Melatonin induces proline, secondary metabolites, sugars and antioxidants activity to regulate oxidative stress and ROS scavenging in salt stressed sword lily.

Sword lily is regarded as a useful and commercially demanding cut flower crop; hence, assessing its responses to abiotic stress, particularly salt stress, is vital. Melatonin (MT) exhibits stress tolerance in crop plants and is an emerging stress relieving alternative to chemicals. Nevertheless, the possible process underlying the effects of MT under salt stress has yet to be fully elucidated in plants. Herein, the salt stress (SS) mitigation potential of MT was assessed in a commercially important cut flower, sword lily. Melatonin, expressed as MT1, MT2, MT3, and MT4, was administered at concentrations of 0.2, 0.4, 0.6, and 0.8 mM. The results revealed that SS (5 dS m-1) restricted the growth and physiological aspects of sword lily. Furthermore, malondialdehyde (MDA), hydrogen peroxide (H2O2), membrane permeability, endogenous proline, and soluble protein contents were enhanced in SS. MT application improved morphological traits, photosynthetic pigments, and corm traits. The application of MT mitigated the effects of SS stress in Gladiolus grandiflorus plants by improving growth and photosynthetic pigments. MT application under SS improved the reducing and non-reducing sugar and NPK contents of the sword lily. Furthermore, MT improved the levels of secondary metabolites, such as anthocyanins, flavonoids, and ascorbic acid, in sword lily. Moreover, MT supplementation ameliorated salt-induced oxidative stress in the gladiolus, as depicted by a decrease in stress markers (EL, MDA, and H2O2) and an increase in defense-related enzymes (POD, CAT, and SOD) with highest increase in the MT3 treatment under salinity stress. The SOD and CAT enzyme activities were 3-3.6-fold higher in the MT3 under stress than the control. In conclusion, MT applications on cut flowers can be an effective strategy to reduce salt stress and can be used to regulate salinity stress in cut flower production. MT can be used as a safe alternative to other agrochemicals to maintain the growth and flower quality of sword lilies, with beneficial effects during vase life.

Resistance to Frankliniella occidentalis during Different Plant Life Stages and under Different Environmental Conditions in the Ornamental Gladiolus.

The defense mechanisms of plants evolve as they develop. Previous research has identified chemical defenses against Western flower thrips (WFT) in Gladiolus (Gladiolus hybridus L.). Consequently, our study aimed to explore the consistency of these defense variations against WFT across the various developmental stages of Gladiolus grown under different conditions. Thrips bioassays were conducted on whole plants at three developmental stages, using the Charming Beauty and Robinetta varieties as examples of susceptible and resistant varieties, respectively. Metabolomic profiles of the leaves, buds and flowers before thrips infestation were analyzed. The thrips damage in Charming Beauty was more than 500-fold higher than the damage in Robinetta at all plant development stages. Relative concentrations of triterpenoid saponins and amino acids that were associated with resistance were higher in Robinetta at all plant stages. In Charming Beauty, the leaves exhibited greater damage compared to buds and flowers. The relative concentrations of alanine, valine and threonine were higher in buds and flowers than in leaves. The Metabolomic profiles of the leaves did not change significantly during plant development. In addition, we cultivated plants under different environmental conditions, ensuring consistency in the performance of the two varieties across different growing conditions. In conclusion, the chemical thrips resistance markers, based on the analysis of vegetative plants grown in climate rooms, were consistent over the plant's lifetime and for plants grown under field conditions.

Laser-induced changes in the gene expression, growth and development of Gladiolus grandiflorus cv. "White Prosperity".

