Genetic variability and inter species relationship between wild and cultivated yams (Dioscorea spp.) from Koraput, India based on molecular and morphological markers.

Wild yams (Dioscorea spp.) are important tuber crops used both as vegetable and medicine by the tribal people of Koraput, India. There is deficiency of documented information on genetic structure and diversity of wild yams and its genetic assessment is necessary for crop improvement program. The present study assessed the level of genetic diversity of eight wild and one cultivated yam species of Koraput by using different morphological and molecular markers. Significant variation in different yield and morphological traits was observed among the studied yam species. The major morphological traits such as branch number, stem thickness, tuber depth, tuber length, number of tubers per plant and yield showed high genetic heritability accompanied with high genetic advance and major determinants of phenotypic diversity. Molecular profiling was carried out by taking five simple sequence repeat markers. A total of 10 polymorphic bands with an average of two were detected at the loci of the five markers across the nine yam species. Genetic similarity analysis revealed that some wild yam species such as D. oppositifolia, D. hamiltonii and D. pubera showed higher genetic similarity with cultivated (D. alata) species. The knowledge of the extent of genetic variations of wild yam species is important for planning of the genetic conservation and the utilization of this resource especially for genetic improvement.

Elicitor-induced defense responses in Solanum lycopersicum against Ralstonia solanacearum.

We investigated on important parameters of induced resistance in hydroponic tomato (Solanum lycopersicum) against Ralstonia solanacearum using the elicitors chitosan (CHT), salicylic acid (SA), and jasmonic acid (JA). The increase in total phenolic content of roots by the elicitors was significantly higher than control. Most pronounced increase in lignin synthesis was triggered by SA followed by CHT. At 24 h post-elicitation (hpe), the activity of phenylalanine ammonia lyase was 4.5 times higher than control elicited by CHT. The peroxidase activity was about 86 nkat/mg protein at 24 hpe in case of SA and 78 nkat/mg protein in case of CHT. The activity of polyphenol oxidase increased several folds by the elicitors. Cinnamyl alcohol dehydrogenase activity increased to the maximum at 48 hpe under the influence of CHT. The results indicate that the elicitors SA and CHT induced effective defense responses in tomato plants against R. solanacearum. This was evident from reduced vascular browning and wilting symptoms of tomato plants treated with SA and CHT and challenged subsequently with R. solanacearum. This reduced disease incidence in tomato by SA and CHT may be a result of cell wall strengthening through deposition of lignin and the coincident induction of defense enzymes.

The soil-borne fungal pathogen Athelia rolfsii: past, present, and future concern in legumes.

Legumes are ubiquitous, low-cost meals that are abundant in protein, vitamins, minerals, and calories. Several biotic constraints are to blame for the global output of legumes not meeting expectations. Fungi, in particular, are substantial restrictions that not only hinder production but also pose a serious health risk to both human and livestock consumption. Athelia rolfsii (Syn. Sclerotium rolfsii) is a dangerous pathogenic fungus that attacks most crops, causing massive yield losses. Legumes are no longer immune to this dreadful fungus, which can potentially result in a 100% yield loss. The initial disease symptoms based on the formation of brown color lesions at the point of infection and further development of mycelia, followed by yellowing and wilting of the whole plant. To tackle such situation, various strategies, i.e., management in cultural practices, disease-free plant growth, genetic changes, crop hybridization and in vitro culture techniques have been undertaken. This present review encapsulates the entire situation, from sclerotial dissemination through infection development and control in legume crops, with the goal of developing a tangible understanding of sustainable legume production improvements. Further study in this area might be led in an integrated manner as a result of this information, which could contribute to a better understanding of the processes of disease incidence, resistance mechanism, and its control, and fostering greater inventiveness in the production of legumes.

Comparative transcriptome profiling reveals the basis of differential sheath blight disease response in tolerant and susceptible rice genotypes.

Rice sheath blight (ShB) disease, caused by the fungal pathogen Rhizoctonia solani AG1-IA, is one of the devastating diseases and causes severe yield losses all over the world. No completely resistant germplasm is known till now, and as a result, the progress in resistance breeding is unsatisfactory. Basic studies to identify candidate genes, QTLs, and to better understand the host-pathogen interaction are also scanty. In this study, we report the identification of a new ShB-tolerant rice germplasm, CR 1014. Further, we investigated the basis of tolerance by exploring the disease responsive differentially expressed transcriptome and comparing them with that of a susceptible variety, Swarna-Sub1. A total of 815 and 551 genes were found to be differentially regulated in CR 1014 and Swarna-Sub1, respectively, at two different time points. The result shows that the ability to upregulate genes for glycosyl hydrolase, secondary metabolite biosynthesis, cytoskeleton and membrane integrity, the glycolytic pathway, and maintaining photosynthesis make CR 1014 a superior performer in resisting the ShB pathogen. We discuss several putative candidate genes for ShB resistance. The present study, for the first time, revealed the basis of ShB tolerance in the germplasm CR1014 and should prove to be particularly valuable in understanding molecular response to ShB infection. The knowledge could be utilized to devise strategies to manage the disease better.

