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24-epibrassinolide ameliorates the adverse effect of salt stress (NaCl) on pepper (Capsicum annuum L.)

24-epibrassinolide ameliorates the adverse effect of salt stress (NaCl) on pepper (Capsicum annuum L.)

Ibn Maaouia-Houimli Samira, Dridi-Mouhandes Bouthaina, Ben Mansour-Gueddes Samia, Denden Mounir

Статья научная

The present study investigates the role of 24-epibrassinolide (EBL) in inducing plant tolerance to salinity. Seedlings of pepper (Capsicum annuum L.) were grown in the presence of 70 mM NaCl and were sprayed with 10-6 M EBL at 7 days after transplantation and were sampled at 28 day. The plants exposed to NaCl exhibited a significant decline in relative growth rate, net CO2 assimilation, stomatal conductance, transpiration and water use efficiency. However, the follow up treatment with EBL significantly improved the above parameters. EBL treated plants had greater relative growth rate compared to untreated plants when exposed to salt stress. Application of EBL increased photosynthesis by increasing stomatal conductance in both control and salt stressed plants and may have contributed to the enhanced growth. The water use efficiency was improved because CO2 assimilation is more important than the transpiration.

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24-epibrassinolide enhances flower and fruit production of pepper (Capsicum annuum L.) under salt stress

24-epibrassinolide enhances flower and fruit production of pepper (Capsicum annuum L.) under salt stress

Ibn Maaouia-Houimli Samira, Ben Mansour-Gueddes Samia, Dridi-Mouhandes Bouthaina, Denden Mounir

Статья научная

Among various environmental stresses, salt stress is extensively damaging to crops production in many areas of the world. The enhancement of plant tolerance is important for plant productivity. Brassinosteroids plays an important role in the plant response to adverse environmental conditions such as salt stress. Therefore, an experiment was conducted to explore the role of exogenously applied 24-epibrassinolide on flower and fruit production of pepper in presence or absence of NaCl-stress (70 mM). The plants were sprayed with 10-6M of 24-epibrassinolide at vegetative, buds formation and early fruiting. 24-epibrassinolide improves flower number, fruit number and yield per plant, but was without effect on fruit mass and size. The detailed measurement of these parameters indicates that the effects of 24-epibrassinolide depend on the development stage of pepper plant and the application frequency of this hormone.

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24-epibrassinolide mediated changes in photosynthetic pigments and antioxidative defence system of radish seedlings under cadmium and mercury stress

24-epibrassinolide mediated changes in photosynthetic pigments and antioxidative defence system of radish seedlings under cadmium and mercury stress

Kapoor Dhriti, Rattan Amandeep, Gautam Vandana, Kapoor Nitika, Bhardwaj Renu

Статья научная

The present work was conducted to study the effects of 24-EBL on photosynthetic pigments (total chlorophyll, chl a, chl b, carotenoid, anthocyanin and flavonoid content) and activities of antioxidative enzymes (guaiacol peroxidase, catalase, superoxide dismutase, ascorbate peroxidase, glutathione reductase, dehydroascorbate reductase, mono-dehydroascorbate reductase, polyphenol oxidase, glutathione peroxidase and glutathione-S- transferase and protein content) in 7-day old seedlings of Raphanus sativus exposed to cadmium and mercury toxicity. Findings of present study were revealed that brassinolide was proved beneficial for amelioration of Cd and Hg stress by altering various metabolic processes of plant.

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28-homobrassinolide modulate antenna complexes and carbon skeleton of Brassica juncea L. under temperature stress

28-homobrassinolide modulate antenna complexes and carbon skeleton of Brassica juncea L. under temperature stress

Kaur Harpreet, Sirhindi Geetika, Bhardwaj Renu, Sharma Poonam, Mudasir Mir

Статья научная

The aim of present study was to explore the ameliorative impact of 28-homoBL on morpho-physiological attributes, photosynthetic pigments and sugars of Brassica juncea L. exposed to oxidative stress caused by extreme temperatures (4 and 44 °C). For this, experiments were carried out at the Plant Physiology Laboratory, Department of Botany, Punjabi University, Patiala. Effect of different degrees of temperature (4 and 44 °C) taking 24 °C as control was studied. 28-homoBL (10 -6, 10 -9 and 10 -12M) primed and unprimed seeds of B. Juncea L. in terms of antenna complexes and end products of photosynthesis that is total carbohydrates and total soluble sugars was investigated. All concentrations of 28-homoBL used in present study showed different effects on morphology and light quenching pigments. All concentrations of 28-homoBL showed promoting effect on growth and light quenching pigments. The carbon makeup ameliorated positively in stressed and non-stressed components of photosynthetic machinery and 10 -9 M 28-homoBL showed best results. In conclusion 28-homoBL showed great potential in protecting the reaction centre of photosynthetic machinery from oxidative stress caused by extreme low and high temperatures but in very dose dependent manner and thus modulate the carbon skeleton of the plant.