Corms of Gladiolus grandiflorus cv. "White Prosperity" was irradiated via red laser at wavelength 635 nm. Various morphological, flowering, elemental and chemical characterizations were studied. Irradiation with different power (5, 20, and 50 mW) and various irradiation time (0.0, 0.5, 1, 3, 5 and 10 min) was studied. Several characters), totaletermined include vegetative growth parameter (spouting days, plant height (cm), leaves number, leaves fresh and dry weights (g/plant), diameter of plant middle part (mm) and leaf area (cm2), floral parameters (flowering days, vase life (day), fresh and dry weights of inflorescence (g/plant), number of flowers per inflorescence, inflorescence length(cm), flowers diameter(cm), number of corms per plant, corms fresh weight(g/plant), circumference/ corms), pigments [total chlorophylls in leaves (SPAD), anthocyanin content (mg/100 g F.W.) in petals], NPK (%) in new corms and chemical composition in corms; total carbohydrates (%),total phenol (µg CE/g (%),total flavonoid (µg CE/g) (%), antioxidant (DPPH IC50 (µg /ml (%), and proline content (µ moles/g). The results showed that the medium level (20 mW) of He-Ne laser at 5 min caused favorable changes in the leaf anatomical structures and other studied characters followed by the low level (5 mW) of He-Ne laser at 5min. 112 bands emerged from 22 SSR primers, ranging between 130 and 540 bp, with 32 bands having polymorphism ranging from 17-100%. Out of the 22 SSR primers, 3 primers exhibited a high polymorphism percentage, i.e., SSR6, SSR16 and SSR22 which exhibited 7 positive markers. These findings revealed the efficiency of SSR primers for differentiating gladiolus plants and revealed that some alleles were affected by laser in their corms and the expression resulted in color or abnormalities in leaves and/or flowers. Mutation in some alleles could result in abnormalities like mutation in the allele with 410 bp revealed by SSR16.

Characterization of a novel high-efficiency cracking Burkholderia gladiolus phage vB_BglM_WTB and its application in black fungus.

Burkholderia gladiolus (B. gladiolus) is foodborne pathogenic bacteria producing bongkrekic acid (BA), which causes food poisoning and has a mortality rate of up to 40 % or more. However, no drugs have been reported in the literature for the prevention and treatment of this infection. In this study, a phage was identified to control B. gladiolus. The novel phage vB_BglM_WTB (WTB), which lyse B. gladiolus with high efficiency, was isolated from sewage of Huaihe Road Throttle Well Sewage Treatment Plant in Hefei. Transmission electron microscopy showed that WTB had an icosahedral head (69 ± 2 nm) and a long retractable tail (108 ± 2 nm). Its optimal temperature and pH ranges to control B. gladiolus were 25 °C -65 °C and 3-11 respectively. The phage WTB was identified as a linear double-stranded DNA phage of 68, 541 bp with 60.04 % G + C content, with a long latent period of 60 min. Phylogenetic analysis and comparative genetic analysis indicated that phage WTB has low identity (<50 %) with other phages, with the highest similarity to Burkholderia phage Maja (25.7 %), which showed that it does not belong to any previous genera recognized by the International Committee on Taxonomy of Viruses (ICTV) and was a candidate for a new genus within the Caudoviricetes. We have submitted a new proposal to ICTV to create a new genus, Bglawtbvirus. No transfer RNA (tRNA), virulence associated and antibiotic resistance genes were detected in phage WTB. Experimental results indicated that WTB at 4 °C and 25 °C had excellent inhibition activity against B. gladiolus in the black fungus, with an inhibition efficiency of over 99 %. The amount of B. gladiolus in the black fungus was reduced to a minimum of 89 CFU/mL when treated by WTB at 25 °C for 2 h. The inhibition rate remained at 99.97 % even after 12 h. The findings showed that the phage WTB could be applied as a food-cleaning agent for enhancing food safety and contributed to our understanding of phage biology and diversity.

Garlic Bulb Decay and Soft Rot Caused by the Cross-Kingdom Pathogen Burkholderia gladioli.

