Modern Research in Botany Vol. 1

Tolerance Potential of Selected Genotypes of Zea mays L. and Pennisetum glaucum (L.) R. Br. under Drought Stress

Emmanuel Iwuala — 2020-01-23

Three genotypes of Pennisetum glaucum (IP14599, IP14787and LRNO3) and Zea mays (DTSYN11, LRNO1 and LRIO1) were screened at specified periods of drought stress and evaluated to detect trait signals for drought tolerance. Specific responses were confirmed against drought among all the selected genotypes of both the species. Genotypes of P. glaucum showed high biomass production, increased tolerance index and high plant nutrient content during drought stress. Conversely, genotype DTSYN11 of Z. mays performed better than the LRNO1 and LRIO1 regarding the evaluated traits. Biochemical studies related to protein content, proline, MDA concentration and antioxidants (CAT, APX) accumulation in both the species highlighted that pearl millet genotypes are better adapted than maize under drought stress. The low concentration of MDA and high accumulation of antioxidant enzymes were observed during increased duration of stress, perhaps this provides some extent of osmotic protection and antioxidant defence modulation to the pearl millet genotypes, thus causing protection to the crop. This work explains the capacity of pearl millet related to the defence mechanism over the maize genotypes under drought condition. Hence, pearl millet would be of great relevance in breeding programmes for crop improvement and can serve as a valued reserve to explicate molecular strategies in manipulating drought tolerance to other susceptible crops.

Analysing Activity of Cellulase and Macerating Enzyme and Effect of Catechin on Cellulase Activity

Karishma Rajbhar — 2020-01-23

Cellulose having linkages like β-1,4-linked homopolymer of glucose is major and primary portion in plant cell wall. Detection of released monosaccharides after enzyme treatment with suitable required cell ambience can unleash the detailed mechanism of optimum activity of cellulase and macerating enzyme. Efficiency of enzyme effect can be studied on sample plant particles. Cellulases and macerating enzyme hydrolyse cellulose into monosaccharides like glucose, dextrose and fructose. Cellulase activity may be affect by presence of catechin, thus catechin effect can be either inhibiting or facilitating the reaction. The detailed mechanism is discussed in the article.

Rapid Multiplication for Producing Quality Planting Material of Olive [Olea europaea L. cv. Barnea] through Plant Tissue Culture and It’s Commercial Application

Susmita Shukla — 2020-01-23

Olea europaea is an economically important tree species native to Mediterranean region. The present study is focused on developing the regeneration protocol and production of true-to-type plants of Olea europaea L. cv Barnea with commercial importance. Olive tree is a long lived evergreen tree but has a long juvenile phase and reaches its full production stage at 25 - 30 years of age. Traditionally, for many years, they have been propagated through cutting, grafting, vegetative propagation and in recent years, through air layering through plant tissue culture methods this problem of long juvenility can be overcome in addition large number of pathogen free plants can be produced. Nodes from adventitious shoots were surface sterilized and inoculated in different tissue culture media such as Woody Plant Media (WPM), Murashige and Skoog media (MS), Rugini Olive media (OM), Schenk and Hildebrandt media (SH) and Multiplication media (Combination of half strength MS and half strength Rugini OM) in which culture establishment was best obtained in multiplication media supplemented with 1 mg/l zeatin. Different treatment was given to induce shoot proliferation and the highest shoot number 4.2±0.4 was obtained in multiplication media enriched with 2.0 mg/l zeatin and silver nitrate at 2.0 mg/l. The in-vitro derived shoots were assessed for genetic fidelity using different molecular markers Random Amplification of Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR). Monomorphic bands were obtained between the mother DNA and in-vitro derived progenies which indicated that there is genetic stability in the in-vitro regenerated plants. Commercial analysis was done with respect to Industrial Use. The global market for olive is estimated to grow at a compound annual growth rate of 4.5% in the forecast period of 2019-2024. In the production of olive, Spain holds the largest share of world production and Italy holds the second largest share. Although in recent years, the world has begun to see olive from various new regions. The present study is the initiative step towards propagation of Olive cultivar in South Asian region.

Seed Germination in Wild and Cultivated Varieties of Withania somnifera (L.) Dunal

Afshan Niyaz — 2020-01-23

Aim: Withania somnifera (L.) Dunal or Ashwagandha is a valuable medicinal plant having a consistent demand in pharmaceutical industries. Traditionally it is propagated from seeds but it’s germination capacity is poor. So in the present study different methods were considered to sort out a suitable method that can be used for its cultivation in the field of Hazaribag, Jharkhand, India.

