Calcium Phosphate Particles Coated with Humic Substances: A Potential Plant Biostimulant from Circular Economy.

Nowadays, the use of biostimulants to reduce agrochemical input is a major trend in agriculture. In this work, we report on calcium phosphate particles (CaP) recovered from the circular economy, combined with natural humic substances (HSs), to produce a plant biostimulant. CaPs were obtained by the thermal treatment of Salmo salar bones and were subsequently functionalized with HSs by soaking in a HS water solution. The obtained materials were characterized, showing that the functionalization with HS did not sort any effect on the bulk physicochemical properties of CaP, with the exception of the surface charge that was found to get more negative. Finally, the effect of the materials on nutrient uptake and translocation in the early stages of development (up to 20 days) of two model species of interest for horticulture, Valerianella locusta and Diplotaxis tenuifolia, was assessed. Both species exhibited a similar tendency to accumulate Ca and P in hypogeal tissues, but showed different reactions to the treatments in terms of translocation to the leaves. CaP and CaP-HS treatments lead to an increase of P accumulation in the leaves of D. tenuifolia, while the treatment with HS was found to increase only the concentration of Ca in V. locusta leaves. A low biostimulating effect on both plants' growth was observed, and was mainly scribed to the low concentration of HS in the tested materials. In the end, the obtained material showed promising results in virtue of its potential to elicit phosphorous uptake and foliar translocation by plants.

Insights on Molecular Characteristics of Hydrochars by 13C-NMR and Off-Line TMAH-GC/MS and Assessment of Their Potential Use as Plant Growth Promoters.

Hydrochar is a carbon-based material that can be used as soil amendment. Since the physical-chemical properties of hydrochar are mainly assigned to process parameters, we aimed at evaluating the organic fraction of different hydrochars through 13C-NMR and off-line TMAH-GC/MS. Four hydrochars produced with sugarcane bagasse, vinasse and sulfuric or phosphoric acids were analyzed to elucidate the main molecular features. Germination and initial growth of maize seedlings were assessed using hydrochar water-soluble fraction to evaluate their potential use as growth promoters. The hydrochars prepared with phosphoric acid showed larger amounts of bioavailable lignin-derived structures. Although no differences were shown about the percentage of maize seeds germination, the hydrochar produced with phosphoric acid promoted a better seedling growth. For this sample, the greatest relative percentage of benzene derivatives and phenolic compounds were associated to hormone-like effects, responsible for stimulating shoot and root elongation. The reactions parameters proved to be determinant for the organic composition of hydrochar, exerting a strict influence on molecular features and plant growth response.

Maleic hydrazide elicits global transcriptomic changes in chemically topped tobacco to influence shoot bud development.

MAIN CONCLUSION: Transcriptomic analysis revealed maleic hydrazide suppresses apical and axillary bud development by altering the expression of genes related to meristem development, cell division, DNA replication, DNA damage and recombination, and phytohormone signaling. Topping (removal of apical buds) is a common agricultural practice for some crop plants including cotton, cannabis, and tobacco. Maleic hydrazide (MH) is a systemic suckercide, a chemical that inhibits shoot bud growth, used to control the growth of apical (ApB) and axillary buds (AxB) following topping. However, the influence of MH on gene expression and the underlying molecular mechanism of controlling meristem development are not well studied. Our RNA sequencing analysis showed that MH significantly influences the transcriptomic landscape in ApB and AxB of chemically topped tobacco. Gene ontology (GO) enrichment analysis revealed that upregulated genes in ApB were enriched for phosphorelay signal transduction, and the regulation of transition timing from vegetative to reproductive phase, whereas downregulated genes were largely associated with meristem maintenance, cytokinin metabolism, cell wall synthesis, photosynthesis, and DNA metabolism. In MH-treated AxB, GO terms related to defense response and oxylipin metabolism were overrepresented in upregulated genes. GO terms associated with cell cycle, DNA metabolism, and cytokinin metabolism were enriched in downregulated genes. Expression of KNOX and MADS transcription factor (TF) family genes, known to be involved in meristem development, were affected in ApB and AxB by MH treatment. The promoters of MH-responsive genes are enriched for several known cis-acting elements, suggesting the involvement of a subset of TF families. Our findings suggest that MH affects shoot bud development in chemically topped tobacco by altering the expression of genes related to meristem development, DNA repair and recombination, cell division, and phytohormone signaling.

