In vitro Seed Germination Behaviour of Simmondsia chinensis | Chapter 15 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Dr. Raman Bala
Department of Environmental Science, Maharshi Dayanand University, Rohtak, India.

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In vitro Propagation of a Medicinal Plant Adhatoda vasica Nees by Shoot Bud Culture | Chapter 14 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Dr. P. Soni
Department of Biotechnology, Spcas College, Raipur, Chhattisgarh, India.

Dr. A. N. Bahadur [Professor]
Department of Botany, Government E. Raghavendra Rao Postgraduate Science College, Bilaspur, Chhattisgarh, India.

Dr. V. K. Kanungo [Assistant Professor]
Government Nagarjuna Post Graduate College of Science, Chhattisgarh, India.

Dr. U. Tiwari [Asst. Professor]
Department of Botany, Government E. Raghavendra Rao Postgraduate Science College, Bilaspur, Chhattisgarh, India.

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Investigating Proteomic Variations in Transgenic Tomatoes Compared to Its Non-transgenic Counterpart | Chapter 13 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Professor Hanaa Abdel-Sadek Oraby
Department of Cell Biology, Genetic Engineering and Biotechnology Research Division, National Research Centre, Dokki, Cairo, Egypt.

Professor Amal A. M. Hassan
Department of Cell Biology, Genetic Engineering and Biotechnology Research Division, National Research Centre, Dokki, Cairo, Egypt.

Mahmoud M. Sakr
Academy of Science and Technology, Cairo, Egypt.

Atef A. A. Haiba
Department of Genetics and Cytology, National Research Centre (NRC), Dokki, Cairo, Egypt.

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Residual Effect of Segregated and Unsegregated Urban Solid Waste Compost on Quality of Cowpea (Vigna unguiculata L.) | Chapter 12 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Roohi
Department of Soil Science and Agricultural Chemistry, UAS, GKVK, Bangalore-65, India.

Hari Mohan Meena
Department of Soil Science and Agricultural Chemistry, UAS, GKVK, Bangalore-65, India.

H. C. Prakasha
Department of Soil Science and Agricultural Chemistry, UAS, GKVK, Bangalore-65, India.

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Rapid and Mass Propagation of Hybanthus enneaspermus (L.) F. Muell. from Shoot Tip and Nodal Explants | Chapter 11 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Dr. P. Velayutham
Krishna College of Arts and Science, Kolluthannipatti, Karur Dt. Tamil Nadu, India.

C. Karthi
ICAR – National Research Centre for Banana, Tiruchirappali, Tamil Nadu, India.

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Determination of the Paper Quality as a Substrate for Oyster Mushroom Cultivation | Chapter 10 | Modern Research in Botany Vol. 1

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%).

Author(s) Details

Dr. Siddhant [M.Sc., Ph.D., FAELS, MIAER, FIARA]
Department of Botany, Durgesh Nandini Degree College, Faizabad (U.P.), India.

Dr. O. P. Ukaogo
Department of Industrial Chemistry, Envirnmental/Analytical Units, Abia State University, Nigeria.

S. S. Walakulu Gamage [BSC, MPhil (Reading)]
Department of Botany, Faculty of Science, University of Ruhana, Matara, Sri Lanka.

Dr. Ruchira Singh [M.Sc., Ph.D.]
Independent Researchers, Ayodhya, India.

Mr. Mahesh Kumar [M.Sc., B.Ed. FIARA]
Independent Researchers, Ayodhya, India.

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Phytochemical, GC/MS Analyses and Cytotoxic Effects of Maerua pseudopetalosa (Gilg and Bened.) De Wolf Tuber Fractions | Chapter 09 | Modern Research in Botany Vol. 1

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 F₈ were 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.

Author(s) Details

Dr. Manal A. Ibrahim
Department of Botany, Faculty of Science and Technology, Omdurman Islamic University, Sudan.

El Bushra E. El Nur
Department of Botany, Faculty of Science, University of Khartoum, Sudan.

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Acacia nilotica, Albizia saman, Azadirachta indica: Ethanobotany and Medicinal Uses | Chapter 08 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Parul Tripathi
Amity Institute of Biotechnology, Amity University, Uttar Pradesh, Lucknow Campus, Gomti Nagar Extension, Lucknow-226028, India.

Aditi Singh
Amity Institute of Biotechnology, Amity University, Uttar Pradesh, Lucknow Campus, Gomti Nagar Extension, Lucknow-226028, India.

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Extraction of High-quality Genomic DNA from Different Plant Orders Applying a Modified CTAB-Based Method | Chapter 07 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Dr. Nadia Aboul-Ftooh Aboul-Maaty
Department of Cell Biology, Genetic Engineering and Biotechnology Research Division, National Research Centre, Dokki, Cairo, Egypt.

Professor Hanaa Abdel-Sadek Oraby
Department of Cell Biology, Genetic Engineering and Biotechnology Research Division, National Research Centre, Dokki, Cairo, Egypt.

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Effect of Light and Temperature through Poly Film Covers on Anthocyanin Content in Rose Cut Flower | Chapter 06 | Modern Research in Botany Vol. 1

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.

Author(s) Details

Dr. G. Oloo-Abucheli
Department of Plant Sciences, Chuka University, P.O.Box 109-60400, Chuka, Kenya and Department of Crops, Horticulture and Soil Sciences, Egerton University, P.O.Box 536, Egerton, Kenya.

Prof. J. N. Aguyoh
Department of Agriculture and Environmental Studies, Rongo University College, P.O.Box 1023-40404, Rongo, Kenya.

G. Liu
Department of Crops, Horticulture and Soil Sciences, Egerton University, P.O.Box 536, Egerton, Kenya.

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