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The basic helix-loop-helix (bHLH) is the second-largest TF family in plants that play important roles in plant growth, development, and stress responses. In this study, a total of 100 bHLHs were identified using Hidden Markov Model profiles in the Nicotiana tabacum genome, clustered into 15 major groups (I–XV) based on their conserved domains and phylogenetic relationships. Group VIII genes were found to be the most abundant, with 27 NtbHLH members. The expansion of NtbHLHs in the genome was due to segmental and tandem duplication. The purifying selection was found to have an important role in the evolution of NtHLHs. Subsequent qRT-PCR validation of five selected genes from transcriptome data revealed that NtbHLH3.1, NtbHLH3.2, NtbHLH24, NtbHLH50, and NtbHLH59.2 have higher expressions at 12 and 24 h in comparison to 0 h (control) of chilling stress. The validated results demonstrated that NtbHLH3.2 and NtbHLH24 genes have 3 and fivefold higher expression at 12 h and 2 and threefold higher expression at 24 h than control plant, shows high sensitivity towards chilling stress. Moreover, the co-expression of positively correlated genes of NtbHLH3.2 and NtbHLH24 confirmed their functional significance in chilling stress response. Therefore, suggesting the importance of NtbHLH3.2 and NtbHLH24 genes in exerting control over the chilling stress responses in tobacco.

Zinc (Zn) is a vital micronutrient in both plants and humans for healthy growth and development. The lesser Zn accessibility causes about 20% yield loss along with low Zn content in grains. About 30% of the human population in the world rely on Zn deficient diets. Dietry Zn deficiency causes impairment of physical growth, immune system functioning, reproductive health, and neurobehavioral development in humans. In various physiological processes, Zn plays a key role and serves as a cofactor for various enzymes and proteins in numerous essential biochemical pathways in both plant and animal. Consequently, it is important to increase Zn content of cereal grains such as rice, maize and wheat. Many investigations have been accomplished to improve Zn deficiency tolerance and improving Zn content in grains. In this regard, improving Zn use efficiency is the most meaningful approach that involves modifying root system architecture, solubilization of Zn complex by organic acids, root exudates, and Zn uptake and translocation mechanism in plants. Here we present an outlook of different biotechnological approaches to improve Zn use efficiency and producing cereals with superior grain quality.

One of the most interesting signaling molecules that regulates a wide array of adaptive stress responses in plants are the micro RNAs (miRNAs) that are a unique class of non-coding RNAs constituting novel mechanisms of post-transcriptional gene regulation. Recent studies revealed the role of miRNAs in several biotic and abiotic stresses by regulating various phytohormone signaling pathways as well as by targeting a number of transcription factors (TFs) and defense related genes. Phytohormones are signal molecules modulating the plant growth and developmental processes by regulating gene expression. Studies concerning miRNAs in abiotic stress response also show their vital roles in abiotic stress signaling. Current research indicates that miRNAs may act as possible candidates to create abiotic stress tolerant crop plants by genetic engineering. Yet, the detailed mechanism governing the dynamic expression networks of miRNAs in response to stress tolerance remains unclear. In this review, we provide recent updates on miRNA-mediated regulation of phytohormones combating various stress and its role in adaptive stress response in crop plants.

Plant responds to Agrobacterium via three-layered immunity that determines its susceptibility or resistance to Agrobacterium infection. Agrobacterium tumefaciens is a soil-borne Gram-negative bacterium that causes crown gall disease in plants. The remarkable feat of interkingdom gene transfer has been extensively utilised in plant biotechnology to transform plant as well as non-host systems. In the past two decades, the molecular mode of the pathogenesis of A. tumefaciens has been extensively studied. Agrobacterium has also been utilised as a premier model to understand the defence response of plants during plant-Agrobacterium interaction. Nonetheless, the threat of Agrobacterium-mediated crown gall disease persists and is associated with a huge loss of plant vigour in agriculture. Understanding the molecular dialogues between these two interkingdom species might provide a cure for crown gall disease. Plants respond to A. tumefaciens by mounting a three-layered immune response, which is manipulated by Agrobacterium via its virulence effector proteins. Comparative studies on plant defence proteins versus the counter-defence of Agrobacterium have shed light on plant susceptibility and tolerance. It is possible to manipulate a plant's immune system to overcome the crown gall disease and increase its competence via A. tumefaciens-mediated transformation. This review summarises the recent advances in the molecular mode of Agrobacterium pathogenesis as well as the three-layered immune response of plants against Agrobacterium infection.

The present study demonstrates the Ni toxicity-ameliorating and growth-promoting abilities of two different bacterial isolates when applied to wheat (Triticum aestivum L.) as the host plant. Two bacterial strains tolerant to Ni stress were isolated from wheat seeds and selected based on their ability to improve the germination of wheat plants, they were identified as Bacillus megaterium AFI1 and Paenibacillus nicotianae AFI2. The protective effects of these epiphytic bacteria against Ni stress were studied in model experiments with two wheat cultivars: Ni stress-tolerant Leningradskaya 6 and susceptible Chinese spring. When these isolates were used as the inoculants applied to Ni-treated wheat plants, the growth parameters and the levels of photosynthetic pigments of the two wheat cultivars both under normal and Ni-stress conditions were increased, though B. megaterium AFI1 had a more pronounced ameliorative effect on the Ni contents in plant tissues due to its synthesis of siderophores. Over the 10 days of Ni exposure, the plant growth promotion bacteria (PGPB) significantly reduced the lipid peroxidation (LPO), ascorbate peroxidase (APX), superoxide dismutase (SOD) activities and proline content in the leaves of both wheat cultivars. The PGPB also increased peroxidase (POX) activity and the levels of chlorophyll a, chlorophyll b, and carotenoids in the wheat leaves. It was concluded that B. megaterium AFI1 is an ideal candidate for bioremediation and wheat growth promotion against Ni-induced oxidative stress, as it increases photosynthetic pigment contents, induces the antioxidant defense system, and lowers Ni metal uptake.

OsGSTU5 interacts and glutathionylates the VirE2 protein of Agrobacterium and its (OsGSTU5) overexpression and downregulation showed a low and high AMT efficiency in rice, respectively. During Agrobacterium-mediated transformation (AMT), T-DNA along with several virulence proteins such as VirD2, VirE2, VirE3, VirD5, and VirF enter the plant cytoplasm. VirE2 serves as a single-stranded DNA binding (SSB) protein that assists the cytoplasmic trafficking of T-DNA inside the host cell. Though the regulatory roles of VirE2 have been established, the cellular reaction of their host, especially in monocots, has not been characterized in detail. This study identified a cellular interactor of VirE2 from the cDNA library of rice. The identified plant protein encoded by the gene cloned from rice was designated OsGSTU5, it interacted specifically with VirE2 in the host cytoplasm. OsGSTU5 was upregulated during Agrobacterium infection and involved in the post-translational glutathionylation of VirE2 (gVirE2). Interestingly, the in silico analysis showed that the 'gVirE2 + ssDNA' complex was structurally less stable than the 'VirE2 + ssDNA' complex. The gel shift assay also confirmed the attenuated SSB property of gVirE2 over VirE2. Moreover, knock-down and overexpression of OsGSTU5 in rice showed increased and decreased T-DNA expression, respectively after Agrobacterium infection. The present finding establishes the role of OsGSTU5 as an important target for modulation of AMT efficiency in rice.

