Global Transcriptomic Analysis Reveals Differentially Expressed Genes Involved in Embryogenic Callus Induction in Drumstick (Moringa oleifera Lam.).

The plant embryogenic callus (EC) is an irregular embryogenic cell mass with strong regenerative ability that can be used for propagation and genetic transformation. However, difficulties with EC induction have hindered the breeding of drumstick, a tree with diverse potential commercial uses. In this study, three drumstick EC cDNA libraries were sequenced using an Illumina NovaSeq 6000 system. A total of 7191 differentially expressed genes (DEGs) for embryogenic callus development were identified, of which 2325 were mapped to the KEGG database, with the categories of plant hormone signal transduction and Plant-pathogen interaction being well-represented. The results obtained suggest that auxin and cytokinin metabolism and several embryogenesis-labeled genes are involved in embryogenic callus induction. Additionally, 589 transcription factors from 20 different families were differentially expressed during EC formation. The differential expression of 16 unigenes related to auxin signaling pathways was validated experimentally by quantitative real time PCR (qRT-PCR) using samples representing three sequential developmental stages of drumstick EC, supporting their apparent involvement in drumstick EC formation. Our study provides valuable information about the molecular mechanism of EC formation and has revealed new genes involved in this process.

Constant theoretical conductance via changes in vessel diameter and number with height growth in Moringa oleifera.

As trees grow taller, hydraulic resistance can be expected to increase, causing photosynthetic productivity to decline. Yet leaves maintain productivity over vast height increases; this maintenance of productivity suggests that leaf-specific conductance remains constant as trees grow taller. Here we test the assumption of constant leaf-specific conductance with height growth and document the stem xylem anatomical adjustments involved. We measured the scaling of total leaf area, mean vessel diameter at terminal twigs and at the stem base, and total vessel number in 139 individuals of Moringa oleifera of different heights, and estimated a whole-plant conductance index from these measurements. Whole-plant conductance and total leaf area scaled at the same rate with height. Congruently, whole-plant conductance and total leaf area scaled isometrically. Constant conductance is made possible by intricate adjustments in anatomy, with conduit diameters in terminal twigs becoming wider, lowering per-vessel resistance, with a concomitant decrease in vessel number per unit leaf area with height growth. Selection maintaining constant conductance per unit leaf area with height growth (or at least minimizing drops in conductance) is likely a potent selective pressure shaping plant hydraulics, and crucially involved in the maintenance of photosynthetic productivity per leaf area across the terrestrial landscape.

Cultivation, Genetic, Ethnopharmacology, Phytochemistry and Pharmacology of Moringa oleifera Leaves: An Overview.

Moringa oleifera is an interesting plant for its use in bioactive compounds. In this manuscript, we review studies concerning the cultivation and production of moringa along with genetic diversity among different accessions and populations. Different methods of propagation, establishment and cultivation are discussed. Moringa oleifera shows diversity in many characters and extensive morphological variability, which may provide a resource for its improvement. Great genetic variability is present in the natural and cultivated accessions, but no collection of cultivated and wild accessions currently exists. A germplasm bank encompassing the genetic variability present in Moringa is needed to perform breeding programmes and develop elite varieties adapted to local conditions. Alimentary and medicinal uses of moringa are reviewed, alongside the production of biodiesel. Finally, being that the leaves are the most used part of the plant, their contents in terms of bioactive compounds and their pharmacological properties are discussed. Many studies conducted on cell lines and animals seem concordant in their support for these properties. However, there are still too few studies on humans to recommend Moringa leaves as medication in the prevention or treatment of diseases. Therefore, further studies on humans are recommended.

Impact of varying levels of soil salinity on emergence, growth and biochemical attributes of four Moringa oleifera landraces.

Salinity in soil and water is one of the environmental factors that severely hinder the crop growth and production particularly in arid and semi-arid regions. A pot experiment was conducted to investigate the impact of salinity levels (1.5 dS m-1, 3.5 dS m-1, 7.5 dS m-1 and 11.5 dS m-1) on emergence, growth and biochemical traits of moringa landraces under completely randomized design having three replications. Four landraces of Moringa oleifera (Faisalabad black seeded moringa [MFB], Patoki black seeded moringa [MPB], Faisalabad white seeded moringa [MFW] and Rahim Yar Khan black seeded moringa [MRB]) were selected for experimentation. All the salinity levels significantly affected the emergence parameters (time to emergence start, time to 50% emergence, mean emergence time, emergence index and final emergence percentage) of moringa landraces. However, 1.5 dS m-1 and 3.5 dS m-1 were found more favorable. Higher salinity levels (7.5 dS m-1 and 11.5 dS m-1) significantly minimized the root surface area, root projected area, root volume and root density as compared to 1.5 dS m-1, 3.5 dS m-1. Number of branches, leaves, leaflets and leaf length were also adversely affected by 7.5 dS m-1 and 11.5 dS m-1. Maximum seedling fresh and dry weights, and seedling length were recorded at 1.5 dS m-1 followed by 3.5 dS m-1. Chlorophyll a and b contents, carotenoids and membrane stability index were also observed highest at salinity level of 1.5 dS m-1. In case of moringa landraces, MRB performed better regarding emergence attributes, growth parameters, and biochemical analysis followed by MFW as compared to MFB and MPB. Moringa landraces i.e. MRB and MFW were found more tolerant to salinity stress as compared to MFB and MPB.

