira de nouvelles possibilités pour l’amélioration de la production agricole. Mots-clés: Plantes, microorganismes, rhizosphère, rétroactions.Plant impacts on soil microbial communities and plant-microbe feedbacks have become the focus of much research. Recent advances in plant-microbe interactions investigations show that plants are able to shape their rhizosphere microbiome through diverse mechanisms. In this review, we gather findings from across multiple studies on the role of plants in altering the structure and functions of microbes in the rhizosphere. In addition, we discuss the roles of diverse phytochemicals in mediating these effects. Finally, we highlight that selective enrichment of specific microorganisms in the rhizosphere has either negative feedbacks, with pathogen accumulation in the rhizosphere; or, perhaps most importantly, positive feedbacks as a result of the recruitment of a beneficial microflora. Insights into the mechanisms that underpin plant selection of microbial communities with positive feedbacks will provide new opportunities to increase crop production. Keywords: Plants, microbes, rhizosphere, feedbacks

Transgenic tobacco plants constitutively expressing Arabidopsis NPR1 show enhanced resistance to root-knot nematode, Meloidogyne incognita

In Arabidopsis, non-expressor of pathogenesis related genes-1, NPR1 has been shown to be a positive regulator of the salicylic acid controlled systemic acquired resistance pathway and modulates the cross talk between SA and JA signaling. Transgenic plants expressing AtNPR1 constitutively exhibited resistance against pathogens as well as herbivory. In the present study, tobacco transgenic plants expressing AtNPR1 were studied further for their response to infection by the sedentary endoparasitic root knot nematode, Meloidogyne incognita. Transgenic plants showed enhanced resistance against the root-knot nematode infection. Prominent differences in the shoot and root weights of wild type and transgenic plants were observed post-inoculation with M. incognita. This was associated with a decrease in the number of root galls and egg masses in transgenic plants compared to WT. The transgenic plants also showed constitutive and induced expression of some PR protein genes, when challenged with M. incognita

First draft genome assembly of the Argane tree (Argania spinosa)

Background: The Argane tree (Argania spinosa L. Skeels) is an endemic tree of southwestern Morocco that plays an important socioeconomic and ecologic role for a dense human population in an arid zone. Several studies confirmed the importance of this species as a food and feed source and as a resource for both pharmaceutical and cosmetic compounds. Unfortunately, the argane tree ecosystem is facing significant threats from environmental changes (global warming, over-population) and over-exploitation. Limited research has been conducted, however, on argane tree genetics and genomics, which hinders its conservation and genetic improvement. Methods: Here, we present a draft genome assembly of A. spinosa. A reliable reference genome of A. spinosa was created using a hybrid de novo assembly approach combining short and long sequencing reads. Results: In total, 144 Gb Illumina HiSeq reads and 7.2 Gb PacBio reads were produced and assembled. The final draft genome comprises 75 327 scaffolds totaling 671 Mb with an N50 of 49 916 kb. The draft assembly is close to the genome size estimated by k-mers distribution and covers 89% of complete and 4.3 % of partial Arabidopsis orthologous groups in BUSCO. Conclusion: The A. spinosa genome will be useful for assessing biodiversity leading to efficient conservation of this endangered endemic tree. Furthermore, the genome may enable genome-assisted cultivar breeding, and provide a better understanding of important metabolic pathways and their underlying genes for both cosmetic and pharmacological purposes

A plant-feeding nematode indirectly increases the fitness of an aphid

Plants suffer multiple, simultaneous assaults from above and below ground. In the laboratory, pests and/or pathogen attack are commonly studied on an individual basis. The molecular response of the plant to attack from multiple organisms and the interaction of different defence pathways is unclear. The inducible systemic responses of the potato (Solanum tuberosum L.) host plant were analysed to characterise the plant-mediated indirect interactions between a sedentary, endoparasitic nematode (Globodera pallida) and a phloem-sucking herbivore (Myzus persicae). The reproductive success of M. persicae was greater on potato plants pre-infected with G. pallida compared to control plants. Salicylic acid (SA) increased systemically in the leaves of potato plants following nematode and aphid infection singly with a corresponding increase in expression of SA-mediated marker genes. An increase in jasmonic acid (JA) associated with aphid infection was suppressed when plants were co-infected with nematodes. Our data suggests a positive, asymmetric interaction between a sedentary endoparasitic nematode and a sap-sucking insect. The systemic response of the potato plant following infection with G. pallida indirectly influences the performance of M. persicae. This work reveals additional secondary benefits of controlling individual crop pests

Significance of Cuscutain, a cysteine protease from Cuscuta reflexa, in host-parasite interactions

<p>Abstract</p> <p>Background</p> <p>Plant infestation with parasitic weeds like <it>Cuscuta reflexa </it>induces morphological as well as biochemical changes in the host and the parasite. These modifications could be caused by a change in protein or gene activity. Using a comparative macroarray approach <it>Cuscuta </it>genes specifically upregulated at the host attachment site were identified.</p> <p>Results</p> <p>One of the infestation specific <it>Cuscuta </it>genes encodes a cysteine protease. The protein and its intrinsic inhibitory peptide were heterologously expressed, purified and biochemically characterized. The haustoria specific enzyme was named cuscutain in accordance with similar proteins from other plants, e.g. papaya. The role of cuscutain and its inhibitor during the host parasite interaction was studied by external application of an inhibitor suspension, which induced a significant reduction of successful infection events.</p> <p>Conclusions</p> <p>The study provides new information about molecular events during the parasitic plant - host interaction. Inhibition of cuscutain cysteine proteinase could provide means for antagonizing parasitic plants.</p

Nematode effector proteins: an emerging paradigm of parasitism

Phytonematodes use a stylet and secreted effectors to modify host cells and ingest nutrients to support their growth and development. The molecular function of nematode effectors is currently the subject of intense investigation. In this review, we summarize our current understanding of nematode effectors, with a particular focus on proteinaceous stylet-secreted effectors of sedentary endoparasitic phytonematodes, for which a wealth of information has surfaced in the past 10 yr. We provide an update on the effector repertoires of several of the most economically important genera of phytonematodes and discuss current approaches to dissecting their function. Lastly, we highlight the latest breakthroughs in effector discovery that promise to shed new light on effector diversity and function across the phylum Nematoda