Dynamics of alkaloid accumulation in Narcissus cv. Hawera: a source of Sceletium-type alkaloids.

The Sceletium-type alkaloids, known for their anxiolytic and antidepressant activities, have been recently found to be biosynthesized in Narcissus cv. Hawera, which is largely used as an ornamental plant. An alkaloid fraction enriched with Sceletium-type alkaloids from the plant has shown promising antidepressant and anxiolytic activities. In the present study, qualitative and quantitative analyses of the alkaloids in the plant organs were performed during one vegetation season by GC-MS. The alkaloid pattern and total alkaloid content was found to depend strongly on the stage of development and plant organ. The alkaloid content of bulbs was found to be highest during the dormancy period and lowest in sprouting bulbs. The leaves showed the highest alkaloid content during the intensive vegetative growth and lowest during flowering. In total, 13 alkaloids were detected in the methanol extracts of Narcissus cv. Hawera, six Sceletium-type and seven typical Amaryllidaceae alkaloids. Major alkaloids in the alkaloid pattern were lycorine, 6-epi-mesembrenol, mesembrenone, sanguinine, and galanthamine. The leaves of flowering plants were found to have the highest amount of 6-epi-mesembrenol. Mesembrenone was found to be dominant alkaloid in the leaves of sprouting bulbs and in the flowers. Considering the biomass of the plant, the dormant bulbs are the best source of alkaloid fractions enriched with 6-epi-mesembrenol. The flowers and the young leaves can be used for preparation of alkaloid fractions enriched with mesembrenone. The results indicates that Narcissus cv. Hawera is an emerging source of valuable bioactive compounds and its utilization can be extended as a medicinal plant.

Range expansion decreases the reproductive fitness of Gentiana officinalis (Gentianaceae).

Plants living at the edge of their range boundary tend to suffer an overall decline in their fitness, including growth and reproduction. However, the reproductive performance of plants in artificially expanded habitats is rarely investigated, although this type of study would provide a better understanding of range limitations and improved conservation of ex situ plants. In the current study, we transplanted a narrowly dispersed species of Gentiana officinalis H. Smith (Gentianaceae) from its natural area of distribution to two different elevations and natural elevation to comprehensively study its pollination biology, including flowering phenology and duration, floral display, reproductive allocation, pollinator activity, and seed production. The findings indicated that the starting point and endpoint of the flowering phenology of G. officinalis were earlier at the low elevation, but the peak flowering periods did not differ significantly between any of the experimental plots. When transplanted, the flowering duration, especially the female phase, was reduced; the floral display, including spray numbers, flower numbers, and flower size (length and width), decreased, especially at high elevations. Moreover, the pollen numbers and pollen-ovule ratio were decreased at both high and low elevations, although the ovule numbers showed no change, and aboveground reproductive allocation was decreased. Furthermore, pollinator richness and activity were significantly decreased, and the seed-set ratio decreased under both natural conditions and with supplemental pollination. Finally, more severe pollen limitation was found in transplanted individuals. These results indicated an overall decrease in reproductive fitness in plants living outside their original area of distribution when the geographical range of G. officinalis was expanded.

An exploratory method to determine the plant characteristics affecting the final yield of Echium amoenum Fisch. & C.A. Mey. under fertilizers application and plant densities.

Employing of advanced statistical methods to quantify agricultural information has helped to carry out targeted planning to alleviate the problems of farmers, researchers and policy section. One of these exploratory methods, is multivariate statistical analysis that examines and models the relationship between variables. Considering the importance of Echium amoenum and its use growing trend in traditional medicine and the pharmaceutical industry, also the lack of information on the correlations between its yield and morpho physiological traits, the objective of this study was to determine the causality path in which the Echium amoenum characteristics affects the yield of Echium amoenum as regards of application of organic and chemical fertilizers under different plant densities. The employed method revealed that organic fertilizers increased flower yield compared with the control. The flower yield as a result of application of compost, vermicompost and cattle manure were increased by 25, 28, and 27% compared with the control, respectively. The results of multiple regression showed that variables of plant height, shoot dry weight, flower number per plant were the main factors affected the flower yield. The relative contribution of shoot dry weight was 16 and 25% more than plant height and flower number per plant, respectively. Causality analysis identified that shoot dry weight per plant had indirect effect on flower yield in different paths, as mainly was imposed through plant height considering the path coefficients. This study suggests that optimum production of Echium amoenum with application of ecological inputs along with effective agronomical managements of the causal paths of flower yield forming, including increase in shoot yield and plant height could be achieved through an ecological cropping system with reduced costs and no health concerning due to agrochemicals residual.