Nickel (Ni) is an essential element for plant growth and is a constituent of several metalloenzymes, such as urease, Ni-Fe hydrogenase, Ni-superoxide dismutase. However, in high concentrations, Ni is toxic and hazardous to plants, humans and animals. High levels of Ni inhibit plant germination, reduce chlorophyll content, and cause osmotic imbalance and oxidative stress. Sustainable plant-bacterial native associations are formed under Ni-stress, such as Ni hyperaccumulator plants and rhizobacteria showed tolerance to high levels of Ni. Both partners (plants and bacteria) are capable to reduce the Ni toxicity and developed different mechanisms and strategies which they manifest in plant-bacterial associations. In addition to physical barriers, such as plants cell walls, thick cuticles and trichomes, which reduce the elevated levels of Ni entrance, plants are mitigating the Ni toxicity using their own antioxidant defense mechanisms including enzymes and other antioxidants. Bacteria in its turn effectively protect plants from Ni stress and can be used in phytoremediation. PGPR (plant growth promotion rhizobacteria) possess various mechanisms of biological protection of plants at both whole population and single cell levels. In this review, we highlighted the current understanding of the bacterial induced protective mechanisms in plant-bacterial associations under Ni stress.

Dehydrins (DHNs) have been reported to act as chaperones to combat drought stress. While in this study, we explored a novel nuclear complex of Oryza sativa FKBP and YSK2-type dehydrin (OsDhn-Rab16D), associated with ABA signaling to impart drought stress tolerance. The transcript levels of OsDhn-Rab16D in rice seedlings were induced in response to drought, abscisic acid (ABA) and H2O2 exposure. Ectopic expression of OsDhn-Rab16D transgenic lines showed enhanced tolerance to both osmotic stresses caused by PEG and drought. Using the yeast two-hybrid (Y2H) assay, O. sativa FKBP (a prolyl cis-trans isomerase) was identified as an interacting partner of OsDhn-Rab16D. Fluorescence signals between OsDhn-Rab16D and OsFKBP were observed in the nucleus. The pull-down assay confirmed the physical interaction between OsDhn-Rab16D and OsFKBP. qRT-PCR of drought and ABA-responsive gene reveals the higher transcript abundance in PEG-treated transgenic lines. Moreover, under drought conditions, transgenic lines maintain membrane integrity and increase the lignification in adventitious rice roots due to the higher expression level of catalase and lignin biosynthesis enzyme, respectively, as compared with wild-type. Overall, our findings suggest that OsDhn-Rab16D and OsFKBP complex involved in ABA-responsive drought stress signaling in rice and probably act as a positive transcriptional co-regulator, in addition, to act as chaperones.

Class III peroxidases are secretory enzymes which belong to a ubiquitous multigene family in higher plants and have been identified to play role in a broad range of physiological and developmental processes. Potentially, it is involved in generation and detoxification of hydrogen peroxide (H2O2), and their subcellular localization reflects through three different cycles, namely peroxidative cycle, oxidative and hydroxylic cycles to maintain the ROS level inside the cell. Being an antioxidant, class III peroxidases are an important initial defence adapted by plants to cope with biotic and abiotic stresses. Both these stresses have become a major concern in the field of agriculture due to their devastating effect on plant growth and development. Despite numerous studies on plant defence against both the stresses, only a handful role of class III peroxidases have been uncovered by its functional characterization. This review will cover our current understanding on class III peroxidases and the signalling involved in their regulation under both types of stresses. The review will give a view of class III peroxidases and highlights their indispensable role under stress conditions. Its future application will be discussed to showcase their importance in crop improvement by genetic manipulation and by transcriptome analysis.

The medicinal plant, Solanum viarum Dunal includes a number of compounds with important pharmacological effects. The effect of temperature (heat and cold) stress on growth, secondary metabolite levels and defense mechanism in two genotypes (prickled and prickleless) of S. viarum was evaluated. The two thermal regimes (4 °C for cold and 35 °C for heat stress) were found to be the most optimum for the accumulation of metabolites (steroidal alkaloids and glycoalkaloids, phenolic acids as well as flavonoids). Prickleless genotype showed 4.08 and 1.51 fold higher total alkaloids/glycoalkaloid content at 35 °C and 4 °C temperatures respectively, in comparison with prickled plants. Prickleless plants also registered 2.03 and 2.28 higher fresh and dry biomass accumulation respectively, under cold stress over the control plants. The accretion of phenolic and flavonoid compounds under heat and cold stress is mainly contributed by gallic and ferulic acid in both the genotypes. The quantitative real-time PCR expression analysis showed the abundance of gene transcript involve in the biosynthesis of alkaloids/glycoalkaloid and phenolics/flavonoids that corroborates with the accumulation of their respective metabolites in prickleless plants under temperature stress. Results revealed that the prickleless plants inhibiting the reactive oxygen species (ROS) mediated oxidative damage by activating the enzymatic (superoxide dismutase, peroxidase, catalase and ascorbate peroxidase) and non-enzymatic (alkaloids, phenols, flavonoids, carotenoids and proline) antioxidants mechanism that confirms its efficiency to tolerate thermal stress under both the thermal regimes. The overall result showed that the prickleless genotype served as a better accumulator of biomass, secondary metabolites with an improved antioxidative mechanism in comparison to prickled genotype. Our findings suggest that thermal stress responses differ significantly between genotypes, emphasizing the necessity of maintaining genotypic diversity in adaptive evolution as in the context of global warming and climate change. The present study also opens the possibility of exploiting improved prickleless genotype of S. viarum as a potential system that can be cultivated under different temperature stress conditions to attain the phytopharmaceutical benefits.

The objective of this work was to study the potential of a constructed bacterial consortium (comprised three strains: Stenotrophomonas maltophilia, Bacillus cereus, and Bacillus anthracis) to treat distillery wastewater via the bioaugmentation process. The discharged wastewater showed elevated total ammonium nitrogen (195.0 ± 1.24 mg L−1), total dissolved solids (25980.6 ± 8.09 mg L−1), chemical oxygen demand (20534.5 ± 3.12 mg L−1), and biological oxygen demand (20534.5 ± 3.12 mg L−1). High concentration of heavy metals, phenolic and organo-metallic compounds were also detected. Results showed that growing the bacterial consortium in the distillery wastewater at 37 °C supplemented with 1% glucose achieved the best color reduction (up to 90%) in 144 h. The physico-chemical quality of the treated wastewater also improved by 50%–70%. Furthermore, many of the major organic pollutants present in the distillery wastewater were degraded by the constructed consortium to below detection limit via active biotransformation and biodegradation. Heavy metals were biosorbed by the bacterial consortium, and the ligninolytic enzymes such as Lip and MnP played an important role in the degradation of the organo-metallic pollutants. The constructed bacterial consortium therefore offered a sustainable and effective solution to treat distillery wastewater.