Toxicity assessment of palladium oxide nanoparticles derived from metallosurfactants using multi assay techniques in Allium sativum.

In today's world, nanotechnology is reaching practically every ground and entering the human lifestyle by becoming a part of it. Thus, it is vital to check the cytotoxic and genotoxic effects of nanosubstances on plants, as they are the base constituent of ecosystem. The present work deals with the toxicity evaluation of metallosurfactant derived palladium oxide nanoparticles towards Allium sativum (Garlic cloves). The nanoparticles were prepared using microemulsion quenching method (a softer approach) using palladium metallosurfactants as precursors. The three ligands used were cetyltrimethylammonium chloride (CTAC), dodecylamine (DDA) and hexadecylamine (HEXA). Further, their characterization was done using TEM, Size Distribution curve, FESEM, EDS, XRD and Zeta potential. Garlic (Allium sativum) cloves were used to investigate the cytotoxicity and genotoxicity of fabricated PdO NPs. To check the cytotoxicity, optical microscopy was employed and for the genotoxic assessment, different parameters such as chromosomal aberrations in the mitosis, circular dichroism, and gel electrophoresis were utilized. From mitosis study, chromosomes aberrations were confirmed such as chromosomes stickiness, breakage, C-Mitosis, delay in anaphase, spindle fibre abnormality, laggard, vagrant and condensed chromosomes. Morphology of A. sativum clove, rooting and shooting pattern in the presence of PdO nanosuspension was observed. From all the experiments, it was concluded that all the three PdO nanosuspension are toxic in nature to both the cells and to genome, although, bishexadecyltrimethyl ammonium palladium tetrachloride (PdCTAC) Ns was found to be the most cytotoxic and genotoxic. Gel electrophoresis also confirmed the complete degradation of DNA in the presence of PdCTAC Ns.

The effects of soil phosphorus and zinc availability on plant responses to mycorrhizal fungi: a physiological and molecular assessment.

The positive effects of arbuscular mycorrhizal fungi (AMF) have been demonstrated for plant biomass, and zinc (Zn) and phosphorus (P) uptake, under soil nutrient deficiency. Additionally, a number of Zn and P transporter genes are affected by mycorrhizal colonisation or implicated in the mycorrhizal pathway of uptake. However, a comprehensive study of plant physiology and gene expression simultaneously, remains to be undertaken. Medicago truncatula was grown at different soil P and Zn availabilities, with or without inoculation of Rhizophagus irregularis. Measures of biomass, shoot elemental concentrations, mycorrhizal colonisation, and expression of Zn transporter (ZIP) and phosphate transporter (PT) genes in the roots, were taken. Mycorrhizal plants had a greater tolerance of both P and Zn soil deficiency; there was also evidence of AMF protecting plants against excessive Zn accumulation at high soil Zn. The expression of all PT genes was interactive with both P availability and mycorrhizal colonisation. MtZIP5 expression was induced both by AMF and soil Zn deficiency, while MtZIP2 was down-regulated in mycorrhizal plants, and up-regulated with increasing soil Zn concentration. These findings provide the first comprehensive physiological and molecular picture of plant-mycorrhizal fungal symbiosis with regard to soil P and Zn availability. Mycorrhizal fungi conferred tolerance to soil Zn and P deficiency and this could be linked to the induction of the ZIP transporter gene MtZIP5, and the PT gene MtPT4.

An efficient micropropagation protocol for an endangered ornamental tree species (Magnolia sirindhorniae Noot. & Chalermglin) and assessment of genetic uniformity through DNA markers.

