Integrative transcriptomics reveals association of abscisic acid and lignin pathways with cassava whitefly resistance.

BACKGROUND: Whiteflies are a global threat to crop yields, including the African subsistence crop cassava (Manihot esculenta). Outbreaks of superabundant whitefly populations throughout Eastern and Central Africa in recent years have dramatically increased the pressures of whitefly feeding and virus transmission on cassava. Whitefly-transmitted viral diseases threaten the food security of hundreds of millions of African farmers, highlighting the need for developing and deploying whitefly-resistant cassava. However, plant resistance to whiteflies remains largely poorly characterized at the genetic and molecular levels. Knowledge of cassava-defense programs also remains incomplete, limiting characterization of whitefly-resistance mechanisms. To better understand the genetic basis of whitefly resistance in cassava, we define the defense hormone- and Aleurotrachelus socialis (whitefly)-responsive transcriptome of whitefly-susceptible (COL2246) and whitefly-resistant (ECU72) cassava using RNA-seq. For broader comparison, hormone-responsive transcriptomes of Arabidopsis thaliana were also generated. RESULTS: Whitefly infestation, salicylic acid (SA), jasmonic acid (JA), ethylene (ET), and abscisic acid (ABA) transcriptome responses of ECU72 and COL2246 were defined and analyzed. Strikingly, SA responses were largely reciprocal between the two cassava genotypes and we suggest candidate regulators. While susceptibility was associated with SA in COL2246, resistance to whitefly in ECU72 was associated with ABA, with SA-ABA antagonism observed. This was evidenced by expression of genes within the SA and ABA pathways and hormone levels during A. socialis infestation. Gene-enrichment analyses of whitefly- and hormone-responsive genes suggest the importance of fast-acting cell wall defenses (e.g., elicitor recognition, lignin biosynthesis) during early infestation stages in whitefly-resistant ECU72. A surge of ineffective immune and SA responses characterized the whitefly-susceptible COL2246's response to late-stage nymphs. Lastly, in comparison with the model plant Arabidopsis, cassava's hormone-responsive genes showed striking divergence in expression. CONCLUSIONS: This study provides the first characterization of cassava's global transcriptome responses to whitefly infestation and defense hormone treatment. Our analyses of ECU72 and COL2246 uncovered possible whitefly resistance/susceptibility mechanisms in cassava. Comparative analysis of cassava and Arabidopsis demonstrated that defense programs in Arabidopsis may not always mirror those in crop species. More broadly, our hormone-responsive transcriptomes will also provide a baseline for the cassava community to better understand global responses to other yield-limiting pests/pathogens.

Genome-wide analyses of cassava Pathogenesis-related (PR) gene families reveal core transcriptome responses to whitefly infestation, salicylic acid and jasmonic acid.

BACKGROUND: Whiteflies are a threat to cassava (Manihot esculenta), an important staple food in many tropical/subtropical regions. Understanding the molecular mechanisms regulating cassava's responses against this pest is crucial for developing control strategies. Pathogenesis-related (PR) protein families are an integral part of plant immunity. With the availability of whole genome sequences, the annotation and expression programs of the full complement of PR genes in an organism can now be achieved. An understanding of the responses of the entire complement of PR genes during biotic stress and to the defense hormones, salicylic acid (SA) and jasmonic acid (JA), is lacking. Here, we analyze the responses of cassava PR genes to whiteflies, SA, JA, and other biotic aggressors. RESULTS: The cassava genome possesses 14 of the 17 plant PR families, with a total of 447 PR genes. A cassava PR gene nomenclature is proposed. Phylogenetic relatedness of cassava PR proteins to each other and to homologs in poplar, rice and Arabidopsis identified cassava-specific PR gene family expansions. The temporal programs of PR gene expression in response to the whitefly (Aleurotrachelus socialis) in four whitefly-susceptible cassava genotypes showed that 167 of the 447 PR genes were regulated after whitefly infestation. While the timing of PR gene expression varied, over 37% of whitefly-regulated PR genes were downregulated in all four genotypes. Notably, whitefly-responsive PR genes were largely coordinately regulated by SA and JA. The analysis of cassava PR gene expression in response to five other biotic stresses revealed a strong positive correlation between whitefly and Xanthomonas axonopodis and Cassava Brown Streak Virus responses and negative correlations between whitefly and Cassava Mosaic Virus responses. Finally, certain associations between PR genes in cassava expansions and response to biotic stresses were observed among PR families. CONCLUSIONS: This study represents the first genome-wide characterization of PR genes in cassava. PR gene responses to six biotic stresses and to SA and JA are demonstrably different to other angiosperms. We propose that our approach could be applied in other species to fully understand PR gene regulation by pathogens, pests and the canonical defense hormones SA and JA.

