Protoplast-mediated transformation in Sporisorium scitamineum facilitates visualization of in planta developmental stages in sugarcane

2021 ◽  
Author(s):  
V. N. Agisha ◽  
N. M. R. Ashwin ◽  
R. T. Vinodhini ◽  
Kumaravel Nalayeni ◽  
Amalraj Ramesh Sundar ◽  
...  
Keyword(s):  
Developmental Stages ◽  
In Planta ◽  
Sporisorium Scitamineum

Factors affecting the infection of sweet cherry (Prunus avium) fruit by Podosphaera cerasi

Plant Disease ◽  
2021 ◽  
Author(s):  
Claudia Probst ◽  
Binod Pandey ◽  
Prashant Swamy ◽  
Gary Glenn Grove
Keyword(s):  
Powdery Mildew ◽  
Prunus Avium ◽  
Developmental Stages ◽  
Disease Incidence ◽  
The United States ◽  
In Planta ◽  
Full Bloom ◽  
Symptomatic Disease ◽  
Conidial Concentration

Powdery mildew caused by Podosphaera cerasi is the most important fungal disease of sweet cherries in the Pacific Northwest of the United States. In this study, several factors related to disease epidemiology were evaluated. The experiments were conducted to investigate flower susceptibility to P. cerasi infection by in planta and in vitro inoculation. The susceptibility of fruit at various developmental stages was investigated using defined concentrations of P. cerasi conidia. Furthermore, the threshold of conidial concentration required for fruit infection was determined. The pathogen activity during full bloom was limited and not related to fruit disease incidence and severity at harvest. Foliar infections always preceded fruit infections by an average of 42 days during the three years of the study. The onset of fruit infection followed, on average, 66 days after full bloom and appeared simultaneously on all susceptible cherry cultivars in the research orchard. Disease symptoms were only observed on fruit in Biologische Bundesanstalt, Bundessortenamt, and Chemical industry (BBCH) scale 8 (maturity) in all cultivars examined. During this stage, a concentration of 500 conidia/ ml was sufficient to cause fruit infection at harvest. Interaction between the inoculation dates and inoculum conidial concentration revealed a dependency of disease development on the host stage at the time of conidial inoculation; the younger the fruit, the more conidia are needed to cause disease at harvest. Molecular studies showed a rapid increase in conidia viability at the transition from asymptomatic to the symptomatic disease of fruit. No evidence of ontogenic resistance of fruit to powdery mildew infection was observed.

scholarly journals Extended Hopanoid Loss Reduces Bacterial Motility and Surface Attachment and Leads to Heterogeneity in Root Nodule Growth Kinetics in a Bradyrhizobium-Aeschynomene Symbiosis

2019 ◽  
Vol 32 (10) ◽  
pp. 1415-1428
Author(s):  
Brittany J. Belin ◽  
Elise M. Tookmanian ◽  
Jaime de Anda ◽  
Gerard C. L. Wong ◽  
Dianne K. Newman
Keyword(s):  
Nitrogen Fixation ◽  
Root Nodule ◽  
Developmental Stages ◽  
Growth Parameters ◽  
Phenotypic Diversity ◽  
Quantitative Model ◽  
Nodule Development ◽  
In Planta ◽  
Surface Attachment

Hopanoids are steroid-like bacterial lipids that enhance membrane rigidity and promote bacterial growth under diverse stresses. Hopanoid biosynthesis genes are conserved in nitrogen-fixing plant symbionts, and we previously found that the extended (C35) class of hopanoids in Bradyrhizobium diazoefficiens are required for efficient symbiotic nitrogen fixation in the tropical legume host Aeschynomene afraspera. Here, we demonstrate that the nitrogen-fixation defect conferred by extended hopanoid loss can be fully explained by a reduction in root nodule sizes rather than per-bacteroid nitrogen-fixation levels. Using a single-nodule tracking approach to quantify A. afraspera nodule development, we provide a quantitative model of root nodule development in this host, uncovering both the baseline growth parameters for wild-type nodules and a surprising heterogeneity of extended hopanoid mutant developmental phenotypes. These phenotypes include a delay in root nodule initiation and the presence of a subpopulation of nodules with slow growth rates and low final volumes, which are correlated with reduced motility and surface attachment in vitro and lower bacteroid densities in planta, respectively. This work provides a quantitative reference point for understanding the phenotypic diversity of ineffective symbionts in A. afraspera and identifies specific developmental stages affected by extended hopanoid loss for future mechanistic work.