In 2021, two gram-negative bacterial strains were isolated from garlic (Allium sativum) bulbs showing decay and soft rot symptoms in Central Iran. The bacterial strains were aggressively pathogenic on cactus, garlic, gladiolus, onion, potato, and saffron plants and induced soft rot symptoms on carrot, cucumber, potato, and radish discs. Furthermore, they were pathogenic on sporophores of cultivated and wild mushrooms. Phylogenetic analyses revealed that the bacterial strains belong to Burkholderia gladioli. Garlic bulb rot caused by B. gladioli has rarely been reported in the literature. Historically, B. gladioli strains had been assigned to four pathovars, namely, B. gladioli pv. alliicola, B. gladioli pv. gladioli, B. gladioli pv. agaricicola, and B. gladioli pv. cocovenenans, infecting onion, Gladiolus sp., and mushrooms and poisoning foods, respectively. Multilocus (i.e., 16S rRNA, atpD, gyrB, and lepA genes) sequence-based phylogenetic investigations including reference strains of B. gladioli pathovars showed that the two garlic strains belong to phylogenomic clade 2 of the species, which includes the pathotype strain of B. gladioli pv. alliicola. Although the garlic strains were phylogenetically closely related to the B. gladioli pv. alliicola reference strains, they possessed pathogenicity characteristics that overlapped with three of the four historical pathovars, including the ability to rot onion (pv. alliicola), gladiolus (pv. gladioli), and mushrooms (pv. agaricicola). Furthermore, the pathotype of each pathovar could infect the hosts of other pathovars, undermining the utility of the pathovar concept in this species. Overall, using phenotypic pathovar-oriented assays to classify B. gladioli strains should be replaced by phylogenetic or phylogenomic analysis.

Cross species/genera transferability of simple sequence repeat markers, genetic diversity and population structure analysis in gladiolus (Gladiolus × grandiflorus L.) genotypes.

Background: Genetic analysis of gladiolus germplasm using simple sequence repeat (SSR) markers is largely missing due to scarce genomic information. Hence, microsatellites identified for related genera or species may be utilized to understand the genetic diversity and assess genetic relationships among cultivated gladiolus varieties. Methods: In the present investigation, we screened 26 genomic SSRs (Gladiolus palustris, Crocus sativus, Herbertia zebrina, Sysirinchium micranthum), 14 chloroplast SSRs (Gladiolus spp., chloroplast DNA regions) and 25 Iris Expressed Sequence Tags (ESTs) derived SSRs across the 84 gladiolus (Gladiolus × grandiflorus L.) genotypes. Polymorphic markers detected from amplified SSRs were used to calculate genetic diversity estimates, analyze population structure, cluster analysis and principal coordinate analysis (PCoA). Results: A total of 41 SSRs showed reproducible amplification pattern among the selected gladiolus cultivars. Among these, 17 highly polymorphic SSRs revealed a total of 58 polymorphic alleles ranging from two to six with an average of 3.41 alleles per marker. Polymorphic information content (PIC) values ranged from 0.11 to 0.71 with an average value of 0.48. A total of 4 SSRs were selectively neutral based on the Ewens-Watterson test. Hence, 66.66% of Gladiolus palustris, 48% of Iris spp. EST, 71.42% of Crocus sativus SSRs showed cross-transferability among the gladiolus genotypes. Analysis of genetic structure of 84 gladiolus genotypes revealed two subpopulations; 35 genotypes were assigned to subpopulation 1, 37 to subpopulation 2 and the remaining 12 genotypes could not be attributed to either subpopulation. Analysis of molecular variance indicated maximum variance (53.59%) among individuals within subpopulations, whereas 36.55% of variation among individuals within the total population. The least variation (9.86%) was noticed between two subpopulations. Moderate (FST = 0.10) genetic differentiation between two subpopulations was observed. The grouping pattern of population structure was consistent with the unweighted pair group method with arithmetic mean (UPGMA) dendrogram based on simple matching dissimilarity coefficient and PCoA. Conclusion: SSR markers from the present study can be utilized for cultivar identification, conservation and sustainable utilization of gladiolus genotypes for crop improvement. Genetic relationships assessed among the genotypes of respective clusters may assist the breeders in selecting desirable parents for crossing.

In Vitro Culture Technology and Advanced Biotechnology Tools Toward Improvement in Gladiolus (Gladiolus species): Present Scenario and Future Prospects.

