Application of an Ovate Leaf Shape Model to Evaluate Leaf Bilateral Asymmetry and Calculate Lamina Centroid Location

Leaf shape is an important leaf trait, with ovate leaves common in many floras. Recently, a new leaf shape model (referred to as the MLRF equation) derived from temperature-dependent bacterial growth was proposed and demonstrated to be valid in describing leaf boundaries of many species with ovate leaf shape. The MLRF model’s parameters can provide valuable information of leaf shape, including the ratio of lamina width to length and the lamina centroid location on the lamina length axis. However, the model wasn’t tested on a large sample of a single species, thereby limiting its overall evaluation for describing leaf boundaries, for evaluating lamina bilateral asymmetry and for calculating lamina centroid location. In this study, we further test the model using data from two Lauraceae species, Cinnamomum camphora and Machilus leptophylla, with >290 leaves for each species. The equation was found to be credible for describing those shapes, with all adjusted root-mean-square errors (RMSE) smaller than 0.05, indicating that the mean absolute deviation is smaller than 5% of the radius of an assumed circle whose area equals lamina area. It was also found that the larger the extent of lamina asymmetry, the larger the adjusted RMSE, with approximately 50% of unexplained variation by the model accounted for by the lamina asymmetry, implying that this model can help to quantify the leaf bilateral asymmetry in future studies. In addition, there was a significant difference between the two species in their centroid ratio, i.e., the distance from leaf petiole to the point on the lamina length axis associated with leaf maximum width to the leaf maximum length. It was found that a higher centroid ratio does not necessarily lead to a greater investment of mass to leaf petiole relative to lamina, which might depend on the petiole pattern.

Ecological Adaptation of Endemic Anthocleista Species under Moisture Gradient in Parts of Niger Delta, Nigeria

Background: Knowledge of the anatomy of Anthocleista species is crucial for understanding how these plants adapt to the environment. Aim: This study was aimed at investigating the adaptive relationship of moisture gradient influence on the anatomy of four species in the genus Anthocleista (A djalonesis A Chev; A. liebrechtsiana De Wild & Th.Dur; A. nobilis G.Don; and A. vogelii Planch) in light of ecological niche adaptation. Place of Study: parts of Akwa-Ibom, Bayelsa, Cross River and Rivers States in the Niger Delta. Methods: Conventional classical anatomical techniques for structural sectioning were used. Results: Though there are similarities and differences in vascular structure among the species, the study has revealed variance in anatomical responses to moisture gradient (ranging from dry mesophytic to mesophytic and wetland conditions) of adaptation. The most important and distinct features observed are the presence of sclerenchymatous idioblast, air sacs and sclereidal idioblast. Sclerenchymatous idioblasts are numerous in A liebrechtsiana, few in A. nobilis and A. vogelii but lacking in A. djalonesis. The sclerenchymatous idioblast in A. liebrechtsiana, A. nobilis, and A. vogelii confirmed these species to be mesophytic to semi-aquatic in their habitat adaptation; while A. djalonesis is dry-mesophytic in adaptation with thicker epidermal layer, multiple hypodermal layers, thicker mesophyll tissues with increased number of palisade layers and thick leaves. The stem and root modification had abundance sclereidal idioblast distribution in A. liebrechtsiana, and A. nobilis, moderate in A. vogelii and very low in A. djalonesis. Conclusion: The variation observed in the leaf, petiole, stem and root anatomical characters are due to moisture gradient influence with the resultant effect of plant species evolving structures such as idioblast and modification to adapt to the niche and environment where they find themselves.

Variations in Morphological Characters and Antioxidant Potential of Different Plant Parts of Four Ziziphus Mill. Species from the Cholistan

