Multiplex CRISPR/Cas9-mediated mutagenesis of alfalfa FLOWERING LOCUS Ta1 (MsFTa1) leads to delayed flowering time with improved forage biomass yield and quality.

Alfalfa (Medicago sativa L.) is a perennial flowering plant in the legume family that is widely cultivated as a forage crop for its high yield, forage quality and related agricultural and economic benefits. Alfalfa is a photoperiod sensitive long-day (LD) plant that can accomplish its vegetative and reproductive phases in a short period of time. However, rapid flowering can compromise forage biomass yield and quality. Here, we attempted to delay flowering in alfalfa using multiplex CRISPR/Cas9-mediated mutagenesis of FLOWERING LOCUS Ta1 (MsFTa1), a key floral integrator and activator gene. Four guide RNAs (gRNAs) were designed and clustered in a polycistronic tRNA-gRNA system and introduced into alfalfa by Agrobacterium-mediated transformation. Ninety-six putative mutant lines were identified by gene sequencing and characterized for delayed flowering time and related desirable agronomic traits. Phenotype assessment of flowering time under LD conditions identified 22 independent mutant lines with delayed flowering compared to the control. Six independent Msfta1 lines containing mutations in all four copies of MsFTa1 accumulated significantly higher forage biomass yield, with increases of up to 78% in fresh weight and 76% in dry weight compared to controls. Depending on the harvesting schemes, many of these lines also had reduced lignin, acid detergent fibre (ADF) and neutral detergent fibre (NDF) content and significantly higher crude protein (CP) and mineral contents compared to control plants, especially in the stems. These CRISPR/Cas9-edited Msfta1 mutants could be introduced in alfalfa breeding programmes to generate elite transgene-free alfalfa cultivars with improved forage biomass yield and quality.

Overexpression of ANAC046 Promotes Suberin Biosynthesis in Roots of Arabidopsis thaliana.

NAC (NAM (no apical meristem), ATAF1/2, and CUC2 (cup-shaped cotyledon)) proteins are one of the largest families of plant-specific transcription factors, and this family is present in a wide range of land plants. Here, we have investigated the role of ANAC046 in the regulation of suberin biosynthesis and deposition in Arabidopsis. Subcellular localization and transcriptional activity assays showed that ANAC046 localizes in the nucleus, where it functions as a transcription activator. Analysis of the PANAC046:GUS lines revealed that ANAC046 is mainly expressed in the root endodermis and periderm, and is also induced in leaves by wounding. The transgenic lines overexpressing ANAC046 exhibited defective surfaces on the aerial plant parts compared to the wild-type (WT) as characterized by increased permeability for Toluidine blue stain and greater chlorophyll leaching. Quantitative RT-PCR analysis showed that the expression of suberin biosynthesis genes was significantly higher in the roots and leaves of overexpression lines compared to the WT. The biochemical analysis of leaf cuticular waxes showed that the overexpression lines accumulated 30% more waxes than the WT. Concurrently, overexpression lines also deposited almost twice the amount of suberin content in their roots compared with the WT. Taken together, these results showed that ANAC046 is an important transcription factor that promotes suberin biosynthesis in Arabidopsis thaliana roots.

Overexpression of OsGATA12 regulates chlorophyll content, delays plant senescence and improves rice yield under high density planting.

Agronomic traits controlling the formation, architecture and physiology of source and sink organs are main determinants of rice productivity. Semi-dwarf rice varieties with low tiller formation but high seed production per panicle and dark green and thick leaves with prolonged source activity are among the desirable traits to further increase the yield potential of rice. Here, we report the functional characterization of a zinc finger transcription factor, OsGATA12, whose overexpression causes increased leaf greenness, reduction of leaf and tiller number, and affects yield parameters. Reduced tillering allowed testing the transgenic plants under high density which resulted in significantly increased yield per area and higher harvest index compared to wild-type. We show that delayed senescence of transgenic plants and the corresponding longer stay-green phenotype is mainly due to increased chlorophyll and chloroplast number. Further, our work postulates that the increased greenness observed in the transgenic plants is due to more chlorophyll synthesis but most significantly to decreased chlorophyll degradation, which is supported by the reduced expression of genes involved in the chlorophyll degradation pathway. In particular we show evidence for the down-regulation of the STAY GREEN RICE gene and in vivo repression of its promoter by OsGATA12, which suggests a transcriptional repression function for a GATA transcription factor for prolonging the onset of senescence in cereals.