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28-homobrassinolide protects photosynthetic machinery in Indian mustard under high temperature stress

28-homobrassinolide protects photosynthetic machinery in Indian mustard under high temperature stress

Fariduddin Qazi, Yusuf Mohammad, Begum Mahmooda, Ahmad Aqil

Статья научная

High temperature is a serious threat to crop production. Brassinosteroids (BRs), a group of plant steroidal hormones, can reduce effects of abiotic stresses. The present study was aimed to study the potency of brassinosteroids on high temperature induced changes in Indian mustard ( Brassica juncea L.) for effects on growth, chlorophyll, photosynthesis, photosystem II, antioxidant system and proline. Surface sterilized seeds of Indian mustard were sown in pots, grown for 21 days and treated with double distilled water or 0.01 µM of 28-homobrassinolide. Treated plants, after 24 h, were exposed to 30°C or 40°C for 48 h. One set of plants were kept at ambient temperature, 25°C, as the control. Plants were harvested at 30 days stage of growth to assess the various parameters. Plants exposed to 40°C had a decline in growth, leaf water potential, chlorophyll, photosynthetic rate, and activities of carbonic anhydrase (E.C.4.2.1.1) and nitrate reductase (E.C.1.6.1.1). The 28-homobrassinolide alone improved growth and photosynthesis responses along with various enzymes activities. Treatment of plants with HBL prior to exposure to 40°C, partially reduced damage and completely controlled damage when exposure was to 30°C. Levels of the antioxidative enzymes catalase (E.C.1.11.1.6), peroxidase (E.C.1.11.1.7), and superoxide dismutase (E.C.1.15.1.1), and the level of proline increased in response to 30 or 40°C and were further enhanced in the presence of 28-homobrassinolide. Plants grown under high temperature had increased levels of H 2O 2; application of HBL before temperature treatment decreased H 2O 2 content compared to the control. Elevated levels of antioxidative enzymes and proline might be responsible for conferring tolerance to high temperature stress in Indian mustard and overcome the loss of productivity of the crop.

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A comparative assessment on physicochemical properties of Senna auriculata leaves and flowers of the natural and polluted sources

A comparative assessment on physicochemical properties of Senna auriculata leaves and flowers of the natural and polluted sources

Nagalakshmi R., Anand S. P.

Статья научная

Background: The standardization of herbal medicines is an essential step in evaluating the quality of medicines based on drug concentration, physical and chemical standards. Due to the fact that the plant of Senna auriculata L. has a great medicinal potential with information about the compounds present in the various parts of the plant, it was considered important to study the preliminary physicochemical properties. Results: Samples A & B were collected from natural habitats and Samples C & D were collected from polluted areas. Physicochemical properties of powdered plant material of S. auriculata include extractive value, ash value, loss on drying, and fluorescence behaviour were determined as per standard methods. The water-soluble extractive value in the leaves and flowers of samples A and B was greater than samples C and D. In terms of alcohol soluble extractive value, both samples A and B had a high percentage yield in their leaves and flowers, whereas samples C and D had a somewhat lower percentage yield. The leaves and flowers of samples A and B had lower moisture content percentages, whereas samples C and D had somewhat higher moisture content percentages. Conclusions: The results of the physicochemical analysis suggest that samples A and B were far preferable to samples C and D in terms of quality. According to the findings of this study, collecting medicinal plants from their natural habitat is preferred since contaminated environments might impact the physicochemical properties of the plant material.

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A comparative study of lichen biochemistry and air pollution status of urban, semi urban and industrial area of Hooghly and Burdwan district, West Bengal

A comparative study of lichen biochemistry and air pollution status of urban, semi urban and industrial area of Hooghly and Burdwan district, West Bengal

Das K., Dey U., Bhaumik R., Datta J.K., Mondal N.K.