Challenges and opportunities for Moringa growers in southern Ethiopia and Kenya.

Moringa oleifera (MO) and M. stenopetala (MS) are two commonly cultivated species of the Moringaceae family. Some households in southern Ethiopia (S. ETH) and Kenya (KEN) plant MS and MO, respectively. The edible parts of these species are rich in amino acids, vitamins and minerals, especially selenium. Despite their nutritional value, Moringa is sometimes considered as a "famine food". The aim of this study was to determine the extent of dietary utilization of these plants by Moringa Growing Households (MGHs). Moringa growing households were surveyed in 2015. Twenty-four and 56 heads of MGHs from S. ETH and KEN, respectively, were interviewed using semi-structured questionnaires. Subsistence agriculture was the main source of livelihood for all MGHs in S. ETH and 71% of those in KEN. All MGHs in S. ETH cultivated MS while those in KEN cultivated MO. Of the MGH heads in S. ETH, 71% had grown MS as long as they remember; the median cultivation period of MO in KEN was 15 years. All MGHs in S. ETH and 79% in KEN used Moringa leaves as a source of food. Forms of consumption of leaves were boiled fresh leaves, and leaf powder used in tea or mixed with other dishes. Other uses of Moringa include as medicine, fodder, shade, agroforestry, and as a source of income. Although MO and MS have multiple uses, MGHs face several challenges, including a lack of reliable information on nutritional and medicinal values, inadequate access to markets for their products, and pest and disease stresses to their plants. Research and development to address these challenges and to promote the use of these species in the fight against hidden hunger are necessary.

Low power continuous wave-laser seed irradiation effect on Moringa oleifera germination, seedling growth and biochemical attributes.

Recently, laser application in agriculture has gained much attention since plant characteristics were improved significantly in response of pre-sowing seed treatment. Pre-sowing laser seed treatment effects on germination, seedling growth and mineral profile were studied in Moringa olifera. M. olifera healthy seeds were exposed to 25, 50, 75mJ low power continuous wave laser light and grown under greenhouse conditions. The seedling growth and biochemical attributes were evaluated from 10-day-old seedlings. The germination parameters (percentage, mean germination time), vigor index, seedling growth (root length, seedling length, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight) enhanced considerably. The laser energy levels used for seed irradiation showed variable effects on germination, seedling growth and mineral profile. The mineral contents were recorded to be higher in seedling raised from laser treated seeds, which were higher in roots versus shoots and leaves. The effect of laser treatment on seedling fat, nitrogen and protein content was insignificant and at higher energy level both nitrogen and protein contents decreased versus control. Results revealed that M. olifera germination, seedling growth and mineral contents were enhanced and optimum laser energy level has more acceleratory effect since at three laser energy levels the responses were significantly different. Overall the laser energy levels effect on germination and seedling growth was found in following order; 75mJ>50mJ>25mJ, where as in case of fat, protein and nitrogen contents the trend was as; 25mJ>50mJ and 75mJ. However, this technique could possibly be used to improve the M. olifera germination, seedling growth, and minerals contents where germination is low due to unfavorable conditions.

Screening Reliable Reference Genes for RT-qPCR Analysis of Gene Expression in Moringa oleifera.

Moringa oleifera is a promising plant species for oil and forage, but its genetic improvement is limited. Our current breeding program in this species focuses on exploiting the functional genes associated with important agronomical traits. Here, we screened reliable reference genes for accurately quantifying the expression of target genes using the technique of real-time quantitative polymerase chain reaction (RT-qPCR) in M. oleifera. Eighteen candidate reference genes were selected from a transcriptome database, and their expression stabilities were examined in 90 samples collected from the pods in different developmental stages, various tissues, and the roots and leaves under different conditions (low or high temperature, sodium chloride (NaCl)- or polyethyleneglycol (PEG)- simulated water stress). Analyses with geNorm, NormFinder and BestKeeper algorithms revealed that the reliable reference genes differed across sample designs and that ribosomal protein L1 (RPL1) and acyl carrier protein 2 (ACP2) were the most suitable reference genes in all tested samples. The experiment results demonstrated the significance of using the properly validated reference genes and suggested the use of more than one reference gene to achieve reliable expression profiles. In addition, we applied three isotypes of the superoxide dismutase (SOD) gene that are associated with plant adaptation to abiotic stress to confirm the efficacy of the validated reference genes under NaCl and PEG water stresses. Our results provide a valuable reference for future studies on identifying important functional genes from their transcriptional expressions via RT-qPCR technique in M. oleifera.

Ecotype variability in growth and secondary metabolite profile in Moringa oleifera: impact of sulfur and water availability.