Novel Insights into Understanding the Molecular Dialogues between Bipolaroxin and the Gα and Gß Subunits of the Wheat Heterotrimeric G-Protein during Host-Pathogen Interaction.

Spot blotch disease of wheat, caused by the fungus Bipolaris sorokiniana (Sacc.) Shoem., produces several toxins which interact with the plants and thereby increase the blightening of the wheat leaves, and Bipolaroxin is one of them. There is an urgent need to decipher the molecular interaction between wheat and the toxin Bipolaroxin for in-depth understanding of host-pathogen interactions. In the present study, we have developed the three-dimensional structure of G-protein alpha subunit from Triticum aestivum. Molecular docking studies were performed using the active site of the modeled G-protein alpha and cryo-EM structure of beta subunit from T. aestivum and 'Bipolaroxin'. The study of protein-ligand interactions revealed that six H-bonds are mainly formed by Glu29, Ser30, Lys32, and Ala177 of G-alpha with Bipolaroxin. In the beta subunit, the residues of the core beta strand domain participate in the ligand interaction where Lys256, Phe306, and Leu352 formed seven H-bonds with the ligand Bipolaroxin. All-atoms molecular dynamics (MD) simulation studies were conducted for G-alpha and -beta subunit and Bipolaroxin complexes to explore the stability, conformational flexibility, and dynamic behavior of the complex system. In planta studies clearly indicated that application of Bipolaroxin significantly impacted the physio-biochemical pathways in wheat and led to the blightening of leaves in susceptible cultivars as compared to resistant ones. Further, it interacted with the Gα and Gß subunits of G-protein, phenylpropanoid, and MAPK pathways, which is clearly supported by the qPCR results. This study gives deeper insights into understanding the molecular dialogues between Bipolaroxin and the Gα and Gß subunits of the wheat heterotrimeric G-protein during host-pathogen interaction.

Improved photosystem II and defense enzymes activity in rice (Oryza sativa) by biopriming against Xanthomonas oryzae pv. oryzae.

Bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo) is a major threat to rice production as it accounts for loss up to 50% of annual rice grain yield. Xoo causes leaf tissue necrosis and as a result there is downregulation of the photosynthetic mechanisms of plant. Measurement of chl a fluorescence is an easy, fast, non-invasive and highly sensitive technique that can be used to understand plant health by measuring alterations in PSII activity, in response to different biotic and abiotic stresses. In the present investigation, rice seeds were bio-primed with two bacterial spp. (e.g. Bacillus subtilis and B. megaterium) and one fungal strain (i.e. Trichoderma erinaceum). The induction of defence against BB disease by bio-priming with biotic agents, was studied as response in alteration of PSII and defence enzymes activity. Field experiment was conducted with the best biotic agent; i.e. B. subtilis obtained from all experiments to study whether defence induction by it have any side effect on rice yield and biomass. Net house and field experiments have suggested that among all the biotic agents studied, bio-priming of rice seeds with B. subtilis has protected the photosynthetic machinery of plants from being damaged by BB without having any detrimental effect on rice grain yield.

Trichoderma-mediated rice straw compost promotes plant growth and imparts stress tolerance.

Rice straw burning is causing huge economic losses and environmental hazards. Microbial mediated ex situ composting could be a viable solution which would not only reduce the straw burning but also will enrich nutrition to the soil. Strains of Trichoderma isolated from tree bark were tested to decompose rice straw efficiently, and the Trichoderma-mediated rice straw compost was used subsequently to improve rice growth. Two isolates of Trichoderma reesei (NRRIT-26 and NRRIT-27) decomposed the straw by producing higher decomposing enzymes, like total cellulase (≥ 1.87 IU mL-1), endoglucanase (≥ 0.75 IU mL-1), xylanase (≥ 163.49 nkat mL-1), and laccase (≥ 11.75 IU mL-1). Trichoderma decomposed rice straw compost had higher nutrient contents (1.97% N, 2.04% K, and 0.88% P) and optimum C/N ratio (28:2) as compared to control. The Trichoderma decomposed rice straw as a nutrient reduced the mean germination time (2.2 days as compared to 4 days in control) and enhanced the seedling vigor and total chlorophyll content in rice. Expression of defense enzymes, like catalase (≥ 200% both in shoot and root), peroxidase (≥ 180% in root and ≥ 300% in shoot), and superoxide dismutase (≥ 160% in root and ≥ 90% in shoot), were higher in treated plants as compared to control indicating higher stress tolerance ability to crops. We conclude that the Trichoderma-mediated rice straw management is a viable option and has the potential to reduce straw burning, and at the same time, the compost could enrich soil fertility and impart intrinsic stress tolerance to rice.