Chemical Changes in the Broccoli Volatilome Depending on the Tissue Treatment.

The storage of plant samples as well as sample preparation for extraction have a significant impact on the profile of metabolites, however, these factors are often overlooked during experiments on vegetables or fruit. It was hypothesized that parameters such as sample storage (freezing) and sample pre-treatment methods, including the comminution technique or applied enzyme inhibition methods, could significantly influence the extracted volatile metabolome. Significant changes were observed in the volatile profile of broccoli florets frozen in liquid nitrogen at -20 °C. Those differences were mostly related to the concentration of nitriles and aldehydes. Confocal microscopy indicated some tissue deterioration in the case of slow freezing (-20 °C), whereas the structure of tissue, frozen in liquid nitrogen, was practically intact. Myrosinase activity assay proved that the enzyme remains active after freezing. No pH deviation was noted after sample storage - this parameter did not influence the activity of enzymes. Tissue fragmentation and enzyme-inhibition techniques applied prior to the extraction influenced both the qualitative and quantitative composition of the volatile metabolome of broccoli.

The Content of Phenolic Compounds and Organic Acids in Two Tagetes patula Cultivars Flowers and Its Dependence on Light Colour and Substrate.

The main focus of the study was to determine the content of phenolic acids, flavonoids, and organic acids in the flowers of Tagetes patula 'Petite Gold' and 'Petite Orange'. The growth of the plants was assessed depending on the cultivation conditions. The above plants were illuminated with white light, whereas the 'Petite Gold' ones with white light enhanced with blue or red light. Both cultivars grew in a two-level-mineral compounds organic substrate. The research showed that the French marigold flowers were rich in phenolic compounds and organic acids. The 'Petite Gold' flowers had more bioactive compounds compared with the 'Petite Orange' flowers. Three flavonoids, 10 phenolic acids and seven organic acids were found in the 'Petite Gold' flowers. The artificial lighting used during the cultivation of the plants showed diversified influence on the content of organic compounds in their flowers. The measurements of the plants' morphological traits and the number of inflorescences showed that illumination with red light resulted in a better effect. Large plants with numerous inflorescences grew in the substrate with a lower content of nutrients.

Beyond vegetables: effects of indoor LED light on specialized metabolite biosynthesis in medicinal and aromatic plants, edible flowers, and microgreens.

Specialized metabolites from plants are important for human health due to their antioxidant properties. Light is one of the main factors modulating the biosynthesis of specialized metabolites, determining the cascade response activated by photoreceptors and the consequent modulation of expressed genes and biosynthetic pathways. Recent developments in light emitting diode (LED) technology have enabled improvements in artificial light applications for horticulture. In particular, the possibility to select specific spectral light compositions, intensities and photoperiods has been associated with altered metabolite content in a variety of crops. This review aims to analyze the effects of indoor LED lighting recipes and management on the specialized metabolite content in different groups of crop plants (namely medicinal and aromatic plants, microgreens and edible flowers), focusing on the literature from the last 5 years. The literature collection produced a total of 40 papers, which were analyzed according to the effects of artificial LED lighting on the content of anthocyanins, carotenoids, phenols, tocopherols, glycosides, and terpenes, and ranked on a scale of 1 to 3. Most studies applied a combination of red and blue light (22%) or monochromatic blue (23%), with a 16 h day-1 photoperiod (78%) and an intensity greater than 200 µmol m-2  s-1 (77%). These treatment features were often the most efficient in enhancing specialized metabolite content, although large variations in performance were observed, according to the species considered and the compound analyzed. The review aims to provide valuable indications for the definition of the most promising spectral components toward the achievement of nutrient-rich indoor-grown products. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Plant age at the time of ozone exposure affects flowering patterns, biotic interactions and reproduction of wild mustard.