Magnolia sirindhorniae Noot. & Chalermglin is an endangered species with high ornamental and commercial value that needs to be urgently protected and judiciously commercialized. In this study, a protocol for efficient regeneration of this species is standardized. The lateral buds of the M. sirindhorniae plant were used as an explant. Half-strength Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzyladenine (BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 2.0 mg/L gibberellic acid (GA3) was found to be the optimal medium for shoot induction. The maximum shoot multiplication rate (310%) was obtained on Douglas-fir cotyledon revised medium (DCR) fortified with 0.2 mg/L BA, 0.01 mg/L NAA, and additives. The half-strength DCR medium supplemented with 0.5 mg/L NAA and 0.5 mg/L indole-3-butyric acid (IBA) supported the maximum rate (85.0%) of in vitro root induction. After a simple acclimatization process, the survival rate of plantlets in a substrate mixture of sterile perlite and peat soil (1:3; v/v) was 90.2%. DNA markers were used for assessment of genetic uniformity, confirming the genetic uniformity and stability of regenerated plants of M. sirindhorniae. Thus, the described protocol can safely be applied for large scale propagation of this imperative plant.

Indirect Regeneration and Assessment of Genetic Fidelity of Acclimated Plantlets by SCoT, ISSR, and RAPD Markers in Rauwolfia tetraphylla L.: An Endangered Medicinal Plant.

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.

Assessment of Cadmium Scavenging Potential of Canna indica L.

The aquatic plant, Canna indica L. (Indian shoot) of Cannaceae family was investigated to assess cadmium scavenging potential at 5, 10, 25, 50, 100 and 150 mg Cd L-1 exposers. The results showed that Canna has considerable potential of cadmium accumulation, which was up to 58.69 and 10.13 mg Cd kg-1 dry weight in root and shoot of Canna, respectively. The effects of different cadmium levels on biomass production of plant tissues were significantly (p = 0.05) showed negative relation due to cadmium toxicity. The root concentration factor was higher than the bioconcentration factor which indicated the lower translocation factor of Canna. Considering the high root concentration factor, average bioconcentration factor, rapid growth and optimum adaptive properties up to 100 mg Cd L-1 level, the Canna could be employed as an eco-friendly and efficient aquatic plant for cadmium scavenging. This study plays a potential role in remediation of cadmium contaminated wastewater.

Evaluation of risk assessment of new industrial pollutant, ionic liquids on environmental living systems.

Ionic liquids (ILs) are much known for their promising alternative for volatile solvents in industries and gained popularity as a greener solvent, however industrial effluent discharge containing ILs are also increasing. There is a scarcity of information on the toxicity of ILs; the present study will explore different facts about their harmfulness. The toxic effects of five different ILs: [C4MIM]Br, [Hx3PC14]N(CN)2, [C10MIM]BF4, [BTDA]Cl and [C4MPY]Cl were analysed on bacteria, fungi, plant and animal cells. Both Gram positive and negative bacteria were found to be more susceptible to [C10MIM]BF4 and [BTDA]Cl than [C4MIM]Br, [Hx3PC14]N(CN)2 and [C4MPY]Cl, whereas fungi revealed quite a resistance to all ILs. All ILs were toxic towards Triticum aestivum affecting their roots and shoots, however [C10MIM]BF4 and [BTDA]Cl were more toxic amongst them. Studies on Allium cepa described their toxic behaviour at the genetic level by altering cell division and nuclear material. Furthermore, studies on human red blood cells described by % haemolysis in which [Hx3PC14]N(CN)2 and [BTDA]Cl exhibited higher toxicity at very lower concentrations. While the genotoxic effect on blood lymphocytes exerted by [Hx3PC14]N(CN)2, [C10MIM]BF4 and [BTDA]Cl confirmed their toxic effects on human cells.

From laboratory to field studies - The assessment of Biscutella laevigata suitability to biological reclamation of areas contaminated with lead and cadmium.