In the world's flower trade, gladiolus (Gladiolus spp.) is ranked first among bulbous flowers and eighth among cut flowers, with more than 30,000 different cultivars being grown. Mass multiplication and commercialization are restricted by the traditional propagation methods. However, the large-scale proliferation and improvement of the gladiolus have been accomplished with the aid of plant tissue culture and other biotechnological techniques. The current review includes a thorough examination of the growth and development parameters required for successful in vitro gladiolus development as well as cormel formation. Moreover, focus is being given to various techniques and methods such as in vitro cytogenetic stability and modification of chromosome number, in vitro mutagenesis and selection of pest resistance, in vitro identification and selection to develop virus-free germplasm, cryopreservation, synthetic seed technology, identifying virus diseases by RT-PCR, somaclonal variation, and protoplast and somatic hybridization. Molecular markers and their applications for genetic diversity analysis, relationships between different genotypes, and clonal stability analysis in Gladiolus species have been conducted by several research groups worldwide and are also being discussed. The article also covers efforts to enhance the functionality of plant phenotypes through genetic transformation. Future prospects for further improvement of ornamental gladiolus are also explored. Overall, the current review provides insight into the applications of basic and advanced biotechnological tools for gladiolus improvement.

GhbZIP30-GhCCCH17 module accelerates corm dormancy release by reducing endogenous ABA under cold storage in Gladiolus.

Gladiolus hybridus is one of the most popular flowers worldwide. However, its corm dormancy characteristic largely limits its off-season production. Long-term cold treatment (LT), which increases sugar content and reduces abscisic acid (ABA), is an efficient approach to accelerate corm dormancy release (CDR). Here, we identified a GhbZIP30-GhCCCH17 module that mediates the antagonism between sugars and ABA during CDR. We showed that sugars promoted CDR by reducing ABA levels in Gladiolus. Our data demonstrated that GhbZIP30 transcription factor directly binds the GhCCCH17 zinc finger promoter and activates its transcription, confirmed by yeast one-hybrid, dual-luciferase (Dual-LUC), chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) and electrophoretic mobility shift assay (EMSA). GhCCCH17 is a transcriptional activator, and its nuclear localisation is altered by surcose and cytokinin treatments. Both GhbZIP30 and GhCCCH17 positively respond to LT, sugars, and cytokinin treatments. Silencing GhbZIP30 or GhCCCH17 resulted in delayed CDR by regulating ABA metabolic genes, while their overexpression promoted CDR. Taken together, we propose that the GhbZIP30-GhCCCH17 module is involved in cold- and glucose-induced CDR by regulating ABA metabolic genes.

Evaluation of phytoremediation potential and resistance of Gladiolus grandiflora L. against cadmium stress.

Although irrigation water is a fundamental need for plant growth, it is also a source of pollutants if contaminated with harmful materials like cadmium (Cd). Irrigation water possessing abundant Cd causes damage to soil, plants, animals and ultimately human beings through the food chain. A pot experiment was conducted to evaluate the gladiolus (Gladiolus grandiflora L.) potential of Cd accumulation and the capability of the plant to be an economically beneficial choice in presence of high Cd irrigation water supply. Artificially prepared four levels of Cd irrigation water were applied to the plants viz., 30, 60, 90 and 120 mg L-1. The results revealed that 30 mg L-1 Cd had no difference in all growth-related parameters when compared to the control. Photosynthesis rate, stomatal conductance and transpiration rate along with plant height and spike length were reduced with high accumulation levels of Cd in plants. The main plant portion for Cd storage found in Gladiolus grandiflora L was corm where the amount of Cd was 10-12 times higher than the amount found in leaves, and 2-4 times more than the stem. This deportment was further established by the translocation factor (TF). In corm to shoot TF and corm to stem TF, the factor reduced with increasing Cd levels, while, in corm to leaves TF, Cd levels were statistically non-significant. From corm to shoot TF value of 0.68 and 0.43 in case of 30 and 60 mg L-1, Cd treatments indicates good phytoremediation potential of Gladiolus in low and moderate Cd-polluted environments. Conclusively, the study reveals the good capability of Gladiolus grandiflora L. to harvest Cd from the soil and water in reasonably good amount with sufficient potential to grow under irrigation-based Cd stress. Under revelations of the study, Gladiolus grandiflora L appeared as a Cd accumulator which could potentially be used as a sustainable approach for phytoremediation of Cd.