Статья научная

Lichens are group of nonvascular plants. Lichen absorbs water and nutrient passively from their environment. Because of this lichens are particularly sensitive to environmental factors such as temperature, water availability, and air pollutants and/or by substances accumulates in lichen from the environment through variety of mechanisms including particulate trapping, ion exchange, extracellular electrolyte adsorption, hydrolysis and intra cellular uptake. Air pollution control is more complex than most other environmental changes. A study has been done to evaluate the air pollution status by means of lichen through the biochemical variability of three macro environment (semiurban area Arambagh, urban area Burdwan and industrial area Durgapur). The study results revealed that there exist inverse relationship between pollutant load and pigment content (chlorophyll and carotenoid content). Average concentration of chlorophyll 'a', 'b', and total chlorophyll was found in following order, Site-II>Site-III>Site-I. From the result of Chlorophyll Stability Index (CSI) it has been found that the chlorophyll degradation is highest in Site-II followed by Site-III and lowest in Site-I. There is a strong inverse relationship of chlorophyll content in lichen with SO2and NOxbut the level of SPM found highest in Site-I followed by Site-III and lowest in Site-II. Moreover, biochemical constituents (protein, sugar, phenol and proline) of all the representative samples were also vary from site to site. The study results reveal that the amount of soluble sugar was highest recorded at Site-III followed by Site-I and lowest in Site-II. Again correlation study indicates that there is positive relationship between soluble sugar and phenol content at 5% level of significance. Therefore, it also indicates that the lichen metabolite and other physiological action monitored by SO2, NOx SPM level in the air. Studies showed that the semiurban area was under more vulnerable condition than industrial and urban area of Hooghly and Burdwan district. Moreover the micrograph study of representative lichen samples directly proved that lichen structure affected by air pollutants, but the extent of deformation exclusively depends on the level of pollutants present in the air. Results also suggest that different level of air pollutants (SO2, NOxand SPM) affect the biochemical parameters of lichen, but more extensive study is needed to confirm the relationship between air pollution status and lichen biochemistry.

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A comprehensive comparative study on Cascabela thevetia (L.) Lippold, seed aqueous extract-mediated escalation of abiotic stress and cellular genotoxicity: insights from multivariate allelochemical analysis vis-a-vis employment of plant bioassays (Lathyrus sativus L., and Allium sativum L., germinating root tip cells)

A comprehensive comparative study on Cascabela thevetia (L.) Lippold, seed aqueous extract-mediated escalation of abiotic stress and cellular genotoxicity: insights from multivariate allelochemical analysis vis-a-vis employment of plant bioassays (Lathyrus sativus L., and Allium sativum L., germinating root tip cells)

Adhikari D., Ghosh T., Ghosh R.

Статья научная

Background : Cascabela thevetia (L.) Lippold, a popular member of family Apocynaceae, (Yellow Oleander), being one of the preferred weapons for suicides in villages of India is grown in garden and roadside ways as a tree a. Pharmacologically active constituents include terpenoids, a-vonoid, steroids and glycosides in seeds.

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A modified D-roms method for determination of lipid peroxidation

A modified D-roms method for determination of lipid peroxidation

Nassar Muammar A.Y., Sharif Saleh H., Hammad Khalda M.A.

Статья научная

The need for simple and low costing and less time consuming methods for the determination of lipid peroxides and oxidative stress is urgent because oxidative stress is still unable to become a routine laboratory biomarker due to the sophisticated procedures and costing, a matter that made oxidative stress a research only biomarker, despite its crucial value as a risk factor and pathologic status, for many inflammatory and metabolic diseases. This work aimed at developing a modified rapid method to determine serum lipid peroxides at wavelength 492 nm and reference wavelength 630 nm. The results showed linear responses with increasing concentration and that the cheap N,N-DM-P-Phenylenediamine diHCl can be used as a chromogen under saving conditions of pH, temperatures and volume of sample.

