Genome-wide transcriptional profiling and physiological investigation elucidating the molecular mechanism of multiple abiotic stress response in Stevia rebaudiana Bertoni.

Considering the major source of plant-derived low/non-calorie steviol glycosides (SGs), comprehensive physiological, biochemical, and deep transcriptional investigations were conducted to explicit deeper insight into multiple abiotic stress responses in Stevia rebaudiana. The physiological indicators including photosynthesis, chlorophyll, relative water content, shoot growth, electrolyte leakage, and SG biosynthesis were negatively impacted under drought (DS), followed by salinity (SS) and waterlogging (WS). Global transcriptional analysis revealed significant upregulated expression of the genes encoding for ROS detoxification (GST, SOD, APX, glutathione peroxidase), osmotic adjustment (alpha-trehalose-phosphate and S-adenosylmethionine decarboxylase), ion transporters (CAX, NHX, CNGS, VPPase, VATPase), water channel (PIP1, TIP) and abiotic stress-responsive candidate genes (LEA, HSPs, and Dehydrins) regulating abiotic stress response in S. rebaudiana. These inferences were complemented with predicted interactome network that revealed regulation of energy metabolism by key stress-responsive genes (GST, HKT1, MAPKs, P5CSs, PIP), transcription factors (HSFA2, DREB1A, DREB2A), and abiotic stress responsive pathways (ABA, ethylene, ion stress). This is the first detailed study to comprehend the molecular regulation of stress response and their interplay under DS, SS, and WS. The key genes and regulators can be functionally validated, and will facilitate targeted gene editing for genetic improvement of crop sustainability under changing environmental conditions in S. rebaudiana.

A comprehensive overview of breeding strategy to improve phenotypic and quality traits in Valeriana jatamansi Jones.

Valeriana jatamansi is a high value perennial herb that grows at an altitude of 1000-3000 MASL in the Indian Himalayan Region and is used in the Ayurvedic, Unani and Chinese systems of medicine. The plant extracts and essential oil (EO) obtained from its roots are used in the pharmaceutical, aromatic and flavouring industries. On account of high global annual demand and lack of organized cultivation of this herb, it is mostly collected from the wild causing depletion of the natural populations and also leading to variable produce making it unsuitable for industrial use. Therefore, to promote its cultivation, it becomes imperative to develop varieties with uniformity. Significant variations have been reported in V. jatamansi populations in terms of underground biomass accumulation, essential oil and valepotriates indicating potential of genotypic variability for effective selection. The review focuses on the diversity and variations at inter- and intra-population levels for phenotypic traits, variations for different active constituents and scope of improvement through selective breeding in V. jatamansi. The species has cross-pollinated breeding behaviour on account of floral dimorphism and presents unique opportunities for development of homozygous progeny lines through controlled self/sib-pollination by applying the breeding methods described in the review for population improvement. The germplasm resources of unique and improved selections can be maintained clonally to ensure their true-to-type identity. This review article was framed in the year 2022 after thoroughly studying the literature from the year 1919-2022. The study focuses on the variations in V. jatamansi which could be used to maximize the production through various breeding techniques for biomass and yield of different active constituents to meet the requirements of pharmaceutical and aroma industries.

Genetic resources and breeding approaches for improvement of amaranth (Amaranthus spp.) and quinoa (Chenopodium quinoa).

Nowadays, the human population is more concerned about their diet and very specific in choosing their food sources to ensure a healthy lifestyle and avoid diseases. So people are shifting to more smart nutritious food choices other than regular cereals and staple foods they have been eating for a long time. Pseudocereals, especially, amaranth and quinoa, are important alternatives to traditional cereals due to comparatively higher nutrition, essential minerals, amino acids, and zero gluten. Both Amaranchaceae crops are low-input demanding and hardy plants tolerant to stress, drought, and salinity conditions. Thus, these crops may benefit developing countries that follow subsistence agriculture and have limited farming resources. However, these are underutilized orphan crops, and the efforts to improve them by reducing their saponin content remain ignored for a long time. Furthermore, these crops have very rich variability, but the progress of their genetic gain for getting high-yielding genotypes is slow. Realizing problems in traditional cereals and opting for crop diversification to tackle climate change, research should be focused on the genetic improvement for low saponin, nutritionally rich, tolerant to biotic and abiotic stresses, location-specific photoperiod, and high yielding varietal development of amaranth and quinoa to expand their commercial cultivation. The latest technologies that can accelerate the breeding to improve yield and quality in these crops are much behind and slower than the already established major crops of the world. We could learn from past mistakes and utilize the latest trends such as CRISPR/Cas, TILLING, and RNA interference (RNAi) technology to improve these pseudocereals genetically. Hence, the study reviewed important nutrition quality traits, morphological descriptors, their breeding behavior, available genetic resources, and breeding approaches for these crops to shed light on future breeding strategies to develop superior genotypes.

