Molecular Characterization and Transmission Study of the Begomovirus on Hollyhock Plant Causing Leaf Curl Disease for the First Time in Province Punjab, Pakistan

I. Background: Hollyhock (Alcea rosea) is an ornamental plant belonging to the Malvaceae family and has a remarkable aesthetic and medicinal value. Previously in Pakistan, the hollyhock plant was not found to be infected by begomovirus and the plant first time showed the symptoms of typical leaf curling, puckering as well as thickened veins. II. Methods and Results: During the year 2018, symptomatic samples of the hollyhock plants were collected that exhibited characteristic typical leaf curling, puckering as well as thickened veins. DNA was extracted from the samples and the PCR technique was optimized for the detection of begomovirus followed by sequencing. The samples were detected to be infected with begomovirus by using Av/Ac core, Begomo 01/02, and CLCV 01/02 primer showed positive results with 579bp, 2.8kb, and 1.1kb nucleotide respectively. The betasatellite was amplified by using beta01/02 and CLCuMuBF11/R33 showed positive results with 1400bp and 481bp respectively. Sequencing results showed that diseased hollyhock plants were associated with Cotton leaf curl Multan virus-Rajasthan strain along with Cotton leaf curl Multan betasatellite. III. Conclusion: Hollyhock plants infected by begomovirus has been reported for the first time as a possible source of virus inoculum from Pakistan.

Roles of two distinct alphasatellites modulating geminivirus pathogenesis

Background Alphasatellites are small coding DNA satellites frequently associated with a begomovirus/betasatellite complex, where they are known to modulate virulence and symptom development. Two distinct alphasatellites, namely, Cotton leaf curl Multan alphasatellite (CLCuMuA), and Gossypium darwinii symptomless alphasatellite (GDarSLA) associated with Cotton leaf curl Multan virus-India (CLCuMuV-IN) and Ludwigia leaf distortion betasatellite (LuLDB) were found to be associated with yellow mosaic disease of hollyhock (Alcea rosea) plants. In this study, we show that alphasatellites CLCuMuA and GDarSLA attenuate and delay symptom development in Nicotiana benthamiana. The presence of either alphasatellites reduce the accumulation of the helper virus CLCuMuV-IN. However, the levels of the associated betasatellite, LuLDB, remains unchanged. These results suggest that the alphasatellites could contribute to the host defence and understanding their role in disease development is important for developing resistance strategies. Methods Tandem repeat constructs of two distinct alphasatellites, namely, CLCuMuA and GDarSLA associated with CLCuMuV-IN and LuLDB were generated. N. benthamiana plants were co-agroinoculated with CLCuMuV and its associated alphasatellites and betasatellite molecules and samples were collected at 7, 14 and 21 days post inoculation (dpi). The viral DNA molecules were quantified in N. benthamiana plants by qPCR. The sequences were analysed using the MEGA-X tool, and a phylogenetic tree was generated. Genetic diversity among the CLCuMuA and GDarSLA was analysed using the DnaSP tool. Results We observed a reduction in symptom severity and accumulation of helper virus in the presence of two alphasatellites isolated from naturally infected hollyhock plants. However, no reduction in the accumulation of betasatellite was observed. The phylogenetic and genetic variability study revealed the evolutionary dynamics of these distinct alphasatellites , which could explain the role of hollyhock-associated alphasatellites in plants. Conclusions This study provides evidence that alphasatellites have a role in symptom modulation and suppress helper virus replication without any discernible effect on the replication of the associated betasatellite.

Nanoliposome-Encapsulated Phenolic Rich Fraction From Alcea Rosea as a Dietary Phytobiotic in Mice Challenged by Escherichia Coli

Purpose This research was performed to evaluate the antibacterial and health-promoting potentials of the nanoliposome-encapsulated phenolic rich fraction (PRF) from Alcea rosea leaves as a dietary phytobiotic in mice challenged by enteropathogenic Escherichia coli (E. coil; O157: H7). Method: The PEF was encapsulated in nanoliposomes (PEF-NLs) and the phenolic profiling of PEF-NLs was confirmed by HPLC. Then 40 white male balb/c mice at four treatment groups were provided and antibacterial potential of PEF-NLs were assessed by measuring the mice blood parameters and liver lipid peroxidation in mice infected by E. coli. Finally, the expression of cyclooxygenase 2 (COX2), inducible nitric oxide synthase (iNOS), superoxide dismutase (SOD) and glutathione peroxidase (GPx) were determined in miceʼs ileum tissues. Meanwhile, relative fold changes in the ileum population of E. coli was analyzed using Real time PCR. Results The overall results demonstrated that the nanoliposome-loaded PRF contained gallic acid, salicylic acid, pyrogallol, cinnamic acid, catechin, naringin, ferulic acid. The E. coil challenge in mice impaired the weight gain, food intake, liver enzymes, lipid peroxidation, morphometric characteristics of the ileum, up-regulated the inflammatory genes (COX2, iNOS), down-regulated the antioxidant-related genes (SOD and GPx) and increased the population of E. coil in the ileum. The dietary inclusion of nonencapsulated PRF and nanoliposome-encapsulated PRF at the concentration of 10 mg TPC/kg BW/day improved these parameters however the nanoliposome-encapsulated PRF appeared to be more effective as compared to nonencapsulated PRF in improving the health parameters in mice. Conclusion Consequently, the nanoliposome-encapsulated PRF could play a critical role as a promising phytobiotic against E. coil infection in mice.

