The mutants resistant to ethylene are helpful in deciphering the role of ethylene in plant development. We isolated an ethylene-resistant tomato (Solanum lycopersicum) mutant by screening for acetylene-resistant (atr-1) seedlings. The atr-1 mutant displayed resistance to kinetin, suggesting attenuation of the ethylene sensing response. atr-1 also exhibited resistance to ABA- and glucose-mediated inhibition of seed germination. Unlike the Never-ripe (Nr) mutant seedlings that were hypersensitive to glucose, atr-1 seedlings were resistant to glucose, indicating ethylene sensing in atr-1 is compromised in a manner distinct from Nr. Metabolically, atr-1 seedlings had lower levels of amino acids but higher levels of several phytohormones, including ABA. atr-1 plants grew faster and produced more flowers, leading to a higher fruit set. However, the atr-1 fruits took a longer duration to reach the red-ripe (RR) stage. The ripened atr-1 fruits retained high ?-carotene and lycopene levels post-RR stage and had longer on-vine longevity. The metabolome profiles of post-RR stage atr-1 fruits revealed increased levels of sugars. The atr-1 had a P279L mutation in the GAF domain of the ETR4, a key ethylene receptor regulating tomato ripening. The atr-1 exhibits phenotypic traits distinct from the Sletr4-1 (G154S) mutant, thus represents a new ETR4 allele named Sletr4-2. Our study highlights that novel alleles in ethylene receptors may aid in enhancing the nutritional quality of tomato.

The tomato (Solanum lycopersicum) ripening inhibitor (rin) mutation is known to completely repress fruit ripening. The heterozygous (RIN/rin) fruits have extended shelf life, ripen normally, but have inferior taste/flavour. To address this, we used genome editing to generate newer alleles of RIN (rin) by targeting the K-domain. Unlike previously reported CRISPR alleles, the rin alleles displayed delayed onset of ripening, suggesting that the mutated K-domain represses the onset of ripening. The rin fruits had extended shelf life and accumulated carotenoids at an intermediate level between rin and progenitor line. Besides, the metabolites and hormonal levels in rin fruits were more akin to rin. To overcome the negative attributes of rin, we crossed the rin alleles with Nps1, a dominant-negative phototropin1 mutant, which enhances carotenoid levels in tomato fruits. The resulting Nps1/rin hybrids had extended shelf life and 4.4–7.1-fold higher carotenoid levels than the wild-type parent. The metabolome of Nps1/rin fruits revealed higher sucrose, malate, and volatiles associated with tomato taste and flavour. Notably, the boosted volatiles in Nps1/rin were only observed in fruits bearing the homozygous Nps1 mutation. The Nps1 introgression into tomato provides a promising strategy for developing cultivars with extended shelf life, improved taste, and flavour.

Induced mutations accelerate crop improvement by providing novel disease resistance and yield alleles. However, the alleles with no perceptible phenotype but have an altered function remain hidden in mutagenized plants. The whole-genome sequencing (WGS) of mutagenized individuals uncovers the complete spectrum of mutations in the genome. Genome-wide induced mutation resources can improve the targeted breeding of tomatoes and facilitate functional genomics. In this study, we sequenced 132 doubly ethyl methanesulfonate (EMS)-mutagenized lines of tomato and detected approximately 41 million novel mutations and 5.5 million short InDels not present in the parental cultivar. Approximately 97% of the genome had mutations, including the genes, promoters, UTRs, and introns. More than one-third of genes in the mutagenized population had one or more deleterious mutations predicted by Sorting Intolerant From Tolerant (SIFT). Nearly one-fourth of deleterious genes mapped on tomato metabolic pathways modulate multiple pathway steps. In addition to the reported GC>AT transition bias for EMS, our population also had a substantial number of AT>GC transitions. Comparing mutation frequency among synonymous codons revealed that the most preferred codon is the least mutagenic toward EMS. The validation of a potato leaf-like mutation, reduction in carotenoids in ?-carotene isomerase mutant fruits, and chloroplast relocation loss in phototropin1 mutant validated the mutation discovery pipeline. Our database makes a large repertoire of mutations accessible to functional genomics studies and breeding of tomatoes.

Plant volatile organic compounds (VOCs) are organic chemicals that plants release as part of their natural biological processes. Various plant tissues produce VOCs, including leaves, stems, flowers, and roots. VOCs are essential in plant communication, defense against pests and pathogens, aroma and flavor, and attracting pollinators. The study of plant volatiles has become an increasingly important area of research in recent years, as scientists have recognized these compounds’ important roles in plant physiology. As a result, there has been a growing interest in developing methods for collecting and analyzing plant VOCs. HS-SPME-GC-MS (headspace solid-phase microextraction-gas chromatography-mass spectrometry) is commonly used for plant volatile analysis due to its high sensitivity and selectivity. This chapter describes an efficient method for extracting and identifying volatile compounds by HS-SPME coupled with GC-MS in tomato fruits.