Genus Ziziphus (Z.) contains various important species in tropical and subtropical regions that are globally famous for their food and medicinal uses. However, no comprehensive study was available on the morphology and phytochemistry of Ziziphus species, mainly under different growth conditions, i.e., irrigated and desert (Cholistan). Therefore, this study was carried out to evaluate the morphological and phytochemical characteristics of Ziziphus species, i.e., Z. jujuba, Z. mauritiana, Z. spina-christi, and Z. nummularia, found in the irrigated and desert conditions. Our results revealed significant variations for most of the measured parameters, showing a large-scale diversity among Ziziphus species under irrigated and desert conditions. Specifically, Ziziphus species showed better morphology of all measured parameters of leaves and fruits under irrigated conditions compared to desert conditions, indicating that the optimum water availability in irrigated conditions improved the morphological parameters of Z. species. Meanwhile, among all Ziziphus species, the maximum leaf length (7.4 cm), leaf width (4.1 cm), leaf area (30.6 cm2), and leaf petiole length (1.3 cm) were observed for Z. jujuba, and the highest leaf dry weight (55.4%) was recorded for Z. mauritiana. Similarly, the highest fruit length (3.9 cm), fruit stalk length (1.5 cm), fruit diameter (3.6 cm), fruit width (3.8 cm), fruit area (66.1 cm2), seed length (2 cm), and seed diameter (1.1 cm) were measured for species Z. jujuba, while the maximum fruit dry weight (49.9%) and seed width (1.4 cm) were recorded for species Z. nummularia. Interestingly, compared to irrigated conditions, higher values of bioactive contents, i.e., phenol, flavonoid, and antioxidant activity, in fruits and leaves of Ziziphus species under desert conditions indicated the positive impact of desert climate on the phytochemistry of the Z. plants. Among Ziziphus species, Z. nummularia accumulated the maximum fruit phenols (304.4 mg GAE/100 g), leaf phenols (314.2 mg GAE/100 g), fruit flavonoids (123.7 mg QE/100 g), and leaf flavonoids (113.4 mg QE/100 g). Overall, this study demonstrated the significant morphological and phytochemical variations of the Ziziphus species under irrigated and desert conditions, which could be utilized for future studies to improve the production and medicinal potential of the Ziziphus, especially in desert areas.

In-vitro propagation and phytochemical profiling of a highly medicinal and endemic plant species of the Himalayan region (Saussurea costus)

Efficient protocols for callus induction and micro propagation of Saussurea costus (Falc.) Lipsch were developed and phytochemical diversity of wild and in-vitro propagated material was investigated. Brown and red compact callus was formed with frequency of 80–95%, 78–90%, 70–95% and 65–80% from seeds, leaf, petiole and root explants, respectively. MS media supplemented with BAP (2.0 mgL−1), NAA (1.0 mgL−1) and GA3 (0.25 mgL−1) best suited for multiple shoot buds initiation (82%), while maximum shoot length was formed on media with BAP (1.5 mgL−1), NAA (0.25 mgL−1) and Kinetin (0.5 mgL−1). Full strength media with IAA (0.5 mgL−1) along with IBA (0.5 mgL−1) resulted in early roots initiation. Similarly, maximum rooting (87.57%) and lateral roots formation (up to 6.76) was recorded on full strength media supplemented with BAP (0.5 mgL−1), IAA (0.5 mgL−1) and IBA (0.5 mgL−1). Survival rate of acclimatized plantlets in autoclaved garden soil, farmyard soil, and sand (2:1:1) was 87%. Phytochemical analysis revealed variations in biochemical contents i.e. maximum sugar (808.32 µM/ml), proline (48.14 mg/g), ascorbic acid (373.801 mM/g) and phenolic compounds (642.72 mgL−1) were recorded from callus cultured on different stress media. Nonetheless, highest flavenoids (59.892 mg/g) and anthocyanin contents (32.39 mg/kg) were observed in in-vitro propagated plants. GC–MS analysis of the callus ethyl acetate extracts revealed 24 different phytochemicals. The variability in secondary metabolites of both wild and propagated plants/callus is reported for the first time for this species. This study may provide a baseline for the conservation and sustainable utilization of S. costus with implications for isolation of unique and pharmacologically active compounds from callus or regenerated plantlets.

Differentiation of valuable for breeding sunflower lines by resistance to Phoma rot

The purpose of the research is differentiation of valuable for breeding sunflower lines by resistance to Phoma rot. The promising constant sunflower lines were estimated by their resistance to Phoma rot using methods of artificial inoculation in the greenhouse and laboratory conditions. In greenhouse conditions, there were used two methods of artificial inoculation with a fungus Plenodomus lindquistii in the bottom of leaf petiole of a sunflower plant at a stage of the first true leaf pair. Application of the pieces of agar nutrient medium with mycelium, pycnidia and picnospores (unbroken) allowed to identify resistant and mid-resistant sunflower lines (VK 900, L 86 and L 107, L 132). At inoculation with 0.2 ml of a fungus spore suspension (concentration of 1 × 106 spores/ml), five susceptible and one very susceptible sunflower lines (L 2385, L 103, L 136, L 131, L 128, and L 82) were determined. In laboratory conditions, the quick test for estimation of susceptibility to Phoma rot (dose of infection – 12 g) allowed to reveal more than 50 % of plants with healthy root system and two constant sunflower lines L 116 and L 136 with the affection degree of one point. Estimation of susceptibility to Phoma rot infection of the different organs (stem and root system) at artificial inoculation showed a possibility to differentiate sunflower lines by resistance of roots and stems and to select the most promising for the further work in breeding.