Exposure of plants to environmental stressors can modify their metabolism, interactions with other organisms and reproductive success. Tropospheric ozone is a source of plant stress. We investigated how an acute exposure to ozone at different times of plant development affects reproductive performance, as well as the flowering patterns and the interactions with pollinators and herbivores, of wild mustard plants. The number of open flowers was higher on plants exposed to ozone at earlier ages than on the respective controls, while plants exposed at later ages showed a tendency for decreased number of open flowers. The changes in the number of flowers provided a good explanation for the ozone-induced effects on reproductive performance and on pollinator visitation. Ozone exposure at earlier ages also led to either earlier or extended flowering periods. Moreover, ozone tended to increase herbivore abundance, with responses depending on herbivore taxa and the plant age at the time of ozone exposure. These results suggest that the effects of ozone exposure depend on the developmental stage of the plant, affecting the flowering patterns in different directions, with consequences for pollination and reproduction of annual crops and wild species.

Transcriptome and Metabolome Analyses Provide Insights into the Stomium Degeneration Mechanism in Lily.

Lily (Lilium spp.) is a widely cultivated horticultural crop that has high ornamental and commercial value but also the serious problem of pollen pollution. However, mechanisms of anther dehiscence in lily remain largely unknown. In this study, the morphological characteristics of the stomium zone (SZ) from different developmental stages of 'Siberia' lily anthers were investigated. In addition, transcriptomic and metabolomic data were analyzed to identify the differentially expressed genes (DEGs) and secondary metabolites involved in stomium degeneration. According to morphological observations, SZ lysis occurred when flower buds were 6-8 cm in length and was completed in 9 cm. Transcriptomic analysis identified the genes involved in SZ degeneration, including those associated with hormone signal transduction, cell structure, reactive oxygen species (ROS), and transcription factors. A weighted co-expression network showed strong correlations between transcription factors. In addition, TUNEL (TdT-mediated dUTP nick-end labeling) assays showed that programmed cell death was important during anther SZ degeneration. Jasmonates might also have key roles in anther dehiscence by affecting the expression of the genes involved in pectin lysis, water transport, and cysteine protease. Collectively, the results of this study improve our understanding of anther dehiscence in lily and provide a data platform from which the molecular mechanisms of SZ degeneration can be revealed.

Pistil Mating Type and Morphology Are Mediated by the Brassinosteroid Inactivating Activity of the S-Locus Gene BAHD in Heterostylous Turnera Species.

Heterostyly is a breeding system that promotes outbreeding through a combination of morphological and physiological floral traits. In Turnera these traits are governed by a single, hemizygous S-locus containing just three genes. We report that the S-locus gene, BAHD, is mutated and encodes a severely truncated protein in a self-compatible long homostyle species. Further, a self-compatible long homostyle mutant possesses a T. krapovickasii BAHD allele with a point mutation in a highly conserved domain of BAHD acyl transferases. Wild type and mutant TkBAHD alleles were expressed in Arabidopsis to assay for brassinosteroid (BR) inactivating activity. The wild type but not mutant allele caused dwarfism, consistent with the wild type possessing, but the mutant allele having lost, BR inactivating activity. To investigate whether BRs act directly in self-incompatibility, BRs were added to in vitro pollen cultures of the two mating types. A small morph specific stimulatory effect on pollen tube growth was found with 5 µM brassinolide, but no genotype specific inhibition was observed. These results suggest that BAHD acts pleiotropically to mediate pistil length and physiological mating type through BR inactivation, and that in regard to self-incompatibility, BR acts by differentially regulating gene expression in pistils, rather than directly on pollen.

Insights into angiosperm evolution, floral development and chemical biosynthesis from the Aristolochia fimbriata genome.