Sustainable production of Saussurea costus under different levels of nitrogen, phosphorus and potassium fertilizers in cold desert region of Western Himalaya.

Introduction: Saussurea costus, an important critically endangered medicinal herb native to the Himalayan region, is commonly used in various ailments, viz. asthma, ulcer, inflammation, and stomach problems. In the international market, the dry roots and essential oil of S. costus has become an important drug. The lack of appropriate fertilizer dose recommendations is one of the limiting factors for its ex-situ conservation and large-scale cultivation, as plant nutrition is vital in determining crop growth and productivity. The study aimed to understand the comparative impact of different levels of fertilizer nutrients on growth, dry root and essential oil yield, and essential oil profile of S. costus. Methods: A field experiment was conducted in Himachal Pradesh, India's cold desert region (Lahaul valley), during 2020-21. The experiment comprised three levels of nitrogen (60, 90, and 120 kg ha-1), three levels of phosphorus (20, 40, 60 kg ha-1), and two levels of potassium (20 and 40 kg ha-1) in a factorial randomized block design. Results: The fertilizer application had an immense effect on growth attributes, root yield attributes, dry root yield, and essential oil yield over control. The treatment combination N120, P60, and K40 had the largest effect on the plant height, number of leaves per plant, leaf length and width, root length and diameter, dry matter per plant, dry root yield, and essential oil yield. However, the results were at par with the treatment comprising N90, P40, and K20. Dry root yield increased by 108.9%, and essential oil yield increased by 210.3% with fertilizer applications over unfertilized plots. The regression curve shows an increasing trend in dry root yield till N90, P40, and K20; after that, it nearly stabilized. The heat map showed that applying fertilizer significantly affected the chemical constituents of S. costus essential oil. Similarly, the plots fertilized with the highest level of NPK recorded the utmost value of available N, P, and K, as compared to unfertilized plots. Discussion: The results emphasize that for sustainable cultivation of S. costus, the application of N90, P40, and K20 combinations is the most suitable one.

Phenotyping floral traits and essential oil profiling revealed considerable variations in clonal selections of damask rose (Rosa damascena Mill.).

Damask rose (Rosa damascena Mill.) is a high-value aromatic plant species belonging to the family Rosaceae. It is being cultivated throughout the world for rose essential oil production. Besides its higher demand in the aromatic and cosmetic industry, the essential oil obtained has many pharmacological and cytotoxic activities. The primary concern of growers with the available varieties of damask rose is short flowering duration, low essential oil content and unstable yield. Thus, there is a requirement for developing new stable varieties with higher flower yield and essential oil content. The present study evaluated the variations in the flower yield parameters, essential oil content, and essential oil compounds in different clonal selections of damask rose. These clonal selections have been developed through a half-sib progeny approach from commercially available varieties 'Jwala' and 'Himroz.' The fresh flower yield varied from 629.57 to 965.7 g per plant, while the essential oil content ranged from 0.030-0.045% among the clonal selections. The essential oil profiling via gas chromatography-mass spectrometry revealed significant variations in the essential oil compounds. Acyclic monoterpene alcohols citronellol (20.35-44.75%) and geraniol (15.63-27.76%) were highest, followed by long-chain hydrocarbons, i.e., nonadecane (13.02-28.78%). The clonal selection CSIR-IHBT-RD-04 was unique in terms of the highest citronellol content (44.75%) and citronellol/geraniol (C/G) ratio of 1.93%. This selection has the potential use as a parental line in future genetic improvement programs of damask rose to achieve higher yield and better quality of rose essential oil.

