Morpho-anatomical characterization of mature embryo-derived callus of rice (Oryza sativa L.) suitable for transformation.

The objective of this study was to morphoanatomically characterize embryogenic rice calli during early induction of somatic embryogenesis of three Brazilian rice cultivars. Herein, we explored embryogenic units (EUs) from 2-week-old cut proliferated calli to verify whether they were suitable for Agrobacterium tumefasciens-mediated transformation. Histological analysis and scanning electron microscopy (SEM) were used to analyze these types of calli during early rice callogenesis in the cultivars BRS Primavera, BRS Bonança, and BRS Caiapó. The characteristics of the embryogenic cells were preserved in the EUs, which showed a globular, compact structure that contained tightly packed cells and thus rendered the cells suitable for transformation. The EUs of BRS Caiapó also maintained the characteristics of the nonembryogenic callus, such as an elongated morphology and a lack of cellular organization. In general, the observations of the histological sections corresponded with those of the SEM images. The histological analysis suggested that all cultivars used in these experiments have morphogenic potential. The EUs from proliferated 2-week-old cut calli maintained their embryogenic features. The EUs were subjected to Agrobacterium-mediated transformation, which exhibited a regeneration frequency of 58 % for transformed hygromycin-resistant cell lines. These results show that EUs from proliferated 2-week-old cut calli are suitable for plant transformation

Arabinogalactan-protein and pectin epitopes in relation to an extracellular matrix surface network and somatic embryogenesis and callogenesis in Trifolium nigrescens Viv

The formation of an extracellular matrix surface network (ECMSN), and associated changes in the distribution of arabinogalactan-protein and pectin epitopes, have been studied during somatic embryogenesis (SE) and callogenesis of Trifolium nigrescens Viv. Scanning electron microscopy observations revealed the occurrence of an ECMSN on the surface of cotyledonary-staged somatic embryos as well as on the peripheral, non-regenerating callus cells. The occurrence of six AGP (JIM4, JIM8, JIM13, JIM16, LM2, MAC207) and four pectin (JIM5, JIM7, LM5, LM6) epitopes was analysed during early stages of SE, in cotyledonary-staged somatic embryos and in non-embryogenic callus using monoclonal antibodies. The JIM5 low methyl-esterified homogalacturonan (HG) epitope localized to ECMSN on the callus surface but none of the epitopes studied were found to localize to ECMSN over mature somatic embryos. The LM2 AGP epitope was detected during the development of somatic embryos and was also observed in the cell walls of meristematic cells from which SE was initiated. The pectic epitopes JIM5, JIM7, LM5 and LM6 were temporally regulated during SE. The LM6 arabinan epitope, carried by side chains of rhamnogalacturonan-I (RG-I), was detected predominantly in cells of embryogenic swellings, whilst the LM5 galactan epitope of RG-I was uniformly distributed throughout the ground tissue of cotyledonary-staged embryoids but not detected at the early stages of SE. Differences in the distribution patterns of low and high methyl-esterified HG were detected: low ester HG (JIM5 epitope) was most abundant during the early steps of embryo formation and highly methyl-esterified form of HG (JIM7 epitope) became prevalent during embryoid maturation

Developmental Localization and Methylesterification of Pectin Epitopes during Somatic Embryogenesis of Banana (Musa spp. AAA)

The plant cell walls play an important role in somatic embryogenesis and plant development. Pectins are major chemical components of primary cell walls while homogalacturonan (HG) is the most abundant pectin polysaccharide. Developmental regulation of HG methyl-esterification degree is important for cell adhesion, division and expansion, and in general for proper organ and plant development.Developmental localization of pectic homogalacturonan (HG) epitopes and the (1→4)-β-D-galactan epitope of rhamnogalacturonan I (RG-I) and degree of pectin methyl-esterification (DM) were studied during somatic embryogenesis of banana (Musa spp. AAA). Histological analysis documented all major developmental stages including embryogenic cells (ECs), pre-globular, globular, pear-shaped and cotyledonary somatic embryos. Histochemical staining of extracellularly secreted pectins with ruthenium red showed the most intense staining at the surface of pre-globular, globular and pear-shaped somatic embryos. Biochemical analysis revealed developmental regulation of galacturonic acid content and DM in diverse embryogenic stages. Immunodots and immunolabeling on tissue sections revealed developmental regulation of highly methyl-esterified HG epitopes recognized by JIM7 and LM20 antibodies during somatic embryogenesis. Cell walls of pre-globular/globular and late-stage embryos contained both low methyl-esterified HG epitopes as well as partially and highly methyl-esterified ones. Extracellular matrix which covered surface of early developing embryos contained pectin epitopes recognized by 2F4, LM18, JIM5, JIM7 and LM5 antibodies. De-esterification of cell wall pectins by NaOH caused a decrease or an elimination of immunolabeling in the case of highly methyl-esterified HG epitopes. However, immunolabeling of some low methyl-esterified epitopes appeared stronger after this base treatment.These data suggest that both low- and highly-methyl-esterified HG epitopes are developmentally regulated in diverse embryogenic stages during somatic embryogenesis. This study provides new information about pectin composition, HG methyl-esterification and developmental localization of pectin epitopes during somatic embryogenesis of banana

Morpho-anatomical characterization of mature embryo-derived callus of rice (Oryza sativa L.) suitable for transformation.

The objective of this study was to morphoanatomically characterize embryogenic rice calli during early induction of somatic embryogenesis of three Brazilian rice cultivars. Herein, we explored embryogenic units (EUs) from 2-week-old cut proliferated calli to verify whether they were suitable for Agrobacterium tumefasciens-mediated transformation. Histological analysis and scanning electron microscopy (SEM) were used to analyze these types of calli during early rice callogenesis in the cultivars BRS Primavera, BRS Bonança, and BRS Caiapó. The characteristics of the embryogenic cells were preserved in the EUs, which showed a globular, compact structure that contained tightly packed cells and thus rendered the cells suitable for transformation. The EUs of BRS Caiapó also maintained the characteristics of the nonembryogenic callus, such as an elongated morphology and a lack of cellular organization. In general, the observations of the histological sections corresponded with those of the SEM images. The histological analysis suggested that all cultivars used in these experiments have morphogenic potential. The EUs from proliferated 2-week-old cut calli maintained their embryogenic features. The EUs were subjected to Agrobacterium-mediated transformation, which exhibited a regeneration frequency of 58 % for transformed hygromycin-resistant cell lines. These results show that EUs from proliferated 2-week-old cut calli are suitable for plant transformation.Made available in DSpace on 2018-07-18T00:55:51Z (GMT). No. of bitstreams: 1 Bevitori2014ArticleMorphoanatomicalCharacterizat.pdf: 1742850 bytes, checksum: 100de21e0b29da2830c09f54e6f5a76a (MD5) Previous issue date: 2014-01-08bitstream/item/179857/1/Bevitori2014-Article-Morpho-anatomicalCharacterizat.pd