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Micropropagation through nodal explants was attempted for a rare and endemic taxon Corynandra chelidonii var. pallae (Cleomaceae) in view of its medicinal uses, bioactive compounds, declining natural populations due to intermittent sterility, low seed set and erratic seed germination. Multiple shoots were regenerated directly from the nodal explants on Murashige and Skoog’s medium containing different concentrations and combinations of cytokinins and auxins. High-frequency multiple shoots of 8.15 ± 0.249 with 1.63 ± 0.031 shoot length (cm) were obtained on MS medium augmented with the combination of 0.5 mg/L of thidiazuron and 1.0 mg/L of indole-3-acetic acid. Directly regenerated shoots were rooted with 95% root induction on a half-strength MS medium supplemented with 1.0 mg/L of indole-3 butyric acid. The in vitro raised plantlets were hardened in plastic pots and acclimatized initially in the greenhouse conditions and later transferred to the field conditions with a 78% of survival rate. The genetic fidelity of in vitro generated and field transferred plantlets with that of the mother plant was checked by carrying ISSR marker-based polymerase chain reaction method. This protocol on direct organogenesis and genetic fidelity analysis in Corynadra chellidonii var. pallae can be successfully employed for large-scale multiplication and conservation.

Rauwolfia tetraphylla L., is an important medicinal plant in Apocynaceae family and is recognized as an alternative source to Rauwolfia serpentina L., in terms of anti-hypertensive alkaloid production i.e. reserpine. In view of this, the present study is conducted to estimate the reserpine content in different parts (leaf, stem and root) of field grown plants (2 years old), tissue cultured plantlets (R1) (two months old) and cell suspensions cultures (two months old with and without precursor feeding) of R. tetraphylla by using high performance liquid chromatography (HPLC) technique. Overall maximum content of reserpine (in %) was estimated from the root samples. Roots of field grown plants has recorded high percent of reserpine (0.39%) followed by roots of tissue cultured plantlets (0.35%) and root callus based cell suspension cultures (0.38 %) which was fed with precursor amino acid (100 mg/L of tryptophan). In control type of root callus based cell suspension cultures, reserpine content was quantified as 0.14%; by precursor feeding (100 mg/L of tryptophan) it was enhanced to 0.38%. In conclusion, the reserpine content (0.35 and 0.38%) produced by the roots of tissue cultured plantlets (R1) and 100 mg/L tryptophan fed root callus based cell suspensions was comparable to that of the reserpine content (0.39%) of root parts of field grown plants. The present study demonstrates the reserpine production by in vitro cell suspension cultures throughout the year without sacrificing the medicinal plants.

An efficient and reproducible protocol was developed for in vitro clonal propagation of Solanum trilobatum L., an ethno-medicinally important plant. Leaf, stem and cotyledon explants were used for callus induction and shoot regeneration via indirect organogenesis. Initially, maximum amounts (mg) of green friable callus (312.86 ± 0.50, 285.3 ± 0.40 and 305.13 ± 0.62) was induced from leaf, stem and cotyledon explants, respectively, on Murashige and Skoog (MS) medium supplemented with 13.57 μM L−1 of 2,4-dichloro phenoxy acetic acid (2,4-D) and 2.21 μM L−1 of 6-benzylaminopurine (BAP), after 4 weeks of culture. Highest mean number of shoots (69.2 ± 0.73, 34.1 ± 0.62 and 57.1 ± 0.62) were differentiated de novo from leaf, stem and cotyledon calluses, respectively, when cultured on MS medium amended with 2.27 μM L−1 of Thidiazuron (TDZ) and 2.68 μM L−1 of Naphthaleneacetic acid (NAA), after 4 weeks of culture. Maximum rooting response (97%) with mean number of roots (31.7 ± 0.61 roots per shoot) was observed from shoots when cultured on half strength MS medium supplemented with 4.90 μM L−1 of indole-3-butyric acid (IBA), after 4 weeks of culture. In vitro raised plantlets (R1) of S. trilobatum were hardened in plastic pots, acclimatized in green house and successfully transferred to field conditions with 82% survivability. ISSR and RAPD based PCR banding profile of the acclimated R1 plantlets was confirmed as synonymous to the mother plant.