Arsenic contamination in agricultural soil leads to the transfer of arsenic into the food-chain and adversely affects the human health. Generation of genetically engineered plants to transform inorganic arsenic to methylated and volatile arsenic species is one of the efficient strategy to lower arsenic contamination. In the present study, we genetically engineered Arabidopsis thaliana with arsenic methyltransferase ( WaarsM ) gene of a fungus Westerdykella aurantiaca , isolated from arsenic-contaminated sites of West Bengal. The WaarsM transgenic A. thaliana plants showed greatly enhanced tolerance to AsV and AsIII compared to wild-type (WT) plants. WaarsM expressing transgenic plants evolved 17.5 ng and 113 ng volatile arsenicals (mg −1 fresh weight) after 48 h of exposure to 250 μM AsV and 50 μM AsIII, respectively. Long-term exposure resulted in 36% and 16% less arsenic accumulation in seeds and shoots, respectively compared to WT plants. Additionally, the S. cerevisiae cells expressing WaarsM showed short lag phase in the presence of arsenic and potentially tolerate up to 5 mM AsV and 1 mM AsIII. In conclusion, WaarsM from arsenic tolerant fungus can be used in a novel biotechnological solution to decrease arsenic accumulation in food crops grows in arsenic affected areas.

Arsenic (As) is an acute poison and class I carcinogen, can cause a serious health risk. Staple crops like rice are the primary source of As contamination in human food. Rice grown on As contaminated areas accumulates higher As in their edible parts. Based on our previous transcriptome data, two rice glutaredoxins (OsGrx_C7 and OsGrx_C2.1) were identified that showed up-regulated expression during As stress. Here, we report OsGrx_C7 and OsGrx_C2.1 from rice involved in the regulation of intracellular arsenite (AsIII). To elucidate the mechanism of OsGrx mediated As tolerance, both OsGrxs were cloned and expressed in Escherichia coli (Δars) and Saccharomyces cerevisiae mutant strains (Δycf1, Δacr3). The expression of OsGrxs increased As tolerance in E. coli (Δars) mutant strain (up to 4 mM AsV and up to 0.6 mM AsIII). During AsIII exposure, S. cerevisiae (Δacr3) harboring OsGrx_C7 and OsGrx_C2.1 have lower intracellular AsIII accumulation (up to 30.43% and 24.90%, respectively), compared to vector control. Arsenic accumulation in As-sensitive S. cerevisiae mutant (Δycf1) also reduced significantly on exposure to inorganic As. The expression of OsGrxs in yeast maintained intracellular GSH pool and increased extracellular GSH concentration. Purified OsGrxs displays in vitro GSH-disulfide oxidoreductase, glutathione reductase and arsenate reductase activities. Also, both OsGrxs are involved in AsIII extrusion by altering the Fps1 transcripts in yeast and protect the cell by maintaining cellular GSH pool. Thus, our results strongly suggest that OsGrxs play a crucial role in the maintenance of the intracellular GSH pool and redox status of the cell during both AsV and AsIII stress and might be involved in regulating intracellular AsIII levels by modulation of aquaporin expression and functions.

Fly-ash (FA) utilization in the agriculture sector is very common practice due to presence of beneficial elements required for plant growth. However, presence of excessive amount of toxic metals in FA may be serious concern for agriculture. In the present study, the effects of FA on soil health, plant growth, toxic metal accumulation and antioxidant responses were investigated in rice (Oryza sativa L.), grown on soil amended with 50 % FA in natural condition. FA application resulted in reduction in soil enzymatic activities viz., dehydrogenase, acid phosphatase, β-glucosidase and urease than garden soil (GS). FA amendments significantly decreased the root, shoot and panicle length and augmented sterility in rice. Interestingly, ICPMS analysis for metal accumulation revealed that the total accumulation of toxic metals, particularly Cd, Cr, Pb and As were 14–15 fold higher in roots and shoots and 4–20 fold higher in grains for the plants grown on FA amended soil than GS. The levels of nutrient elements viz., Mn, Co, Cu and Se were lesser in grains of FA treated soil than GS. Lipid peroxidation was increased in root and shoot of FA treated plants indicating oxidative stress. Activities of various antioxidant enzymes viz., SOD, APX, GPX, GR and their isozymes were correlated to each other and also increased against heavy metal induced toxicity. Human associated risk analysis revealed that the calculated maximum tolerable daily intake values for toxic metals (µg d−1) viz., Cr, As, Cd, Hg and Pb was beyond the safe limit in the rice grown in FA implicated soil. In conclusion, 50 % FA implication deteriorates the soil quality, rice production and elevates the toxic metals in grains, which may be a concern for safer rice consumption.

Abiotic stresses adversely affect cellular homeostasis, impairing overall growth and development of plants. These initial stress signals activate downstream signalling processes, which, subsequently, activate stress-responsive mechanisms to re-establish homeostasis. Dehydrins (DHNs) play an important role in combating dehydration stress. Rice (Oryza sativa L.), which is a paddy crop, is susceptible to drought stress. As drought survival in rice might be viewed as a trait with strong evolutionary selection pressure, we observed DHNs in the light of domestication during the course of evolution. Overall, 65 DHNs were identified by a genome-wide survey of 11 rice species, and 3 DHNs were found to be highly conserved. The correlation of a conserved pattern of DHNs with domestication and diversification of wild to cultivated rice was validated by synonymous substitution rates, indicating that Oryza rufipogon and Oryza sativa ssp. japonica follow an adaptive evolutionary pattern; whereas Oryza nivara and Oryza sativa ssp. indica demonstrate a conserved evolutionary pattern. A comprehensive analysis of tissue-specific expression of DHN genes in japonica and their expression profiles in normal and PEG (poly ethylene glycol)-induced dehydration stress exhibited a spatiotemporal expression pattern. Their interaction network reflects the cross-talk between gene expression and the physiological processes mediating adaptation to dehydration stress. The results obtained strongly indicated the importance of DHNs, as they are conserved during the course of domestication.

Amino acids (AAs) play significant roles in metal binding, antioxidant defense, and signaling in plants during heavy metal stress. In the present study, the essential amino acids (EAAs), non-essential amino acids (NEAAs), as well as the enzymes of proline and cysteine biosynthetic pathways were studied in contrasting arsenic accumulating rice genotypes grown in hydroponic solutions with addition of arsenate (AsV) or arsenite (AsIII). Under a mild As stress, the total AAs content significantly increased in both the rice genotypes with a greater increase in a low As accumulating rice genotype (LAARG; IET-19226) than in a high As accumulating rice genotype (HAARG; BRG-12). At the equimolar concentration (10 μM), AsIII had a greater effect on EAAs than AsV. Conversely, AsV was more effective in inducing a proline accumulation than AsIII. Among NEAAs, As significantly induced the accumulation of histidine, aspartic acid, and serine. In contrast, a higher As concentration (50 μM) reduced the content of most AAs, the effect being more prominent during AsIII exposure. The inhibition of glutamate kinase activity was noticed in HAARG, conversely, serine acetyltransferase and cysteine synthase activities were increased which was positively correlated with the cysteine synthesis.