Seed Biopriming With Trichoderma Strains Isolated From Tree Bark Improves Plant Growth, Antioxidative Defense System in Rice and Enhance Straw Degradation Capacity.

This study is a unique report of the utilization of Trichoderma strains collected from even tree barks for rice plant growth, its health management, and paddy straw degradation. Seven different spp. of Trichoderma were characterized according to morphological and molecular tools. Two of the isolated strains, namely Trichoderma hebeiensis and Trichoderma erinaceum, outperformed the other strains. Both of the strains controlled four important rice pathogens, i.e., Rhizoctonia solani (100%), Sclerotium oryzae (84.17%), Sclerotium rolfsii (66.67%), and Sclerotium delphinii (76.25%). Seed bio-priming with respective Trichoderma strains reduced the mean germination time, enhanced the seedling vigor and total chlorophyll content which could be related to the higher yield observed in two rice varieties; Annapurna and Satabdi. All the seven strains accelerated the decomposition of rice straw by producing higher straw degrading enzymes like total cellulase (0.97-2.59 IU/mL), endoglucanase (0.53-0.75 IU/mL), xylanase (145.35-201.35 nkat/mL), and laccase (2.48-12.60 IU/mL). They also produced higher quantities of indole acetic acid (19.19-46.28 µg/mL), soluble phosphate (297.49-435.42 µg/mL), and prussic acid (0.01-0.37 µg/mL) which are responsible for plant growth promotion and the inhibition of rice pathogen populations. Higher expression of defense enzymes like catalase (≥250% both in shoot and root), peroxidase (≥150% in root and ≥100% in shoot), superoxide dismutase (≥ 150% in root and ≥100% in shoot), polyphenol oxidase (≥160% in shoot and ≥120% in shoot), and total phenolics (≥200% in root and ≥250% in shoot) as compared to the control indicates stress tolerance ability to rice crop. The expression of the aforementioned enzymes were confirmed by the expression of corresponding defense genes like PAL (>3-fold), DEFENSIN (>1-fold), POX (>1.5-fold), LOX (>1-fold), and PR-3 (>2-fold) as compared to the non-treated control plants. This investigation demonstrates that Trichoderma strains obtained from tree bark could be considered to be utilized for the sustainable health management of rice crop.

Genetic relationships among 22 taxa of bamboo revealed by ISSR and EST-based random primers.

Genetic relationships among 22 taxa of bamboo were evaluated using 12 inter simple sequence repeats (ISSR) and four expressed sequence tag (EST)-based random primers, resulting in amplification of 220 loci. The grouping of species based on Jaccard's similarity matrix using UPGMA and principal coordinate analysis agreed with earlier published reports on molecular phylogenetic studies in bamboos with few deviations. In the dendrogram, however, species of one genus were placed in different clusters along with members of other genera. This calls for correct taxonomic delineation at the genus and species level using both vegetative and reproductive characters and correlation of molecular data with morphologically definable taxonomic groupings at the proper taxonomic level.

A compatible interaction of Alternaria brassicicola with Arabidopsis thaliana ecotype DiG: evidence for a specific transcriptional signature.

BACKGROUND: The interaction of Arabidopsis with Alternaria brassicicola provides a model for disease caused by necrotrophs, but a drawback has been the lack of a compatible pathosystem. Infection of most ecotypes, including the widely-studied line Col-0, with this pathogen generally leads to a lesion that does not expand beyond the inoculated area. This study examines an ecotype, Dijon G (DiG), which is considered sensitive to A. brassicicola. RESULTS: We show that the interaction has the characteristics of a compatible one, with expanding rather than limited lesions. To ask whether DiG is merely more sensitive to the pathogen or, rather, interacts in distinct manner, we identified genes whose regulation differs between Col-0 and DiG challenged with A. brassicicola. Suppression subtractive hybridization was used to identify differentially expressed genes, and their expression was verified using semi-quantitative PCR. We also tested a set of known defense-related genes for differential regulation in the two plant-pathogen interactions. Several known pathogenesis-related (PR) genes are up-regulated in both interactions. PR1, and a monooxygenase gene identified in this study, MO1, are preferentially up-regulated in the compatible interaction. In contrast, GLIP1, which encodes a secreted lipase, and DIOX1, a pathogen-response related dioxygenase, are preferentially up-regulated in the incompatible interaction. CONCLUSION: The results show that DiG is not only more susceptible, but demonstrate that its interaction with A. brassicicola has a specific transcriptional signature.