Advancement in plant biology using high-throughput molecular tools has offered several exciting opportunities to gain new insight towards mechanisms controlling growth and developmental processes. In the past few years, omics technology has shown tremendous potential in the field of plant tissue culture. This chapter focuses on the adoption of omics-based techniques such as genomics, transcriptomics and proteomics to stimulate research interest in model plants and agriculturally important crops through tissue culture. It provides information about new candidate genes which encode for signalling and regulatory proteins that control early embryogenesis and morphogenesis. Omics has given an alternative mean for tissue culture to characterize the genomic data for marker development related to somaclonal variants. The large amount of data generated using these tools provide a quantum increase in knowledge related to differentiation, redifferentiation, somaclonal variation and somatic embryogenesis and lay foundation for analysing tissue culture samples at cellular and molecular levels.

Mulberry (Morus alba L.) is the sole food source for the mulberry silkworm, Bombyx mori and therefore important for sericulture industry. Different abiotic stress conditions like drought, salt, heat and cold stress adversely affect the productivity and quality of mulberry leaves. Quantitative real time PCR (qPCR) is a reliable and widely used method to identify abiotic stress responsive genes and molecular mechanism in different plant species. Selection of suitable reference genes is important requirement for normalizing the expression of genes through qRT-PCR study. In the present study, we have selected eight candidate reference genes in mulberry for analyzing their expression stability in different abiotic stress treatments including drought, salt, heat and cold stresses. The expression stability of these reference genes was determined using geNorm, NormFinder and RefFinder statistical algorithms. The results showed that Ubiquitin and protein phosphatase 2A regulatory subunit A (PP2A) were the most stable genes across all the treatment samples. However, analysis of individual stresses revealed different expression profiles and stability of reference genes. Actin3 and PP2A were most stable in drought and salt conditions respectively. RPL3 most preferred in heat stress and Ubiquitin was most stable in cold stress. We propose the ubiquitin and PP2A are the preferred reference genes for normalization of gene expression data from abiotic stresses. In addition, Actin3 are preferred for drought stress, PP2A for salt stress, RPL3 for heat stress and Ubiquitin for cold stress studies.

The coronavirus disease (COVID-19) is highly pathogenic viral infection caused by SARS-CoV-2. Currently, COVID-19 has caused global health concern. It is assumed that COVID-19 has zoonotic origin based on the large number of infected people who were exposed to the wet market in Wuhan City, China. The phylogenetic analysis has revealed that SARS-CoV-2 has significant sequence similarity with severe acute respiratory syndrome-like (SARS-like) bat viruses, thus bats could be primary possible reservoir. The intermediate host and there subsequent transfer is not known yet, although human to human transfer is widely confirmed. The transmission of COVID-19 infection from one person to another resulted in the isolation of patients who were subsequently given a variety of treatments. To monitor the current outbreak, robust steps have been taken around the globe to reduce the transmission of COVID-19 infection particularly banning international and domestic flights, inducting lockdowns in vulnerable areas, social distancing etc. No clinically approved antiviral drug or vaccine against COVID-19 is reported yet. However, in clinical trials, few broad-spectrum antiviral drugs were evaluated against COVID-19 infection which resulted in clinical recovery. In this article emergence and pathogenicity of COVID-19 infection along with potential therapeutic strategies are analyzed to combat the COVID-19 pandemic.