Moringa oleifera is widely cultivated in plantations in the tropics and subtropics. Previous cultivation studies with M. oleifera focused primarily only on leaf yield. In the present study, the content of potentially health-promoting secondary metabolites (glucosinolates, phenolic acids, and flavonoids) were also investigated. Six different ecotypes were grown under similar environmental conditions to identify phenotypic differences that can be traced back to the genotype. The ecotypes TOT4880 (origin USA) and TOT7267 (origin India) were identified as having the best growth performance and highest secondary metabolite production, making them an ideal health-promoting food crop. Furthermore, optimal cultivation conditions-exemplarily on sulfur fertilization and water availability-for achieving high leaf and secondary metabolite yields were investigated for M. oleifera. In general, plant biomass and height decreased under water deficiency compared to normal cultivation conditions, whereas the glucosinolate content increased. The effects depended to a great extent on the ecotype.

Analytical characterization of Moringa oleifera seed oil grown in temperate regions of Pakistan.

The hexane-extracted oil content of Moringa oleifera seeds ranged from 38.00 to 42.00%. Protein, fiber, and ash contents were found to be 26.50-32.00, 5.80-9.29, and, 5.60-7.50%, respectively. Results of physical and chemical parameters of the extracted oil were as follows: iodine value, 68.00-71.80; refractive index (40 degrees C), 1.4590-1.4625; density (24 degrees C), 0.9036-0.9080 mg/mL; saponification value, 180.60-190.50; unsaponifiable matter, 0.70-1.10%; and color (1 in. cell), 0.95-1.10 R + 20.00-35.30 Y. Tocopherols (alpha, gamma, and delta) in the oil were up to 123.50-161.30, 84.07-104.00, and 41.00-56.00 mg/kg, respectively. The oil was found to contain high levels of oleic acid (up to 78.59%) followed by palmitic, stearic, behenic, and arachidic acid up to levels of 7.00, 7.50, 5.99, and 4.21%, respectively. The induction period (Rancimat, 20 L/h, 120 degrees C) of the crude oil was 9.99 h and reduced to 8.63 h after degumming. Specific extinctions at 232 and 270 nm were 1.70 and 0.31, respectively. Many parameters of M. oleifera oil indigenous to Pakistan were comparable to those of typical Moringa seed oils reported in the literature. The results of the present analytical study were also compared with those of different vegetable oils.

Anti-nutritional factors in the roots of a local cultivar of Moringa oleifera (Lam).

The evergreen plant, Moringa oleifera (Lam) has been known to have both medicinal and nutritional properties, thus its wide use in traditional medicine in Africa and Asia. The roots, in particular, have been reported to possess antibiotic, anti-tumour and anti-oxidative activities. This study therefore seeks to determine the levels of the anti-nutritional factors and other proximate analyses in the roots of a local cultivar of Moringa oleifera which might be responsible for such activities. The concentrations of oxalates and phytates in the roots were determined using the methods of Munro and Bassir and Griffith and Thomas, respectively. The methods of Association of Analytical Chemists (AOAC) were used to estimate the amount of tannins, saponins and cyanogenic glycosides while the Technicon sequential Multi-sample amino acid analyzer (TSM) was used determine the amino acid concentration in the roots. Tannins (45 mg 100 g(-1)) and oxalates (17.08 mg 100 g(-1)) were present in the roots at higher levels while saponins (4.20mg 100 g(-1)), cyanogenic glycosides (2.72 mg 100 g(-1)) and phytates (0.07 mg 100 g(-1)) occurred at much lower levels. The roots contained (Mean +/- Standard Error of mean) %crude lipid (6.33 +/- 1.64), %crude proteins (5.02 +/- 1.52), %carbohydrates (76.75), %ash (4.97 +/- 0.53) and %moisture (6.93 +/- 0.58). The roots lacked the water-soluble vitamins pyridoxine, riboflavin and thiamine but contained ascorbic acid (48.13 mg 100 g(-1)) and niacin (5.83 mg 100 g(-1)). This study has shown that Moringa oleifera roots are rich in anti-nutritional factors and that is why they are widely used in traditional medicine in Africa, Asia and Americas for its medicinal importance.

Arsenic removal from the aqueous system using plant biomass: a bioremedial approach.

Metal species released into the environment by technological activities tend to persist indefinitely, circulating and eventually accumulating throughout the food chain, thus becoming a serious threat to the environment. Environment pollution by toxic metals occurs globally through military, industrial, and agricultural processes and waste disposal. Bioremediation processes are the target of recent research and are considered low-cost, ecofriendly methods to alleviate the current problems of water decontamination, particularly for remote and rural areas. The present piece of work reports the unexploited sorption properties of the powdered seed of the plant Moringa oleifera (SMOS) for the removal of Arsenic [As(III) and As(V)] from aqueous solutions. Sorption studies, using standard practices, result in the standardization of optimum conditions such as biomass dosages (2.0 g), metal concentrations (25 ppm), contact time (60 min) and volume of the test solutions (200 ml) at pH 7.5, for As(III) and pH 2.5 for As(V). Maximum sorption for As(III) and As(V) species is 60.21 and 85.6%, respectively. Protein/Amino acid-Arsenic interactions are found to play an important role in the biosorption process using plant biomass SMOS.