Investigation on pollination approaches, reproductive biology and essential oil variation during floral development in German chamomile (Matricaria chamomilla L.).

German chamomile is an important medicinal and aromatic herb known for its blue essential oil. It lacks studies on anthesis, breeding systems and floral development with their impact on the essential oil. Therefore, the study investigated floral development and divided it into six reproductive stages (RS-1 to RS-6). The first four stages (5-6 days long) were identified as the floral enlargement and differentiation, followed by the fifth stage (10 days long) of three anthesis flushes, i.e., anther dehiscence, ray and disc florets' style branches flush. Anther dehiscence started 1-2 days before style branches flushes showed protandry and overlapped later with style branches flushes. Pollen production started from RS-3 and showed maximum viability (89%) at anther dehiscence (RS-5.1). Pollen showed dispersal through the air up to 0.7 m distance. Seed setting in controlled pollination experiments showed that removing disc florets could be successfully used as the emasculation alternate in German chamomile. The maximum essential oil content (0.40%) at the full blossomed floral stage (RS-4 &-5) suggested the right time for capitula harvesting. The findings on reproductive biology and breeding systems would offer several tools and techniques to support future breeding programs for genetic improvement of German chamomile.

Clonal Propagation of Valeriana jatamansi Retains the Essential Oil Profile of Mother Plants: An Approach Toward Generating Homogenous Grade of Essential Oil for Industrial Use.

Valeriana jatamansi Jones (Syn. V. wallichii DC.) is an aromatic, medicinal herb used as a tranquilizer and in treating sleep disorders. Rhizome is mainly used to extract essential oil (EO) and valepotriates. High quality and economic yield of rhizomes are available in the third year of growth. Therefore, the cultivation of V. jatamansi is not picking up, and over-exploitation of this plant from wild habitats to meet the increasing demand of the pharmaceutical industry is the cause of threat to the genetic diversity of the species. Further, collections from the wild are heterogeneous, resulting in variable produce. The development of clonal lines can ensure uniform quality and yield of rhizome biomass. An effective clonal propagation method was standardized using different hormonal concentrations of naphthalene acetic acid (NAA) on apical shoot cuttings from the selected clone CSIR-IHBT-VJ-05 for different time durations and raised over various planting media. NAA treatment of 50 ppm concentration for 30 min was found optimum for root induction in apical shoots of V. jatamansi. Variations for EO composition within the clone were non-significant, while samples of the control population were variable. The best quality EO (patchouli alcohol ∼62%) was available during the third year of plant growth. A propagation technique for large-scale quality plant material (QPM) production has been standardized to reduce the stress over natural resources and promote V. jatamansi for use in the aromatic and pharmaceutical industry.

Understanding the Effect of Different Abiotic Stresses on Wild Marigold (Tagetes minuta L.) and Role of Breeding Strategies for Developing Tolerant Lines.

Wild marigold has a growing demand for its essential oil in the flavor and fragrance industries. It can be grown over a broad range of climates, but the changing climatic conditions lead to abiotic stresses, thus restricting its productivity. Abiotic stresses at elevated levels result in the reduction of germination, growth, and essential oil quality of wild marigold leading to heterogeneous and inferior grades of "Tagetes oil." Drought, salinity, and heavy metal stress at elevated levels have common effects in terms of ROS formation, which are the major cause of growth deterioration in wild marigold. Temperatures above 35°C inhibit seed germination. Irradiance stress reduces the biomass and essential oil yield. Waterlogging adversely affects the survival of wild marigold in high rainfall regions. The application of plant nutrients (fertilizers) modulates the biomass and essential oil yield. Wild marigold employs multiple tolerance mechanisms to cope up with the adverse effects of abiotic stresses such as the increased activity of antioxidants to maintain cellular redox homeostasis, enhanced lipid peroxidation in the cell membrane to maintain cell wall architecture, production of secondary metabolites, and accumulation of osmolytes. In this review, we tried to understand how abiotic stresses affect wild marigold. Understanding the physiological changes and biochemical characteristics of stress tolerance will contribute to the development of stress-tolerant lines of wild marigold.

A Comprehensive Review on Biology, Genetic Improvement, Agro and Process Technology of German Chamomile (Matricaria chamomilla L.).

