NUE of major nutrients like N, P and K are very low within the plant systems. In the enhancement of NUE in the plant system miRNA can play a very important role.
miRNA diversity related to NUE indicates different miRNA sequences and their target sequence in respect of Nutrient uptake ,translocation and their assimilation in plants. miRNAs are vital for maintaining nutrient homeostasis in plants by regulating the expression of transporters that are involved in nutrient uptake and mobilization. The alteration of nutrient levels in soil trigger specific signaling molecules that act as repressors of target nutrient responsive- miRNAs . The decreased accumulation of miRNAs subsequently stabilizes the expression of transporters. The identification of promoter regions of specific up- or down- regulated miRNAs that are responsive to micronutrient stresses and the subsequent development of knock-out mutants by inducing mutation in cis-acting elements using targeted genome- editing technologies ,such as TALEN or CRISPR-Cas9 techniques, may lead to essential crop-improvement strategies in the future.
Ribosomal Proteins and their Extra Ribosomal Functions in Abiotic Stress Tole...CrimsonpublishersMCDA
Ribosomal proteins (RPs) that include both small (RPS) and large subunit (RPL) proteins have been known to be involved in several very important functions in ribosome assembly, protein synthesis and other cellular functions in association with several other components [1]. The composition of ribosomal protein subunits that are involved in ribosome assembly is heterogeneous [2] indicating clearly that individual subunit protein components have functions also in phenomena like stress tolerance. Although each RP gene has multiple paralogs, the expression of all of them is differentially required for normal development with some of them functioning in spatio-temporal and signal-induced manner while others exhibit binding properties. The expression of ribosomal proteins has been shown to be regulated by various environmental cues and treatments with signaling molecules [3,4]. The involvement of ribosomal proteins in extra ribosomal functions in animal systems has been well documented [5].
https://crimsonpublishers.com/mcda/fulltext/MCDA.000591.php
For more open access journals in Crimson Publishers please click on link: https://crimsonpublishers.com
For more articles on journal of agronomy and crop science please click on below link: https://crimsonpublishers.com/mcda/
Post-translational modifications play an important role in cells, such as DNA recognition, protein-protein interactions, catalytic activity, and protein stability. Protein acetylation/deacetylation is a histone covalent modification that is mainly catalyzed by histone acetylase and histone deacetylase, respectively.
Ribosomal Proteins and their Extra Ribosomal Functions in Abiotic Stress Tole...CrimsonpublishersMCDA
Ribosomal proteins (RPs) that include both small (RPS) and large subunit (RPL) proteins have been known to be involved in several very important functions in ribosome assembly, protein synthesis and other cellular functions in association with several other components [1]. The composition of ribosomal protein subunits that are involved in ribosome assembly is heterogeneous [2] indicating clearly that individual subunit protein components have functions also in phenomena like stress tolerance. Although each RP gene has multiple paralogs, the expression of all of them is differentially required for normal development with some of them functioning in spatio-temporal and signal-induced manner while others exhibit binding properties. The expression of ribosomal proteins has been shown to be regulated by various environmental cues and treatments with signaling molecules [3,4]. The involvement of ribosomal proteins in extra ribosomal functions in animal systems has been well documented [5].
https://crimsonpublishers.com/mcda/fulltext/MCDA.000591.php
For more open access journals in Crimson Publishers please click on link: https://crimsonpublishers.com
For more articles on journal of agronomy and crop science please click on below link: https://crimsonpublishers.com/mcda/
Post-translational modifications play an important role in cells, such as DNA recognition, protein-protein interactions, catalytic activity, and protein stability. Protein acetylation/deacetylation is a histone covalent modification that is mainly catalyzed by histone acetylase and histone deacetylase, respectively.
The Electrophoretic Profile Myofibrillar Proteins Extracted From Camel Muscle...IJEAB
Changes in electrophoretic profiles of myofibrillar protein (MFP) in the Longissimus thoracis (LD) of young camels (2 to 4 years), preserved by refrigeration has been treated or not by lactic acid solution 4% or citric acid 1%, were followed during the post-mortem time at the following times: 1, 2, 4, 6, 8, 10, 12, 24 and 48 hours. The cold preservation for 48 hours has not shown any particular distinctions in the protein profiles of this muscle. Changes related to the type of treatment were recorded during the storage time. Proteolysis of the myofibrillar fraction was earlier in this muscle in the case of treatment with one of two solutions of organic acids used, particularly in the case of using lactic acid. Indeed, these changes have affected at the first hour after slaughter the proteolysis of the myofibrillar proteins. Fragments of low molecular weight (42, 36, 33, 26, 23, 18, 16, 14 and 13 kDa) have been identified. The electrophoretic analysis showed that during refrigeration, LD treated with a solution of lactic acid is more sensitive to disruption phenomena and muscle protein proteolysis that lots of this muscle that even in the case of preservation by refrigeration only or by refrigeration
''Translational Repression and eIF4A2 Activity Are Critical for MicroRNA-Medi...Wisdom Deebeke Kate
Translational Repression and eIF4A2 Activity Are Critical for MicroRNA-Mediated Gene Regulation.
This assessed presentation looked at the effects of miRNA-mediated repression on mRNA. These effects include deadenylation, translational inhibition and degradation. It was observed that the eIF4A2 is a required component in the repression pathway mediated by the miRNA let-7. Presentation by KATE, Wisdom Deebeke; Mene Nkue; and Michael Thirlwell. Dated: Thursday, December 5th, 2013. (Phase 2, stage 3, semester 1).