A metabolomics characterisation of natural variation in the resistance of cassava to whitefly.

BACKGROUND: Cassava whitefly outbreaks were initially reported in East and Central Africa cassava (Manihot esculenta Crantz) growing regions in the 1990's and have now spread to other geographical locations, becoming a global pest severely affecting farmers and smallholder income. Whiteflies impact plant yield via feeding and vectoring cassava mosaic and brown streak viruses, making roots unsuitable for food or trading. Deployment of virus resistant varieties has had little impact on whitefly populations and therefore development of whitefly resistant varieties is also necessary as part of integrated pest management strategies. Suitable sources of whitefly resistance exist in germplasm collections that require further characterization to facilitate and assist breeding programs. RESULTS: In the present work, a hierarchical metabolomics approach has been employed to investigate the underlying biochemical mechanisms associated with whitefly resistance by comparing two naturally occurring accessions of cassava, one susceptible and one resistant to whitefly. Quantitative differences between genotypes detected at pre-infestation stages were consistently observed at each time point throughout the course of the whitefly infestation. This prevalent differential feature suggests that inherent genotypic differences override the response induced by the presence of whitefly and that they are directly linked with the phenotype observed. The most significant quantitative changes relating to whitefly susceptibility were linked to the phenylpropanoid super-pathway and its linked sub-pathways: monolignol, flavonoid and lignan biosynthesis. These findings suggest that the lignification process in the susceptible variety is less active, as the susceptible accession deposits less lignin and accumulates monolignol intermediates and derivatives thereof, differences that are maintained during the time-course of the infestation. CONCLUSIONS: Resistance mechanism associated to the cassava whitefly-resistant accession ECU72 is an antixenosis strategy based on reinforcement of cell walls. Both resistant and susceptible accessions respond differently to whitefly attack at biochemical level, but the inherent metabolic differences are directly linked to the resistance phenotype rather than an induced response in the plant.

SPX1 is a phosphate-dependent inhibitor of Phosphate Starvation Response 1 in Arabidopsis.

To cope with growth in low-phosphate (Pi) soils, plants have evolved adaptive responses that involve both developmental and metabolic changes. Phosphate Starvation Response 1 (PHR1) and related transcription factors play a central role in the control of Pi starvation responses (PSRs). How Pi levels control PHR1 activity, and thus PSRs, remains to be elucidated. Here, we identify a direct Pi-dependent inhibitor of PHR1 in Arabidopsis, SPX1, a nuclear protein that shares the SPX domain with yeast Pi sensors and with several Pi starvation signaling proteins from plants. Double mutation of SPX1 and of a related gene, SPX2, resulted in molecular and physiological changes indicative of increased PHR1 activity in plants grown in Pi-sufficient conditions or after Pi refeeding of Pi-starved plants but had only a limited effect on PHR1 activity in Pi-starved plants. These data indicate that SPX1 and SPX2 have a cellular Pi-dependent inhibitory effect on PHR1. Coimmunoprecipitation assays showed that the SPX1/PHR1 interaction in planta is highly Pi-dependent. DNA-binding and pull-down assays with bacterially expressed, affinity-purified tagged SPX1 and ΔPHR1 proteins showed that SPX1 is a competitive inhibitor of PHR1 binding to its recognition sequence, and that its efficiency is highly dependent on the presence of Pi or phosphite, a nonmetabolizable Pi analog that can repress PSRs. The relative strength of the SPX1/PHR1 interaction is thus directly influenced by Pi, providing a link between Pi perception and signaling.

Antagonistic role of 9-lipoxygenase-derived oxylipins and ethylene in the control of oxidative stress, lipid peroxidation and plant defence.