ROS Induces Anthocyanin Production Via Late Biosynthetic Genes and Anthocyanin Deficiency Confers the Hypersensitivity to ROS-Generating Stresses in Arabidopsis.

Anthocyanins are known to have antioxidant activities. Their accumulation can be triggered by many chemical and environmental factors, including reactive oxygen species (ROS). However, the mechanism of ROS-induced anthocyanin accumulation and the role of anthocyanins in the response of Arabidopsis (Arabidopsis thaliana) to different stresses are largely unknown. Here, we study the cross-regulation between ROS and anthocyanin production. Ten Arabidopsis mutants covering the main anthocyanin regulatory and biosynthetic genes are systematically analyzed under ROS-generating stresses. We find that ROS triggers anthocyanin accumulation by up-regulating the anthocyanin late biosynthetic and the corresponding regulatory genes. The anthocyanin-deficient mutants have more endogenous ROS and are more sensitive to ROS-generating stresses while having decreased antioxidant capacity. Supplementation with cyanidin makes them less susceptible to ROS, with increased anthocyanin and reduced ROS accumulation. In contrast, pap1-D, which overaccumulates anthocyanins, shows the opposite responses. Gene expression analysis reveals that photosynthetic capacity is more impaired in anthocyanin-deficient mutants under high-light stress. Expression levels of ROS-scavenging enzyme genes are not correlated with the radical-scavenging activity in different mutants. We conclude that ROS are an important source signal to induce anthocyanin accumulation by up-regulating late biosynthetic and the corresponding regulatory genes and, as a feed-back regulation, anthocyanins modulate the ROS level and the sensitivity to ROS-generating stresses in maintaining photosynthetic capacity.

OsPIN5b modulates rice (Oryza sativa) plant architecture and yield by changing auxin homeostasis, transport and distribution.

Plant architecture attributes such as tillering, plant height and panicle size are important agronomic traits that determine rice (Oryza sativa) productivity. Here, we report that altered auxin content, transport and distribution affect these traits, and hence rice yield. Overexpression of the auxin efflux carrier-like gene OsPIN5b causes pleiotropic effects, mainly reducing plant height, leaf and tiller number, shoot and root biomass, seed-setting rate, panicle length and yield parameters. Conversely, reduced expression of OsPIN5b results in higher tiller number, more vigorous root system, longer panicles and increased yield. We show that OsPIN5b is an endoplasmic reticulum (ER) -localized protein that participates in auxin homeostasis, transport and distribution in vivo. This work describes an example of an auxin-related gene where modulating its expression can simultaneously improve plant architecture and yield potential in rice, and reveals an important effect of hormonal signaling on these traits.

ANAC032 Positively Regulates Age-Dependent and Stress-Induced Senescence in Arabidopsis thaliana.

Members of the NAC transcription factor family have been implicated in the regulation of different processes of plant development including senescence. In this study, the role of ANAC032 is analyzed in Arabidopsis thaliana (Col-0). ANAC032 is shown to act as a transcriptional activator and its expression is induced in senescing leaves as well as in dark-treated detached leaves. Analysis of transgenic overexpressors (OXs) and chimeric repressors (SRDXs) of ANAC032 indicates that ANAC032 positively regulates age-dependent and dark-induced leaf senescence. Quantitative real-time PCR analysis showed that ANAC032 regulates leaf senescence mainly through the modulation of expression of the senescence-associated genes AtNYE1, SAG113 and SAUR36/SAG201, which are involved in Chl degradation, and ABA and auxin promotion of senescence, respectively. In addition, ANAC032 expression is induced by a range of oxidative and abiotic stresses. As a result, ANAC032 overexpression lines exhibited enhanced leaf senescence when challenged with different oxidative (3-aminotriazole, fumonisin B1 and high light) and abiotic stress (osmotic and salinity) conditions compared with the wild type. In contrast, ANAC032 SRDX lines displayed the opposite phenotype. ANAC032 transgenic lines showed altered 2,4-D-mediated root tip swelling and root inhibition responses when compared with the wild type. The altered response to auxin, oxidative and abiotic stress treatments in ANAC032 transgenic lines involves differential accumulation of H2O2 compared with the wild type. Taken together, these results indicate that ANAC032 is an important transcription factor that positively regulates age-dependent and stress-induced senescence in A. thaliana by modulating reactive oxygen species production.