scholarly journals Genome-wide identification and characterization of the GDP-L-galactose phosphorylase gene family in bread wheat

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Ronan C. Broad ◽  
Julien P. Bonneau ◽  
Jesse T. Beasley ◽  
Sally Roden ◽  
Joshua G. Philips ◽  
...  
Keyword(s):  
Transposable Element ◽  
Bread Wheat ◽  
Developmental Stages ◽  
Phylogenetic Analyses ◽  
Expression Patterns ◽  
Leaf Tissue ◽  
Upstream Open Reading Frame ◽  
Transient Transformation ◽  
In Planta ◽  
Coding Sequences

Abstract Background Ascorbate is a powerful antioxidant in plants and an essential micronutrient for humans. The GDP-L-galactose phosphorylase (GGP) gene encodes the rate-limiting enzyme of the L-galactose pathway—the dominant ascorbate biosynthetic pathway in plants—and is a promising gene candidate for increasing ascorbate in crops. In addition to transcriptional regulation, GGP production is regulated at the translational level through an upstream open reading frame (uORF) in the long 5′-untranslated region (5’UTR). The GGP genes have yet to be identified in bread wheat (Triticum aestivum L.), one of the most important food grain sources for humans. Results Bread wheat chromosomal groups 4 and 5 were found to each contain three homoeologous TaGGP genes on the A, B, and D subgenomes (TaGGP2-A/B/D and TaGGP1-A/B/D, respectively) and a highly conserved uORF was present in the long 5’UTR of all six genes. Phylogenetic analyses demonstrated that the TaGGP genes separate into two distinct groups and identified a duplication event of the GGP gene in the ancestor of the Brachypodium/Triticeae lineage. A microsynteny analysis revealed that the TaGGP1 and TaGGP2 subchromosomal regions have no shared synteny suggesting that TaGGP2 may have been duplicated via a transposable element. The two groups of TaGGP genes have distinct expression patterns with the TaGGP1 homoeologs broadly expressed across different tissues and developmental stages and the TaGGP2 homoeologs highly expressed in anthers. Transient transformation of the TaGGP coding sequences in Nicotiana benthamiana leaf tissue increased ascorbate concentrations more than five-fold, confirming their functional role in ascorbate biosynthesis in planta. Conclusions We have identified six TaGGP genes in the bread wheat genome, each with a highly conserved uORF. Phylogenetic and microsynteny analyses highlight that a transposable element may have been responsible for the duplication and specialized expression of GGP2 in anthers in the Brachypodium/Triticeae lineage. Transient transformation of the TaGGP coding sequences in N. benthamiana demonstrated their activity in planta. The six TaGGP genes and uORFs identified in this study provide a valuable genetic resource for increasing ascorbate concentrations in bread wheat.

scholarly journals Differential Activity of Carboxylic Acid Amide Fungicides Against Various Developmental Stages of Phytophthora infestans

2007 ◽  
Vol 97 (10) ◽  
pp. 1274-1283 ◽  
Author(s):  
Yigal Cohen ◽  
Ulrich Gisi
Keyword(s):  
Carboxylic Acid ◽  
Late Blight ◽  
Phytophthora Infestans ◽  
Developmental Stages ◽  
Acid Amide ◽  
Intrinsic Activity ◽  
Dependent Manner ◽  
Carboxylic Acid Amide ◽  
In Planta ◽  
Mycelium Growth