Complete Chloroplast Genome of Gladiolus gandavensis (Gladiolus) and Genetic Evolutionary Analysis.

Gladiolus is an important ornamental plant that is one of the world's four most-grown cut flowers. Gladiolus gandavensis has only been found in the Cangnan County (Zhejiang Province) of China, which is recorded in the "Botanical". To explore the origin of G. gandavensis, chloroplast genome sequencing was conducted. The results indicated that a total of 151,654 bp of circular DNA was obtained. The chloroplast genome of G. gandavensis has a quadripartite structure (contains a large single-copy (LSC) region (81,547 bp), a small single-copy region (SSC) (17,895 bp), and two inverted repeats (IRs) (IRa and IRb, 52,212 bp)), similar to that of other species. In addition, a total of 84 protein-coding genes, 8 rRNA-encoding genes, and 38 tRNA-encoding genes were present in the chloroplast genome. To further study the structural characteristics of the chloroplast genome in G. gandavensis, a comparative analysis of eight species of the Iridaceae family was conducted, and the results revealed higher similarity in the IR regions than in the LSC and SSC regions. In addition, 265 simple sequence repeats (SSRs) were detected in this study. The results of the phylogenetic analysis indicated that the chloroplast genome of G. gandavensis has high homology with the Crocus cartwrightianus and Crocus sativus chloroplast genomes. Genetic analysis based on the rbcl sequence among 49 Gladiolus species showed that samples 42, 49, 50, and 54 had high homology with the three samples from China (64, 65, and 66), which might be caused by chance similarity in genotypes. These results suggest that G. gandavensis may have originated from South Africa.

Impact of Foliar Fertilization on Growth, Flowering, and Corms Production of Five Gladiolus Varieties.

Degraded and salt affected soils are appearing more often in cultivated areas. These specific problems could reduce nutrient uptake, which can result in quality and yield loss of the cultivated plants. In order to cope with this pedo-climatic condition growers are applying fertilizers; however, due to inadequate application, soil degradation will continue. Five Gladiolus varieties were subjected to foliar fertilization treatments to assess the effect on the plant's growth parameters, vase durability and daughter corm production. Our results indicate that plants treated with foliar fertilization show significant increase in the measured parameters, flower stem length, vase durability and daughter corm production. In conclusion, our study suggests that application of foliar fertilization can increase Gladiolus plants decoration and propagation, even with a smaller footprint on nature.

Morphological and Chemical Factors Related to Western Flower Thrips Resistance in the Ornamental Gladiolus.

Understanding the mechanisms involved in host plant resistance opens the way for improved resistance breeding programs by using the traits involved as markers. Pest management is a major problem in cultivation of ornamentals. Gladiolus (Gladiolus hybridus L.) is an economically important ornamental in the Netherlands. Gladiolus is especially sensitive to attack by western flower thrips (Frankliniella occidentalis (Pergande) (Thysanoptera:Thripidae)). The objective of this study was, therefore, to investigate morphological and chemical markers for resistance breeding to western flower thrips in Gladiolus varieties. We measured thrips damage of 14 Gladiolus varieties in a whole-plant thrips bioassay and related this to morphological traits with a focus on papillae density. Moreover, we studied chemical host plant resistance to using an eco-metabolomic approach comparing the 1H NMR profiles of thrips resistant and susceptible varieties representing a broad range of papillae densities. Thrips damage varied strongly among varieties: the most susceptible variety showed 130 times more damage than the most resistant one. Varieties with low thrips damage had shorter mesophylls and epidermal cells, as well as a higher density of epicuticular papillae. All three traits related to thrips damage were highly correlated with each other. We observed a number of metabolites related to resistance against thrips: two unidentified triterpenoid saponins and the amino acids alanine and threonine. All these compounds were highly correlated amongst each other as well as to the density of papillae. These correlations suggest that papillae are involved in resistance to thrips by producing and/or storing compounds causing thrips resistance. Although it is not possible to distinguish the individual effects of morphological and chemical traits statistically, our results show that papillae density is an easy marker in Gladiolus-breeding programs targeted at increased resistance to thrips.