Study Design: The study was done in the laboratory and field of Hazaribag, Jharkhand, India.

Place and Duration of Study: Department of Botany, Vinoba Bhave University, Hazaribag, Jharkhand, India. The study was carried out from July 2012 to August 2012 and again from mid June 2013 to August 2013.

Methodology: Different pre germination treatments including 24 hours water soaking, 48 hours water soaking, mechanical scarification, heat treatment at 50 degree Celsius (5 min, 10 min, 15 min) and Gibberellic acid [GA3] (250 µg/l, 500 µg/l, 1000 µg/l) treatments were proposed. The mean germination percentage and the mean germination time were calculated for each of the treatments.

Results: The results revealed that GA3 500 µg/l treated seeds showed increased germination percentage in laboratory (86±0.34%) and in soil (84.1±0.36%) as well as reduced mean germination time in laboratory (5.8±0.41 days) and in soil (10.6±0.17 days). Whereas heat treatment at 50 degree Celsius (5 min, 10 min, 15 min) drastically reduced germination percentage in laboratory (32±0.47%, 16±0.36%, 12±0.31%) and in soil (51±0.49%, 49.6±0.49%, 32.2±0.46%), respectively in wild variety. In cultivated variety the mean germination time was also more (10.36±0.18 days) in laboratory and (13.75±0.50 days) in field as was found for wild variety. Gibberellic acid treatment improved (83.9± 0.95%) in laboratory and (82.9±.64%) in field at 500 µg/l.

Conclusion: Pre treating fresh seeds of Ashwagandha with GA3 500 µg/l for 24 hours before sowing can be adopted to overcome dormancy of seeds with good germination percentage. Likewise mechanical scarification of seeds can also be taken into consideration as an alternative, cost-effective and eco-friendly way to break seed dormancy which has also given favourable results.

Role of Aerosolized Coal Fly Ash in the Global Plankton Imbalance: Case of Florida's Toxic Algae Crisis

Mark Whiteside — 2020-01-23

Red tide is the term used in Florida (USA) and elsewhere to describe a type of marine harmful algal bloom (HAB) that grows out of control and produces neurotoxins that adversely affect humans, birds, fish, shellfish, and marine mammals. HABs are becoming more abundant, extensive, and closer to shore, and longer in duration than any time in recorded history. Our objective is to review the effects the multifold components of aerosolized coal fly ash as they relate to the increasing occurrences of HABs. Aerosolized coal fly ash (CFA) pollutants from non-sequestered coal-fired power plant emissions and from undisclosed, although “hidden in plain sight,” tropospheric particulate geoengineering operations are inflicting irreparable damage to the world’s surface water-bodies and causing great harm to human health (including lung cancer, respiratory and neurodegenerative diseases) and environmental health (including major die-offs of insects, birds and trees). Florida’s ever-growing toxic nightmare of red tides and blue-green algae is a microcosm of similar activity globally. Atmospheric deposition of aerosol particulates, most importantly bioavailable iron, has drastically shifted the global plankton community balance in the direction of harmful algae and cyanobacterial blooms in fresh and salt water. Proposed geoengineering schemes of iron fertilization of the ocean would only make a bad situation unimaginably worse. Based on the evidence presented here, the global spread of harmful algae blooms will only be contained by rapidly reducing particulate air pollution both by implementation of universal industrial particulate-trapping and by the immediate halting of jet-sprayed particulate aerosols. Corrective actions depend not only on international cooperation, but on ending the deadly code of silence throughout government, academe, and media on the subject of ongoing tropospheric aerosol geoengineering. Long-standing weather control, climate intervention, and geoengineering operations have come to threaten not only all humans but the entire web of life on Earth.