Transcriptional regulation of functional genes involved in cuticular wax bios...Agriculture Journal IJOEAR
Abstract— MYB transcription factor family is one of the largest families in the plant kingdom, specifically characterized by a Helix-turn-helix domain in their structural configuration with two repeats of R2R3 MYB domain and are known for regulating plant stress tolerance (abiotic and biotic) through ABA dependent signaling systems. R2R3- MYB family transcription factors plays a role in the regulation of specific downstream genes of Very Long Chain Fatty Acid Biosynthesis like KCS1, KCS2, KCS6 and KCR1, related to water use efficiency traits like cuticular wax biosynthesis. Analysis of R2R3-MYB family transcription factors regulating the production of cuticular wax emphasizes the value of the family outside of traditionally accepted roles in stress tolerance. Some MYB transcription factors like MYB96, MYB94, MYB41, MYB30, MYB106 and MYB16 isolated from Arabidopsis thaliana (Col-0) are studied with respect to their role in biosynthesis of cuticular waxes under environmental stress signals. In the present review we intensely focused on the elucidation of R2R3- MYB family transcription factors MYB96, MYB94, MYB41, MYB30, MYB106 and MYB16 role in cuticular wax biosynthesis and aid in the development of transgenics for enhanced stress tolerance.
Activation of p53 mediated glycolytic inhibition-oxidative stressapoptosis pa...rkkoiri
There is a general agreement that most of the cancer cells switch over to aerobic glycolysis (Warburg
effect) and upregulate antioxidant enzymes to prevent oxidative stress induced apoptosis. Thus, there is
an evolving view to target these metabolic alterations by novel anticancer agents to restrict tumor
progression in vivo. Previously we have reported that when a non toxic dose (10 mg/kg bw i.p.) of a novel
anticancer ruthenium(II)-complex containing 4-carboxy N-ethylbenzamide; Ru(II)-CNEB, was administered
to the Dalton's lymphoma (DL) bearing mice, it regressed DL growth by inducing apoptosis in the
DL cells. It also inactivated M4-LDH (M4-lactate dehydrogenase), an enzyme that drives anaerobic
glycolysis in the tumor cells. In the present study we have investigated whether this compound is able to
modulate regulation of glycolytic inhibition-apoptosis pathway in the DL cells in vivo. We observed that
Ru(II)-CNEB could decline expression of the inducible form of 6-phosphofructo-2-kinase (iPFK2:
PFKFB3), the master regulator of glycolysis in the DL cells. The complex also activated superoxide dismutase
(the H2O2 producing enzyme) but declined the levels of catalase and glutathione peroxidase (the
two H2O2 degrading enzymes) to impose oxidative stress in the DL cells. This was consistent with the
enhanced p53 level, decline in Bcl2/Bax ratio and activation of caspase 9 in those DL cells. The findings
suggest that Ru(II)-CNEB is able to activate oxidative stress-apoptosis pathway via p53 (a tumor
supressor protein) mediated repression of iPFK2, a key glycolytic regulator, in the DL cells in vivo.
Brian Covello: Diabetes Research ProposalBrian Covello
Brian Covello's diabetes research proposal. Type 2 diabetes mellitus consists of an array of dysfunctions characterized by hyperglycemia and resulting from the combination of resistance to insulin action, inadequate insulin secretion, and excessive or inappropriate glucagon secretion.
Essential update: FDA approves subcutaneous albiglutide for management of DM2
The FDA has approved once-weekly injectable albiglutide (Tanzeum), a glucagonlike peptide 1 (GLP-1) receptor agonist, along with diet and exercise for the treatment of type 2 diabetes.[1, 2] This agent may be used either as monotherapy or in combination with metformin, glimepiride, pioglitazone, or insulin.
Albiglutide should not be used for the following[1, 2] :
Patients with type 1 diabetes
Patients with diabetic ketoacidosis
First-line therapy in patients who can’t be managed with diet and exercise
Patients who have a personal or family history of medullary thyroid carcinoma (MTC)
Patients who have multiple endocrine neoplasia syndrome type 2
The most common adverse reactions associated with albiglutide were nausea/diarrhea and injection-site reactions.
There will be a boxed warning on albiglutide’s labeling about thyroid C-cell tumors being observed in rodent studies with this class of drugs; it is currently unknown whether albiglutide causes these tumors in humans, including MTC.[1, 2] Moreover, the FDA is also requiring a number of postmarketing studies, including a pediatric trial; an MTC case registry (≥15 y); and a cardiovascular (CV)-outcomes trial in patients with a baseline high risk of CV disease.
Signs and symptoms
Many patients with type 2 diabetes are asymptomatic. Clinical manifestations include the following:
Classic symptoms: Polyuria, polydipsia, polyphagia, and weight loss
Blurred vision
Lower-extremity paresthesias
Yeast infections (eg, balanitis in men)
See Presentation for more detail.
Diagnosis
Diagnostic criteria by the American Diabetes Association (ADA) include the following[3] :
A fasting plasma glucose (FPG) level of 126 mg/dL (7.0 mmol/L) or higher, or
A 2-hour plasma glucose level of 200 mg/dL (11.1 mmol/L) or higher during a 75-g oral glucose tolerance test (OGTT), or
A random plasma glucose of 200 mg/dL (11.1 mmol/L) or higher in a patient with classic symptoms of hyperglycemia or hyperglycemic crisis
Whether a hemoglobin A1c (HbA1c) level of 6.5% or higher should be a primary diagnostic criterion or an optional criterion remains a point of controversy.
Indications for diabetes screening in asymptomatic adults includes the following[4, 5] :
Sustained blood pressure >135/80 mm Hg
Overweight and 1 or more other risk factors for diabetes (eg, first-degree relative with diabetes, BP >140/90 mm Hg, and HDL < 35 mg/dL and/or triglyceride level >250 mg/dL)
ADA recommends screening at age 45 years in the absence of the above criteria
See Workup for more detail.