Aristolochia, a genus in the magnoliid order Piperales, has been famous for centuries for its highly specialized flowers and wide medicinal applications. Here, we present a new, high-quality genome sequence of Aristolochia fimbriata, a species that, similar to Amborella trichopoda, lacks further whole-genome duplications since the origin of extant angiosperms. As such, the A. fimbriata genome is an excellent reference for inferences of angiosperm genome evolution, enabling detection of two novel whole-genome duplications in Piperales and dating of previously reported whole-genome duplications in other magnoliids. Genomic comparisons between A. fimbriata and other angiosperms facilitated the identification of ancient genomic rearrangements suggesting the placement of magnoliids as sister to monocots, whereas phylogenetic inferences based on sequence data we compiled yielded ambiguous relationships. By identifying associated homologues and investigating their evolutionary histories and expression patterns, we revealed highly conserved floral developmental genes and their distinct downstream regulatory network that may contribute to the complex flower morphology in A. fimbriata. Finally, we elucidated the genetic basis underlying the biosynthesis of terpenoids and aristolochic acids in A. fimbriata.

Cross-pollination affects fruit colour, acidity, firmness and shelf life of self-compatible strawberry.

Cross-pollination affects the fruit characteristics of many crops but the effects of cross-pollination on fruit quality of strawberry (Fragaria × ananassa Duch.) are poorly known. This study determined how cross-pollination affects fruit quality of the strawberry cultivar, Redlands Joy, under controlled environment conditions. Plants were allocated to one of four treatments, with all flowers on each plant receiving either: (1) unassisted self-pollination (Autogamy); (2) hand-pollination with Redlands Joy pollen (Self); (3) hand-pollination with cross-pollen from a small-fruited cultivar (Sugarbaby); or (4) hand-pollination with cross-pollen from a large-fruited cultivar (Rubygem). Cross-pollination did not significantly affect plant yield or fruit mass, size, shape, firmness or shelf life. However, cross-pollination affected fruit colour and taste attributes. Cross-pollinated fruit were 3%-5% darker than self-pollinated fruit. They also had 26%-34% lower acidity and 43%-58% higher Brix:acid ratio. Cross-pollination by Sugarbaby increased fruit P, K, Ca, Fe and Mn, but decreased B, Cu and Zn, concentrations. Cross-pollination by Rubygem increased fruit Mn, but decreased K and Na, concentrations and reduced shelf life. Fruit mass, length, diameter and firmness within all treatments increased with increasing numbers of fertilized seeds per fruit. Hand self-pollinated fruit had a higher percentage of fertilized seeds than fruit arising from autogamy and they were also darker, redder, firmer, and had a longer shelf life, higher protein concentration, and lower Al and Na concentrations. The results indicate that strawberry fruit quality can be affected by both the source of pollen and the number of stigmas pollinated.

Genetic and Transcriptomic Analysis Reveal the Molecular Basis of Photoperiod-Regulated Flowering in Xishuangbanna Cucumber (Cucumis sativus L. var. xishuangbannesis Qi et Yuan).

Xishuangbanna (XIS) cucumber (Cucumis sativus L. var. xishuangbannesis Qi et Yuan), is a botanical variety of cucumber cultivars native to southwest China that possesses excellent agronomic traits for cucumber improvement. However, breeding utilization of XIS cucumber is limited due to the current poor understanding of its photoperiod-sensitive flowering characteristics. In this study, genetic and transcriptomic analysis were conducted to reveal the molecular basis of photoperiod-regulated flowering in XIS cucumber. A major-effect QTL locus DFF1.1 was identified that controls the days to first flowering (DFF) of XIS cucumbers with a span of 1.38 Mb. Whole-genome re-sequencing data of 9 cucumber varieties with different flowering characteristics in response to photoperiod suggested that CsaNFYA1 was the candidate gene of DFF1.1, which harbored a single non-synonymous mutation in its fifth exon. Transcriptomic analysis revealed the positive roles of auxin and ethylene in accelerating flowering under short-day (SD) light-dark cycles when compared with equal-day/night treatment. Carbohydrate storage and high expression levels of related genes were important reasons explaining early flowering of XIS cucumber under SD conditions. By combining with the RNA-Seq data, the co-expression network suggested that CsaNFYA1 integrated multiple types of genes to regulate the flowering of XIS cucumber. Our findings explain the internal regulatory mechanisms of a photoperiodic flowering pathway. These findings may guide the use of photoperiod shifts to promote flowering of photoperiod-sensitive crops.