The aim of the work was to evaluate the usefulness of the in vitro multiplication of Biscutella laevigata calamine ecotype for in situ reclamation of post-flotation wastes polluted with Pb and Cd. The experiment was conducted on three steps: (i) plant shoots' production under in vitro condition, (ii) establishment of the material in greenhouse experiment, and finally (iii) field cultivation directly on the mining-waste heap of Olkusz Ore-Bearing Region, Poland. This region is known to be one of the most chemically-degraded area in central Europe. The laboratory-set in vitro analysis enabled to obtain the high-quality plant shoots, which multiply the most effectively (with growth tolerance index 130-150%) on medium containing 5.0µM CdCl2 and 0.5mM Pb(NO3)2. These plants were used for the next two ex vitro experiments. Several biometric and physiological analysis (i.e. of photosystem II activity Fv/Fm and PI, photosynthetic pigment contents) were done to indicate plant physiological status during these experiments. The main novelty of the work was to prove that in vitro-multiplied shoots of B. laevigata - the representative of native flora from Olkusz Ore-Bearing Region - can be successfully implemented in situ for the restoration of these degraded area. Moreover, the addition of sewage sludge as a source of organic compounds significantly improved plants' growth and development what is especially important due to the lack of other legal solutions for the management of the sewage sludge in some countries.

Effects of Copper Exposure on Photosynthesis and Growth of the Seagrass Cymodocea nodosa: An Experimental Assessment.

Seagrasses form some of the most important coastal habitats. They may be negatively affected by trace metal contamination in certain coastal areas. In this study we experimentally assessed selected morphological and physiological traits of the seagrass Cymodocea nodosa, with increasing concentrations of copper (Cu) under controlled laboratory conditions. Short term (21 days) sub-lethal effects such as decreased maximum quantum yield, increased leaf necrosis and decreased shoot growth and shoot recruitment were clearly observed at the highest Cu exposure (5 mg L(-1)), while the effects were weaker at the intermediate concentration (2.5 mg L(-1)) and almost absent at the lowest concentration (1 mg L(-1)), indicating that this species is highly tolerant to copper exposure, at least in the short term. This fact could help to explain its distribution in relatively polluted coastal waters.

Influence of different formulations on chlorpyrifos behavior and risk assessment in bamboo forest of China.

The effects of two formulations (emulsifiable concentrate (EC) and granule (G)) on the distribution, degradation, sorption, and residue risk of chlorpyrifos (CHP) were investigated in two producing areas of bamboo shoot. The results showed that CHP was mainly distributed in the topsoil (0-5 cm, P < 0.05), with the proportion of CHP in the total quantity ranging from 76.0 to 100.0 % (G) and 12.0 to 98.1 % (EC), respectively. The degradation of CHP-EC in soils (half-life 27.7-36.4 days) was faster than that of CHP-G in soils (half life above 120-150 days). The main metabolite of CHP, 3,5,6-trichloro-2-pyridinol (TCP), was found in soil samples. CHP showed good sorption ability in the two tested soils, with the sorption coefficient (KF) of 43.76 and 94.43 mg/kg. The terminal residues of CHP in bamboo shoots were in the range of 15.2-75.6 (G) and 10.4-35.7 µg/kg (EC), respectively. The soil type had a notable effect on the CHP behaviors in soil (P < 0.05, especially for CHP-G), but it did not affect the metabolite of CHP. Although some positive bamboo shoot samples (CHP residue exceeding maximum residue limits) were found, the hazard quotients did not exceed 7 %, which meant there was a negligible risk associated with the exposure to CHP via the consumption of bamboo shoots.

Interactive Effects of Growth Regulators, Carbon Sources, pH on Plant Regeneration and Assessment of Genetic Fidelity Using Single Primer Amplification Reaction (SPARS) Techniques in Withania somnifera L.