Production of Genetically Modified Grape (Vitis vinifera L.) PlantsAI Publications
Grape (Vitis vinifera L.) is one of the most economically important fruits in the world. High salinity stress adversely affects plant growth and limits agricultural production worldwide. This study describes a successful method of somatic embryogenesis using in vitro-derived leaf explants and introduction of a vacuolar-type Na+/H+ antiporter gene from a halophytic plant, Atriplex gmelini (AgNHX1) confers salt tolerance to grape cv. Superior Seedless using the Agrobacterium-mediated transformation. Callus embryogenic was induced on NN medium 2.0 mgL-1 2,4-D, 0.5 mgL-1 BAP and 0.5 mgL-1 NAA. Subsequent subculture of callus on NN medium containing 1.5 mgL-1 BAP, 0.5 mgL-1kinetin and 0.5 mgL-1NAA induced shoot organogenesis after eight weeks of culture. The leaf explants were co-cultivated with Agrobacterium strain LBA4404 harbouring the binary vector pBI121 which contained the AgNHX1 and nptII genes and putative transgenic plants were produced. The presence and stable integration of AgNHX1 gene in transgenic plants was confirmed by PCR and northern blot hybridization. The transgenic grape plants overexpressing the AgNHX1 gene showed a strong tolerance to salt stress under 250 mM NaCl, whereas non-transgenic plants died under the same conditions. Salt tolerance assays followed by salt treatments showed that the transgenic plants overexpressing AgNHX1 could survive under conditions of 250 mM NaCl for 4 weeks while the non-transgenic plants died under the same conditions. These results indicate that overexpression of the Na+/H+ antiporter gene in grape plants significantly improves their salt tolerance.
NAD-Glycohydrolase Depletes Intracellular NAD+ and Inhibits Acidification of Autophagosomes to Enhance Multiplication of Group A Streptococcus in Endothelial Cells
Extraction of β-galactosidase and β-glucosidase from the seeds of Tamarindus ...Innspub Net
The enzymes β–galactosidase and β–glucosidase were extracted from the tamarind seeds using different buffers at different pH. Highest activity was obtained with 10 mM sodium acetate buffer, pH 5.6 and 10 mM tris buffer, pH 7.4. The effect of NaCl and Triton X–100 at different concentrations on the extraction of the enzymes indicated 10 mM sodium acetate buffer, pH 5.6 containing 1 M NaCl as a better extractant of the enzyme. The enzyme assay was carried out using p–nitrophenyl–β–D–galactoside and p–nitrophenyl–β–D–glucoside as substrates. Highest enzyme activities were observed on 6th and 24th day of germination. The protein content gradually decreased upto 5th day of germination and suddenly increased on 6th day. However, on subsequent days of germination, the protein content greatly decreased upto 11th day. During the latter period of germination (18th day onwards) the content remained almost constant. The kinetic parameters varied for both β–galactosidase and β–glucosidase. The activity of β–galactosidase was show to have an optimal operating condition at pH 5.5 and a temperature of 500C. The thermostability of the enzyme was in the range of 400C – 700C with the pH stability in the range of 5.0 – 7.0. The Km and Vmax values for pNPGal were determined as 66μM and 2.27nmolesmin-1. In contrast the activity of β–glucosidase was shown to have an optimal operating condition at pH 5.0 and a temperature of 300C. The thermostability of the enzyme was in the range of 270C – 350C with the pH stability in the range of 4.0 – 7.0. The Km and Vmax values for pNPGlu were determined as 121μM and 5.26nmolesmin-1. The presented study is a preliminary work carried out for the standardization of protocols. The purification and characterization of β–galactosidase and β–glucosidase is under progress.
The Electrophoretic Profile Myofibrillar Proteins Extracted From Camel Muscle...IJEAB
Changes in electrophoretic profiles of myofibrillar protein (MFP) in the Longissimus thoracis (LD) of young camels (2 to 4 years), preserved by refrigeration has been treated or not by lactic acid solution 4% or citric acid 1%, were followed during the post-mortem time at the following times: 1, 2, 4, 6, 8, 10, 12, 24 and 48 hours. The cold preservation for 48 hours has not shown any particular distinctions in the protein profiles of this muscle. Changes related to the type of treatment were recorded during the storage time. Proteolysis of the myofibrillar fraction was earlier in this muscle in the case of treatment with one of two solutions of organic acids used, particularly in the case of using lactic acid. Indeed, these changes have affected at the first hour after slaughter the proteolysis of the myofibrillar proteins. Fragments of low molecular weight (42, 36, 33, 26, 23, 18, 16, 14 and 13 kDa) have been identified. The electrophoretic analysis showed that during refrigeration, LD treated with a solution of lactic acid is more sensitive to disruption phenomena and muscle protein proteolysis that lots of this muscle that even in the case of preservation by refrigeration only or by refrigeration
''Translational Repression and eIF4A2 Activity Are Critical for MicroRNA-Medi...Wisdom Deebeke Kate
Translational Repression and eIF4A2 Activity Are Critical for MicroRNA-Mediated Gene Regulation.
This assessed presentation looked at the effects of miRNA-mediated repression on mRNA. These effects include deadenylation, translational inhibition and degradation. It was observed that the eIF4A2 is a required component in the repression pathway mediated by the miRNA let-7. Presentation by KATE, Wisdom Deebeke; Mene Nkue; and Michael Thirlwell. Dated: Thursday, December 5th, 2013. (Phase 2, stage 3, semester 1).
Transcriptional regulation of functional genes involved in cuticular wax bios...Agriculture Journal IJOEAR
Abstract— MYB transcription factor family is one of the largest families in the plant kingdom, specifically characterized by a Helix-turn-helix domain in their structural configuration with two repeats of R2R3 MYB domain and are known for regulating plant stress tolerance (abiotic and biotic) through ABA dependent signaling systems. R2R3- MYB family transcription factors plays a role in the regulation of specific downstream genes of Very Long Chain Fatty Acid Biosynthesis like KCS1, KCS2, KCS6 and KCR1, related to water use efficiency traits like cuticular wax biosynthesis. Analysis of R2R3-MYB family transcription factors regulating the production of cuticular wax emphasizes the value of the family outside of traditionally accepted roles in stress tolerance. Some MYB transcription factors like MYB96, MYB94, MYB41, MYB30, MYB106 and MYB16 isolated from Arabidopsis thaliana (Col-0) are studied with respect to their role in biosynthesis of cuticular waxes under environmental stress signals. In the present review we intensely focused on the elucidation of R2R3- MYB family transcription factors MYB96, MYB94, MYB41, MYB30, MYB106 and MYB16 role in cuticular wax biosynthesis and aid in the development of transgenics for enhanced stress tolerance.