9-lipoxygenases (9-LOXs) initiate fatty acid oxygenation in plant tissues, with formation of 9-hydroxy-10,12,15-octadecatrienoic acid (9-HOT) from linolenic acid. A lox1 lox5 mutant, which is deficient in 9-LOX activity, and two mutants noxy6 and noxy22 (non-responding to oxylipins), which are insensitive to 9-HOT, have been used to investigate 9-HOT signalling. Map-based cloning indicated that the noxy6 and noxy22 mutations are located at the CTR1 (CONSTITUTIVE ETHYLENE RESPONSE1) and ETO1 (ETHYLENE-OVERPRODUCER1) loci, respectively. In agreement, the noxy6 and noxy22 mutants, renamed as ctr1-15 and eto1-14, respectively, showed enhanced ethylene (ET) production. The correlation between increased ET production and reduced 9-HOT sensitivity indicated by these results was supported by experiments in which exogenously added ethylene precursor ACC (1-aminocyclopropane-1-carboxylic acid) impaired the responses to 9-HOT. Moreover, a reciprocal interaction between ET and 9-HOT signalling was indicated by results showing that the effect of ACC was reduced in the presence of 9-HOT. We found that the 9-LOX and ET pathways regulate the response to the lipid peroxidation-inducer singlet oxygen. Thus, the massive transcriptional changes seen in wild-type plants in response to singlet oxygen were greatly affected in the lox1 lox5 and eto1-14 mutants. Furthermore, these mutants displayed enhanced susceptibility to both singlet oxygen and Pseudomonas syringae pv. tomato, in the latter case leading to increased accumulation of the lipid peroxidation product malondialdehyde. These findings demonstrate an antagonistic relationship between products of the 9-LOX and ET pathways, and suggest a role for the 9-LOX pathway in modulating oxidative stress, lipid peroxidation and plant defence.

Tamoxifeno como supresor de la carcinogenesis mamaria inducida por 7,12-dimetil-benzoantraceno y nitroso-metilurea, en ratas hembras con hiperprolactinemia provocada / Tamoxifen as a suppressor of mammary carcinogenesis induced by 7,12-dimetil-benzanthracene and methylnitrosourea, in female rats with induced hyperprolactinemia

El efecto del tamoxifeno sobre tumores mamarios inducidos con DMBA (7, 12-dimetil-benzoantraceno) y NMU (nitroso-metilurea) se analiza. De 7 lotes de ratas Sprague-Dayley, 4 (A-B-C-D) reciben adiconalmene sulpirida 0,5 mg/100g durante 5 días de la semana y hasta finalizar el experimento. Dos lotes (C-D) recibieron una sola inyección intraperitoneal de 900 microcuries por 100g de peso de I. Tres lotes (E-F-G) recibieron sólo I para activar secreción de prolactina, y de ellos, 2(F-G)recibieron tamoxifeno. Los tumores que se desarrollaron fueron todos del tupo adenocarcinoma papilífero. Los receptores hormonales estrogénicos fueron positivos. El estudio por microscopia electrónica mostró vacuolas acuosas grasosas en el citoplasma y la falta de organoides y mitocondrias en algunas células. Los lotes que recibieron tamoxifeno (F-G) no desarrollaron tumores

Tamoxifeno como supresor de la carcinogenesis mamaria inducida por 7,12-dimetil-benzoantraceno y nitroso-metilurea, en ratas hembras con hiperprolactinemia provocada / Tamoxifen as a suppressor of mammary carcinogenesis induced by 7,12-dimetil-benzanthracene and methylnitrosourea, in female rats with induced hyperprolactinemia

El efecto del tamoxifeno sobre tumores mamarios inducidos con DMBA (7, 12-dimetil-benzoantraceno) y NMU (nitroso-metilurea) se analiza. De 7 lotes de ratas Sprague-Dayley, 4 (A-B-C-D) reciben adiconalmene sulpirida 0,5 mg/100g durante 5 días de la semana y hasta finalizar el experimento. Dos lotes (C-D) recibieron una sola inyección intraperitoneal de 900 microcuries por 100g de peso de I. Tres lotes (E-F-G) recibieron sólo I para activar secreción de prolactina, y de ellos, 2(F-G)recibieron tamoxifeno. Los tumores que se desarrollaron fueron todos del tupo adenocarcinoma papilífero. Los receptores hormonales estrogénicos fueron positivos. El estudio por microscopia electrónica mostró vacuolas acuosas grasosas en el citoplasma y la falta de organoides y mitocondrias en algunas células. Los lotes que recibieron tamoxifeno (F-G) no desarrollaron tumores (AU)