An improved and methodical in vitro shoot morphogenic approach through axillary bud multiplication was established in a drug yielding plant, Withania somnifera L. Effects of plant growth regulators [6-benzyladenine (BA), kinetin (Kin), 2-isopentenyladenine (2iP), and thidiazuron (TDZ)] either singly or in combination with α-napthalene acetic acid (NAA), indole-3-butyric acid (IBA), and indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium were tested. The highest regeneration frequency (90 %) with optimum number of shoots (32 ± 0.00)/explant were obtained on MS medium fortified with 2.5 µM 6-benzyladenine (BA) and 0.5 µM NAA and 30 g/l sucrose at pH 5.8. Among the tried TDZ concentrations, 0.5 µM resulted in maximum number of shoots (20.4 ± 0.40)/explant after 4 weeks of exposure. The proliferating shoot cultures established by repeated subculturing of the mother explants on the hormone-free medium produced the highest shoot number (29.4 ± 0.40) with shoot length (6.80 ± 0.12 cm)/explant at fourth subculture passage, which a decline in shoot proliferation was recorded. Different concentrations of NAA were tested for ex vitro rooting of microshoots. The maximum percentage of rooting 100 % with maximum roots (18.3 ± 0.1) was achieved in soilrite when basal portion of the microshoots were treated with 200 µM (NAA) for 15 min per shoot. The plantlets went through hardening phase in a growth chamber, prior to ex vitro transfer. The PCR-based single primer amplification reaction (SPAR) methods which include random amplified polymorphic DNA (RAPD) and direct amplification of minisatellite DNA (DAMD) markers has been used for assessment of genetic stability of micropropagated plantlets. No variation was observed in DNA fingerprinting patterns among the micropropagated and the donor plants illustrating their genetic uniformity.

Impact assessment of mercury accumulation and biochemical and molecular response of Mentha arvensis: a potential hyperaccumulator plant.

The present study was focused on examining the effect of Hg oxidative stress induced physiochemical and genetic changes in M. arvensis seedlings. The growth rate of Hg treated seedlings was decreased to 56.1% and 41.5% in roots and shoots, respectively, compared to the control. Accumulation of Hg level in both roots and shoots was increased with increasing the concentration of Hg. Superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities were found to be increased with increasing the Hg concentration up to 20 mg/L; however, it was decreased at 25 mg/L Hg concentration. The POX enzyme activity was positively correlated with Hg dose. The changes occurring in the random amplification of ploymorphic DNA (RAPD) profiles generated from Hg treated seedlings included variations in band intensity, disappearance of bands, and appearance of new bands compared with the control seedlings. It was concluded that DNA polymorphisms observed with RAPD profile could be used as molecular marker for the evaluation of heavy metal induced genotoxic effects in plant species. The present results strongly suggested that Mentha arvensis could be used as a potential phytoremediator plant in mercury polluted environment.

Evaluation of biological endpoints in crop plants after exposure to non-steroidal anti-inflammatory drugs (NSAIDs): implications for phytotoxicological assessment of novel contaminants.

Human pharmaceuticals have been detected in the terrestrial environment at µg to mg kg(-1) concentrations. Repeated application of sewage sludge (biosolids) and increasing reclaimed wastewater use for irrigation could lead to accumulation of these novel contaminants in soil systems. Despite this, potential phytotoxicological effects on higher plants have rarely been evaluated. These studies aimed to test effects upon germination, development, growth and physiology of two crop plants, namely radish (Raphanus sativus Spakler 3) and lettuce (Lactuca sativa All Year Around), after exposure to different, but structurally related non-steroidal anti-inflammatory drugs (NSAIDs) at environmentally relevant concentrations. A range of biological endpoints comprising biomass, length, water content, specific root and shoot length, root to shoot ratio, daily progress of stages of cell elongation and organ emergence (primary root, hypocotyl elongation, cotyledon emergence, cotyledon opening, and no change), as well as photosynthetic measurements were evaluated. Compounds from the fenamic acid class were found to affect R. sativus root endpoints (root length and water content), while ibuprofen affected early root development of L. sativa. In general, phytotoxicological effects on root endpoints demonstrated that impacts upon higher plants are not only compound specific, but also differ between plant species. It was found that the usage of a wide range of biological endpoints (all simple, cost-effective and ecologically relevant) were beneficial in detecting differences in plant responses to NSAID exposure. Due to paucity and discrepancy within the few previously available phytotoxicological studies with pharmaceuticals, it is now essential to allocate time and resources to consider development of suitable chronic toxicity tests, and some suggestions regarding this are presented.