A reliable protocol was developed for in vitro micro propagation of Morus alba L.cv. Chinese white. Initially, friable callus was induced (242.8 and 128.5 mg) from in vivo leaf and nodal explants on Murashige and Skoog’s (MS) medium amended with 4.0 μM/L of 2,4-Dichlorophenoxyacetic acid (2,4-D) and 3.0 μM/L of Naphthalene acetic acid (NAA) respectively within 3 weeks. Shoot regeneration (12.2 and 8.6) was obtained from leaf and node derived callus on 6-benzylaminopurine (BAP) + Thidiazuron (TDZ) at 2.5 + 2.0 and 7.5 + 2.0 μM/L concentrations respectively, after 4 weeks of incubation. In vitro shoots were rooted (90 %) on half strength MS medium with 7.5 μM/L indole-3 butyric acid (IBA) and plantlets were hardened in plastic pots contained farmyard manure, sand and garden soil in 1:1:2 ratio. The genetic stability of plantlets were confirmed by start codon targeted (SCoT) and inter simple sequence repeats (ISSR) primers based molecular analysis. Keywords: Mulberry, Callus mediated regeneration, Acclimatization, Start Codon targeted and inter simple sequence repeats

Introduction:Nanotechnology, an advanced science discipline has proved to be vital in solving the major issues or problems, the present world is facing. Nanotechnology has already proved to be the science of revolutionizing agriculture, energy conversions, applied & medical science and other domains.Conclusion:In this paper, we present the recent developments taken place in bio-nanotechnology related to different forms of nanomaterial’s developed along with their application; overcoming hazardous effects of chemical pesticides, fertilizers and herbicides by promoting green and sustainable agriculture through the use of nanofertilizers, nano pesticides and detection & control of plant diseases by using nanoparticles; development of diagnostic tools for detection and control of human diseases; targeted delivery of drugs by using nanomaterial’s, protection of the environment through nanoparticles based pollution checking, bioremediation, renewable energy production and role of nanotechnology in applied sciences.

Mulberry is a tree species; hence, it is difficult to carry out the indirect generation. In view of this limitation, in present study an indirect regeneration protocol was developed for in vitro clonal propagation of sericulturally important superior temperate mulberry variety cv. Pampore-1 (PPR-1) by using leaf, petiole and nodal based callus. Initially friable callus (mean fresh weight) was induced in maximum amounts (288.2 ± 21.09; 248.2 ± 16.52 & 138.4 ± 04.25) from leaf, petiole and nodal explants on Murashige and Skoog (MS) media supplemented with 5.0 μM/L, 7.5 μM/L of 2,4-Dichlorophenoxyacetic acid (2,4-D) and 12.5 μM/L of Naphthalene acetic acid (NAA), respectively, after 3 weeks of culture. Best response of shoot regeneration from the induced callus with maximum frequency (14.8 ± 0.82) was observed in leaf callus on MS media supplemented with 2.5 μM/L of Thidiazuron (TDZ) + 2.5 μM/L of 6-Benzyl amino purine (BAP). The regenerated shoots were rooted with maximum rooting frequency (96 ± 02.2) on MS media with 7.5 μM/L concentration of Indole-3-Butyric acid (IBA). The raised plantlets were then hardened using 1:1:2 ratio of farmyard manure, sand and garden soil, then gradually they were transferred and acclimatized to field conditions. The survival rate of in vitro raised PPR-1 plantlets in field conditions is about 82%. Genetic homogeneity between the micropropagated plants and mother plant was confirmed by carrying out the inter simple sequence repeats (ISSR) primer based polymerase chain reaction (PCR) analysis. Among the ten ISSR primers used, two primers, i.e., M5 and M8 has given good amplification with clear, distinct and scorable DNA bands which were monomorphic across the micropropagated plantlets and the mother plant. Hence, the in vitro regenerated PPR-1 mulberry plantlets were confirmed as clonally uniform and genetically stable.

The present world is facing environmental management problem as the major issue in almost every country across the globe; this is due to increased human population which in turn has led to the rapid urbanization and industrialization which has delivered lot of non-biodegradable chemical substances in to the environment. Similarly, to feed the increased human population by doubling the agricultural productivity, all countries have adopted usage of highly aromatic and toxic pesticides in agriculture sector. This might have solved the problem of food security to some extent, but usage of high amounts of organic pesticides has contaminated the environmental sites such as water bodies, air and soil systems. Due to all of these anthropogenic stresses, now the environment has changed drastically and posed biodiversity loss and public health issues which has attracted the attention of environmentalists for the cleanup and restoration of environment at an earliest mode. As compared to conventional and natural methods; by using the biotechnological tools and applications it is an easier, time saving, safer, and sustainable way to remediate the contaminated sites. In this chapter, we represent an overview on role of biotechnology in pesticide remediation from contaminated freshwater and soils.