Three carboxylic acid amide (CAA) fungicides, mandipropamid (MPD), dimethomorph (DMM) and iprovalicarb (IPRO) were examined for their effects on various asexual developmental stages of Phytophthora infestans in vitro and in planta. Germination of cystospores and direct germination of sporangia were inhibited with nanomole concentrations of MPD (0.005 μg/ml) and micromole concentrations of DMM (0.05 μg/ml) or IPRO (0.5 μg/ml). A temporary exposure of 1 h to CAAs was not detrimental to germination and infectivity of sporangia or cystospores. CAAs applied to cystospores at 1 h after the onset of germination did not prevent the emergence of germ tubes, but inhibited their further growth and deformed their shape. None of the fungicides affected discharge of zoospores from sporangia or the encystment (cell wall formation/assembly) of the zoospores. Mycelium growth in solid or liquid media was inhibited with micromole concentrations. CAAs mixed with sporangia and drop inoculated onto detached leaves strongly suppressed infection. Curative application at 1 day postinoculation (dpi) required higher concentrations of CAAs than preventive application to inhibit infection and lost its effectiveness at 2 dpi. When sprayed on established late blight lesions 4 days after inoculation, CAAs reduced sporangial production in a dose-dependent manner. Trans-laminar protection of potato or tomato leaves, although achieved with higher doses, was more effective with MPD than with DMM or IPRO. Shade house studies demonstrated superior control of late blight epidemics by MPD compared with the other molecules. The data suggest that germ tube formation by cystospores or sporangia is the most sensitive stage in the life cycle of P. infestans to CAAs. Of the three CAAs, MPD had the highest intrinsic activity against spore germination. This property, together with its better trans-laminar activity, makes MPD more effective than DMM or IPRO in controlling epidemics caused by P. infestans.

scholarly journals Ovule Development and in Planta Transformation of Paphiopedilum Maudiae by Agrobacterium-Mediated Ovary-Injection

2020 ◽  
Vol 22 (1) ◽  
pp. 84
Author(s):  
Bai-Xue Luo ◽  
Li Zhang ◽  
Feng Zheng ◽  
Kun-Lin Wu ◽  
Lin Li ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Developmental Stages ◽  
Embryo Sac ◽  
Mature Embryo ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Pcr Analysis ◽  
In Planta Transformation ◽  
In Planta ◽  
Hygromycin Selection

In this paper, the development of the Paphiopedilum Maudiae embryo sac at different developmental stages after pollination was assessed by confocal laser scanning microscopy. The mature seeds of P. Maudiae consisted of an exopleura and a spherical embryo, but without an endosperm, while the inner integument cells were absorbed by the developing embryo. The P. Maudiae embryo sac exhibited an Allium type of development. The time taken for the embryo to develop to a mature sac was 45-50 days after pollination (DAP) and most mature embryo sacs had completed fertilization and formed zygotes by about 50–54 DAP. In planta transformation was achieved by injection of the ovaries by Agrobacterium, resulting in 38 protocorms or seedlings after several rounds of hygromycin selection, corresponding to 2, 7, 5, 1, 3, 4, 9, and 7 plantlets from Agrobacterium-mediated ovary-injection at 30, 35, 42, 43, 45, 48, 50, and 53 DAP, respectively. Transformation efficiency was highest at 50 DAP (2.54%), followed by 2.48% at 53 DAP and 2.45% at 48 DAP. Four randomly selected hygromycin-resistant plants were GUS-positive after PCR analysis. Semi-quantitative PCR and quantitative real-time PCR analysis revealed the expression of the hpt gene in the leaves of eight hygromycin-resistant seedlings following Agrobacterium-mediated ovary-injection at 30, 35, 42, 43, 45, 48, 50, and 53 DAP, while hpt expression was not detected in the control. The best time to inject P. Maudiae ovaries in planta with Agrobacterium is 48-53 DAP, which corresponds to the period of fertilization. This protocol represents the first genetic transformation protocol for any Paphiopedilum species and will allow for expanded molecular breeding programs to introduce useful and interesting genes that can expand its ornamental and horticulturally important characteristics.