German chamomile (M. chamomilla) is recognized as a star herb due to its medicinal and aromatic properties. This plant is found across a wide range of climatic and soil conditions. Both the flower heads and blue essential oils of German chamomile possess several pharmacological properties of an anti-inflammatory, antimicrobial, antiseptic, antispasmodic and sedative, etc., nature, which makes it a highly sought after herb for use in many pharma and aroma industries. Chamomile tea, prepared from its flower heads, is also a well-known herbal tea for mind and body relaxation. Though it is a high-demand herb, farmers have not adopted this plant for large scale cultivation as a crop, which could improve their livelihood, due to the high cost in flower heads harvesting, loss in over mature and immature flower heads picking during harvesting, unavailability of varieties and agrotechnologies for machine harvesting, a lack of efficient process development of oil extraction and in the lack of improved stable varieties. There are many studies that have reported on the phytochemistry and pharmacological uses of chamomile, which further explore its importance in the medicine industry. Several studies are also present in the literature on its cultivation practices and plant ecology. However, studies on breeding behavior, genetic improvement, varietal development and mechanical harvesting are scarce in German chamomile. Hence, keeping in mind various aspects of farmers' and researchers' interest, earlier reports on taxonomy, floral biology, processing of oil extraction, active constituents, uses, agronomy, breeding challenges and opportunities in German chamomile are summarized in this review.

Comparative transcriptome analysis revealed gamma-irradiation mediated disruption of floral integrator gene(s) leading to prolonged vegetative phase in Stevia rebaudiana Bertoni.

Stevia rebaudiana Bert. is getting global attention because of its ability to synthesize commercially important low/no calorie natural sweeteners (LNCSs) steviol glycosides (SGs). Considering, higher accumulation of SGs in vegetative phase followed by decrement during reproductive phase necessitate the understanding of different molecular components of floral transition to develop superior varieties/cultivars with prolonged vegetative phase in Stevia. Current comparative transcriptional analysis of low dose (5 kR) gamma-irradiated mutant genotype (SMG) with prolonged vegetative phase vis-à-vis background genotype (SBG) identified DGEs of major floral transition pathways, and expressed according to their physiological fate irrespective to SMG & SBG. Contrarily, reduced expression of floral integrator genes (FT and LEAFY) in mutant genotype suggests their involvement in prolonged vegetative phase phenotype. Likewise, GO and KEGG enrichment of photosynthesis and carbon assimilation efficiency might be associated with prolonged vegetative phase and higher accumulation of Stevioside content in mutant genotype. Furthermore, deviation of flowering related transcription factors (higher expressions except MIKS-type MADS-box SMG_PV compared to SBG_F) may possibly be correlated with low expression of floral integrator genes. Findings of current studies will facilitate the genetic manipulations and crop improvement efforts in Stevia through conventional breeding and genome editing approaches for increased SGs biosynthesis.

Molecular dissection of transcriptional reprogramming of steviol glycosides synthesis in leaf tissue during developmental phase transitions in Stevia rebaudiana Bert.

Stevia is a natural source of commercially important steviol glycosides (SGs), which share biosynthesis route with gibberellic acids (GAs) through plastidal MEP and cytosolic MVA pathways. Ontogeny-dependent deviation in SGs biosynthesis is one of the key factor for global cultivation of Stevia, has not been studied at transcriptional level. To dissect underlying molecular mechanism, we followed a global transcriptome sequencing approach and generated more than 100 million reads. Annotation of 41,262 de novo assembled transcripts identified all the genes required for SGs and GAs biosynthesis. Differential gene expression and quantitative analysis of important pathway genes (DXS, HMGR, KA13H) and gene regulators (WRKY, MYB, NAC TFs) indicated developmental phase dependent utilization of metabolic flux between SGs and GAs synthesis. Further, identification of 124 CYPs and 45 UGTs enrich the genomic resources, and their PPI network analysis with SGs/GAs biosynthesis proteins identifies putative candidates involved in metabolic changes, as supported by their developmental phase-dependent expression. These putative targets can expedite molecular breeding and genetic engineering efforts to enhance SGs content, biomass and yield. Futuristically, the generated dataset will be a useful resource for development of functional molecular markers for diversity characterization, genome mapping and evolutionary studies in Stevia.