The microscopic diagnostic signs of some Fabaceae L. genus representatives. Message ІІ. Plant conducting system

The world plant products market is expanded rapidly and trade in them tends to grow by 15–25 % annually. The number of reports is increased proportionally about accidental contamination or deliberate, economically motivated falsification of plant raw materials. 27 % of the nearly 6.000 herbal preparations that are sold in 37 countries have contained undeclared contaminants, substitutes, or other components, according to the literature. We have conducted a plant conduction system microscopic analysis of the individual members’ genus Clover (Trifoliae L.), Fabaceae L. to identify morphoanatomical characteristics. Clover has anti-inflammatory, antiseptic, choleretic, diaphoretic, diuretic, hemostatic, expectorant, astringent properties and is used in many diseases. Aim. The finding common features and those that differ and can be used as diagnostic during studying the conducting system structure of genus Trifolium L. leaves and stems. Materials and methods. Plant material (herb) from Trifolium pratense L., T. incarnatum L., T. repens L. and T. fragiferum L. was harvested during the active flowering period – (May – June) and was dried in a well-ventilated place. Leaves and stems preparations were pre-boiled in 5 % sodium hydroxide water solution and fixed in chloral hydrate solution. Cross-sections were made with a microtome. The BIOLAM LOMO light microscope (Russia) and OLYMPUS SH-21 digital camera were used to record the data about identify the conducting apparatus of the plant’s leaf, petiole, and stem. Results. It has been examined the central vein structure of T. pratense L. and T. fragiferum L. leaves, it was determined that the conductive system is covered with a crystalline coating and there is one closed collateral bundle in the center, which is not typical for dicotyledonous plants. The petioles of T. incarnatum L., T. fragiferum L., and T. repens L. in cross-section are several different shapes. There are kidney-shaped and round. The conducting apparatus T. incarnatum L. and T. repens L. have arranged in a circle, closed and collateral. The leafstalk structure type of T. fragiferum L. is bunchles. It contradicts too the information about the structure conducting system of dicotyledonous plants. The stem’s conducting bundles are collateral and open. Conclusions. We have paid attention to the structural peculiarities of the conductive system of the central vein and petiole of objects for study: Trifolium pratense L., T. incarnatum L., T. repens L., and T. fragiferum L. when searching for differentiating features in some members of the genus Trifoliae L. in pharmacognostic analysis. The Dicotyledonae representatives are not characteristic of the closed type of conductive bundles, which we observed during microscopic examination.

An Alternative to Increase Accumulation of Phenolic Compound in Grapevine Callus Cultures: Chemical Mutagen Applications

This research was carried out to investigate the effects of chemical mutagen applications on producing phenolic compounds in calli obtained from leaf petiole explants of the Royal grape variety.Petioles were cultured in B5 medium containing 0.5 mg/L benzyl amino purine and 0.5 mg/L indole acetic acid to obtain callus. Calli obtained during the period following planting were subcultured in the same culture conditions three times at one month intervals. In the fourth subculture, the calli were transferred to nutrient mediums with the same composition containing ethyl methanesulfonate, sodium azide, azacitidine and acridine orange in three different doses, 0.5 mM, 1.0 mM, and 2.0 mM, and cultured in these mediums for 60, 120 and 180 minutes. Then, they were cultured again for four weeks in mutagen-free nutrient mediums with the same content as their previous mediums. At the end of this period, calli were taken from their nutrient medium, and total phenolic compound, total flavanol, total flavonol and anthocyanin analyzes were performed.As a result of the research, it has been determined that all mutagens applied are effective in increasing the production of phenolic compounds depending on the dose and time.

Ectoedemia argyropeza (Zeller, 1839) sur Peuplier Tremble : traits de comportement inférés à partir de la distribution des traces d’activités subsistant sur les feuilles-hôtes (Lepidoptera, Nepticulidae)

Ectoedemia argyropeza on Aspen leaves: deciphering retrospectively some behavioural traits from the distribution of traces of the insect activities subsisting on host-leaves (Lepidoptera, Nepticulidae). Despite their usually small size, endotrophic insects —especially those whose larvae are mining into their host-leaves— exhibit patterns of behaviour that are often more elaborate than what is usually seen in many ectotrophic herbivorous insects. However, since it is generally difficult to capture properly these behaviours in the field, precisely due to the small size of these insects, it turns out to be more convenient attempting to uncover retrospectively some of these behaviours on the basis of their resulting traces which subsist on the host-leaves. In order to be able to infer reliable information from this retrospective approach, the examination of a substantial number of host leaves and the support of appropriate statistical tests are required. The present study concerns a species of microlepidoptera, Ectoedemia argyropeza, whose caterpillars are exclusively mining the leaves of Aspen (Populus tremula) and which is still further distinguished, in a remarkable manner, by the induction of a preliminary galling (“cecidian”) development stage. This unusual combination of life traits contributes to enrich the insect’s behavioural repertoire and therefore offers a more promising field of investigation. For this species, I more particularly focus on the way the artefacts resulting from the insect activities are distributed spatially, on (or in) the host leaf, namely: (i) the spatial distribution of eggs deposited on the host-leaf petiole, (ii) the hierarchy of preferential positioning of the caterpillar corridor in the section of the petiole, hypertrophied by the cecidogenic reaction, (iii) the hierarchy of preferential locations of mines in the host-leaf blade. Were also tested, on the one hand, the existence (or not) of paired relationships between each of the three categories of distributions mentioned above and, on the other hand, the degree of conformation of each of these different distributions to the bilateral symmetry of the leaf support. The behavioural aspects that can be tentatively inferred from the above information are subsequently discussed.