Activation of p53 mediated glycolytic inhibition-oxidative stressapoptosis pa...rkkoiri
There is a general agreement that most of the cancer cells switch over to aerobic glycolysis (Warburg
effect) and upregulate antioxidant enzymes to prevent oxidative stress induced apoptosis. Thus, there is
an evolving view to target these metabolic alterations by novel anticancer agents to restrict tumor
progression in vivo. Previously we have reported that when a non toxic dose (10 mg/kg bw i.p.) of a novel
anticancer ruthenium(II)-complex containing 4-carboxy N-ethylbenzamide; Ru(II)-CNEB, was administered
to the Dalton's lymphoma (DL) bearing mice, it regressed DL growth by inducing apoptosis in the
DL cells. It also inactivated M4-LDH (M4-lactate dehydrogenase), an enzyme that drives anaerobic
glycolysis in the tumor cells. In the present study we have investigated whether this compound is able to
modulate regulation of glycolytic inhibition-apoptosis pathway in the DL cells in vivo. We observed that
Ru(II)-CNEB could decline expression of the inducible form of 6-phosphofructo-2-kinase (iPFK2:
PFKFB3), the master regulator of glycolysis in the DL cells. The complex also activated superoxide dismutase
(the H2O2 producing enzyme) but declined the levels of catalase and glutathione peroxidase (the
two H2O2 degrading enzymes) to impose oxidative stress in the DL cells. This was consistent with the
enhanced p53 level, decline in Bcl2/Bax ratio and activation of caspase 9 in those DL cells. The findings
suggest that Ru(II)-CNEB is able to activate oxidative stress-apoptosis pathway via p53 (a tumor
supressor protein) mediated repression of iPFK2, a key glycolytic regulator, in the DL cells in vivo.
Brian Covello: Diabetes Research ProposalBrian Covello
Brian Covello's diabetes research proposal. Type 2 diabetes mellitus consists of an array of dysfunctions characterized by hyperglycemia and resulting from the combination of resistance to insulin action, inadequate insulin secretion, and excessive or inappropriate glucagon secretion.
Essential update: FDA approves subcutaneous albiglutide for management of DM2
The FDA has approved once-weekly injectable albiglutide (Tanzeum), a glucagonlike peptide 1 (GLP-1) receptor agonist, along with diet and exercise for the treatment of type 2 diabetes.[1, 2] This agent may be used either as monotherapy or in combination with metformin, glimepiride, pioglitazone, or insulin.
Albiglutide should not be used for the following[1, 2] :
Patients with type 1 diabetes
Patients with diabetic ketoacidosis
First-line therapy in patients who can’t be managed with diet and exercise
Patients who have a personal or family history of medullary thyroid carcinoma (MTC)
Patients who have multiple endocrine neoplasia syndrome type 2
The most common adverse reactions associated with albiglutide were nausea/diarrhea and injection-site reactions.
There will be a boxed warning on albiglutide’s labeling about thyroid C-cell tumors being observed in rodent studies with this class of drugs; it is currently unknown whether albiglutide causes these tumors in humans, including MTC.[1, 2] Moreover, the FDA is also requiring a number of postmarketing studies, including a pediatric trial; an MTC case registry (≥15 y); and a cardiovascular (CV)-outcomes trial in patients with a baseline high risk of CV disease.
Signs and symptoms
Many patients with type 2 diabetes are asymptomatic. Clinical manifestations include the following:
Classic symptoms: Polyuria, polydipsia, polyphagia, and weight loss
Blurred vision
Lower-extremity paresthesias
Yeast infections (eg, balanitis in men)
See Presentation for more detail.
Diagnosis
Diagnostic criteria by the American Diabetes Association (ADA) include the following[3] :
A fasting plasma glucose (FPG) level of 126 mg/dL (7.0 mmol/L) or higher, or
A 2-hour plasma glucose level of 200 mg/dL (11.1 mmol/L) or higher during a 75-g oral glucose tolerance test (OGTT), or
A random plasma glucose of 200 mg/dL (11.1 mmol/L) or higher in a patient with classic symptoms of hyperglycemia or hyperglycemic crisis
Whether a hemoglobin A1c (HbA1c) level of 6.5% or higher should be a primary diagnostic criterion or an optional criterion remains a point of controversy.
Indications for diabetes screening in asymptomatic adults includes the following[4, 5] :
Sustained blood pressure >135/80 mm Hg
Overweight and 1 or more other risk factors for diabetes (eg, first-degree relative with diabetes, BP >140/90 mm Hg, and HDL < 35 mg/dL and/or triglyceride level >250 mg/dL)
ADA recommends screening at age 45 years in the absence of the above criteria
See Workup for more detail.
Production of Genetically Modified Grape (Vitis vinifera L.) PlantsAI Publications
Grape (Vitis vinifera L.) is one of the most economically important fruits in the world. High salinity stress adversely affects plant growth and limits agricultural production worldwide. This study describes a successful method of somatic embryogenesis using in vitro-derived leaf explants and introduction of a vacuolar-type Na+/H+ antiporter gene from a halophytic plant, Atriplex gmelini (AgNHX1) confers salt tolerance to grape cv. Superior Seedless using the Agrobacterium-mediated transformation. Callus embryogenic was induced on NN medium 2.0 mgL-1 2,4-D, 0.5 mgL-1 BAP and 0.5 mgL-1 NAA. Subsequent subculture of callus on NN medium containing 1.5 mgL-1 BAP, 0.5 mgL-1kinetin and 0.5 mgL-1NAA induced shoot organogenesis after eight weeks of culture. The leaf explants were co-cultivated with Agrobacterium strain LBA4404 harbouring the binary vector pBI121 which contained the AgNHX1 and nptII genes and putative transgenic plants were produced. The presence and stable integration of AgNHX1 gene in transgenic plants was confirmed by PCR and northern blot hybridization. The transgenic grape plants overexpressing the AgNHX1 gene showed a strong tolerance to salt stress under 250 mM NaCl, whereas non-transgenic plants died under the same conditions. Salt tolerance assays followed by salt treatments showed that the transgenic plants overexpressing AgNHX1 could survive under conditions of 250 mM NaCl for 4 weeks while the non-transgenic plants died under the same conditions. These results indicate that overexpression of the Na+/H+ antiporter gene in grape plants significantly improves their salt tolerance.