Plant-growth-promoting rhizobacteria (PGPR) improve plant growth by altering the root architecture, although the mechanisms underlying this alteration have yet to be unravelled. Through microarray analysis of PGPR-treated rice roots, a large number of differentially regulated genes were identified. Ectopic expression of one of these genes, OsASR6 (ABA STRESS RIPENING6), had a remarkable effect on plant growth in Arabidopsis. Transgenic lines over-expressing OsASR6 had larger leaves, taller inflorescence bolts and greater numbers of siliques and seeds. The most prominent effect was observed in root growth, with the root biomass increasing four-fold compared with the shoot biomass increase of 1.7-fold. Transgenic OsASR6 over-expressing plants showed higher conductance, transpiration and photosynthesis rates, leading to an ˜30% higher seed yield compared with the control. Interestingly, OsASR6 expression led to alterations in the xylem structure, an increase in the xylem vessel size and altered lignification, which correlated with higher conductance. OsASR6 is activated by auxin and, in turn, increases auxin responses and root auxin sensitivity, as observed by the increased expression of auxin-responsive genes, such as SAUR32 and PINOID, and the key auxin transcription factor, ARF5. Collectively, these phenomena led to an increased root density. The effects of OsASR6 expression largely mimic the beneficial effects of PGPRs in rice, indicating that OsASR6 activation may be a key factor governing PGPR-mediated changes in rice. OsASR6 is a potential candidate for the manipulation of rice for improved productivity.

Nitric oxide (NO) and salicylic acid (SA) are important signaling molecules in plant system. In the present study both NO and SA showed a protective role against arsenite (AsIII) stress in rice plants when supplied exogenously. The application of NO and SA alleviated the negative impact of AsIII on plant growth. Nitric oxide supplementation to AsIII treated plants greatly decreased arsenic (As) accumulation in the roots as well as shoots/roots translocation factor. Arsenite exposure in plants decreased the endogenous levels of NO and SA. Exogenous supplementation of SA not only enhanced endogenous level of SA but also the level of NO through enhanced nitrate reductase (NR) activity, whether AsIII was present or not. Exogenously supplied NO decreased the NR activity and level of endogenous NO. Arsenic accumulation was positively correlated with the expression level of OsLsi1, a transporter responsible for AsIII uptake. The endogenous level of NO and SA were positively correlated to each other either when AsIII was present or not. This close relationship indicates that NO and SA work in harmony to modulate the signaling response in AsIII stressed plants.

Along with many adaptive strategies, dynamic changes in protein abundance seem to be the common strategy to cope up with abiotic stresses which can be best explored through proteomics. Understanding of drought response is the key to decipher regulatory mechanism of better adaptation. Rice (Oryza sativa L.) proteome represents a phenomenal source of proteins that govern traits of agronomic importance, such as drought tolerance. In this study, a comparison of root cytoplasmic proteome was done for a drought tolerant rice (Heena) cultivar in PEG induced drought conditions. A total of 510 protein spots were observed by PDQuest analysis and 125 differentially regulated spots were subjected for MALDI-TOF MS-MS analysis out of which 102 protein spots identified which further led to identification of 78 proteins with a significant score. These 78 differentially expressed proteins appeared to be involved in different biological pathways. The largest percentage of identified proteins was involved in bioenergy and metabolism (29%) and mainly consists of malate dehydrogenase, succinyl-CoA, putative acetyl-CoA synthetase, and pyruvate dehydrogenase etc. This was followed by proteins related to cell defense and rescue (22%) such as monodehydroascorbate reductase and stress-induced protein sti1, then by protein biogenesis and storage class (21%) e.g. putative thiamine biosynthesis protein, putative beta-alanine synthase, and cysteine synthase. Further, cell signaling (9%) proteins like actin and prolyl endopeptidase, and proteins with miscellaneous function (19%) like Sgt1 and some hypothetical proteins were also represented a large contribution toward drought regulatory mechanism in rice. We propose that protein biogenesis, cell defense, and superior homeostasis may render better drought-adaptation. These findings might expedite the functional determination of the drought-responsive proteins and their prioritization as potential molecular targets for perfect adaptation.

Glutaredoxins (Grxs) are a family of small multifunctional proteins involved in various cellular functions, including redox regulation and protection under oxidative stress. Despite the high number of Grx genes in plant genomes (48 Grxs in rice), the biological functions and physiological roles of most of them remain unknown. Here, the functional characterization of the two arsenic-responsive rice Grx family proteins, OsGrx_C7 and OsGrx_C2.1 are reported. Over-expression of OsGrx_C7 and OsGrx_C2.1 in transgenic Arabidopsis thaliana conferred arsenic (As) tolerance as reflected by germination, root growth assay, and whole plant growth. Also, the transgenic expression of OsGrxs displayed significantly reduced As accumulation in A. thaliana seeds and shoot tissues compared to WT plants during both AsIII and AsV stress. Thus, OsGrx_C7 and OsGrx_C2.1 seem to be an important determinant of As-stress response in plants. OsGrx_C7 and OsGrx_C2.1 transgenic showed to maintain intracellular GSH pool and involved in lowering AsIII accumulation either by extrusion or reducing uptake by altering the transcript of A. thaliana AtNIPs. Overall, OsGrx_C7 and OsGrx_C2.1 may represent a Grx family protein involved in As stress response and may allow a better understanding of the As induced stress pathways and the design of strategies for the improvement of stress tolerance as well as decreased As content in crops.

Glutathione S-transferases (GSTs; EC 2.5.1.18) are members of an isozyme family and catalyze the conjugation of the reduced tripeptide glutathione to a variety of hydrophobic and electrophilic substrates. Though members of different classes of the GST superfamily have been identified and characterized from many plant species including rice, no detailed information is available for the Lambda class gene family in rice. In this study, a genome-wide analysis was carried out to investigate expression patterns of three Lambda class GST members of rice including OsGSTL1, OsGSTL2 and OsGSTL3 in seedlings, at different growth and developmental stages as well as in response to various biotic and abiotic stresses. Expression analysis using microarray datasets and quantitative real-time reverse transcriptase polymerase chain reaction suggests that this gene family express differentially in various tissues, in response to hormones and during different biotic and abiotic stresses including heavy metals, cold, drought and salt stress. Massively Parallel Signature Sequencing (MPSS) analysis also showed differential expression of OsGSTLs during plant growth and development and under different stresses. Out of three members, maximum expression of OsGSTL2 was observed for the MPSS libraries in comparison to other members. We conclude that members of rice Lambda class GST family play an important role in plant growth and development and in combating different biotic and abiotic stresses.