Riboflavin (Vitamin B2) mediated defence induction against bacterial leaf blight: probing through chlorophyll a fluorescence induction O-J-I-P transients.

Bacterial leaf blight (BLB) is a serious threat for rice (Oryza sativa L.) cultivation caused by the bacterial pathogen Xanthomonas oryzae pv. oryzae. The pathogen mainly damages the leaf chlorophyllous tissue, resulting in poor photosynthesis and causing up to 50% reductions in grain yield. In the present work, we have compared the structural and functional ability of the chloroplast of three varieties of rice with different degrees of susceptibility (TN1, highly susceptible; IR-20, moderately resistant; DV-85, resistant to BLB) treated with riboflavin (1 and 2mM) and infected with BLB, with chlorophyll fluorescence as a tool. As indicated by the chlorophyll fluorescence technique, the disease progress curve and yield data, riboflavin acted as an effective vitamin for inducing resistance against BLB. Plants treated with riboflavin showed improved PSII activity, more chlorophyll content and higher yield than the diseased plants.

Novel Trichoderma strains isolated from tree barks as potential biocontrol agents and biofertilizers for direct seeded rice.

This study is the first time report of utilization of Trichoderma spp. isolated from different tree bark from Odisha state of India for rice crop health management and higher productivity. Six isolates of Trichoderma spp. were identified based on the morphological characteristics and species determination was performed by molecular assays. One of the isolated strains determined as Trichoderma erinaceum outperformed others. Trichoderma erinaceum controlled three soil borne plant pathogens i.e. Rhizoctonia solani, Sclerotium rolfsii and Sclerotium oryzae effectively under controlled condition and R. solani and Helminthosporium oryzae under filed condition. Seed treatments with the formulated isolates improved the germination rate of rice and enhanced vigour. These parameters along with higher chlorophyll content could be related to higher yield observed in two rice varieties; Karuna and Sahabhagidhan. Among the six isolates tested, Trichoderma erinaceum treatment recorded highest yield. Significantly higher expression of some stress related enzymes was observed in Trichoderma treated plants which helped in better crop growth both under biotic and abiotic stresses. These isolates helped both the varieties to accumulate more nutrients. This study proves that Trichoderma erinaceum obtained from tree bark may be incorporated in integrated rice crop management both as biocontrol agent and biofertilizer.

Analysis of genetic diversity among selected grasspea (Lathyrus sativus L.) genotypes using RAPD markers.

Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variability among five selected genotypes of grasspea. Out of 30 random decamer primers tested for the present investigation 20 showed reproducible DNA amplification. A total of 257 loci were amplified of which 159 were polymorphic including 57 genotype-specific unique bands. Amplicons had molecular weights ranging from 3.0 kb to 0.1 kb. Majority amplicons were shared by most of the genotypes which indicated a very narrow genetic gap between them. The dendrogram constructed on the basis of RAPD data showed two clusters. The local genotype collected from Nayagarh was grouped along with IC-120451 and IC-120453, sharing a common node at an 82% similarity level. The other genotypes, IC-120478 and IC-120487, were located in the second clade having a common node at 84% similarity level. The investigation showed that though all the genotypes of grasspea were of apparently similar morphology there exists polymorphism at the molecular level, which can be exploited in breeding programmes aimed at crop improvement.

Phenotypic and Marker-Assisted Genetic Enhancement of Parental Lines of Rajalaxmi, an Elite Rice Hybrid.