NAD-Glycohydrolase Depletes Intracellular NAD+ and Inhibits Acidification of Autophagosomes to Enhance Multiplication of Group A Streptococcus in Endothelial Cells
Extraction of β-galactosidase and β-glucosidase from the seeds of Tamarindus ...Innspub Net
The enzymes β–galactosidase and β–glucosidase were extracted from the tamarind seeds using different buffers at different pH. Highest activity was obtained with 10 mM sodium acetate buffer, pH 5.6 and 10 mM tris buffer, pH 7.4. The effect of NaCl and Triton X–100 at different concentrations on the extraction of the enzymes indicated 10 mM sodium acetate buffer, pH 5.6 containing 1 M NaCl as a better extractant of the enzyme. The enzyme assay was carried out using p–nitrophenyl–β–D–galactoside and p–nitrophenyl–β–D–glucoside as substrates. Highest enzyme activities were observed on 6th and 24th day of germination. The protein content gradually decreased upto 5th day of germination and suddenly increased on 6th day. However, on subsequent days of germination, the protein content greatly decreased upto 11th day. During the latter period of germination (18th day onwards) the content remained almost constant. The kinetic parameters varied for both β–galactosidase and β–glucosidase. The activity of β–galactosidase was show to have an optimal operating condition at pH 5.5 and a temperature of 500C. The thermostability of the enzyme was in the range of 400C – 700C with the pH stability in the range of 5.0 – 7.0. The Km and Vmax values for pNPGal were determined as 66μM and 2.27nmolesmin-1. In contrast the activity of β–glucosidase was shown to have an optimal operating condition at pH 5.0 and a temperature of 300C. The thermostability of the enzyme was in the range of 270C – 350C with the pH stability in the range of 4.0 – 7.0. The Km and Vmax values for pNPGlu were determined as 121μM and 5.26nmolesmin-1. The presented study is a preliminary work carried out for the standardization of protocols. The purification and characterization of β–galactosidase and β–glucosidase is under progress.
Abstract— Storage roots are important for the growth and development in plants because they provide nutrients, water, and energy storage. Storage roots are also modulating growth direction, disease resistance, and root formation at the cellular and molecular level through interactions of genes and gene networks. However, molecular mechanisms regulating storage root formation in plants are not fully understood. In this review, we have overviewed transcriptional regulation of storage root formation, proteomic regulation of storage root formation, ethylene regulation of storage root formation, auxin regulation of storage root formation, gene expression regulation of storage root formation, and metabolism regulation of storage root formation. We have reviewed the basic regulatory principles of storage root formation from the network of genomics to proteomics and metabolism in plants that will be valuable to research work in storage root growth and development regulation at the molecular level.
In vitro callus induction of Melothria purpusilla, a traditional medicinal pl...IJERA Editor
Melothria purpusilla, a member of Cucurbitaceae, is an endemic species found in North-Eastern part of India.
The plant is used traditionally by the people of Manipur in the treatment of jaundice and its roots in fever and
diarrhoea. Tissue culture of medicinal plants was performed as a measure for the conservation of endangered
medicinal plants, Melothria perpusilla. Morphogenetic changes were observed in Melothria perpusilla
explants in the MS medium supplemented with different concentrations of PGRs. Different colours of callus
formation were observed in MS supplemented with BAP, kinetin and IBA. The best callus induction was
observed with MS media supplemented with combination of 1BAP mg/l + 1 IBA mg/l and combination of
1Kinetin mg/l + 1 IBA mg/l.
Hybrid rice breeding problems, prospects and future strategies by Deepak SharmaDeepak Sharma
The presentation describes all the constraints worldwide regarding hybrids in rice and potential solutions. The material includes all the findings and the researches going on in the world. Material collection is surely going to be very helpful from conventional and molecular point of view and having all the recent achievement and work done .
In silico Study of Oyster Mushroom Pleurotus Ostreatus Targeting PARP Protein...ijtsrd
Oyster mushroom Pleurotus ostreatus is belong to the group of healthy foods, as they contain high levels of proteins, vitamins and different classes of compounds, it is discovered that oyster mushrooms could play a key role in maintaining good health. Oyster mushroom Pleurotus sp. Class Basidiomycetes and Family Agaricaceae are widely known as ‘dhingri' in India. Pleurotus Ostreatus have several medicinal properties including , antitumor, immune modulatory, anti inflammatory, anticancer, antigenotoxic, anti arthritic, hypo cholesterolaemic, antihyperglycaemic, antioxidant, antihypertensive, antiplatelet aggregating, antiviral and antimicrobial activities.. In this paper studied that effects of chemical constituents of oyster mushroom Pleurotus sp. on DNA damaging protein which analyzed its activity of PARP inhibiting or vice - versa.For this analysis we choose the molecular docking technique to check the effects of different chemical constituents of oyster mushroom Pleurotus sp. on DNA damaging protein and compare their results to PARP inhibitory drugs which taken as standard. We perform the molecular docking in between chemical constituents of oyster mushroom Pleurotus sp. and 4UND protein compare to performance of molecular docking in between standard PARP inhibitory drugs and 4UND protein with the help of PyRx and BIOVIA Discovery studio software.The analysis of molecular docking shows that some chemical constituents of oyster mushroom Pleurotus sp. having more binding affinity than standard PARP inhibitory drugs. The Rutin shows better binding affinity than PARP inhibitory drugs on the same protein. Kajal S. Nadaf | Dr. Pravin Badhe "In-silico Study of Oyster Mushroom (Pleurotus Ostreatus) Targeting PARP Protein (4UND)" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-6 , October 2021, URL: https://www.ijtsrd.com/papers/ijtsrd47680.pdf Paper URL : https://www.ijtsrd.com/pharmacy/other/47680/insilico-study-of-oyster-mushroom--pleurotus-ostreatus-targeting-parp-protein-4und/kajal-s-nadaf
Abstract— MicroRNAs (miRNAs) function on post-transcriptional gene silencing and regulate the gene expression by degrading the transcripts of their targets, leading to down-regulation of the target genes. Plant miRNAs have been reported to play important roles in developmental control, hormone secretion, cell proliferation, and response to environmental stresses. In this review, we have reviewed miRNA expression and its potential role in regulating cell differentiation in Arabidopsis and summarized the miRNAs regulated cell differentiation during root, shoot, leave, and embryo development. We have further described practical application of expression of miRNAs in plant molecular breeding.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
3. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Nutrient Use Efficiency (NUE) may be defined
as yield per unit input. In agriculture this is usually
related to the input of fertilizer, whereas in
scientific literature the NUE is often expressed as
fresh weight or product yield per content of
nutrient.
