Xylella fastidiosa : An in vivo system to study possible survival strategies within citrus xylem vessels based on global gene expression analysis

Abstract Xylella fastidiosa inhabits the plant xylem, a nutrient-poor environment, so that mechanisms to sense and respond to adverse environmental conditions are extremely important for bacterial survival in the plant host. Although the complete genome sequences of different Xylella strains have been determined, little is known about stress responses and gene regulation in these organisms. In this work, a DNA microarray was constructed containing 2,600 ORFs identified in the genome sequencing project of Xylella fastidiosa 9a5c strain, and used to check global gene expression differences in the bacteria when it is infecting a symptomatic and a tolerant citrus tree. Different patterns of expression were found in each variety, suggesting that bacteria are responding differentially according to each plant xylem environment. The global gene expression profile was determined and several genes related to bacterial survival in stressed conditions were found to be differentially expressed between varieties, suggesting the involvement of different strategies for adaptation to the environment. The expression pattern of some genes related to the heat shock response, toxin and detoxification processes, adaptation to atypical conditions, repair systems as well as some regulatory genes are discussed in this paper. DNA microarray proved to be a powerful technique for global transcriptome analyses. This is one of the first studies of Xylella fastidiosa gene expression in vivo which helped to increase insight into stress responses and possible bacterial survival mechanisms in the nutrient-poor environment of xylem vessels

Recurrent de novo SPTLC2 variant causes childhood-onset amyotrophic lateral sclerosis (ALS) by excess sphingolipid synthesis

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease of the upper and lower motor neurons with varying ages of onset, progression and pathomechanisms. Monogenic childhood-onset ALS, although rare, forms an important subgroup of ALS. We recently reported specific SPTLC1 variants resulting in sphingolipid overproduction as a cause for juvenile ALS. Here, we report six patients from six independent families with a recurrent, de novo, heterozygous variant in SPTLC2 c.778G>A [p.Glu260Lys] manifesting with juvenile ALS. METHODS: Clinical examination of the patients along with ancillary and genetic testing, followed by biochemical investigation of patients' blood and fibroblasts, was performed. RESULTS: All patients presented with early-childhood-onset progressive weakness, with signs and symptoms of upper and lower motor neuron degeneration in multiple myotomes, without sensory neuropathy. These findings were supported on ancillary testing including nerve conduction studies and electromyography, muscle biopsies and muscle ultrasound studies. Biochemical investigations in plasma and fibroblasts showed elevated levels of ceramides and unrestrained de novo sphingolipid synthesis. Our studies indicate that SPTLC2 variant [c.778G>A, p.Glu260Lys] acts distinctly from hereditary sensory and autonomic neuropathy (HSAN)-causing SPTLC2 variants by causing excess canonical sphingolipid biosynthesis, similar to the recently reported SPTLC1 ALS associated pathogenic variants. Our studies also indicate that serine supplementation, which is a therapeutic in SPTLC1 and SPTCL2-associated HSAN, is expected to exacerbate the excess sphingolipid synthesis in serine palmitoyltransferase (SPT)-associated ALS. CONCLUSIONS: SPTLC2 is the second SPT-associated gene that underlies monogenic, juvenile ALS and further establishes alterations of sphingolipid metabolism in motor neuron disease pathogenesis. Our findings also have important therapeutic implications: serine supplementation must be avoided in SPT-associated ALS, as it is expected to drive pathogenesis further

Metarhizium brunneum Blastospore Pathogenesis in Aedes aegypti Larvae: Attack on Several Fronts Accelerates Mortality