Comparison of Soaking Corms with Moringa Leaf Extract Alone or in Combination with Synthetic Plant Growth Regulators on the Growth, Physiology and Vase Life of Sword Lily.

Gladiolus is in demand worldwide as a cut-flower or landscaping plant, because of its superior commercial and ornamental value. Application of plant-based biostimulants has gained interest in the horticulture industry as an innovative and promising approach to ensure enhanced and sustainable yields along with better product quality. The influence of pre-plant corm soaks supplemented to 5% (v/v) with an aqueous extract from Moringa oleifera leaves (MLE) either alone or in combination with 50 mg/L salicylic acid (SA) or 50 mg/L gibberellic acid (GA) on the vegetative, physiological, and ornamental characteristics of potted gladiolus (Gladiolus grandiflorus) was investigated. In general, the treatment order for greatest horticultural value for all the parameters examined was: MLE + SA + GA > MLE + GA or SA individually > MLE alone > water-only control. Compared to other treatments, corms soaked in MLE + SA + GA had the earliest sprout time (3.7 days earlier than control), shortest production time (11 days earlier than control), tallest plant (159.5 cm), greatest number of leaves per plant (8.85 leaves), greatest maximum leaf area (66 cm2), highest SPAD reading (112) and photosynthetic activity (6.7 mmol m-2 s-1), longest spike length (91 cm), greatest number of florets per spike (20 florets), longest vase life (14.8 days), greatest N (1.53%), P (0.28%), and K (0.64%) concentrations, and largest corm diameter (4.68 cm) and mass (22.25 g). The highest total protein and proline concentrations were observed with the combined application of MLE + GA + SA. Our findings suggested that MLE either alone or in combination with other plant growth regulators not only increased the yield and quality of cut spikes, but also prolonged the vase life of cut gladiolus.

The role of multiple reproductive barriers: strong post-pollination interactions govern cytotype isolation in a tetraploid-octoploid contact zone.

BACKGROUND AND AIMS: Polyploidy is an important contributor to sympatric speciation and assortative mating is a key mechanism driving cytotype interactions in contact zones. While strong reproductive barriers can mediate the coexistence of different cytotypes in sympatry, positive frequency-dependent mating disadvantage ultimately drives the transition to single-ploidy populations. However, comprehensive estimates of reproductive isolation among cytotypes and across multiple barriers are rare. We quantify the strength of isolation across multiple reproductive stages in a tetraploid-octoploid contact zone to understand the potential for coexistence. METHODS: Assortative mating due to flowering asynchrony, pollinator behaviour, morphological overlap, self-fertilization and gametic competition between tetraploid and octoploid Gladiolus communis in a contact zone in the Western Iberian Peninsula were assessed in natural and experimental populations to quantify reproductive isolation (RI) between cytotypes. KEY RESULTS: Tetraploids and octoploids have a high degree of overlap in flowering time and similar floral morphology, and are visited by generalist insects without cytotype foraging preferences, resulting in weak pre-pollination RI (from 0.00 to 0.21). In contrast, post-pollination isolation resulting from gametic selection was a strong barrier to inter-cytotype mating, with ploidy composition in stigmatic pollen loads determining the levels of RI (from 0.54 to 1.00). Between-cytotype cross-incompatibility was relatively high (RI from 0.54 to 0.63) as was isolation acquired through self-pollination (RI of 0.59 in tetraploids and 0.39 in octoploids). CONCLUSIONS: Total RI was high for both tetraploids (from 0.90 to 1.00) and octoploids (from 0.78 to 0.98). Such high rates of assortative mating will enable cytotype coexistence in mixed-ploidy populations by weakening the impacts of minority cytotype exclusion. This study reveals the key role of gametic selection in cytotype siring success and highlights the importance of comprehensive estimates across multiple reproductive barriers to understand cytotype interactions at contact zones.

Isolation and characterization of 15 SSR loci for the endangered European tetraploid species Gladiolus palustris (Iridaceae).