Nutrient Use Efficiency can be designated as
nutrient efficiency ratio, agronomic efficiency,
physiological efficiency, agrophysiological
efficiency, apparent recovery efficiency, and
utilization efficiency.
The nutrients most commonly limiting plant
growth are N, P, K and S. NUE depends on the
ability to efficiently take up the nutrient from the
soil, but also on transport, storage, mobilization,
usage within the plant, and even on the
environment.
NUTRIENT USE EFFICIENCY
Fig: The Uptake of nutrient by a plant
5. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Import of fertilizers of value Rs. 34600 crores (2012-2013)
Loss of 2900 Cr. In India due to losses of N Fertilizer
Human health issue- in Punjab and Haryana ground water samples had
Nitrate-N of >22mg/l.
1% increase in NUE in N and P will save Rs. 10056 million
Need To Enhance NUE
P. fixen , 2014
7. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
1st mi-RNA to be discovered in 1993 in
Lin-4 gene of C.elegans
By Victor Ambros and Gary Ruvkun
miRNAs that are20 -24nucleotides long
bind to specific complementary transcripts,
attenuating gene expression at the post-
transcriptional level or via translational
inhibition.
miRNAs are vital for maintaining nutrient
homeostasis in plants by regulating the
expression of transporters that are involved
in nutrient uptake and mobilization.
ICAR-INDIANAGRICULTURALRESEARCHINSTITUTE
Role of miRNA in NUE
Victor Ambros and Gary Ruvkun
10. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
miRNA diversity related to NUE
Plant
tissue
Plant
species(r
eference)
Plant
tissue
Plant
species(r
eference)
156 SQUAMOSA
PROMOTER
BINDING PROTEIN-
LIKE (SPL)
transcription
factors
Shoot development .
Delayed vegetative phase
change
R (+) maize R (+) Arabidopsis Zhao et
al,2012
160 Auxin response
factors
Reduce auxin responsive
activities and the vegetative
growth.
Lateral and adventitious root
development ,
signal transduction
R (+) Maize R(+),L(−) (White
Lupin)
Hsieh et
al,2009
164 NAC transcription
factors
Accelerate senescence,
N remobilization
L(+) R(-)
S(-)
Maize R(+) SM(-)
L(-)
White lupin Liang et al
,2012
miRNA
family
Target gene
or protein
Description of
function
Involvement
under low N
Involvement
under low Pi
Referen
ce
11. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
169 HAP2 transcription
factors, CAAT binding
factor/NFYA
Nitrogen homeostasis, stress response
Nitrogen homeostasis, N uptake
Antioxidant
R(-) S(-)
L(-)
Maize SD(-)
R(-) S(-)
Arabido
psis
172 AP2 like transcription
factors
Ethylene-responsive pathway,
N remobilization ,Flower development
L(+) S(+)
R(-)
Maize
Arabid
opsis
L(-) Tomato Zhu et al
,2010
395 ATP sulfurylase; sulfate
transporters
Sulfate homeostasis R(-) Arabid
opsis
R(-) Arabido
psis
Zeng et
al,2010
397 Laccases Laccases Reduce root growth,
Copper homeostasis
L(-) R(-)
S(-)
Maize L(-) White
Lupin
Zhao et
al,2013
399 Ubiquitin conjugase
E2/UBC24
Phosphate homeostasis, uptake and
translocation
L(-) R(-) Maize R(+)
S(+)
SD(+)
Arabido
psis
Wang et
al,2013
Contd...
12. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
780 Na+/H+ antiporter Sodium ion export R(+) Arabid
opsis
Valdes et
al,2008
827 Ubiquitin E3 ligase with
RING and SPX
Nitrogen/phosphorus metabolism
Accelerate leaf senescence,
P homeostasis, P uptake
R(-)
L(-)
R(-)
Arabid
opsis
Maize
SD(+)
R(+)
R(+)
S(+)
Arabido
psis
Rice
Hackenberg
et al,2013
444 MADS-box Root development R(+) Rice R(+) Rice Lundmark et
al,2010
398 COX5b-1; CCS1 COX Copper homeostasis, oxidative stress
Enhanced to produce ATP under stress
R(-)
SD (-)
Arabid
opsis
SD(-)
R(-) S(-)
Arabido
psis
Xu et al,2011
171 SCARECROW-like
transcription factors
Root development R(+) Arabid
opsis
SM(+)
L(+)
White
lupin
Pant et
al,2012
Contd…
13. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Role of miRNA in Nitrogen use efficiency
miR156 and Squamosa Promoter Binding Protein Like
(SPL)
SPL proteins play critical roles in maintaining normal growth throughout
plant life cycle .
These miRNAs are involved in phase change via their targets, members of
the SPL transcription factors.
miR156 targets, SPL3, is downregulated , suggesting that a miR156/SPL3
module might act by repressing vegetative phase change under limiting N
availability.
miR156 acts as a negative regulator of miR172 by controlling miR172
expression via its targets SPL9 and SPL15.
14. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
miR160 and Auxin Response Factor 16 (ARF16)
ARFs are DNA binding proteins that control auxin -regulated
transcription and are only present in plants.
ARF16, being this auxin responsive factor already characterized to
control root cap formation.
N-deficiency induces expression of miR160, which increases
ARF16 degradation and consequently supports lateral root formation.
miR169 and Nuclear Factor Y Subunit A (NFYA)
NFYA also called heme -activated protein (HAP) or CCAAT- box binding-factor(CBF),
NFYA have being associated with nodule differentiation and drought tolerance.