scholarly journals Patterns of Expression of a Lolitrem Biosynthetic Gene in the Epichloë festucae–Perennial Ryegrass Symbiosis

2008 ◽  
Vol 21 (2) ◽  
pp. 188-197 ◽  
Author(s):  
Kimberley J. May ◽  
Michelle K. Bryant ◽  
Xiuwen Zhang ◽  
Barbara Ambrose ◽  
Barry Scott
Keyword(s):  
Gene Expression ◽  
Developmental Stages ◽  
Expression Patterns ◽  
In Planta ◽  
Temporal Expression ◽  
Epichloë Festucae ◽  
Gusa Reporter Gene ◽  
Reporter Gene Analysis ◽  
High Level ◽  
Germinating Seeds

Lolitrem B is synthesized by Epichloë festucae in associations with Pooid grasses. A complex cluster of at least 10 genes (ltm genes) is required for its synthesis. An early step in this pathway is catalyzed by ltmM, a symbiosis-expressed gene. PltmM-gusA reporter gene analysis was used to monitor ltmM gene expression patterns in planta. The minimum promoter length required for high-level gusA expression in infected seedlings is in the range of 480 to 782 bp. gusA was expressed by the endophyte in all infected vegetative plant tissues and in epiphyllous hyphae. Spikelets from reproductive tillers were analyzed at different developmental stages. During pre-anthesis, gusA expression was observed in all infected floral organs except the immature gynoecium. In post-anthesis florets, gene expression occurred almost exclusively in the gynoecium. Expression of gusA by the endophyte was observed in germinating seeds 24 h postimbibition and seedlings older than 6 days postimbibition in hyphae from the mesocotyl to the tip of the emerging first leaf. This work provides a detailed analysis of the spatial and temporal expression patterns of a symbiosis-expressed gene in planta.

scholarly journals Harnessing Pseudomonas protegens to Control Bacterial Panicle Blight of Rice

2020 ◽  
Vol 110 (10) ◽  
pp. 1657-1667
Author(s):  
Laura Ortega ◽  
Katherine A. Walker ◽  
Casey Patrick ◽  
Yeshi Wamishe ◽  
Alejandro Rojas ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Developmental Stages ◽  
In Planta ◽  
Burkholderia Glumae ◽  
Continuous Increase ◽  
Disease Symptoms ◽  
Seedborne Disease ◽  
Antagonistic Microorganisms ◽  
Panicle Blight

Bacterial panicle blight of rice is a seedborne disease caused by the bacterium Burkholderia glumae. This disease has affected rice production worldwide and its effects are likely to become more devastating with the continuous increase in global temperatures, especially during the growing season. The bacterium can cause disease symptoms in different tissues and at different developmental stages. In reproductive stages, the bacterium interferes with grain development in the panicles and, as a result, directly affects rice yield. Currently, there are no methods to control the disease because chemical control is not effective and completely resistant cultivars are not available. Thus, a promising approach is the use of antagonistic microorganisms. In this work, we identified one strain of Pseudomonas protegens and one strain of B. cepacia with antimicrobial activity against B. glumae in vitro and in planta. We further characterized the antimicrobial activity of P. protegens and found that this activity is associated with bacterial secretions. Cell-free secretions from P. protegens inhibited the growth of B. glumae in vitro and also prevented B. glumae from causing disease in rice. Although the specific molecules associated with these activities have not been identified, these findings suggest that the secreted fractions from P. protegens could be harnessed as biopesticides to control bacterial panicle blight of rice.