Effect of alpha-naphthalene acetic acid and thidiazuron on seedling of economic crops grown in endosulfan sulfate-spiked sand.

The effect of two plant growth regulators, alpha-naphthalene acetic acid (NAA) and thidiazuron (TDZ) on the growth of sweet corn (Zea mays), cowpea (Vigna sinensis) and cucumber (Cucurmis sativus) seedling planted in 1-100 mg kg(-1) of endosulfan sulfate spiked sand was investigated. Endosulfan sulfate had no apparent toxicity as seedlings of these crop plants grew normally in endosulfan sulfate spiked sand. Concentration of endosulfan sulfate in sand affected the response of seedling induction by NAA or TDZ. Induction of crop seeds by NAA or TDZ did not promote growth of sweet corn, cowpea and cucumber to an appreciable extent. Both plant regulators at concentration of 10 mg l(-1) seemed to exert adverse effect on crop seedling. TDZ decreased shoot length, root length and chlorophyll contents in leaves of sweet corn and cowpea growing in endosulfan sulfate spiked sand. In contrast, NAA was not toxic and promoted growth of sweet corn and cowpea seedling. However, cucumber was affected by NAA and TDZ more than other plants. TDZ significantly decreased biomass and root length of cucumber. Also, NAA significantly decreased cucumber root length and tended to increase cucumber root dried weight when grown in 100 mg kg(-1) of endosulfan sulfate spiked sand.

Assessment of plant uptake models used in exposure assessment tools for soils contaminated with organic pollutants.

The aim of this study was to evaluate and improve the accuracy of plant uptake models for neutral hydrophobic organic pollutants (1 < logK(OW) < 9, -8 < logK(AW) < 0) used in regulatory exposure assessment tools, using uncertainty and sensitivity analyses. The models considered were RAIDAR, EUSES, CSOIL, CLEA, and CalTOX. In this research, CSOIL demonstrated the best performance of all five exposure assessment tools for root uptake from polluted soil in comparison with observed data, but no model predicted shoot uptake well. Recalibration of the transpiration and volatilisation parameters improved the performance of CSOIL and CLEA. The dominant pathway for shoot uptake simulated differed according to the properties of the chemical under consideration; those with a higher air-water partition coefficient were transported into shoots via the soil-air-plant pathway, while chemicals with a lower octanol-water partition coefficient and air-water partition coefficient were transported via the root. The soil organic carbon content was a particularly sensitive parameter in each model and using a site specific value improved model performance.

Cryopreservation of Thymus lotocephalus shoot tips and assessment of genetic stability.

BACKGROUND: Thymus lotocephalus is a rare endemic species from the Algarve, Portugal, and is legally protected by Portuguese and European legislation. OBJECTIVE: The aim is to develop a cryopreservation protocol for T. lotocephalus shoot tips, as an alternative approach for the long-term conservation of this species. METHODS: Several methods (droplet-vitrification, vitrification and encapsulation-dehydration) were tested. Conditions regarding the subculture period, cold-hardening and preculture were optimized. Cryopreserved shoot tips were also assessed for their genetic stability using RAPD markers. RESULTS: Droplet-vitrification presented the best results. The best regrowth of cryopreserved shoot tips obtained eight weeks after rewarming was 67%. This was accomplished with four weeks subculture period of in vitro-donor plants at 25 degree C, preculture of excised shoot tips for one day on MS medium containing 0.3 M sucrose, treatment in PVS2 for 60 min, and MS supplemented with 0.2 mg per L zeatin as recovery medium. The assessment using RAPD markers observed variation at a low frequency and shoots regenerated from cryopreserved apices showed normal development compared to the regular in vitro-grown shoots. CONCLUSION: Droplet-vitrification is thus a viable method for the cryopreservation of T. lotocephalus shoot tips.