The present study reports an efficient protocol for indirect shoot organogenesis and plantlets regeneration of Withania somnifera (L.) Dunal. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP) and indole-3-acetic acid (IAA). The highest callus induction rate (89.5 %) and shoot regeneration rate (92 %) were obtained when 2 mg dm−3 BAP was combined with 0.5 mg dm−3 IAA. Three major withanolides (withaferine A, 12-deoxywithastramonolide and withanolide A) were investigated in different plant organs from in vitro and greenhouse grown plants. Leaves contained higher contents of withanolides and phenolics than roots or stems, whereas roots contained the highest contents of flavonoids and polysacharides. In vitro grown plants contained greater contents of phenolics, flavonoids and polysaccharides while lower contents of withanolides than greenhouse grown plants.

The following study was carried out to investigate the cadmium (Cd) accumulating potential of Vallisneria. After subjecting plants to different concentrations of Cd, it was observed that plants are able to accumulate ample amount of metal in their roots (5,542 microg g(-1) dw) and leaves (4,368 microg g(-1) dw) in a concentration- and duration-dependent manner. Thus, it is evident that the accumulation in roots was 1.3 times higher than the shoots. It was also noted that with increasing Cd accumulation, roots of the plant appeared darker in color and harder in texture. In response to metal exposure, amount of low molecular weight antioxidants such as cysteine and nonprotein thiols (NP-SH) and activity of enzymes such as APX and GPX were significantly enhanced at lower concentrations of Cd, followed by decline at higher doses. It was also observed that in exposed plants, activity of APX enzyme was higher in roots (ca. 3 times) as compared to leaves. However, chlorophyll and protein content was found to decline significantly in a dose-dependent manner. Results suggested that due to its high accumulation potential, Vallisneria may be effectively grown in water bodies moderately contaminated with Cd.

Arsenic is commonly present in subsoil and is a carcinogen in humans. Rice takes up arsenic and it accumulates in different plant parts, including grains, at levels several-fold higher than the soil. In high arsenic regions, rice can contribute substantially to arsenic intake by the human population. Arsenic in rice grains is present in the carcinogenic inorganic or the relatively safer organic (methylated) form. A wide variation is noticed in different rice genotypes with respect to the proportion of arsenic in these forms in grains. Mechanisms involved in arsenic uptake, efflux from roots, loading into xylem, transport, partitioning, arsenate reduction, arsenic sequestration in vacuoles, volatilization from leaves, accumulation in grains etc. are poorly understood. Selection of cultivars accumulating low inorganic arsenic is an important trait to be used by breeders to develop rice varieties safer for cultivation in arsenic-contaminated regions. Systematic efforts have not been made to screen rice genotypes for mining the genes involved in arsenic uptake, transport and accumulation in grains. Identification of rice germplasm with varying arsenic uptake and partitioning, and development of mapping populations with contrasting grain arsenic, are required for association studies and QTL mapping for accelerating rice improvement. Efforts on gene expression profiling, deep transcriptome sequencing, high throughput metabolomics and phenotyping of contrasting arsenic accumulating lines need to be increased to develop strategies for design of safer rice varieties. Network research projects need to be developed along these approaches to accelerate the development of crop varieties safer for farming in arsenic-contaminated environments.

Very little is known about molecular events associated with callus differentiation in indica rice. The genes expressed differentially during shoot meristem initiation were identified on genomic arrays applied to efficiently regenerating rice calli. A thidiazuron (TDZ; N-phenyl-N-thiadiazol-1,2,3-5,ylurea)-dependent regeneration protocol was developed for efficient embryogenesis in indica rice. The regenerating embryogenic calli induced by TDZ for 10 days showed transcriptional modulation of a number of genes associated with photosynthesis, hormone metabolism, plant development, signal transduction, light response, and plant defense. Eighteen candidate miRNAs were predicted to target the genes expressed differentially in the embryogenic calli grown in TDZ-containing medium. The majority of the photosynthesis-related genes up-regulated in differentiating calli were not expressed or were down-regulated in developing seeds and inflorescences. Most of the genes down-regulated in differentiating calli were up-regulated in developing seeds. The transcriptome of differentiating callus most closely resembled that of the germinating whole seed.

An in vitro process for rapid clonal propagation of Clerodendrum serratum (Linn.) Moon, a rare and threatened medicinal shrub, has been developed. Nodal stem segments having axillary bud, taken from field-grown plant, showed bud-break within 15 days of culture on modified Murashige and Skoog (MS) (Physiol Plant 15:473–497, 1962) medium supplemented with 0.25 mg/l each of 6-benzylaminopurine and indole-3-acetic acid along with 15 mg/l adenine sulphate (AdS). Regenerated shoots could be further multiplied on the same agarified morphogenetic medium in presence of 0.5 mg/l 2-chloroethyltrimethyl ammonium chloride with increased concentration of AdS, i.e., 30 mg/l. A group of five shoots used as inoculum produced on an average 4.98 new shoots per original shoot after 4 weeks of subculture. Shoots excised from cultures of proliferating shoots were rooted in half-strength MS medium having 1 mg/l indole-3-propionic acid. In vitro rooted shoots—plantlets—grew luxuriantly under field conditions and came to flowering after 10 months of transplantation. The genetic fidelity of in vitro-raised field-grown plants and their mother plant was ascertained by random amplified polymorphic DNA markers. The protocol developed holds good for in vitro cloning of C. serratum.

Nitric oxide is a gaseous signalling molecule and has a profound impact on plant growth and development. It is reported to serve as pro oxidant as well as antioxidant in plant system. In present study, we evaluated the protective role of nitric oxide against AsV toxicity in rice plants. Arsenate exposure has hampered the plant growth, reduced the chlorophyll content and enhanced the oxidative stress while the exogenous NO supplementation has reverted these symptoms. Nitric oxide supplementation has reduced the As accumulation in root as well as shoot. Nitric oxide supplementation to AsV exposed plants has reduced the gene expression level of OsLsi1 and OsLsi2. Arsenate stress significantly impacted thiol metabolism, it reduced GSH content and GSH/GSSG ratio and enhanced the level of PCs. Nitric oxide supplementation maintained the GSH/GSSG ratio and reduced the level of PCs. Nitric oxide supplementation reverted AsV induced iron deficiency in shoot and had significant impact of gene expression level of various iron transporters (OsYSL2, OsFRDL1, OsIRT1 and OsIRO2). Conclusively, exogenous application of nitric oxide could be advantageous against AsV toxicity and could confer the tolerance to AsV stress in rice.