Phenolics are one of the most abundant groups of secondary metabolites found throughout the plant kingdom. Phenolic compounds are widely distributed in plant tissues, particularly contributing to color, flavor, taste, and astringency to fruits. Phenolics are aromatic benzene ring compounds with one or more hydroxyl groups attached to it. Phenolics constitute most likely the largest group of plant secondary metabolites, varying in size from a simple structure with an aromatic ring to complex ones such as lignins. Most phenolic compounds are thought to be by-products of the metabolism of the aromatic amino acid phenylalanine. Phenolics are considered to play a vital role in the evolution of plants by providing an adaptive advantage to cope up with the changing environments. Furthermore, they are believed to be involved in leaf and petal senescence. Change in the leaf or petal color during senescence is also due to anthocyanin pigments, a kind of plant phenolics.

In plants among the different phytochemicals, phenolic and polyphenolic compound-based research has gained rapid momentum during the present decade. Even though the phenolic compounds are not required in primary processes such as growth and development of a plant, these compounds which are considered as secondary metabolites have gained a lot of attention of researchers across the world for their role in diversified functions of a plant system. Majorly phenolic compound-based research is being carried out to understand the role of various classes of phenols right from their role in adapting a plant species to different environments which encompasses biotic and abiotic factors to understanding their role in reproduction, in interaction with other biomolecules, and in specified function of every individual phenolic compound. More advanced research is also being carried to know the action of phenolic compounds on the enzymes involved in the epigenetic regulation of gene expression. From the 1970s, the plant phenolic-based research has gained importance, and researchers have started identifying the new phenolic compounds from several plant species. Now, through this book chapter, we put together the newly identified phenolic compounds of the present decade (i.e., 2010–April 2019) which were identified from different plant families.

Highlights • A reliable protocol developed for micropropagation of Morus alba L.cv. Chinese white through indirect regeneration using different explants. • Friable callus was induced in maximum amounts from leaf and nodal explants on MS media supplemented with 2,4-D and NAA respectively. • Highest frequency of regeneration was obtained from the leaf and node derived callus on MS media with BAP + TDZ and plantlets were rooted on IBA. • Genetic homogeneity of in vitro raised plants was confirmed by SCoT and ISSR primers based molecular analysis., A reliable protocol was developed for in vitro micro propagation of Morus alba L.cv. Chinese white. Initially, friable callus was induced (242.8 and 128.5 mg) from in vivo leaf and nodal explants on Murashige and Skoog’s (MS) medium amended with 4.0 μM/L of 2,4-Dichlorophenoxyacetic acid (2,4-D) and 3.0 μM/L of Naphthalene acetic acid (NAA) respectively within 3 weeks. Shoot regeneration (12.2 and 8.6) was obtained from leaf and node derived callus on 6-benzylaminopurine (BAP) + Thidiazuron (TDZ) at 2.5 + 2.0 and 7.5 + 2.0 μM/L concentrations respectively, after 4 weeks of incubation. In vitro shoots were rooted (90 %) on half strength MS medium with 7.5 μM/L indole-3 butyric acid (IBA) and plantlets were hardened in plastic pots contained farmyard manure, sand and garden soil in 1:1:2 ratio. The genetic stability of plantlets were confirmed by start codon targeted (SCoT) and inter simple sequence repeats (ISSR) primers based molecular analysis.