Transgenic Arabidopsis plants over expressing miR169a ,they showed a decrease in N
accumulation .These plants showed higher sensitivity to N limitation comparing to the wild
type, since NFYA regulates the nitrate transporters NRT1 and NRT2(Zhao etal.,2011) .
15. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
miR393 and Auxin Signaling F-Box Protein 3 (AFB3)
Nitrate is able to transcriptionally induce expression of AFB3 in roots and that N
metabolites produced after nitrate reduction and assimilation lead to a down regulation of
AFB3 levels due to miR393 induction .
miR393/ARF3 is the responsible mechanism to repress primary root elongation and
induce lateral root emergence under the presence of N.
The monocot specific miR444 has been demonstrated to regulate four MIKC-type
MADS-box transcriptional factor genes in rice (OsMADS23 , OsMADS27a , OsMADS27b
, and OsMADS57 .
miR444 targets with Arabidopsis ANR1clade, which is a pivotal regulator in NO3−
signaling pathway in lateral root growth.
Plants overexpressing this miRNA presented a decrease in the expression of the four
MADS- box genes, and a reduced nitrate induced lateral root growth .
miR444 and MADS-Box Transcription Factor
17. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Role of miRNA in P use Efficiency
The uptake and assimilation of P in plants is orchestrated by two key proteins , phosphate
transporter1 (PHT1) and phosphate 1 (PHO1).
PHT1 proteins are able to use energy to co-transport Pi and H+ and are therefore involved in Pi
acquisition .
PHO1, is involved in the loading of acquired Pi into xylem, facilitating therefore the root-to-shoot
transport of this macronutrient in plant.
Upon Pi stress , plants trigger Pi Starvation Responses (PSR). Since,PHT1 and PHO1 proteins are
central in the assimilation and allocation of Pi in plants, PSR try to maximize their expression .
The MYB transcription factors Phosphate Starvation Regulator 1 (PHR1) and Phosphate Starvation
Regulator 1- like (PHR1-LIKE1) leads to PHT1 and PHO1 over-accumulation.
PHR1 and PHR1- LIKE1 induce the expression of Phosphate Transporter Traffic Facilitator 1
(PHF1), a protein that facilitates the transport of PHT1 to membranes, increasing therefore their
availability for Pi assimilation.
miR399, miR 827/PHT1,PHO1,PHO2
18. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
An E2 ligase, PHO2 mediates the
ubiquitination of the Pi/H+ transporters during
normal Pi conditions ,preventing their
trafficking to membranes.
PHO2 also mediates a post-translational
inhibition of PHO1 .Therefore, by repressing
PHO2, miR399 contributes to the
accumulation of both PHT1 and PHO1 during
Pi starvation .
PHT1 may also suffer ubiquitination by the
Nitrogen Limitation Adaptation(NLA) protein
, an E3 RING ubiquitin enzyme. NLA-
mediated ubiquitination of PHT1 leads to
endocytosis and degradation of the protein.
The post-transcriptional regulation of the
NLA encoding gene by miR827 , therefore ,
helps to stabilize PHT1 levels during stress
conditions.
Figure: Hypothetical model for the role of miR399
& miR827 to maintain Pi homeostasis in plants.
Nguyen et al,2015
20. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Role of miRNA in K Homeostasis
MADS-box genes encode a family of transcription factors and are associated with several
developmental regulatory pathways, from root to flower and fruit development .
The unique miRNA investigated with respect to K+ signaling is the monocots specific
miR444a .The expression profile of this miRNA and its respective targets(MADS-
23,MADS-27a,MADS-27b,and MADS-57) during K+ deprivation in rice roots was explored
and it was found that this condition caused as lightly decrease of miR444a levels .
MADS-23 target was strongly induced compared to the control situation.
The presence of NH4+ in potassium containing medium favors nitrogen uptake by HAK5
protein , which is suggested to be a high affinity K+ transporter . Under this nutrition
circumstance ,the protein AKT1 turns to be the main K+ uptake protein even in low K+
concentrations .
22. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Role of miRNA in S homeostasis
In Arabidopsis, sulfate is transported through xylem or phloem via cell-specific transporters
such as sulfate transporters1;1 (SULTR1;1),SULTR2;1,andSULTR2;2.
sulfate limitation induces the expression of miR395 and its low affinity sulfate transporter
SULTR2;1,
restriction of SULTR2;1 expression by miR395 in the xylem parenchyma facilitates the
translocation of sulfate ions from the roots to the shoots.
miR395 has also been elucidated in S assimilation by sup- pressing the expression of ATP
sulfurylase genes, such as APS1, APS3, and APS4, which catalyze the first step of S
assimilation.
S deficiency leads to the elevated synthesis of SULFUR LIMITATION1 (SLIM1) protein in
the roots, which in turn activates various sulfate transporters to enhance S uptake.
24. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Role of miRNA in Copper homeostasis
Cu plays an important role against oxidative stress responses by acting as a
cofactor of Copper/Zinc superoxide dismutase (CSD).
superoxidedismutase SOD1 (CCS1) 1 is a chaperone protein that delivers the Cu
ions to CSD1 and CSD2 apoprotein .
During Cu limitation , the induction of miR398 downregulates ,SOD1 (CCS1)
gene for the expression of CSD1 and CSD 2 protein.
miR397, miR408, andmiR857, have been found to be up regulated during Cu
starvation, which in turn suppresses the expression of laccase and plastocyanin
genes.
27. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
To characterize the small RNAs that are
responsive to C, N, or S deficiency, 10-day-
old seedlings grown on full nutrient (FN),
sucrose-free (–C), nitrogen-free (–N), or
sulfate-free (–S) MS medium were used to
construct small RNA libraries.
Figure: Response of seedling to Nutrient
Deficiency
By sequencing they identified the most
abundant size of small RNAs in the FN library
and –N Library was 21 nucleotides . In
contrast, 20 and 24-nucleotide-length small
RNAs were the most abundant in –C and –S,
respectively . These results suggested that
different nutrient supplies caused differential
distribution patterns of the sizes of small
RNAs.