The cytoplasmic male sterile line system comprising CRMS 32A and its maintainer line CRMS 32B is a popular choice for the development of new hybrids in India as CRMS 32A, having Kalinga 1 cytoplasm (other than WA), is a viable alternative to WA cytoplasm. However, both lines are susceptible to bacterial blight (BB), a major disease on rice. As enhancement of host plant resistance is the most effective and economical strategy to control this disease, four resistance genes (Xa4, xa5, xa13, and Xa21) were transferred from a BB pyramid line of IR64, into the A and B lines using a marker-assisted backcrossing (MAB) breeding strategy. During the transfer of genes into CRMS 32B, foreground selection was applied using markers associated with the genes, and plants having resistance alleles of the donor, are selected. Selection for morphological and quality traits was practiced to select plants similar to the recurrent parent. The four gene and three gene pyramid lines exhibited high levels of resistance against the BB pathogen when challenged with eight virulent isolates. Using genome wide based SSR markers for background selection, pyramids having >95% of the recurrent parent genome were identified. With CRMS 32B gene pyramid as donor, the four resistance genes were transferred into the A line through repeated backcrosses and the A line pyramids also exhibited high level of resistance against BB. Through a combination of selection at phenotypic and molecular levels, four BB resistance genes were successfully introduced into two parental lines (CRMS 32 B and A) of Rajalaxmi, an elite popular hybrid. The pyramided B lines did exhibit high levels of resistance against BB. Selection for morphological and quality traits and background selection hastened the recovery of the recurrent parent genome in the recombinants. Through repeated backcrosses, all the four resistance genes were transferred to CRMS 32A and test crosses suggest that the maintenance ability of the improved CRMS 32B lines is intact. These improved maintainer and CMS lines can directly be used in hybrid rice breeding and the new hybrids can play an important role in sustainable rice production in India.

Assessment of genetic diversity among 16 promising cultivars of ginger using cytological and molecular markers.

Ginger (Zingiber officinale Roscoe) is an economically important plant, valued all over the world. The existing variation among 16 promising cultivars as observed through differential rhizome yield (181.9 to 477.3 g) was proved to have a genetic basis using different genetic markers such as karyotype, 4C nuclear DNA content and random amplified polymorphic DNA (RAPD). The karyotypic analysis revealed a differential distribution of A, B, C, D and E type of chromosomes among different cultivars as represented by different karyotype formulas. A significant variation of 4C DNA content was recorded in ginger at an intraspecific level with values ranging from 17.1 to 24.3 pg. RAPD analysis revealed a differential polymorphism of DNA showing a number of polymorphic bands ranging from 26 to 70 among 16 cultivars. The RAPD primers OPC02, OPA02, OPD20 and OPN06 showing strong resolving power were able to distinguish all 16 cultivars. The extent of genetic diversity among these cultivars was computed through parameters of gene diversity, sum of allele numbers per locus and Shannon's information indices. Cluster analysis, Nei's genetic similarity and genetic distances, distribution of cultivars into special distance classes and principal coordinate analysis and the analysis of molecular variance suggested a conspicuous genetic diversity among different cultivars studied. The genetic variation thus detected among promising cultivars of ginger has significance for ginger improvement programs.

Electrostatic contribution to DNA condensation--application of 'energy minimization' in a simple model in the strong Coulomb coupling regime.

The process of bending of straight DNA to a circular form in the presence of any of the mono-, di-, tri- or tetravalent counterions has been simulated in a strong Coulomb coupling environment, employing a previously developed energy minimization simulation technique. The inherent characteristics of the simulation technique allow the monitoring of the required electrostatic contribution to the bending. The curvature of the bending has been found to play a crucial role in facilitating the electrostatic attractive potential energy. The total electrostatic potential energy has been found to decrease with bending, which indicates that bending straight DNA to a circular form or to a toroidal form in the presence of neutralizing counterions is energetically favourable and is practically a spontaneous phenomenon.

Proteomics of the response of Arabidopsis thaliana to infection with Alternaria brassicicola.

We have studied the proteome of the model plant Arabidopsis thaliana infected with a necrotrophic fungal pathogen, Alternaria brassicicola. The Arabidopsis-A. brassicicola host-pathogen pair is being developed as a model genetic system for incompatible plant-fungal interactions, in which the spread of disease is limited by plant defense responses. After confirming that a defense response was induced at the transcriptional level, we identified proteins whose abundance on 2-DE gels increased or decreased in infected leaves. At least 11 protein spots showed reproducible differences in abundance, increasing or decreasing during the progress of the infection. The pathogenesis-related protein PR4, a glycosyl hydrolase, and the antifungal protein osmotin are strongly up-regulated. Two members of the Arabidopsis glutathione S-transferase (GST) family increased in abundance in infected leaves. The spots in which these GST proteins were identified contain additional members of the GST family. Representation of GST family members in several protein spots migrating at similar molecular weight suggests post-translational modifications. The signature of GST regulation may be specific for the type of plant-pathogen interaction. The proteomic view of the defense response to A. brassicicola can be compared with other types of plant-pathogen interactions, and to leaf senescence, identifying unique regulatory patterns.