Metabolites are intermediate products formed during metabolism. Metabolites play different roles, including providing energy, supporting structure, transmitting signals, catalyzing reactions, enhancing defense, and interacting with other species. Plant metabolomics research aims to detect precisely all metabolites found within tissues of plants through GC-MS. This chapter primarily focuses on extracting metabolites using chemicals such as methanol, chloroform, ribitol, MSTFA, and TMCS. The metabolic analysis method is frequently used according to the specific kind of sample or matrix being investigated and the analysis objective. Chromatography (LC, GC, and CE) with mass spectrometry and NMR spectroscopy is used in modern metabolomics to analyze metabolites from plant samples. The most frequently used method for metabolites analysis is the GC-MS. It is a powerful technique that combines gas chromatography’s separation capabilities with mass spectrometry, offering detailed information, including structural identification of each metabolite. This chapter contains an easy-to-follow guide to extract plant-based metabolites. The current protocol provides all the information needed for extracting metabolites from a plant, precautions, and troubleshooting.

Annexins are a ubiquitous, evolutionarily conserved group of Ca2+-dependent phospholipid-binding proteins. They are a family of less numerous and more varied proteins that form a unique monophyletic group. They play an important role in various abiotic and biotic stress responses through Ca2+-mediated signaling. Chickpea (Cicer arietinum L.) is one of the most widely grown legume crops in the world. In recent years, intensive research has been carried out to identify and elucidate genes and molecular pathways that control stress responses in plants. The availability of the chickpea genome has hastened the functional genomics of chickpea. In the current study, we attempted Genome-wide identification and in silico analysis of Annexins in chickpea. Thirteen annexin sequences have been identified in the chickpea genome. Four conserved annexin domains were found in ten annexin members, while three annexins CaAnn5, CaAnn12, and CaAnn13, showed three, two, and one conserved domain, respectively. The gene structure analysis showed the presence of multiple exons in all thirteen annexins. Most Annexin genes are composed of 3–5 introns. Their chromosomal locations showed that out of thirteen genes, ten could be mapped on four chromosomes. Three genes were placed on the scaffold regions. The promoter sequence analysis of all thirteen annexins showed the presence of various elements related to growth and development and response to different phytohormones and abiotic stress. The gene expression data of different annexins in various tissues like leaf, shoot, root, flower bud, and young pod showed their differential expression. Analysis of expression data of roots in drought stress showed their differential expression with the different stages of plant growth. Overall, the current findings show the possible role of CaAnns in different stages of plant growth and development in normal and stressful conditions. Moreover, these findings will be helpful in the further characterization of CaAnn genes and their promoters.

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Brecht in India analyses the dramaturgy and theatrical practices of the German playwright Bertolt Brecht in post-independence India.The book explores how post-independence Indian drama is an instance of a cultural palimpsest, a site celebrating a dialogue between Western and Indian theatrical traditions, rather than a homogenous and isolated canon. Analysing the dissemination of a selection of Brecht’s plays in the Hindi belt between the 1960s and the 1990s, this study demonstrates that Brecht’s work provided aesthetic and ideological paradigms to modern Hindi playwrights, helping them develop and stage a national identity. The book also traces how the reception of Brecht was mediated in India, how it helped post-independence Indian playwrights formulate a political theatre, and how the dissemination of Brechtian aesthetics in India addressed the anxiety related to the stasis in Brechtian theatre in Europe.Tracking the dialogue between Brechtian aesthetics in India and Europe and a history of deliberate cultural resistance, Brecht in India is an invaluable resource for academics and students of theatre studies and theatre historiography, as well as scholars of post-colonial history and literature.

This article investigates the crucial role of operatic theatre tradition in representing the mountain culture of Uttarakhand, a region that became the twenty-seventh state of the Republic of India in 2000. Uttarakhand culture is extremely diverse, so in this article I solely examine the performance practices of the Kumaoni community and, in particular, a Kumaoni opera called Rajula Malushahi, which is based on a folk mountain legend. This article problematizes and expands the range of influence of mountain cultures by drawing on a version of this story by Indian director Amit Saxena, performed on 17 January 2017, at the Sri Ram Centre for Performing Arts, Delhi. The first part of the article traces the emergence of the Kumaoni opera tradition and its ability to articulate the voice of these mountain people. In the second and third parts, I examine the dramaturgical practices of Rajula Malushahi and demonstrate how it diverged from mainstream Indian dramaturgies, and how this voiced the anxieties and dilemmas of peripheral mountain cultures. These strategic diversions reimagined the idea of mountains and the female gender. I argue that Kumaoni operatic theatre tradition, which is based on folk narratives, continually connects mountains, human bodies and modern environmental discourses, and thus offers a critique of entrenched modern divisions between humans and non-humans.

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