There are several studies about the phytochemical investigations and pharmacological activities of Andrographis echioides (L.), but there is scanty information about its in vitro propagation. Hence the present study is undertaken for in vitro micropropagation of Andrographis echioides (L.) through direct organogenesis. Initially, aseptic leaf explants of in vitro seedlings were excised and inoculated on to plant growth regulators (PGR’s) supplemented in the Murashige and Skoog (MS) media. Among the individual and combinations of PGR’s tested, the combination of thidiazuron (TDZ) along with indole-3-acetic acid (IAA) at 0.5 and 0.2 mg/L concentrations respectively, was proved to be best for high frequency (24.48 ± 1.54) of direct shoot regeneration of A. echioides. Rooting of in vitro raised shoots with the highest rooting percent of 90.42 was carried on IAA (1.0 mg/L), and indole-3-butyric acid (IBA) (1.0 mg/L) amended MS media. in vitro plantlets of A. echioides were hardened in poly cups possessing a 1:2 ratio of compost and garden soil. Initially, hardened plantlets were maintained in a greenhouse for four weeks by irrigating with half-strength MS salt solution and later transferred to field conditions with 80 % survivability. Genetically clonal nature of regenerated plantlets was confirmed by carrying polymerase chain reaction (PCR) based molecular analysis using inter simple sequence repeats (ISSR) and ribulose bisphosphate carboxylase large chain (rbcLa) primers.

Rauwolfia tetraphylla L. is an important medicinal plant species which is well known for its pharmaceutically important alkaloids. In the present study, we are reporting about its conservation by in vitro clonal multiplication through the standardized protocol of indirect regeneration by using leaf and stem based callus and assessment of genetic fidelity of acclimated plantlets by start codon targeted (SCoT), inter simple sequence repeats (ISSR), and randomly amplified polymorphic DNA (RAPD) marker based analysis. Initially friable callus was induced in maximum amounts (378.7, 323.8, and 412.8 in mg) from leaf, root, and stem explants on Murashige and Skoog (MS) media supplemented with 5.0 mg/L, 3.0 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.0 mg/L of naphthalene acetic acid (NAA), respectively. Shoot regeneration with the maximum number of shoot buds (25 and 20) was obtained from leaf and stem calluses on MS media supplemented with TDZ (0.25 mg/L) + BAP (2 mg/L). The regenerated shoots were rooted successfully with maximum rooting percentage of 98.0 on full strength MS media amended with IAA (1.0 mg/L) and IBA (1.0 mg/L). The regenerated plantlets were hardened using 2:1 ratio of sterile garden soil and sand, followed by acclimatization in field conditions with 86% of survival. SCoT, ISSR, and RAPD primers based polymerase chain reaction (PCR) analysis was carried out to check possible genetic variations in micro propagated plants in comparison with mother plant. Among the ten SCoT (S), ISSR (R), and RAPD (OPA) primers used, S2, R10, and OPA3 has given good amplification with scorable DNA bands. The results revealed that the regenerated plants did not have any polymorphism with mother plant. Hence, the in vitro regenerated R. tetraphylla plantlets were confirmed as true-to-type.

Rauwolfia tetraphylla L. is an important medicinal plant species which is well known for its pharmaceutically important alkaloids. In the present study, we are reporting about its conservation by in vitro clonal multiplication through the standardized protocol of indirect regeneration by using leaf and stem based callus and assessment of genetic fidelity of acclimated plantlets by start codon targeted (SCoT), inter simple sequence repeats (ISSR), and randomly amplified polymorphic DNA (RAPD) marker based analysis. Initially friable callus was induced in maximum amounts (378.7, 323.8, and 412.8 in mg) from leaf, root, and stem explants on Murashige and Skoog (MS) media supplemented with 5.0 mg/L, 3.0 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) and 5.0 mg/L of naphthalene acetic acid (NAA), respectively. Shoot regeneration with the maximum number of shoot buds (25 and 20) was obtained from leaf and stem calluses on MS media supplemented with TDZ (0.25 mg/L) + BAP (2 mg/L). The regenerated shoots were rooted successfully with maximum rooting percentage of 98.0 on full strength MS media amended with IAA (1.0 mg/L) and IBA (1.0 mg/L). The regenerated plantlets were hardened using 2:1 ratio of sterile garden soil and sand, followed by acclimatization in field conditions with 86% of survival. SCoT, ISSR, and RAPD primers based polymerase chain reaction (PCR) analysis was carried out to check possible genetic variations in micro propagated plants in comparison with mother plant. Among the ten SCoT (S), ISSR (R), and RAPD (OPA) primers used, S2, R10, and OPA3 has given good amplification with scorable DNA bands. The results revealed that the regenerated plants did not have any polymorphism with mother plant. Hence, the in vitro regenerated R. tetraphylla plantlets were confirmed as true-to-type.