Assessment of growth and yield losses in two Zea mays L. cultivars (quality protein maize and nonquality protein maize) under projected levels of ozone.

Rapid industrialization and economic developments have increased the tropospheric ozone (O3) budget since preindustrial times, and presently, it is supposed to be a major threat to crop productivity. Maize (Zea mays L.), a C4 plant is the third most important staple crop at global level with a great deal of economic importance. The present study was conducted to evaluate the performance of two maize cultivars [HQPM1: quality protein maize (QPM)] and [DHM117: nonquality protein maize (NQPM)] to variable O3 doses. Experimental setup included filtered chambers, nonfiltered chambers (NFC), and two elevated doses of O3 viz. NFC+15 ppb O3 (NFC+15) and NFC+30 ppb O3 (NFC+30). During initial growth period, both QPM and NQPM plants showed hormetic effect that is beneficial due to exposure of low doses of a toxicant (NFC and NFC+15 ppb O3), but at later stages, growth attributes were negatively affected by O3. Growth indices showed the variable pattern of photosynthate translocation under O3 stress. Foliar injury in the form of interveinal chlorosis and reddening of leaves due to increased production of anthocyanin pigments was observed at higher concentrations of O3. One-dimensional gel electrophoresis of leaves taken from NFC+30 showed reductions of major photosynthetic proteins, and differential response was observed between the two test cultivars. Decline in the number of male flowers at elevated O3 doses suggested damaging effect of O3 on reproductive structures which might be a cause of productivity losses. Variable carbon allocation pattern particularly to husk leaves, foliar injury, and damage of photosynthetic proteins led to significant reductions in economic yield at higher O3 doses. PCA showed that both the cultivars responded more or less similarly to O3 stress in their respective groupings of growth and yield parameters, but magnitude of their response was variable. It is further supported by difference in the significance of correlations between variables of yield and AOT40. Cultivar response reflects that QPM performed better than NQPM against elevated O3.

In vitro propagation and assessment of the genetic fidelity of Musa acuminata (AAA) cv. Vaibalhla derived from immature male flowers.

An efficient in vitro propagation method has been developed for the first time for Musa acuminata (AAA) cv. Vaibalhla, an economically important banana cultivar of Mizoram, India. Immature male flowers were used as explants. Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) were used for the regeneration process. Out of different PGR combinations, MS medium supplemented with 2 mg L(-1) 6-benzylaminopurine (BAP) + 0.5 mg L(-1) α-naphthalene acetic acid (NAA) was optimal for production of white bud-like structures (WBLS). On this medium, explants produced the highest number of buds per explant (4.30). The highest percentage (77.77) and number (3.51) of shoot formation from each explants was observed in MS medium supplemented with 2 mg L(-1) kinetin + 0.5 mg L(-1) NAA. While MS medium supplemented with a combination of 2 mg L(-1) BAP + 0.5 mg L(-1) NAA showed the maximum shoot length (14.44 cm). Rooting efficiency of the shoots was highest in the MS basal medium without any PGRs. The plantlets were hardened successfully in the greenhouse with 96% survival rate. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were employed to assess the genetic stability of in vitro regenerated plantlets of M. acuminata (AAA) cv. Vaibalhla. Eight RAPD and 8 ISSR primers were successfully used for the analysis from the 40 RAPD and 30 ISSR primers screened initially. The amplified products were monomorphic across all the regenerated plants and were similar to the mother plant. The present standardised protocol will find application in mass production, conservation and genetic transformation studies of this commercially important banana.