Aedes aegypti is the vector of a wide range of diseases (e.g. yellow fever, dengue, Chikungunya and Zika) which impact on over half the world's population. Entomopathogenic fungi such as Metarhizium anisopliae and Beauveria bassiana have been found to be highly efficacious in killing mosquito larvae but only now are the underlying mechanisms for pathogenesis being elucidated. Recently it was shown that conidia of M. anisopliae caused stress induced mortality in Ae. aegypti larvae, a different mode of pathogenicity to that normally seen in terrestrial hosts. Blastospores constitute a different form of inoculum produced by this fungus when cultured in liquid media and although blastospores are generally considered to be more virulent than conidia no evidence has been presented to explain why. In our study, using a range of biochemical, molecular and microscopy methods, the infection process of Metarhizium brunneum (formerly M. anisopliae) ARSEF 4556 blastospores was investigated. It appears that the blastospores, unlike conidia, readily adhere to and penetrate mosquito larval cuticle. The blastospores are readily ingested by the larvae but unlike the conidia are able infect the insect through the gut and rapidly invade the haemocoel. The fact that pathogenicity related genes were upregulated in blastospores exposed to larvae prior to invasion, suggests the fungus was detecting host derived cues. Similarly, immune and defence genes were upregulated in the host prior to infection suggesting mosquitoes were also able to detect pathogen-derived cues. The hydrophilic blastospores produce copious mucilage, which probably facilitates adhesion to the host but do not appear to depend on production of Pr1, a cuticle degrading subtilisin protease, for penetration since protease inhibitors did not significantly alter blastospore virulence. The fact the blastospores have multiple routes of entry (cuticle and gut) may explain why this form of the inoculum killed Ae. aegypti larvae in a relatively short time (12-24hrs), significantly quicker than when larvae were exposed to conidia. This study shows that selecting the appropriate form of inoculum is important for efficacious control of disease vectors such as Ae. aegypti

The Southern Ocean Exchange: Porous boundaries between humpback whale breeding populations in southern polar waters

Humpback whales (Megaptera novaeangliae) are a cosmopolitan species and perform long annual migrations between low-latitude breeding areas and high-latitude feeding areas. Their breeding populations appear to be spatially and genetically segregated due to long-term, maternally inherited fidelity to natal breeding areas. In the Southern Hemisphere, some humpback whale breeding populations mix in Southern Ocean waters in summer, but very little movement between Pacific and Atlantic waters has been identified to date, suggesting these waters constituted an oceanic boundary between genetically distinct populations. Here, we present new evidence of summer co-occurrence in the West Antarctic Peninsula feeding area of two recovering humpback whale breeding populations from the Atlantic (Brazil) and Pacific (Central and South America). As humpback whale populations recover, observations like this point to the need to revise our perceptions of boundaries between stocks, particularly on high latitude feeding grounds. We suggest that this “Southern Ocean Exchange” may become more frequent as populations recover from commercial whaling and climate change modifies environmental dynamics and humpback whale prey availability

Draft genome sequence of Bradyrhizobium elkanii TnphoA 33, a producer of polyhydroxyalkanoates

The genus Bradyrhizobium comprises bacteria with the ability to form nitrogen-fixing symbioses with legumes. They are of great interest in agriculture, as well as for the production of biopolymers such as polyhydroxyalkanoates. Here, we report the draft genome assembly of Bradyrhizobium elkanii TnphoA 33 comprising 9 Mb, 1,124 contigs, and 9,418 open reading frames

Xylella fastidiosa: An in vivo system to study possible survival strategies within citrus xylem vessels based on global gene expression analysis