Effect of Light and Temperature through Poly Film Covers on Anthocyanin Content in Rose Cut Flower

G. Oloo-Abucheli — 2020-01-23

Quality is the most important attribute in rose cut flowers for both export and domestic market. Quality in cut flowers may be defined by many attributes however, among the most important is colour. A group of pigments commonly known as anthocyanins determine colour in plants. Anthocyanins play a significant role by ameliorate the effect of high irradiance in plants under stressful environment. They also play a key role in delaying senescence hence enhancing the cut flower vase life. Despite the advantages anthocyanins are affected by the preharvest conditions mainly light and temperature interfering with their stability. An experiment was set up to investigate the effect of light and temperature through selected coloured poly film covers on rose petal anthocyanin content. The greenhouse structure was covered by poly films of different colours that were compartmentalized i.e. UV-A clear, IR504 with yellow tint and UV-A 205/N with green tint replicated three times. Two rose cultivars Red calypso and Furiosa were established and maintained, upon maturity the flower heads were plucked and oven dried at 60°C to constant weight. 5 g of the crushed petals was used in anthocyanin extraction. The anthocyanins were extracted and quantified in comparison with commercial standards using HPLC machine. The data obtained from the chromatogram as peak areas was subjected to analysis of variance (ANOVA) using SAS statistical package (SAS Inst., Inc., Cary, NC) at P = .05. Where there were treatment differences, mean separation was done using Tukey’s procedure. Poly films significantly affected the quantity and quality of anthocyanin accumulation in rose petals. Cyanidin 3-0-glucoside was the most prevalent anthocyanin across all poly film covers and it was noted to be high under the UV-A 205/N (110.95±8.26 µg ^_ 5 g–1 DW) and IR504 (109.69±8.26 µg ^_ 5 g–1 DW) compared to UV-A clear (84.56±8.26 µg ^_ 5 g–1 DW). The quantity of anthocyanins was low under the UV-A clear poly film that was characterized by high light transmission and day temperature. Combination of high irradiance and temperature affect the quality and quantity of anthocyanin in rose cut flowers.

Extraction of High-quality Genomic DNA from Different Plant Orders Applying a Modified CTAB-Based Method

Nadia Aboul-Ftooh Aboul-Maaty — 2020-01-23

Background: Reliable measurement of DNA concentration and purity is important for almost all molecular genetics studies. Different plant species have varying levels of polysaccharides, polyphenols and other secondary metabolites which combine with nucleic acids during DNA isolation and further affect the quality of the extracted DNA. The current extraction protocol is based-upon the conventional CTAB method with further modifications for the extraction of DNA from variable plant seeds and crops belong to seven different orders. The principle modifications currently employed for DNA extraction involved the use of higher CTAB concentration and higher levels of β-Mercaptoethanol. Additionally, higher concentrations of sodium chloride and potassium acetate were added simultaneously with absolute ice cold isopropanol for the precipitation of DNA free from polysaccharides.

Results and Conclusion: The prescribed modifications in the present method establish a quick and efficient standardized protocol for DNA extraction from different plant orders. The current extraction protocol, therefore, can be of great value for molecular analysis involving large numbers of different plant samples from different orders. These modifications consistently produced pure and high quality DNA suitable for further molecular analysis. Successful PCR amplification with RAPD primer, the complete digestion of the isolated DNA with the HindIII restriction enzyme and amplification of nptII gene applying both conventional and real-time PCR (RT-PCR) in presence of SYBR Green 1 dye pursued by the analysis of melting curve analysis validated the quality of the isolated DNA. Moreover, it reflects the efficiency of the protocol and proves its suitability for further applications for the assessment of food safety, detection of genetically modified (GM) crops and conservation of biodiversity.

Acacia nilotica, Albizia saman, Azadirachta indica: Ethanobotany and Medicinal Uses

Parul Tripathi — 2020-01-23

Ethno-botany accounts for the study of relationship between people and plants for their use as medicines, food, shelter, clothing, fuel, fodder and other household purposes. India is a repository of medicinal plants. Different medicinal plant parts and their products have been used in traditional medicine since time immemorial. They still play an important role in effective disease control with no side effect and have natural origin. They are considered as the great source of unique compounds for the development of medicines for the cure of various diseases. The present article is an attempt to explore and comprehensively highlight the biological activities, pharmacological actions and medicinal applications of three plants namely Acacia nilotica, Albizia saman, Azadirachta indica.

Phytochemical, GC/MS Analyses and Cytotoxic Effects of Maerua pseudopetalosa (Gilg and Bened.) De Wolf Tuber Fractions

Manal A. Ibrahim — 2020-01-23

Tuber extracts were subjected to column chromatography technique. Eight fractions were obtained for ethyl acetate and twelve for ethanol. The brine shrimp lethality assay was used for assessment of toxicity. For the first time promising result was shown for ethanolic extract. The fractions F₈, F₉, F₁₁ and F₁₂ were represented high toxicity equal to 1.25, 7.98, 0.185, 0.041 µg/ml respectively. Also F₇ and F₁₀ showed toxic effects (89.9, 30.6 µg/ml) whereas F₅ (LC₅₀ 807 µg/ml) was weakly toxic.