Indirect somatic embryogenesis and indirect regeneration protocol was developed for the first time from leaf and nodal explants of Mirabilis jalapa Linn. Initially callus was induced in leaf and nodal explants cultured on MS medium supplemented with various concentrations of 2, 4-D, NAA and Kn, after 3 weeks of culture. Maximum amount of mean fresh and dry weight (mg) of callus(189.6 ± 0.25, 7.6 ± 0.23); (198.4± 0.34, 9.2 ± 0.23)was obtained from leaf and nodal explants, respectively when cultured on MS media supplemented with combinations of 2,4-D(3 mg/L) and NAA(5mg/L).At high concentrations of NAA (5 mg/L), somatic embryos are induced from the nodal explants after 4 weeks of culture. Shoot buds were induced from the leaf and nodal calluses when cultured on combinations of TDZ & BAP after 3 weeks of culture. Maximum number of shoot buds (08 ± 0.17 & 06 ± 0.09) were produced from leaf and nodal calluses cultured on MS + TDZ (0.5mg/L) +BAP (2 mg/L).When micro shoots were transferred to rooting media, the best rooting percentage (100%) was obtained on MS + IAA (1.0 mg/L) + IBA (2.0 mg/L). The raised plantlets when transferred to soil for acclimatization, 75% plantlets survived in field conditions.

Leaf and stem explants of Rauwolfia tetraphylla (syn.R. canescens), an alternative of endangered medicinal plant, R. serpentina cultured on MS + 2,4-D (2.26 μM.L−1) + Kn (9.29 μM.L−1) produced green and compact callus after 4 weeks of culture. When these primary calluses were sub-cultured successively for 3 passages, they became light green and fragile and when they were grown on MS + TDZ (2.27 μM.L−1), they produced highest no of multiple shoot buds. Individual microshoots were successfully rooted on MS+IAA (5.37 μM.L−1), and 88% of the plantlets survived in field conditions. ISSR-specific primer HY3 banding profile of the plantlets are similar to the mother plant.

In this present study, a protocol was developed for the DNA profiling of medicinally important Rauwolfia species (05 species) by ISSR-PCR method. Initially genomic DNA was isolated from the fresh leaves (1 gram) of 24 months old Rauwolfia tetraphylla, R.serpentina, R.pentaphylla , R.vomitaria and R.micrantha. The content of genomic DNA isolated from 1 gram of fresh leaves of these five species ranges from 1.6 to 3.4 µg. Out of six inter simple sequence repeats (ISSR) primers used, HY3 primer has given the good amplification with distinct bands which were reproducible. The range of DNA bands produced by the HY3 primer were between 300 bp to 1500 bp among the five species. A dendrogram was constructed on the basis of similarity matrix data by unweighted pair group method with arithmetic average (UPGMA) cluster analysis using indigenous software. Average genetic similarity generated by HY3 has revealed 38% similarity between species of Sub cluster-A and species of Sub cluster-B which is consistent with their geographical distribution i.e. Sub cluster-A species R.tetraphylla and R.serpentina inhabit peninsular India, Sub cluster- B species R.micrantha and R. vomitoria inhabit the Himalayan foot hills. The developed protocol of DNA profiling in Rauwolfia species by ISSR-PCR method could be used efficiently for the differentiation and identification of the Rauwolfia species at molecular level.