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A novel index to estimate the stress-physiological status in wheat and barley against Fusarium head blight pathogens

A novel index to estimate the stress-physiological status in wheat and barley against Fusarium head blight pathogens

Sakr Nachaat

Статья научная

Wheat and barley can be massively invaded with Fusarium head blight (FHB) which results in decrease in grain weight and quality. Accurate evaluation of the stress-physiological status including quantitative resistance (QR) and aggressiveness in breeding programs is a key to selecting resistant cultivars. Out of several analyzed components, one or two metric components composing a disease index are commonly employed as a typical methodology to score FHB; however, this resistance index can not describe correctly and accurately the whole resistance in host plants. We have therefore assessed a novel index to estimate the physiological and pathogenic status in a set of diverse bread and durum wheat and barley cultivars under artificial infection with four Fusarium species. Eighteen QR components obtained under in vitro , growth camber and field conditions were evaluated and nine components at the earliest and latest growth stages were selected and combined in one index to score QR. This novel index was further investigated for its stability in the same fungi-seedling and -adult plants treated with silicon that reduced these nine pathogenic components forming this index. The scores of our index were highly significant and correlated with data of reductions resulted in silicon treatments across the bread and durum wheat and barley cultivars, thus it is suitable to be used for quantification of resistance in host plants and aggressiveness of Fusarium isolates. Findings show that the novel index analyzed in the present report captured key components of quantitative wheat and barley resistance. This index can be applied for screening the most aggressive isolates of different FHB species for FHB resistance breeding of wheat and barley. The mean of nine disease components is introduced in this paper as a new index for QR in plants and aggressiveness of fungi. Moreover, the novel index could be a starting point for more correct techniques for assessing on of the most damaging biotic stresses on cereals, FHB.

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A review of lysimeter studies and experiments by considering agricultural production

A review of lysimeter studies and experiments by considering agricultural production

Shahrajabian M.H., Sun W.

Статья обзорная

Lysimeters have been used to obtain accurate information for developing calibrating, and validating crop evapotranspiration and crop coefficients for many plants and crops. Lysimeters are also an unique equipment for studying the transport of solutes when saline waters are used, and therefore, for assessing alkalization and salinization hazards. Three main types of lysimeters are used: constatnt water-table lysimeters, drainage lysimeters, and weighing lysimeters. The weighing lysimeters provide scientist the basic information for research related to the evapotranspiration, and they are commonly divided into two types, continuous weighing and intermittent weighing. Lysimeters are foremost devices, typically tanks or containers, that define a specific boundary to contain soil water and permit measurement of either the soil-water balance or the volume of water percolating vertically and its quality. The limitations are expense which depends on design, variable experimental conditions such as climatic/environmental factors which are usually not controlled, the spatial variability is normally less, they are not appropriate for every plant species and even every soil type. The main goal of lysimeter is defining the crop coefficient (Kc) which used to convert Etr to equivalent crop ET (Etc) values, and determining agronomical parameters of crops which are planted on the field of lysimeter. All weather data like air temperature, humidity, solar radiation, and potential evaporation should be obtained onsite, and the frequency and time of measurements should be at least daily. For crop products, the management such as fertilization, sowing tillage, seed bed preparation, and harvest of the lysimeter including its surrounding area is carried out on the basis of good agricultural practice. it may be required to complement natural precipitation by irrigation. The main purpose of this literature review is to give a brief summary about lysimeters, and survey the impacts of lysimeter studies and crops production. The information provides is obtained from randomized control experiments, review articles, and analytical studies and observations which were gathered from numerous literature sources such as Scopus, PubMed, Google Scholar,and Science Direct.

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A study of the relationship between serum sialic acid and urine microalbumin in patients of type 2 diabetes mellitus nephropathy

A study of the relationship between serum sialic acid and urine microalbumin in patients of type 2 diabetes mellitus nephropathy

Patel Darshan J., Chakrabarti Chandan, Sharma Dinesh P., Maheshwari Amit V.

Статья научная

Background and Aim: Laboratory assessment of sialic acid as a marker of microvascular complications can be a diagnostic tool for diabetes mellitus. Microalbumin is an established predictor of diabetic nephropathy. The study was undertaken to evaluate serum sialic acid and microalbumin levels and to correlate it with glycemic control in diabetic nephropathy patients. Material and Methods: Clinically diagnosed diabetic nephropathy patientsvisiting the medicine OPD (outdoor patient department) and NCD (Non-Communicable Disease) were selected for the study. This study included 75 subjectswith diabetes types 2 of which 44 were males and 31were females. Control includes a total of 75 out of which 49 Males and 26 females. Blood samples were analysed for the glucose, C-reactive protein, and sialic acid. Urine sample was analyzed for microalbumin and sialic acid.