Ethyl acetate fractions showed moderate toxicity for F₇ and F₄ (299.7 and 375.4 µg/ml), while F₂, F₆ and F₈were weakly toxic. However F₁ exhibited high toxic effect.

Ethanolic extract which is the highest bioactive extract was subjected to TLC analysis. Tests for secondary metabolites proved the presence of tannins, sterols and alkaloids. Also detection of triterpenes, sterols and flavonoid represented positive results.

The fractions F₈, F₉, F₁₁ and F₁₂ with high cytotoxic values were identified by gas chromatography/ mass spectrometry analysis. Thirty three compounds were detected; which were not recorded in any previous work in the available literature. Fraction 8 and 9 were found to be cytotoxic due to the presence of oleate and linoleate compounds; with more cytotoxicity in fraction 8 as a result of the additional presence of decenoic acid. Also, fraction 12 was more cytotoxic than fraction 11 and this was attributed to the presence of a proline derivative (Proline-N-methyl- butyl ester). This compound might be considered as the cause of the high toxicity of the fraction; since free proline was used as an inhibitor of breast cancer development. Surprisingly, M. pseudopetalosa tubers were used in the folkloric medicine of the natives of the South Blue Nile State in Sudan for the treatment of breast cancer growth without any knowledge of their chemical constituents.

Determination of the Paper Quality as a Substrate for Oyster Mushroom Cultivation

Siddhant . — 2020-01-23

Eight different kinds of papers, viz., glaze paper, brown paper, news paper, magazine paper, chart paper, kite paper, rough copy paper and A-4 size printing paper, and two types of cardboard viz., corrugated cardboard and card board were evaluated for different manifestations of white oyster mushroom Pleurotus florida Strain-P1. Among them news paper was later treated as a control. The mushroom was utilized all the substrates for their growth and sporophore formation. Majority of substrates were taken almost equal time for spawn run, primordial development and fruiting bodies maturation. The measured parameters were net yield (Weight of fresh mushrooms), biological efficiency, number of fruiting bodies produced and average weight of sporophores varied among themselves. The crop of mushroom was harvested in three flushes where yield and biological efficiency ranged 190-495 gm, 38-99% for the substrate used. Magazine paper (450 gm; 90%) and card board (495 gm; 99%) produced significant (P=0.05) yield and biological efficiency over control. They also produced significant number of mushroom fruit bodies (56 and 64, respectively). Corrugated cardboard (10.29 gm) was found significant in terms of average weight per sporocarp. The percentage yield of different substrates was also evaluated. Among the substrates, card board contributed 14% of total mushroom production followed by magazine paper (13%) and news paper (12%).

Rapid and Mass Propagation of Hybanthus enneaspermus (L.) F. Muell. from Shoot Tip and Nodal Explants

P. Velayutham — 2020-01-23

A rapid and efficient protocol for in vitro propagation of Hybanthus enneaspermus (L.) F. Muell. (Violaceae) has been developed from the shoot tip and nodal explants. The explants were cultured on MS basal medium supplemented with different concentrations of cytokinins, viz., BAP and Kin, ranging from 5 µM to 25 µM, either individually or in combinations of both these cytokinins for shoot induction. Shoot buds of both the explants proliferated on MS medium supplemented with both cytokinins. The best response was observed on MS medium containing 15 µM BAP. Subsequently the optimum concentration of BAP (15 µM) was combined with different concentrations of Kin ranging from 2 µM to 10 µM. Maximum number of 28.6 ± 0.90 and 36.8 ± 1.54 shoots were produced on MS medium containing 15 µM BAP + 6 µM Kin from the shoot tip and nodal explants respectively. The regenerated shoots were transferred to rooting medium containing auxins at different concentrations ranging from 2 µM to 10 µM of IAA, IBA or NAA. The highest number of roots were observed on half strength MS medium fortified with 4 µM IBA. The plantlets were then hardened and acclimatized in soil. About 80% of plantlets were survived in the field condition. A completely randomized design was used in all experiments and analysis of variance and mean separations were carried out using Duncan’s Multiple Range Test. Each treatment factor consists of 10 replicates repeated for 5 times. This protocol would help ex situ conservation of this medicinal plant.