PREMISE: Gladiolus palustris (Iridaceae) is an endangered European perennial tetraploid herb with special conservation interest in the European Union. Microsatellite markers can serve as effective tools for the conservation genetics of this species. METHODS AND RESULTS: We utilized a 454 pyrosequencing approach to identify simple sequence repeat (SSR) regions in a microsatellite-enriched library. Of all SSR regions, 46 were screened for specific PCR amplification, and 15 were found to be applicable in the target species. We found 1.62-3.08 alleles per population (effective alleles: 1.58-2.08) that indicated moderate to high genetic diversity values (0.28-0.44) in three pilot populations. Cross-species amplification was less effective in G. imbricatus and G. tenuis. CONCLUSIONS: The primers reported here can be used for the population genetic characterization of G. palustris. They will help us to better understand the conservation genetics of this highly endangered species.

Elimination of Bean yellow mosaic virus from infected cormels of three cultivars of gladiolus using thermo-, electro- and chemotherapy.

Bean yellow mosaic virus (BYMV) is a prevalent virus and major threat to gladiolus cultivation the world over. In the gladiolus repository at CSIR-NBRI, Lucknow, several plants (82-88%) of three economically important cultivars were found infected by BYMV showing severe mosaic and stripe symptoms. Affected plants exhibit diminished quality and quantity of florets and corms, thus reducing their value. Attempts were made to eliminate BYMV from the infected gladiolus cormel explants in vitro through thermotherapy (37 °C for 30 days), chemotherapy (30 mg/L ribavirin for 30 days), and electrotherapy (30 mA for 20 min), either alone and in different combinations. The in vitro regenerated plants were free from BYMV infection when checked by RT-PCR using BYMV-specific primers. The combination of electro- and chemotherapies has given the best response as compared to other treatments. Among the individual therapies, electrotherapy (30 mA/20 min) was found to be the best for and production of BYMV-free gladiolus plants (44-46%) with moderate regeneration efficiency (54-58%) followed by chemotherapy and thermotherapy. However, the cormels obtained from a combination of electro- and chemotherapy treatment (30 mA/20 min + 30 mg/L) has given highest virus free (46-52%) and highest therapy efficiency indices (56%) as compared to other treatments. Further, these cormels showed better developed root systems and produced more cormels which were larger in size as compared to the other treatments and control when grown in tissue culture media.

GhNAC83 inhibits corm dormancy release by regulating ABA signaling and cytokinin biosynthesis in Gladiolus hybridus.

Corm dormancy is an important trait for breeding in many bulb flowers, including the most cultivated Gladiolus hybridus. Gladiolus corms are modified underground stems that function as storage organs and remain dormant to survive adverse environmental conditions. Unlike seed dormancy, not much is known about corm dormancy. Here, we characterize the mechanism of corm dormancy release (CDR) in Gladiolus. We identified an important ABA (abscisic acid) signaling regulator, GhPP2C1 (protein phosphatase 2C1), by transcriptome analysis of CDR. GhPP2C1 expression increased during CDR, and silencing of GhPP2C1 expression in dormant cormels delayed CDR. Furthermore, we show that GhPP2C1 expression is directly regulated by GhNAC83, which was identified by yeast one-hybrid library screening. In planta assays show that GhNAC83 is a negative regulator of GhPP2C1, and silencing of GhNAC83 promoted CDR. As expected, silencing of GhNAC83 decreased the ABA level, but also dramatically increased cytokinin (CK; zeatin) content in cormels. Binding assays demonstrate that GhNAC83 associates with the GhIPT (ISOPENTENYLTRANSFERASE) promoter and negatively regulates zeatin biosynthesis. Taken together, our results reveal that GhNAC83 promotes ABA signaling and synthesis, and inhibits CK biosynthesis pathways, thereby inhibiting CDR. These findings demonstrate that GhNAC83 regulates the ABA and CK pathways, and therefore controls corm dormancy.

Computational modeling and functional characterization of a GgChi: A class III chitinase from corms of Gladiolus grandiflorus.