Immunolocalization of a putative cuticular collagen protein in several developmental stages of Meloidogynearenaria, Globodera pallida and G. rostochiensis

2002 ◽  
Vol 76 (1) ◽  
pp. 1-6 ◽  
Author(s):  
I.M. de O. Abrantes ◽  
R.H.C. Curtis
Keyword(s):  
Developmental Stages ◽  
Globodera Pallida ◽  
Parasitic Nematodes ◽  
Cdna Expression Library ◽  
In Planta ◽  
Cyst Nematodes ◽  
Expression Library ◽  
Second Stage ◽  
Collagen Protein ◽  
Cuticular Surface

AbstractThe monoclonal antibody IACR-CCNj.3d has previously been used to isolate a gene (gp-col-8) with strong similarity to cuticular collagen from a mixed stage Globodera pallida cDNA expression library. The antibody has also been shown to label specifically the amphidial canal of pre-parasitic second stage juveniles (J2) of several plant nematode species without any reactivity on the cuticular surface, indicating that this protein is either not present or is inaccessible on the cuticular surface. This paper investigates the cross-reactivity of Mab IACR-CCNj.3d with Meloidogyne arenaria and the localization of the putative collagen protein on the cuticular surface of parasitic stages in planta and on the cuticular surface of juveniles inside eggs. The antigen was shown to be present in all developmental stages of the two species of potato cyst nematodes and M. arenaria. The antibody bound strongly to the amphidial canal and hypodermis of pre-parasitic J2 and adult females. The antigen was present on the cuticular surface of the sausage-shaped J2 in planta and of first stage juveniles (J1) inside the eggs. The presence of collagen on the surface of the cuticle of moulting stages of plant parasitic nematodes has been observed for the first time. It is clear that this protein has a role in the construction of the cuticle of the first stage juveniles and parasitic second stage juveniles, during moulting inside the eggs and in the root tissue, respectively.

scholarly journals In silico identification, characterization and expression profile of WUSCHEL-related homeobox (WOX) gene family in Vanilla planifolia

2020 ◽  
Vol 7 (2) ◽  
pp. 206-213
Author(s):  
Thiveyarajan Victorathisayam ◽  
Madhvi Kanchan ◽  
` Himani ◽  
Thandullu R. Suriyanarayanan ◽  
Jaspreet K. Sembi ◽  
...  
Keyword(s):  
Gene Family ◽  
Expression Profile ◽  
Developmental Stages ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Alpha Helix ◽  
Vanilla Planifolia ◽  
In Planta ◽  
Specific Expression ◽  
Spatio Temporal

Vanilla planifolia is an economically important orchid, which is being commercially exploited by the food industry for the highly valued secondary metabolite vanillin. WUSCHEL-related homeobox (WOX) gene family encodes for WUSCHEL-related homeobox (WOX) transcription factors that participate in embryogenesis, organogenesis and florigenesis and in diverse plant developmental processes as well. In the present study, we analysed V. planifolia transcriptome and identified 6 WOX (VpWOX) transcripts, that encode putative WOX (VpWOX) transcription factor proteins. Domain analysis was done which indicates the presence of helix-loop-helix-turn-helix which is identifying feature of WOX gene family proteins. We executed phylogenetic clustering for the VpWOX proteins with their counterpart from the model plant Arabidopsis thaliana (AtWOX) and other closely related orchid species, Phalaenopsis equestris (PeWOX), Dendrobium catenatum (DcWOX) and Apostasia shenzhenica (AsWOX) and established their clade specific grouping. Spatio-temporal expression profile for VpWOX genes was analysed for different plant developmental stages which shows that VpWOX13 is expressing uniformly in all the developmental stages whereas, other genes have tissue specific expression. Based on gene expression patterns, we selected four VpWOX proteins and carried out secondary and tertiary structural analysis which indicates the presence of alpha helix and beta turn in the protein structure. The present study provides basic understanding of the functioning of WOX gene family in V. planifolia and paves the path for functional characterization of selected VpWOX genes in planta and in heterologous system in future for commercial utilization.

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