Residual Effect of Segregated and Unsegregated Urban Solid Waste Compost on Quality of Cowpea (Vigna unguiculata L.)

Roohi . — 2020-01-23

A greenhouse experiment was conducted to study the residual effect of segregated and unsegregated urban solid waste compost on chlorophyll content, crude fiber and crude protein in cowpea (Vigna unguiculata L.) during kharif-2016 in Department of Soil Science and Agricultural Chemistry, University of Agricultural Sciences, Gandhi Krishi Vignana Kendra, Bangalore (India). The results revealed that chlorophyll content, crude fiber and crude protein content were significantly improved by the application of 100% NPK + segregated urban solid waste compost (10 t ha^-1) followed by 100% NPK + unsegregated urban solid waste compost (10 t ha^-1) as compared to the treatment which includes only inorganics.

Investigating Proteomic Variations in Transgenic Tomatoes Compared to Its Non-transgenic Counterpart

Hanaa Abdel-Sadek Oraby — 2020-01-23

Three complementary approaches were used for the assessment of proteomic variations due to genetic transformation. These approaches were gel electrophoresis, Gas chromatography coupled with mass spectrometry (GC/MS) and amino acids analysis. First we assessed proteomic variations applying gel electrophoresis analysis in two different transformed plants (GM-potato and GM-Tomato) along with their non-transformed counterparts. Further we pursued with our analysis using one of the tested GM-plants (transgenic tomato) and its non-transgenic counterpart applying Gas chromatography coupled with mass spectrometry (GC/MS) and amino acids analysis.

The results indicated proteomic variations between both transgenic plants (GM-potato and GM-tomato) compared to their non-transgenic counterparts, where the protein patterns separation on the 1D SDS-PAGE were not similar in both cases. Results of the amino acid concentrations of the transformed tomato were also different compared to its non-transgenic counterpart. These detected differences are most likely due to transformation process.

Results also revealed that the efficiency of GC/MS approach to identify a mixture of unknown proteins was limited. GC/MS analysis was only able to identify few number of protein molecules. Therefore, more advanced and specific technologies like LC/MS/MS and MALDI-TOF-MS are recommended to be employed for the identification of unintended effects due to genetic transformation in plants.

In vitro Propagation of a Medicinal Plant Adhatoda vasica Nees by Shoot Bud Culture

P. Soni — 2020-01-23

Mass propagation of plant species through in vitro culture is one of the useful and most successful examples of commercial application of plant tissue culture technology. Recently, much progress has been done in this technology for regenerating medicinal plants. In present study an efficient protocol is devised for a rapid in vitro propagation through shoot bud culture of a valuable medicinal plant Adhatoda vasica. In present investigation the proliferating auxiliary shoot cultures were established on MS medium and Gamborg B5 medium supplemented with different concentrations of BAP, NAA, kinetin and 2,4-D using nodal explants from the field grown mature healthy plant of Adhatoda vasica. After 30 days of culture raised from nodal explants of Adhatoda vasica, maximum number of shoots was produced, on MS medium supplemented with 1.0 mg/l BAP. These explants had developed more than two shoots per nodes, while in other concentration of Kn, NAA and 2,4-D developed either two or less than two shoots/explants. Highest frequency of shoot formation and maximum number of shoots per explants were obtained on MS medium supplemented with 1 mg/I IBA.

In vitro Seed Germination Behaviour of Simmondsia chinensis

Raman Bala — 2020-01-23

Simmondsia chinensis (Link) Schneider (Simmondsiaceae) is a evergreen shrub of desert and marginal land of India. It produces a unique liquid-wax which has commercial potential in pharmaceutical industry. It is difficult to propagate vegetatively as it is a seasonal procedure and grows slowly. Therefore, there is a need to improve the propagation method through seed germination and seedling behaviour. The present investigation was carried out to assess viability and in vitro germination tests of fresh and old seeds. It was found that light (16/8-h day/night photoperiod) conditions resulted in early and higher seed germination percentage as compared to total dark period under in vitro culture conditions. Improvement in germination percentage of one-year-old seeds was observed using Gibberellic acid (GA₃), Benzylaminopurine (BAP) and Thidiazuron (TDZ) under given photoperiod conditions. Interestingly, TDZ 1.0 mg/L^-1 were produced highest germination rate of seeds (92%), as well as better seedling growth, followed by BAP with 70% germination rate at 0.5 mg·L^-1 concentration. The method of propagation through in vitro seed germination could be effectively employed for large scale production of plant material.