Episodes of water deficit (WD) during the crop cycle of tomato may negatively impact plant growth and fruit yield, but they may also improve fruit quality. Moreover, a moderate WD may induce a plant "memory effect" which is known to stimulate plant acclimation and defenses for upcoming stress episodes. The objective of this study was to analyze the positive and negative impacts of repeated episodes of WD at the plant and fruit levels. Three episodes of WD (-38, -45, and -55% of water supply) followed by three periods of recovery ("WD treatments"), were applied to the eight parents of the Multi-Parent Advanced Generation Inter-Cross population which offers the largest allelic variability observed in tomato. Predawn and midday water potentials, chlorophyll a fluorescence, growth and fruit quality traits [contents in sugars, acids, carotenoids, and ascorbic acid (AsA)] were measured throughout the experiment. Important genotypic variations were observed both at the plant and fruit levels and variations in fruit and leaf traits were found not to be correlated. Overall, the WD treatments were at the origin of important osmotic regulations, reduction of leaf growth, acclimation of photosynthetic functioning, notably through an increase in the chlorophyll content and in the quantum yield of the electron transport flux until PSI acceptors (J 0 (RE1)/J (ABS)). The effects on fruit sugar, acid, carotenoid and AsA contents on a dry matter basis ranged from negative to positive to nil depending on genotypes and stress intensity. Three small fruit size accessions were richer in AsA on a fresh matter basis, due to concentration effects. So, fruit quality was improved under WD mainly through concentration effects. On the whole, two accessions, LA1420 and Criollo appeared as interesting genetic resources, cumulating adaptive traits both at the leaf and fruit levels. Our observations show that the complexity involved in plant responses, when considering a broad range of physiological traits and the variability of genotypic effects, represent a true challenge for upcoming studies aiming at taking advantage of, not just dealing with WD.

Gerbera jamesonii H. Bolus ex Hook (Family: Asteraceae) has been successfully acclimatized from temperate to subtropical North Indian plains of Lucknow through in vitro propagation. Flower heads were collected from greenhouse, segmented into 4–16 pieces and cultured in Murashige and Skoog’s medium (MS) (Physiol Plant 15:472–497, 1962) supplemented with 2.87 μM indole-3-acetic acid (IAA) and 8.88 μM N6-benzyladenine (BA) for shoot regeneration. Shoots were subcultured on growth regulator free MS medium. Apical shoot meristems from in vitro plantlets of gerbera were tested in MS medium with different combination of cytokinins [BA, kinetin, and thidiazuron (TDZ)] alongwith 2.68 μM 1-naphthaleneacetic acid (NAA) for shoot multiplication. The optimum results were obtained with 8.88 μM BA. Regenerated plants with well-established root system were transferred to pots containing soil and sand (1:1 v/v) and were kept in humidity chamber with 80–90% relative humidity for 0, 5, 10, 15, 20, and 25 days before they were transferred to field (during October, 2005 to February, 2006). Survival percentage was higher when regenerated plantlets were kept under humidity chamber for 15 days. An attempt was made to obtain basic information on different biochemical changes during acclimatization process of in vitro raised plantlets. Increased lipid peroxidation and high H2O2 content in early stages of acclimatization process reflected a similar process of oxidative stress. Our work suggests that tissue-cultured plants develop antioxidant enzymatic protective system which determine the ability to survive in oxidative stress and up regulation of these enzymes would help to reduce the built up of reactive oxygen species (ROS).

The present study reports on the kinetics of nutrient utilization during in vitro flowering of Spathiphyllum in air-lift bioreactor cultures. Levels of electrical conductivity (EC), anions and cations, pH, ethylene, sugar content and photosynthetic enzymes were determined in bioreactor cultures of both flowering (FPs) and non-flowering (NFPs) plantlets over a growth period of 12 weeks. A decrease in ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity with a corresponding increase in phosphoenolpyruvate carboxylase (PEPcase) activity occurred during floral induction of Spathiphyllum in vitro. Sucrose concentration decreased significantly in FPs, while no changes in glucose, fructose and total sugars were observed in both FPs and NFPs up to 8 weeks of culture. There were significant variations in mineral nutrient utilization between FP and NFP cultures. These results provide an insight to the physiological processes involved in inflorescence formation in Spathiphyllum.

Stems of chrysanthemum (Chrysanthemum morifolium Ramat.) cv. Maghi were harvested when half of the buds showed colour and were put in distilled water at 21°C. Flowers showed visible senescence symptoms after 12–15 d. Reactive oxygen species (ROS) concentration and lipid peroxidation increased from young floret stage to the senescent stage. Activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD) and catalase (CAT) showed uniform increases from young floret through to the mature stage and thereafter, declined. Among the SOD isoforms, Fe-SOD and Cu/Zn-SOD were induced during the onset of senescence. Similarly different isoforms of APX and glutathione reductase (GR) also appeared during the senescence process. The capacity of the antioxidative defence system increased during the onset of senescence but the imbalance between ROS production and antioxidant defences ultimately led to oxidative damage. It is proposed that a decrease in the activity of a number of antioxidant enzymes that normally prevent the build up of free radicals can at least partially account for the observed senescence of chrysanthemum florets.

Accumulation of arsenic (As) in rice (Oryza sativa L.) grain is a serious concern worldwide. Long-term exposure to As affects nutritional status in rice grain and is associated with higher rates of skin, bladder, and lung cancers, and heart disease. Genotypic variations in rice for As accumulation or tolerance are prevalent and are regulated by genetic and environmental factors. To understand molecular networks involved in As accumulation, genome-wide expression analysis was performed in roots of low- and high-As accumulating rice genotypes (LARGs and HARGs). Six rice genotypes with contrasting As accumulation potential and tolerance were used in this study. Genome-wide expression analysis suggested their differential response against As stress. This study suggests up- and downregulation of a number of unique genes involved in various pathways and biological processes in response to As stress in rice genotypes. A comparison of gene expression profiles, principal component analysis, and K-means clustering suggests that an independent pathway is operating during As stress tolerance or accumulation in contrasting genotypes. It was also observed that the differential behavior of aus genotype, Nayanmoni, from other LARGs might be due to its different genetic background. Cis-motif profiling of As-induced coexpressed genes in diverse rice genotypes led to the identification of unique cis-motifs present in differentially expressed genes. This study suggests that the genetic mechanism regulating the differential As accumulation in different genotypes may not be dependent on gene expression at the transcriptional level. However, many genes identified in this study can be analyzed and used for marker-trait associations related to As accumulation in diverse genotypes around the world.

Arsenic (As) is posing serious health concerns in South East Asia where rice, an efficient accumulator of As, is prominent crop. Salicylic acid (SA) is an important signalling molecule and plays a crucial role in resistance against biotic and abiotic stress in plants. In present study, ameliorative effect of SA against arsenate (AsV) toxicity has been investigated in rice (Oryza sativa L.). Arsenate stress hampered the plant growth in terms of root, shoots length and biomass as well as it enhanced the level of H2O2 and MDA in dose dependent manner in shoot. Exogenous application of SA, reverted the growth and oxidative stress caused byAsV and significantly decreased As translocation to the shoots. Level of As in shoot was positively correlated with the expression of OsLsi2, efflux transporter responsible for root to shoot translocation of arsenic in the form of arsenite (AsIII). Salicylic acid also overcame AsV induced oxidative stress and modulated the activities of antioxidant enzymes in a differential manner in shoots. Arsenic treatment hampered the translocation of Fe in the shoot which was compensated by the SA treatment. The level of Fe in root and shoot was positively correlated with the transcript level of transporters responsible for the accumulation of Fe,OsNRAMP5 and OsFRDL1, in the root and shoot respectively. Co-application of SA was more effective than pre-treatment for reducing As accumulation as well as imposed toxicity.