Xylella fastidiosa inhabits the plant xylem, a nutrient-poor environment, so that mechanisms to sense and respond to adverse environmental conditions are extremely important for bacterial survival in the plant host. Although the complete genome sequences of different Xylella strains have been determined, little is known about stress responses and gene regulation in these organisms. In this work, a DNA microarray was constructed containing 2,600 ORFs identified in the genome sequencing project of Xylella fastidiosa 9a5c strain, and used to check global gene expression differences in the bacteria when it is infecting a symptomatic and a tolerant citrus tree. Different patterns of expression were found in each variety, suggesting that bacteria are responding differentially according to each plant xylem environment. The global gene expression profile was determined and several genes related to bacterial survival in stressed conditions were found to be differentially expressed between varieties, suggesting the involvement of different strategies for adaptation to the environment. The expression pattern of some genes related to the heat shock response, toxin and detoxification processes, adaptation to atypical conditions, repair systems as well as some regulatory genes are discussed in this paper. DNA microarray proved to be a powerful technique for global transcriptome analyses. This is one of the first studies of Xylella fastidiosa gene expression in vivo which helped to increase insight into stress responses and possible bacterial survival mechanisms in the nutrient-poor environment of xylem vessels.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Efeitos de herbicidas no consórcio de milho com Brachiaria brizantha Effects of herbicides on corn and Brachiaria brizantha intercropping

A eficiência de herbicidas aplicados em pós-emergência foi avaliada para o controle de plantas daninhas no consórcio de milho com Brachiaria brizantha e verificando seus efeitos sobre as espécies consorciadas. Os tratamentos foram representados pelos herbicidas atrazine (1.500 g ha-1), foramsulfuron + iodosulfuron methyl sodium + atrazine (15 + 1 + 1.500 e 30 + 2 + 1.500 g ha-1), foramsulfuron + iodosulfuron methyl sodium (45 + 3 g ha¹), nicosulfuron + atrazine (2 + 1.500; 4 + 1.500; 8 + 1.500; 12 + 1.500 g ha-1) mais a testemunha capinada (milho solteiro) e sem capina, arranjados em blocos ao acaso, com três repetições. Foram avaliados a porcentagem de controle de Digitaria horizontalis e de espécies do gênero Ipomoea, a toxicidade à cultura do milho, a cobertura vegetal do solo, o estado nutricional e a produtividade do milho e da forrageira. Verificou-se que para D. horizontalis os herbicidas foramsulfuron + iodosulfuron methyl sodium + atrazine, foramsulfuron + iodosulfuron methyl sodium + espalhante e nicosulfuron + atrazine nas duas maiores doses proporcionaram controle semelhante (acima de 70%) aos 28 dias após a aplicação (DAA). Para Ipomoea spp. houve diferença dos tratamentos apenas em relação à testemunha sem capina. Foramsulfuron + iodosulfuron methyl sodium + espalhante mostrou-se tóxico ao milho até 28 DAA, enquanto para B. brizantha somente o atrazine não causou dano. Leituras correspondentes ao teor de clorofila total, N, K e a produtividade do milho foram inferiores na testemunha sem capina e no tratamento com atrazine, sendo os demais não influenciados pelos herbicidas. Ao contrário, no período de convivência com o milho, depois da colheita deste e após período de pastejo, B. brizantha foi mais produtiva nos tratamentos em que foi aplicado atrazine e na testemunha sem capina, apresentando menor desenvolvimento nas maiores doses das sulfoniluréias.<br>The objective of this study was to evaluate the efficiency of herbicides applied in post-emergence on weeds and corn - Brachiaria brizantha intercropping. The treatments were applications of atrazine (1.500 g ha-1), foramsulfuron + iodosulfuron methyl sodium + atrazine (15 + 1 + 1.500 and 30 + 2 + 1.500 g ha-1) foramsulfuron + iodosulfuron methyl sodium (45 + 3 g ha-1), nicosulfuron + atrazine (2 + 1.500; 4 + 1.500; 8 + 1.500; 12 + 1.500 g ha-1), besides the weeded corn and intercropping without weeding, arranged in randomized blocks, with three replications. Digitaria horizontalis and Ipomoea spp. control percentage, maize crop toxicity degree, soil plant cover, nutritional state, and maize and forage yields were evaluated. It was verified that the herbicides foramsulfuron + iodosulfuron methyl sodium + atrazine, foramsulfuron + iodosulfuron methyl sodium + sulfactant, and nicosulfuron + atrazine applied at the two highest doses provided a similar control percentage (above 70%) at 28 days after application (DAA) of herbicides for D. horizontalis. For Ipomoea spp., a difference in the treatments was found only in relation to the control without weeding. Foramsulfuron + iodosulfuron methyl sodium + sulfactant proved toxic for corn up to 28 DAA, while only atrazine proved not harmful for B. brizantha. Readings corresponding to total chlorophyll, N, and K contents and corn yield were lower in the control without weeding and atrazine treatment. The others were not influenced by the use of the herbicides. On the other hand, B. brizantha was more productive in the treatments and control without weeding, presenting a reduced development under the highest sulfonylurea doses during the intercropping with corn, after harvest and pasture