ZmFAR1 and ZmABCG26 Regulated by microRNA Are Essential for Lipid Metabolism in Maize Anther.

The function and regulation of lipid metabolic genes are essential for plant male reproduction. However, expression regulation of lipid metabolic genic male sterility (GMS) genes by noncoding RNAs is largely unclear. Here, we systematically predicted the microRNA regulators of 34 maize white brown complex members in ATP-binding cassette transporter G subfamily (WBC/ABCG) genes using transcriptome analysis. Results indicate that the ZmABCG26 transcript was predicted to be targeted by zma-miR164h-5p, and their expression levels were negatively correlated in maize B73 and Oh43 genetic backgrounds based on both transcriptome data and qRT-PCR experiments. CRISPR/Cas9-induced gene mutagenesis was performed on ZmABCG26 and another lipid metabolic gene, ZmFAR1. DNA sequencing, phenotypic, and cytological observations demonstrated that both ZmABCG26 and ZmFAR1 are GMS genes in maize. Notably, ZmABCG26 proteins are localized in the endoplasmic reticulum (ER), chloroplast/plastid, and plasma membrane. Furthermore, ZmFAR1 shows catalytic activities to three CoA substrates in vitro with the activity order of C12:0-CoA > C16:0-CoA > C18:0-CoA, and its four key amino acid sites were critical to its catalytic activities. Lipidomics analysis revealed decreased cutin amounts and increased wax contents in anthers of both zmabcg26 and zmfar1 GMS mutants. A more detailed analysis exhibited differential changes in 54 monomer contents between wild type and mutants, as well as between zmabcg26 and zmfar1. These findings will promote a deeper understanding of miRNA-regulated lipid metabolic genes and the functional diversity of lipid metabolic genes, contributing to lipid biosynthesis in maize anthers. Additionally, cosegregating molecular markers for ZmABCG26 and ZmFAR1 were developed to facilitate the breeding of male sterile lines.

Complete Mitochondrial Genome Sequence and Identification of a Candidate Gene Responsible for Cytoplasmic Male Sterility in Celery (Apium graveolens L.).

Celery (Apium graveolens L.) is an important leafy vegetable worldwide. The development of F1 hybrids in celery is highly dependent on cytoplasmic male sterility (CMS) because emasculation is difficult. In this study, we first report a celery CMS, which was found in a high-generation inbred line population of the Chinese celery "tanzhixiangqin". Comparative analysis, following sequencing and assembly of the complete mitochondrial genome sequences for this celery CMS line and its maintainer line, revealed that there are 21 unique regions in the celery CMS line and these unique regions contain 15 ORFs. Among these ORFs, only orf768a is a chimeric gene, consisting of 1497 bp sequences of the cox1 gene and 810 bp unidentified sequences located in the unique region, and the predicted protein product of orf768a possesses 11 transmembrane domains. In summary, the results of this study indicate that orf768a is likely to be a strong candidate gene for CMS induction in celery. In addition, orf768a can be a co-segregate marker, which can be used to screen CMS in celery.

Building a Flower: The Influence of Cell Wall Composition on Flower Development and Reproduction.

Floral patterning is a complex task. Various organs and tissues must be formed to fulfill reproductive functions. Flower development has been studied, mainly looking for master regulators. However, downstream changes such as the cell wall composition are relevant since they allow cells to divide, differentiate, and grow. In this review, we focus on the main components of the primary cell wall-cellulose, hemicellulose, and pectins-to describe how enzymes involved in the biosynthesis, modifications, and degradation of cell wall components are related to the formation of the floral organs. Additionally, internal and external stimuli participate in the genetic regulation that modulates the activity of cell wall remodeling proteins.

Caraway (Carum carvi L.): Anther Culture and Production of DH Plants Caraway.

We describe the production of doubled haploids through anther culture in caraway. Induction conditions for the cultivation of donor plants, anther collection, composition of culture media, and physical induction conditions for embryogenesis have been described. As a result, responsive lines with numerous haploid embryo production were obtained, which after colchicine treatment became fertile. From a practical point of view, two doubled haploid populations are tested under field conditions.