The present study investigated the dynamics of nutrient utilization and various growth and physiological parameters during in vitro proliferation of apple root stock ‘M9 EMLA’ in two different bioreactor systems, i.e. temporary and continuous immersions. Individual shoots obtained from temporary immersion system had higher dry mass and were of better quality than those obtained from continuous immersion. In continuous immersion bioreactor, apple shoots appeared to utilize more nutrients from liquid culture medium than that from temporary immersion. The shoot growth was limited by the availability of phosphate and nitrogen in continuous immersion system. The shoots produced in temporary immersion bioreactor showed higher photosynthetic rate, maximum quantum yield of photosystem-II and slow but steady rate of nutrient absorption, indicating the occurrence of higher photomixotrophic metabolism. The study also showed that high level of antioxidant scavenging enzymes in shoots grown in continuous immersion system induced physiological changes to foster adaptation to stresses.

A large number of micropropagated Euphorbia millii shoots from temporary immersion bioreactor showed thick broad leaves that were translucent, wrinkled and/or curled and brittle, symptoms of hyperhydricity. The environment inside bioreactor normally used in plant micropropagation is characterised by high relative humidity, poor gaseous exchange between the internal atmosphere of the bioreactor and its surrounding environment, and the accumulation of ethylene, conditions that may induce physiological disorders. A comparison of hyperhydric shoots (HS) with normal plants shows marked increase in malondialdehyde (MDA) content in HS plants. MDA, a decomposition product of polyunsaturated fatty acids hydroperoxides, has been utilized very often as a suitable biomarker for lipid peroxidation, which is an effect of oxidative damage. This hypothesis is also confirmed by the higher lipoxygenase (LOX) activity in HS plants. The potential role of antioxidant enzymes in protecting hyperhydric shoots from oxidative injury was examined by analyzing enzyme activities and isozyme profiles of hyperhydric and non-hyperhydric leaves of E. millii. Superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activity were significantly higher in hyperhydric tissue as compared to non-hyperhydric normal leaf tissue. After native polyacrylamide gel electrophoresis (PAGE) analysis, seven SOD isoenzymes were detected and the increase in SOD activity observed in hyperhydric tissue seemed to be mainly due to Mn-SOD and Cu/Zn-SOD. The activity of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) was proportionally increased in HS tissue compared to normal leaves indicating a crucial role in eliminating toxic H2O2 from plant cells. The depletion of GSH and total glutathione in spite of higher GR activities observed in HS tissue indicates that mechanism of antioxidant defense was by enhanced oxidation of GSH to GSSG by DHAR yielding ascorbate (AA). The antioxidant metabolism has been shown to be important in determining the ability of plants to survive in hyperhydric stress and the up regulation of these enzymes would help to reduce the build up of ROS.

A method for the micropropagation of Spathiphyllum cannifolium is presented using shoot tip proliferation onto Murashige and Skoog (MS) medium supplemented with different plant growth regulator concentrations and combinations. The proliferation responses were significantly influenced by the cytokinin type and concentrations. Supplementation of the medium with benzyladenine (BA; 4.44–13.32 μM) increased the shoot proliferation rate significantly as compared to other treatments. When cytokinins were used with auxin (indole-3-butyric acid, IBA and naphthalene acetic acid. NAA), the number of shoots per explant increased in comparison with treatments with BA alone. The largest number of shoots, 9.3 per explant, was obtained with 13.32 μM BA and 4.9 μM IBA. Different MS medium strengths and sucrose concentrations were used with the aim to stimulate in vitro shoot proliferation. Full MS medium with 30 gl−1 sucrose was found to be suitable for shoot tip culture of Spathiphyllum. Comparative studies between gelled medium and bioreactor culture [continuous immersion (with or without net) and temporary immersion in liquid media using ebb and flood] revealed that shoot multiplication and growth were more efficient in continuous immersion (with net) bioreactor with low cytokinin-supplemented media. Plantlets from the bioreactor were cultured hydroponically for 30 d and 100% of plants were rooted and acelimatized successfully. Rapid and efficient multiplication rate in bioreactor and successful transfer to greenhouse makes this protocol suitable for large-scale multiplication of this important foliage plant.

In vitro regenerated shoots of Spathiphyllum from bioreactor were hydroponically cultured for 30 days. The response of plant growth and photosynthesis to different substrates, photosynthetic photon flux (PPF), nutrient scheduling and electrical conductivity (EC) of hydroponic solution were studied. The best plant growth response was observed in perlite based substrates with moderate PFF (70–100μmol m−2 s−1). Highest fresh weight, dry weight, shoot length, root length, root number and photosynthetic characteristics (chlorophyll, carotenoids and Fv/Fm) was observed in continuous immersion system. Plant growth responses, photosynthetic rate, stomatal conductance and transpiration rate were also found to be affected by EC levels. The optimum EC of a balanced nutrient solution was recorded as 1.2 dS m−1. Photosynthetic activity was also characterized in terms of photochemical efficiency using measurements of chlorophyll fluorescence. Fv/Fm (it is a measure of the intrinsic or maximum efficiency of PSII i.e. the quantum efficiency if all PSII centers were open) also decreased significantly in plants grown under higher EC level; a decrease in this parameter indicates down regulation of photosynthesis or photoinhibition. Antioxidant defense enzymes such as catalase (CAT), ascorbate peroxidase (APX), peroxidase (POD), glutathione reductase (GR) and monodehydroascorbate reductase (MDHAR) significantly elevated in the leaves and roots of plantlets at higher EC levels. This increase could reflect a defense response to the cellular damage provoked by higher EC levels in the nutrient solution.

A rapid and efficient procedure is outlined for in vitro clonal propagation of an elite cultivar of jewel orchid (Anoectochilus formosanus). Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with 1 mg dm−3 benzyladenine or 1 – 2 mg dm−3 thidiazuron (TDZ). Addition of activated charcoal (1 g dm−3) to the TDZ containing medium promoted multiple shoot formation (11.1 shoots per explant). However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 medium supplemented with 2 % sucrose and 0.5 g dm−3 activated charcoal. Rooting was induced in 100 % of the regenerated shoots in the same media. The plantlets were acclimatized and established in greenhouse.

Cut flowering stems (45 cm long) of carnation (cv. Master) and rose (cv. Red Sandra) were placed in test tubes containing distilled water with or without commercial preservative (20 ml l−1), under fluorescent tubes of 50 ± 10 μmol m−2 s−1 Photosynthetic photon flux (PPF). A mixed radiation from fluorescent tubes with red light provided by light emitting diodes (LEDs) and monochromic red light of low 50 or high 90 ± 10 μmol m−2 s−1 was also tested in the absence of preservative solution. Both red light with high PPF and the mixed radiation under low PPF extended the vase life of cut carnations, and flower freshness could be maintained for 10.9 days compared with a water/fluorescent light control. In cut rose, the treatment containing the preservative solution under fluorescent tubes alone and in red alone, regardless of light intensity, prolonged vase life for 4.6 and 4.2 days longer than the control, respectively. Treatment with red LEDs plus high PPF resulted in complete petal opening in carnation flowers.