The present study describes the predicted model and functional characterization of an endochitinase (30 kDa) from corms of Gladiolus grandiflorus. ESI-QTOF-MS generated peptide showed 96% sequence homology with family 18, Class III acidic endochitinase of Gladiolus gandavensis. Purified G. grandiflorus chitinase (GgChi) hydrolyzed 4-methylumbelliferyl ß-d-N,N',N''-triacetylchitotriose substrate showing specific endochitinase activity. Since no structural details of GgChi were available in the Protein Data Bank (PDB), a homology model was predicted using the coordinate information of Crocus vernus chitinase (PDB ID: 3SIM). Ramachandran plot indicated 84.5% in most favored region, 14.8% in additional and 0.6% in generously allowed region while no residue in disallowed region. The predicted structure indicated a highly conserved (ß/α)8 (TIM barrel) structure similar to the family 18, class III chitinases. The GgChi also showed sequence and structural homologies with other active chitinases. The GgChi (50 µg/disc) showed no antibacterial activity, but did provide mild growth inhibition of phytopathogenic fungus Fusarium oxysporum at a concentration of 500 µg/well Similarly, insect toxicity bioassays of GgChi (50 µg) against nymphs of Bemisia tabaci showed 14% reduction in adult emergence and 14% increase in mortality rate in comparison to control values. The GgChi (1.5 mg) protein showed significant reduction in a population of flour beetle (Tribolium castaneum) after 35 days, but lower reactivity against rice weevil (Sitophilus oryzae). The results of this study provide detai.led insight on functional characterization of a family 18 class III acidic plant endochitinase.

Assessment of genetic diversity and population structure in gladiolus (Gladiolus hybridus Hort.) by ISSR markers.

ISSR (Inter simple sequence repeat) markers were used to assess the genetic diversity and population structure in 53 indigenous and exotic genotypes of gladiolus (Gladiolus hybridus Hort.). Molecular markers analysis showed PIC ranges from 0.42 (ISSR 861) to 0.99 (ISSR 855, ISSR 856 and ISSR 889) with an average 0.812, marker index ranged from 0.99 (ISSR 889) to 9.26 (ISSR 851) with an average 4.66 and resolving power of the primers ranged from 0.03 (ISSR 889) to 11.58 (ISSR 861) with an average value 3.80. The dendrogram based UPGMA clustering showed that all the 53 genotypes grouped into three main clusters. Nei's gene diversity (Na) varied from 0.929 to 1.717, effective number of alleles (Ne) varied from 1.262 to 1.369, Shannon's information index (I) ranged from 0.251 to 0.359 and gene diversity (He) was in the range from 0.167 to 0.229. Population structure analysis revealed three groups in which 32 genotypes were admixture types.

Complex cytogeographical patterns reveal a dynamic tetraploid-octoploid contact zone.

The distribution of cytotypes in mixed-ploidy species is crucial for evaluating ecological processes involved in the establishment and evolution of polyploid taxa. Here, we use flow cytometry and chromosome counts to explore cytotype diversity and distributions within a tetraploid-octoploid contact zone. We then use niche modelling and ploidy seed screening to assess the roles of niche differentiation among cytotypes and reproductive interactions, respectively, in promoting cytotype coexistence. Two cytotypes, tetraploids and octoploids, were dominant within the contact zone. They were most often distributed parapatrically or allopatrically, resulting in high geographic isolation. Still, 16.7 % of localities comprised two or more cytotypes, including the intermediate hexaploid cytotype. Tetraploids and octoploids had high environmental niche overlap and associated with similar climatic environments, suggesting they have similar ecological requirements. Given the geographical separation and habitat similarity among cytotypes, mixed-ploidy populations may be transitional and subject to the forces of minority cytotype exclusion which lead to pure-ploidy populations. However, seed ploidy analysis suggests that strong reproductive barriers may enforce assortative mating which favours stable cytotype coexistence. High cytogenetic diversity detected in the field suggests that unreduced gamete formation and hybridization events seem frequent in the studied polyploid complex and might be involved with the recurrent polyploid formation, governing, as well, the gene flow between cytogenetic entities.