29. ICAR-IndianAgriculturalResearchInstitute,NewDelhi To explore the miRNAs that were differentially expressed in response to a specific nutrient deficiency, They
compared the read counts of miRNAs under nutrient-deficient conditions with those under FN. 92, 79,
and 59 differentially expressed miRNAs, which are clustered into 40, 41 and 31 miRNA families, were
obtained for –C, –N, and –S, respectively.
32. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
In addition, some miRNAs were positively
correlated with their targets, such as
miR395-APS3 in –S, miR397-LAC2 in –C,
and miR160-ARF17 in –C and –S. It is
likely that these targets are also regulated
by other transcription factors or their
expression does not completely overlap
with miRNAs Spatiotemporally.
Figure : Phenotype of Transgenic plants under the
Nutrient starvation conditions
Conclusion: A large number of miRNAs were differentially expressed in response to nutrient deficiencies,
some miRNAs were specifically responsive to specific nutrient depletions. miR169b/c, miR826, and miR395
showed the largest changes in response to –C, –N, and –S, respectively.
To investigate the functions of nutrient-
responsive miRNAs in nutrient starvation
adaptation, miRNA overexpression plants
(miR160a-ox, miR395a-ox, and miR399b-
ox) and miR160 suppression plants
(STTM160) were used to evaluate
phenotypes in nutrient starvation conditions.
33. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Case study-2
Polymorphism of flax (Linum usitatissimum L.) genome, genotype CDC Bethune, under nutrient stress in
vitro, was analyzed by newly developed type of molecular markers based on microRNA molecules.
Two types of stress-sensitive miRNAs , miR395 and miR399 were evaluated. The miR395 loci profile has
shown to be more polymorphic and more specific in comparison to miR399 loci pattern.
They supported the role of miRNA molecules as potential biomarkers of abiotic stress.
34. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Flax is known by its phytoremediation capabilities . In addition, flax is of research interest
because some lines undergo phenotype and genome changes in response to environmental
conditions (Cullis 2004).
They applied nutritional stress under in vitro conditions on the genotype CDC Bethune &
were interested at the level of miRNA-based markers, to record the polymorphism of nutrition
stress-sensitive miRNA, miR395 and miR399, in the genome of flax.
Seeds of flax genotype CDC Bethune were cultivated on four different nutritional variants of
the MS basal medium as follows:
1 – full-strength of microelements and vitamins, half-strength of macroelements,
2 – full-strength of macroelements and vitamins, half-strength of microelements,
3 – full-strength of microelements and macroelements, half-strength of vitamins,
4 – half-strength of microelements, macroelements and vitamins,
C – control variant – basal MS medium.
In vitro cultivation was carried out during the period of six weeks at 22°C under photoperiod
16 h light/8 h dark cycle & then Total genomic DNA was extracted .
35. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
The primers for the miRNA-based markers were designed according to the mature
miRNAs sequences, originated from the miRNA database (http://www.mirbase.org/) .
A total of 2 miRNA-based forward primers and 1 universal miRNA reverse primer
were used and randomly combined together to perform a marker assay.
Combination of primer pairs used for miRNA-based marker assay.
36. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
They found variability not in no. of miRNA Loci
but also in their size.
highest number of miR395 loci (9) has been
recorded at the MS basal medium having half-
strength of macroelements .
Within the macroelements of MS medium
sulfur is present only in one component and that is
magnesium sulphate while, four microelements
are in the form of sulphates (copper, iron,
manganese and zinc).
Figure : Representative gel showing amplification profiles of
CDC Bethune generated by primer pair miR 395_F / miR_R.
Their results supports the possibility that
miR395 response varies between different
plant species and some plants that are
adapted to inferior growing conditions
might evolved constitutive adaptive
mechanisms.
37. ICAR-IndianAgriculturalResearchInstitute,NewDelhi The pattern of miR399 is less polymorphic than the
miR395 pattern.
They observed significantly different miR399 loci
pattern in the case of stress variant having half-
strength of all components of MS medium.
Within the macroelements of MS medium
phosphate is present only in one component and that
is potassium phosphate- while in the microelements
there are no phosphates.
Figure: Representative gel showing amplification profiles of
CDC Bethune generated by primer pair miR 399_F / miR_R.
miRNA based molecular markers are sufficient for evaluation of flax genome polymorphism under specific
condition of abiotic stress . It shows the capability of miRNA molecule as potential biomarkers of
environmental stress.
That means that the stress variant number 1
represents, in this case, the conditions of low
phosphate characterized by miR399 up-regulation.
In this variant the highest number of miRNA loci in
comparison to control has been observed.
38. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Conclusion
The alteration of nutrient levels in soil can trigger specific signaling molecules that can act as
repressors of target nutrient responsive- miRNAs . The decreased accumulation of miRNAs
subsequently stabilizes the expression of transporters .
majority of miRNAs involved in NPK deprivation are associated with mechanisms involved
in the adaption to stress conditions by affecting root architecture ,controllingNO3− or Pi
transporters , controlling shoot growth, affecting vegetative phase transition , and managing
these nutrients leakage.
On contrary, the optimal conditions or higher amounts of nutrients can trigger a specific
group of miRNAs that directly affect the transporter or induce other miRNA s that suppress the
expression of repressor genes.
39. ICAR-IndianAgriculturalResearchInstitute,NewDelhi
Future Prospective
Investigation of novel miRNAs and their role in phytate biosynthesis like regulation of
different inositol phosphate kinase genes, alteration of specific miRNA expression by
overexpression or genome editing.
Role of miRNAs in regulation of nitrate transporters and metabolic enzymes such as aspartate
amino transferase, glutamine synthase, glutamate dehydrogenase .
Role of miRNAs in different groups of Fe and Zn transporters from roots to seed, miRNA
promoter/ genome editing .
Identification of root specific novel miRNAs under nutrient stress and investigation the role of
miRNA- mediated miRNA activation or removal of supressors of transporters .
Role of miRNAs in down-regulation of heavy metal transporters .