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A study on serum electrolyte pattern during ambient stress in Murrah buffalo of arid tracts in India

A study on serum electrolyte pattern during ambient stress in Murrah buffalo of arid tracts in India

Joshi A., Kataria N., Kataria A.K., Pandey N., Khan S.

Статья научная

A study was undertaken to assess the effect of ambient stress on serum electrolyte pattern of Murrah buffaloes of arid tracts in India. Healthy adult female Murrah buffaloes were grouped according to physiological states and screened during moderate, hot and cold ambiences. The mean values (m mol L-1) of serum inorganic phosphorus, calcium and magnesium during moderate ambience were 1.80± 0.01, 2.80± 0.01 and 0.90± 0.002, respectively. During hot and cold ambiences significant (p≤0.05) decrease was observed in the mean value of each electrolyte. Decline was higher during hot ambience as compared to cold ambience. In each ambience it was noticed that serum electrolyte concentration of pregnant dry animals was lowest significantly (p≤0.05) as compared to non pregnant milch followed by pregnant milch. Multipara animals showed significantly (p≤0.05) lower value of each electrolyte as compared to primipara. It was concluded that ambient stress during hot and cold ambiences resulted in depletion of electrolyte levels of the buffaloes of all physiological states.

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ABA biosynthesis defective mutants reduce some free amino acids accumulation under drought stress in tomato leaves in comparison with arabidopsis plants tissues

ABA biosynthesis defective mutants reduce some free amino acids accumulation under drought stress in tomato leaves in comparison with arabidopsis plants tissues

AL.asbahi Adnan ali, AL.maqtari Maher ali, Naji Khalid Mohammed

Статья научная

The ability of plants to tolerate drought conditions is crucial for plant survival and crop production worldwide. The present data confirm previous findings reported existence of a strong relation between abscisic acid (ABA) content and amino acid accumulation as response water stress which is one of the most important defense mechanism activated during water stress in many plant species. Therefore, free amino acids were measured to determine any changes in the metabolite pool in relation to ABA content. The ABA defective mutants of Arabidopsis plants were subjected to leaf dehydration for Arabidopsis on Whatman 3 mm filter paper at room temperature while, tomato mutant plants were subjected to drought stresses for tomato plants by withholding water. To understand the signal transduction mechanisms underlying osmotic stress-regulating gene induction and activation of osmoprotectant free amino acid synthesizing genes, we carried out a genetic screen to isolate Arabidopsis mutants defective in ABA biosynthesis under drought stress conditions. The present results revealed an accumulation of specific free amino acid in water stressed tissues in which majority of free amino acids are increased especially those playing an osmoprotectant role such as proline and glycine. Drought stress related Amino acids contents are significantly reduced in the mutants under water stress condition while they are increased significantly in the wild types plants. The exhibited higher accumulation of other amino acids under stressed condition in the mutant plants suggest that, their expressions are regulated in an ABA independent pathways. In addition, free amino acids content changes during water stress condition suggest their contribution in drought toleration as common compatible osmolytes.

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Abiotic stress-induced changes in antioxidative system and secondary metabolites production in Andrographis paniculata

Abiotic stress-induced changes in antioxidative system and secondary metabolites production in Andrographis paniculata

Mourya Suraj Kumar, Mohil Praveen, Kumar Anil

Статья обзорная

Andrographis paniculata is an annual herbaceous plant of the family Acanthaceae. Andrographolide (a bicycle diterpene) is the main component of A. paniculata and responsible for its bitter taste. Abiotic stress considerably affects the life cycle of plants through morphological, biochemical, and physiological changes. These stresses also act as elicitors for the synthesis of secondary metabolites (SMs) in plants. Secondary metabolites help plants to survive in adverse conditions via alleviating osmotic stress and reducing Reactive Oxygen Species (ROS). In this review, we discuss the response of the antioxidative system and biosynthesis of secondary metabolites in A. paniculata under abiotic stress. The mechanism involve in biosynthesis of ent-labdane diterpenoids is also discussed.

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Absorption of power plants СО 2 emissions by coniferous tree stands

Absorption of power plants СО 2 emissions by coniferous tree stands

Suvorova G.G.