Controle de plantas daninhas na cultura do milho-pipoca com herbicidas aplicados em pós-emergência Weed control in popcorn maize using post emergence herbicide

Objetivou-se avaliar a eficiência de controle de plantas daninhas, a tolerância e a produção de milho-pipoca UFVM2 aos herbicidas atrazine (1.500 g ha-1), foramsulfuron + iodosulfuron methyl sodium + atrazine (15+1+1.500 e 30+2+1.500 g ha-1), foramsulfuron + iodosulfuron methyl sodium (15+1, 30+2, 45+3 e 60+4 g ha-1), nicosulfuron + atrazine (8+1.500 e 16+1.500 g ha-1) e nicosulfuron (16 g ha-1). A aplicação dos herbicidas foi realizada aos 25 dias após a emergência da cultura. As plantas daninhas predominantes na área foram Brachiaria plantaginea, Brachiaria decumbens e Ipomoea spp. Os herbicidas foramsulfuron + iodosulfuron methyl sodium, em todas as doses testadas, proporcionaram maiores índices de toxidez à cultura, aos 7, 14 e 28 dias após a aplicação dos tratamentos (DAA). Todas as combinações de herbicidas proporcionaram controle acima de 90% para B. plantaginea e B. decumbens, aos 28 DAA. O nicosulfuron aplicado isoladamente proporcionou controle de apenas 80% e o atrazine não controlou essas espécies daninhas. Para Ipomoea spp., nenhum tratamento herbicida proporcionou controle equivalente ao da testemunha capinada. A competição das plantas daninhas resultou em menores teores de clorofila total, N, P e K no tecido foliar das plantas de milho-pipoca e menor rendimento de grãos. Para o cultivar UFVM2, as sulfoniluréias isoladas ou associadas ao atrazine não afetaram o estado nutricional da cultura, o rendimento e a capacidade de expansão dos grãos de milho-pipoca.<br>The efficiency of different herbicide combinations for weed control and tolerance and yield of UFVM2 popcorn maize were evaluated. The herbicide treatments and respective doses were: atrazine (1.500 g ha-1); foramsulfuron + iodosulfuron methyl sodium + atrazine (15+1+1.500 and 30+2+1.500 g ha-1); foramsulfuron + iodosulfuron methyl sodium (15+1, 30+2, 45+3, and 60+4 g ha-1); nicosulfuron + atrazine (8+1.500 and 16+1.500 g ha-1) and nicosulfuron (16 g ha-1). The herbicides were applied 25 days after crop emergence. The predominant weeds in the area were Brachiaria plantaginea, Brachiaria decumbens, and Ipomoea spp. The herbicides foramsulfuron + iodosulfuron methyl sodium in all doses provided the highest rates of toxicity in popcorn maize 7, 14, and 28 days after application (DAA). All herbicide combinations controlled over 90% of B. plantaginea and B. decumbens up to the 28 DAA. Nicosulfuron controlled only 80% with no atrazine control being observed. The competition with the weeds resulted in lower contents of total chlorophyll, N, P, and K in the leaf tissue of the popcorn maize plants and smaller grain yield. For the UFVM2 cultivar, the sulfonylureias isolated or associated to atrazine had no effect on the nutritional state of the culture, yield and kernel expansion volume of the maize-popcorn