Microsporogénesis y micromorfología del polen de la planta Alcea rosea (Malvaceae)

Introducción: Los estudios sobre microsporogénesis, micromorfología y estructura de los granos de polen en Malvaceae son escasos. Objetivos: Describir el proceso de microsporogénesis y aspectos micromorfológicos de los granos de polen en A. rosea. Métodos: Se procesaron más de 30 andróforos de acuerdo con los protocolos estándar para incrustar y seccionar en parafina. Las secciones obtenidas se tiñeron con azul de Safranina-Alcian, las anteras inmaduras y no fijadas se tiñeron con azul de anilina. Se procesaron secciones de resina adicionales de los andróforos y se tiñeron con azul de toluidina. Se observaron secciones ultrafinas con microscopía electrónica de transmisión (TEM). Para la observación con microscopía electrónica de barrido (SEM), el material se fijó y deshidrató en 2,2 dimetoxipropano, luego se secó hasta un punto crítico y se recubrieron con oro. Resultados: las anteras se diferencian de una masa celular en los extremos distales de los filamentos del estambre. La pared de la antera madura presenta una capa externa de células epidérmicas y una capa interna, el endotecio. Las células madre de microesporas se dividen por mitosis y luego experimentan meiosis para formar tétradas. El tapete es inicialmente celular y forma una sola capa de células y luego pierde integridad celular al invadir el lóculo de microsporangio, formando un periplasmodio. Durante la formación de la esporodermis, primero se deposita la exina y luego la intina. Para el momento de la liberación de los granos de polen, el tapete se ha degenerado por completo. Los granos de polen son pantoporados, apolares, con simetría radial, esferoidales, con espinas, bacula, gránulos y microgránulos. El tectum está perforado con foveolea dispuesta homogéneamente en toda la superficie y con polenkit. La exina es ancha (5-6 µm) y consta de una endexina gruesa de 3.5 a 4 µm y una ektexina fina (0.6-0.7 µm). La ultraestructura muestra columelas claramente definidas formando el infratectum. Se aprecian tricomas nectaríferos unicelulares glandulares capitados (TG) cubriendo toda la superficie de los filamentos de los estambres. Conclusiones: La estructura y desarrollo de las anteras sigue los patrones conocidos de las angiospermas. La microsporogénesis simultánea y el depósito centrípeto de la esporodermis se han descrito previamente para Malvaceae.

Nanoliposome-encapsulated Phenolic Rich Fraction From Alcea Rosea as a Dietary Phytobiotic in Mice Challenged by Escherichia Coli

Background: This research was performed to evaluate the antibacterial and health-promoting potentials of the nanoliposome-encapsulated phenolic rich fraction (PRF) from Alcea rosea leaves as a dietary phytobiotic in mice challenged by enteropathogenic Escherichia coli (E. coil; O157: H7). Results: The overall results demonstrated that the nanoliposome-loaded PRF contained gallic acid, salicylic acid, pyrogallol, cinnamic acid, catechin, naringin, ferulic acid. The E. coil challenge in mice impaired the weight gain, food intake, liver enzymes, lipid peroxidation, morphometric characteristics of the ileum, up-regulated the inflammatory genes (COX2, iNOS), down-regulated the antioxidant-related genes (SOD and GPx) and increased the population of E. coil in the ileum. The dietary inclusion of nonencapsulated PRF and nanoliposome-encapsulated PRF at the concentration of 10 mg TPC/kg BW/day improved these parameters however the nanoliposome-encapsulated PRF appeared to be more effective as compared to nonencapsulated PRF in improving the health parameters in mice. Conclusion: Consequently, the nanoliposome-encapsulated PRF could play a critical role as a promising phytobiotic against E. coil infection in mice.

Estudio de especies no leñosas de la provincia de Loja (Ecuador) como potenciales materias primas para la fabricación de papel artesanal

Quince especies no leñosas de Loja, Ecuador: Linum usitatissimun L., Sida rhombifolia L., Sida poeppigiana (K.Schum.) Fryxell., Hibiscus rosa-sinensis L., Alcea rosea L., Lavatera arborea L., Cortaderia jubata Stapf., Andropogon gayanus Kunth., Guadua angustifolia Kunth., Saccharum sinense Roxb., Arundo donax L., Yucca guatemalensis Baker., Agave americana L., Agave americana var. marginata Trel in L.H.Bailey., y Saccharum officinarum L., fueron analizadas para estudiar el potencial de las fibras extraídas en manufactura de papel. Se realizó la selección de materia prima, extracción mecánica de fibras, separación y caracterización de sus componentes utilizando las normas TAPPI. Con las fibras se elaboró papel artesanal, usando el método alcalino de cocción, preparación de la pulpa, formación y secado. El control de calidad en el papel final, se ejecutó mediante determinación del número de Kappa, gramaje, pH, resistencia a la explosión y humedad del papel. El rendimiento de las especies en cuanto a obtención de fibra, varía de 4% en Agave americana L., a 42% en Guadua angustifolia Kunth., con los siguientes intervalos en el análisis químico: celulosa total 16% a 59%; -celulosa 67% a 84% -celulosa 3% a 19%; y -celulosa 1% a 28%. El análisis multicriterio indica que las especies de Hibiscus rosa-sinensis L., Saccharum officinarum L., Arundo donax L., Agave americana var. marginata Trol., y Saccharum sinense Roxb., representan las mejores alternativas para fabricación de papel.