Статья научная

The article reviews the ability of coniferous (common pine, siberian larch and siberian spruce) stands growing in 9 municipal districts of the Irkutsk region to absorb СО 2 technogenic emission of heat power plants. (EIGAF) index is suggested to characterize gas-absorbing (СО 2-absorbing) activity; the index reflects proportion between СО 2 technogenic emission and photosynthetic productivity (GPP) of coniferous tree stands. СО 2-absorbing capacity in 8 of the monitored districts has been shown to significantly exceed the amount of carbon dioxide emission from heat power sector. The index values EIGAF=0.01-0.97 demonstrate that СО 2 technogenic emission amounts to 1-97% of coniferous stands photosynthetic productivity in the areas under study. At the same time, the most industrially developed Angarsk district shows СО 2 photosynthetic absorption to be 8-12 times lower than technogenic СО 2 emission. Reasons of low gas-absorbing capacity of coniferous tree stands of this area are discussed.

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Abuse of anabolic androgenic steroids

Abuse of anabolic androgenic steroids

Yavari Abbas

Статья научная

According to the International Olympic Committee, the abuse of anabolic androgenic steroids (AASS) is found in over 50% of positive doping tests. AASS abuse is not restricted to the organized sports and widespread use. It remains as an unsolved public-health problem. Lower black market price, easier access to AASS, bodybuilding clubs and internet advertising are factors of this increasingly misuse. There is not real data about the prevalence of AASS abuse in various populations or countries, because most of athletes or students, due to their prohibition or ethical aspects do not admit to AASS abuse. Often they are aware of the risks of their choice and yet, are eager to put themselves at risk without deeper consideration. The abusers use them to improve their physical fitness and appearance. Present article has been collected to elucidate the risks and adverse effects of AASS and explanation of mechanisms of these events.

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Accumulation of heavy metals in soil-crop systems

Accumulation of heavy metals in soil-crop systems

Prajapati Karishma, Patel Ruby

Статья обзорная

The health concerns associated with heavy metal pollution (HMP) in agricultural soils have garnered attention on a global scale, and evaluations of the dangers to human health are based on research on the buildup of heavy metals in soil-plant systems. The pedosphere, hydrosphere, atmosphere, lithosphere, and biosphere all suffer from environmental pollution. As a result of the industrialization of many nations, heavy metal pollution is one of the most important environmental problems today. Heavy metal pollutants in the environment have been removed using a variety of ways, however these procedures have drawbacks such high cost, lengthy process times, logistical issues, and mechanical complexity. To use phytoremediation, which uses plants to remove, transport, and stabilize heavy metals from soil and water, one must understand the accumulation of heavy metals in plants and the function of plants in eliminating contaminants. Physiological and metabolic processes that enable plants to phytoremediator heavy metal-contaminated locations are now being optimized via genetic engineering techniques. This review investigated the accumulation of seven common heavy metals in soil agriculture systems: Cd, Cr, As, Pb, Hg, Cu, and Zn. It is reported that wheat was generally more prone than corn to acquire heavy metals. The seven heavy metals were ranked as follows, in ascending order of accumulation in grains: Pb

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Accumulation of osmoprotectants acclimating proso millet (Panicum miliaceum L.) to drought stress tolerance

Accumulation of osmoprotectants acclimating proso millet (Panicum miliaceum L.) to drought stress tolerance

Mir Reyaz Ahmad, Somasundaram R.

Статья научная

Drought is one of the environmental stressors that limits plant growth, production, and is a great threat to the rising population and security throughout the globe. In the present work, we aimed to check the impact of drought stress given at different days interval drought (DID) on proso millet and its tolerance capability. The pot culture experiment was conducted and drought stress was given at 3 DID, 5 DID, and 7 DID respectively, and sampling was done on 15, 30, and 60 DAS, and unstressed plants were irrigated regularly. Drought stress results in decreased protein contents in both the roots and shoots of P. miliaceum on all sampling days. However, with increasing drought intensity and duration there was a tremendous increase in amino acids, and proline contents as well. Therefore, increasing osmolytes like proline with an increased drought period proves the role of proline in stress tolerance of proso millet and helps it to survive by maintaining osmotic potential, water influx, and detoxifying ROS. Thus, proso millet can be cultivated in drought-prone areas, and used for the breeding program.

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