Doubled Haploid Broccoli (Brassica olearacea var. italica) Plants from Anther Culture.

Broccoli (Brassica olearecea var. italica) is a cole crop grown for its floral heads and stalks. It is rich in bioactive chemicals good for human health. Broccoli has been consumed as a vegetable since Roman times, but its production and consumption have increased significantly over the past few decades. Breeders try to develop new broccoli varieties with high yield, improved quality, and resistance to biotic and abiotic stresses. Almost all new broccoli varieties are F1 hybrids. Development of inbred broccoli lines that can be used as parents in hybrid production is a time-consuming and difficult process. Haploidization techniques can be utilized as a valuable support in broccoli breeding programs to speed up the production of genetically pure genotypes. Haploid plants of broccoli can be produced from immature male gametophytes via anther and microspore cultures with similar success rates. The most important parameters affecting the success of haploidization in broccoli are the genetic background (genotype) and the developmental stage of the microspores. Broccoli genotypes differ in their responses to androgenesis induction. The highest androgenesis response could be induced from microspores in late uninucleate and early binucleate stages. Recovery of diploid broccoli plants from haploids is possible via spontaneous and induced doubling. Doubled haploid (DH) broccoli lines are considered to be fully homozygous. Therefore, the production of DH lines is an alternative way to obtain pure inbred lines that can be utilized as parents in the development of new F1 hybrid varieties showing high levels of heterosis, high-quality heads, and uniform harvestable crop. We are using an anther culture-based haploid plant production system to develop DH broccoli lines in our broccoli breeding program. DH broccoli lines are produced from different genetic backgrounds within a year and handed to broccoli breeders.

Anther and Isolated Microspore Culture in Eggplant (Solanum melongena L.).

Eggplant is one of the five important, worldwide-distributed solanaceous crops. The use of anther culture technology to produce pure, 100% homozygous doubled haploid lines for hybrid seed production is possible since 1982, where the first protocol of wide application to different eggplant materials was published. From then on, different improvements and adaptations to different materials have been made. In parallel, protocols to implement isolated microspore culture technology in eggplant have been developed principally in the last decade, which opens the door for a more efficient DH production in this species. In this chapter, two protocols, one for anther and other for isolated microspore culture in eggplant, are described. Some steps and materials are common to both approaches. A detailed description of each step from is provided.

Shed-Microspore Culture in Ornamental Peppers for Doubled Haploid Plant Production.

The shed-microspore culture technique is an alternative sub-method combining anther and isolated microspore culture to induce microspore embryogenesis. Recently, its effective use in different types of peppers has drawn attention, because it has a higher embryo yield potential compared to anther culture and is more practical than isolated microspore culture. In this chapter, a stepwise protocol for shed-microspore culture of ornamental pepper is described. This protocol includes the steps of donor plant growth conditions, the choice of suitable flower buds based on DAPI staining of microspores, application of a cold pretreatment to flower buds, surface sterilization of the buds, shed-microspore culture of anthers, stress treatments, regeneration of androgenic in vitro plantlets, their acclimatization and ploidy analysis, and in vivo chromosome doubling of the haploid plants.

Optimized Protocol for Development of Androgenic Haploids and Doubled Haploids in FCV Tobacco (Nicotiana tabacum).

Haploids are plants with gametophytic chromosome number, which upon chromosome duplication results in production of doubled haploids (DHs). There are several methods to obtain haploids and DHs, of which in vitro anther culture is the most effective and widely used method in tobacco. The production of haploids and DHs through androgenesis allows for a single-step development of complete homozygous lines from heterozygous genotypes, shortening the time required to produce homozygous genotypes in comparison to the conventional breeding scheme. The DH development process comprises two main steps: induction of androgenesis and duplication of the haploid genome. The critical stages of DH protocol in tobacco are determining the bud stage for anther culture, pretreatment, anther culture media, detection and identification of haploids, and chromosome doubling. Here we present an efficient anther culture protocol to get haploids and DHs in flue-cured virginia (FCV) tobacco. This optimized protocol can be used as a potential tool for generation of haploids and DHs for genetic improvement of tobacco.