In vitro bulblet formation was studied using solid, liquid and bioreactor culture (immersion and periodic immersion in liquid media using ebb and flood) in order to develop a cost effective method for the mass propagation of Lilium oriental hybrid ‘Casablanca’. Although the percent of bulblet formation was higher in solid culture, the increased growth rate and production of large number of bulblets in bioreactor makes it suitable for mass propagation. Four times per day and 15 min of medium supply was optimal for bulblet formation in ebb and flood bioreactor. Bulblet formation was also found to be effective in 16-h photoperiod. It was also observed that bulblet formation in the medium with 1.0 mg dm−3 BA and 0.3 mg dm−3 NAA was higher than in the medium without growth regulators, but formation of abnormal bulblets was higher in medium with BA and NAA.

Flowers of Chrysanthemum morifolium Ramat cv. Lalima were greyed red and florets were flat spoon shaped. Ray florets after inoculation on the Murashige and Skoog's medium supplemented with 1.07 μM α-naphthaleneacetic acid and 8.87 μM benzyladenine were irradiated with γ-radiation (0.5 Gy and 1 Gy). All the regenerated shoots either from control or from γ-irradiated florets were isolated, rooted and transplanted in the field after hardening. Two mutants were obtained in the γ-irradiated plants (0.5 Gy). Both the mutants were yellow coloured but one having flat spoon shaped ray florets similar to the original cultivar, while the other having tubular florets. Both the mutants were propagated vegetatively and have produced true-to-type flowers.

Adverse environmental conditions including heavy metal stress impose severe effects on the plant growth and development limiting productivity and yield. Studies demonstrated that changes in genome-wide expression modulate various biochemical processes and molecular components in response to heavy metal stress in plants. Some of the key components involved in such a regulation are the transcription initiation machinery, nucleotide sequence of promoters and presence of cis-acting elements. Therefore, identification of the putative cis-acting DNA sequences involved in gene regulation and functional characterization of promoters are important steps in understanding response of plants to heavy metal stress. In this study, comprehensive analysis of the proximal promoters of members of rice sulfate transporter gene family which is an essential component of stress response has been carried out. Analysis suggests presence of various common stress related cis-acting elements in the promoters of members of this gene family. In addition, transcriptional regulation of the arsenic-responsive high affinity sulfate transporter, OsSul1;1, has been studied through development of Arabidopsis transgenic lines expressing reporter gene encoding β-glucuronidase under the control of OsSul1;1 promoter. Analysis of the transgenic lines suggests differential response of the OsSul1;1 promoter to various heavy metals as well as other abiotic stresses.

We have established cell-suspension cultures of mountain ginseng (Panax ginseng G A. Mayer), and have attempted to increase the yield of saponin by manipulating our processing method and culturing factors (e.g., media strengths; the presence of plant growth regulators or sucrose; ratios of NO+ 3/ NH- 4). Maximum biomass yield was obtained in media containing 2,4-D. However, saponin productivity was much higher in a medium comprising either IBA or NAA; 7.0 mg/L IBA was optimal for promoting both cell growth (10.0 g/L dry weight) and saponin production (7.29 mg/g DW total ginsenoside). Although the addition of cytokinins (BA and kinetin) did not affect cell growth, the level of saponin (particularly in the Rb group) was enhanced when the media were supplemented with either 0.5 mg/L BA or 0.5 mg/L kinetin. Half- and full-strength MS media were equally suitable for inducing both biomass as well as saponin production. We also investigated the effect of various concentrations of sucrose and nitrogen, and found that 30 g/L sucrose enhanced biomass yield as well as saponin content However, further increases (i.e., up to 70 g/L) led to a decrease in saponin accumulation and biomass production. Maximum growth and saponin productivity were reported from treatments with an initial nitrogen concentration of 30 mM. In general, the amount of saponin increased when the test media had high NO+ 3/ NH- 4 ratios; in fact, saponin production was greatest when nitrate was the sole nitrogen source.

High-frequency protocorm-like body (PLB) formation directly from thin leaf sections of Doritaenopsis hybrid was achieved in order to develop a mass-scale propagation system. Concentrated efforts were made to study the effects of different cytokinins on in vitro PLB induction from thin leaf sections. Among the cytokinins tested, thidiazuron (TDZ) was found to be a more effective inducer of PLBs than benzyladenine and zeatin. A modified Murashige and Skoog medium supplemented with 9.0 µM TDZ was found to be the optimum concentration for PLB development from thin leaf sections of Doritaenopsis hybrid. Of the two different explant types used in the present experiment, the highest percentage of PLB formation (72.3%) and highest number of PLBs (18) per explant were observed on thin leaf sections (1 mm thick), while only 20% (4.3 per explant) of comparatively large leaf segments (5 mm thick) were able to produce PLBs under the same culture conditions. Light microscopy observations indicated that the initial cell divisions for PLB formation occurred on the region near the cut surface and that an intact epidermal layer appeared to play an important role in PLB formation. Proembryo initiation occurred from several cells just beneath the intact epidermal cell, and globular PLBs were clearly visible after 3 weeks of culture and subsequently developed into mature PLBs.

The effects of light generated by monochromic blue, red or mixed radiation from a fluorescent lamp (FL) with light emitting diodes (LEDs) (blue, red, or far-red) on growth and morphogenesis of marigold and salvia seedlings were investigated and the responses compared with those of plantlets grown under a broad spectrum conventional fluorescent lamp (a 16 h photoperiod per day). Dry weight of marigold seedlings was significantly increased in monochromic red light (R), fluorescent light plus red LED (FLR) or fluorescent light (FL) but reduced when monochromic blue light (B) was used, whereas in salvia dry weight was significantly greater under fluorescent light plus blue LED (FLB), fluorescent light plus red LED (FLR) and fluorescent light plus far-red LED (FLFr) as compared to other treatments. Stem length in marigold was greatest in monochromic blue light, being three times greater than in FLR or FL treatments. In salvia, FLFr increased stem length but this was significantly decreased by R as compared to other treatments. The number of visible flower buds in marigold was much higher in FLR as well as in the control (FL), and it was about five times greater than in B or R. However, the number of open flowers in salvia varied slightly in all the treatments. Different light qualities also influenced the duration of the blooming period in both the species. No flower buds were formed when monochromic B or R was used in salvia and FLFr inhibited flower bud formation in marigold. In comparison with monochromic blue or red light, the number of stomata was greater in mixed radiation of FL with LEDs in both the plants. Our study demonstrates the effectiveness of a LED system for plantlet growth and morphogenesis in space-based plant research chambers.