Genome-Wide Analysis of the R2R3-MYB Gene Family in Fragaria × ananassa and Its Function Identification During Anthocyanins Biosynthesis in Pink-Flowered Strawberry

The strawberry (Fragaria × ananassa) is an economically important fruit throughout the world. The large R2R3-MYB gene family participates in a variety of plant functions, including anthocyanin biosynthesis. The present study is the first genome-wide analysis of the MYB gene family in the octoploid strawberry and describes the identification and characterization of the family members using the recently sequenced F. × ananassa genome. Specifically, we aimed to identify the key MYBs involved in petal coloration in the pink-flowered strawberry, which increases its ornamental value. A comprehensive, genome-wide analysis of F. × ananassa R2R3-FaMYBs was performed, investigating gene structures, phylogenic relationships, promoter regions, chromosomal locations, and collinearity. A total of 393 R2R3-FaMYB genes were identified in the F. × ananassa genome and divided into 36 subgroups based on phylogenetic analysis. Most genes with similar functions in the same subgroup exhibited similar exon-intron structures and motif compositions. These R2R3-FaMYBs were unevenly distributed over 28 chromosomes. The expansion of the R2R3-FaMYB gene family in the F. × ananassa genome was found to be caused mainly by segmental duplication. The Ka/Ks analysis indicated that duplicated R2R3-FaMYBs mostly experienced purifying selection and showed limited functional divergence after the duplication events. To elucidate which R2R3-FaMYB genes were associated with anthocyanin biosynthesis in the petals of the pink-flowered strawberry, we compared transcriptional changes in different flower developmental stages using RNA-seq. There were 131 differentially expressed R2R3-FaMYB genes identified in the petals, of which three genes, FaMYB28, FaMYB54, and FaMYB576, appeared likely, based on the phylogenetic analysis, to regulate anthocyanin biosynthesis. The qRT-PCR showed that these three genes were more highly expressed in petals than in other tissues (fruit, leaf, petiole and stolon) and their expressions were higher in red compared to pink and white petals. These results facilitate the clarification on the roles of the R2R3-FaMYB genes in petal coloration in the pink-flowered strawberry. This work provides useful information for further functional analysis on the R2R3-FaMYB gene family in F. × ananassa.

Specific Physiological and Anatomical Traits Associated With Polyploidy and Better Detoxification Processes Contribute to Improved Huanglongbing Tolerance of the Persian Lime Compared With the Mexican Lime

Huanglongbing (HLB) is presently a major threat to the citrus industry. Because of this disease, millions of trees are currently dying worldwide. The putative causal agent is a motile bacteria belonging to Candidatus Liberibacter spp., which is transmitted by psyllids. The bacteria is responsible for the synthesis of callose at the phloem sieve plate, leading to the obstruction of the pores that provide connections between adjacent sieve elements, thus limiting the symplastic transport of the sugars and starches synthesized in leaves to the other plant organs. The Persian triploid lime (Citrus latifolia) is one of the most HLB-tolerant citrus varieties, but the determinants associated with the tolerance are still unknown. HLB-infected diploid Mexican lime (Citrus aurantiifolia) and Persian lime were investigated. The leaf petiole was analyzed using scanning electron microscopy (SEM) to observe callose deposition at the phloem sieve plate. Leaf starch contents and detoxification enzyme activities were investigated. In the field, Persian lime leaves present more limited symptoms due to HLB than the Mexican lime leaves do. Photosynthesis, stomatal conductance, and transpiration decreased compared with control plants, but values remained greater in the Persian than in the Mexican lime. Analysis of the petiole sieve plate in control petiole samples showed that pores were approximately 1.8-fold larger in the Persian than in the Mexican lime. SEM analyses of petiole samples of symptomatic leaves showed the important deposition of callose into pores of Mexican and Persian limes, whereas biochemical analyses revealed better detoxification in Persian limes than in Mexican limes. Moreover, SEM analyses of infected petiole samples of asymptomatic leaves showed much larger callose depositions into the Mexican lime pores than in the Persian lime pores, whereas biochemical traits revealed much better behavior in Persian limes than in Mexican limes. Our results reveal that polyploids present specific behaviors associated with important physiological and biochemical determinants that may explain the better tolerance of the Persian lime against HLB compared with the Mexican lime.