Difficulties in maintenance of clubfoot abduction brace and solutions - maintenance of clubfoot abduction brace, locks and keys.

OBJECTIVE: To determine the frequency of early relapse after achieving good initial correction in children who were on clubfoot abduction brace. METHODS: The cross-sectional study was conducted at the Jinnah Postgraduate Medical Centre, Karachi, and included parents of children of either gender in the age range of 6 months to 3years with idiopathic clubfoot deformities who had undergone Ponseti treatment between September 2012 and June 2013, and who were on maintenance brace when the data was collected from December 2013 to March 2014. Parents of patients with follow-up duration in brace less than six months and those with syndromic clubfoot deformity were excluded. The interviews were taken through a purposive designed questionnaire. SPSS 16 was used for data analysis. RESULTS: The study included parents of 120 patients. Of them, 95(79.2%) behaved with good compliance on Denis Browne Splint, 10(8.3%) were fair and 15(12.5%)showed poor compliance. Major reason for poor and non-compliance was unaffordability of time and cost for regular follow-up. Besides, 20(16.67%) had inconsistent use due to delay inre-procurement of Foot Abduction Braceonce the child had outgrown the shoe. Only 4(3.33%) talked of cultural barriers and conflict of interest between the parents. Early relapse was observed in 23(19.16%) patients and 6(5%) of them responded to additional treatment and were put back on brace treatment; 13(10.83%) had minor relapse with forefoot varus, without functional disability, and the remaining 4(3.33%) had major relapse requiring extensive surgery. Overall success was recorded in 116(96.67%) cases. CONCLUSIONS: The positioning of shoes on abduction brace bar, comfort in shoes, affordability, initial and subsequent delay in procurement of new shoes once the child's feet overgrew the shoe, were the four containable factors on the part of Ponseti practitioner.

Mutating alfalfa COUMARATE 3-HYDROXYLASE using multiplex CRISPR/Cas9 leads to reduced lignin deposition and improved forage quality.

Alfalfa (Medicago sativa L.) forage quality is adversely affected by lignin deposition in cell walls at advanced maturity stages. Reducing lignin content through RNA interference or antisense approaches has been shown to improve alfalfa forage quality and digestibility. We employed a multiplex CRISPR/Cas9-mediated gene-editing system to reduce lignin content and alter lignin composition in alfalfa by targeting the COUMARATE 3-HYDROXYLASE (MsC3H) gene, which encodes a key enzyme in lignin biosynthesis. Four guide RNAs (gRNAs) targeting the first exon of MsC3H were designed and clustered into a tRNA-gRNA polycistronic system and introduced into tetraploid alfalfa via Agrobacterium-mediated transformation. Out of 130 transgenic lines, at least 73 lines were confirmed to contain gene-editing events in one or more alleles of MsC3H. Fifty-five lines were selected for lignin content/composition analysis. Amongst these lines, three independent tetra-allelic homozygous lines (Msc3h-013, Msc3h-121, and Msc3h-158) with different mutation events in MsC3H were characterized in detail. Homozygous mutation of MsC3H in these three lines significantly reduced the lignin content and altered lignin composition in stems. Moreover, these lines had significantly lower levels of acid detergent fiber and neutral detergent fiber as well as higher levels of total digestible nutrients, relative feed values, and in vitro true dry matter digestibility. Taken together, these results showed that CRISPR/Cas9-mediated editing of MsC3H successfully reduced shoot lignin content, improved digestibility, and nutritional values without sacrificing plant growth and biomass yield. These lines could be used in alfalfa breeding programs to generate elite transgene-free alfalfa cultivars with reduced lignin and improved forage quality.

Exploring vegetation trends and restoration possibilities in Pakistan by using Hurst exponent.

Vegetation cover change and its interaction with climate are significant to study as it has impact on ecosystem stability. We used the Normalized Difference Vegetation Index (NDVI) and climatic factors (temperature and rainfall) for investigating the relationship between vegetation and climate. We also traced spatiotemporal changes in the vegetation in Pakistan from 2000 to 2020; we used the Hurst exponent to estimate future vegetation trends in Pakistan. Our results show an increase in vegetation throughout Pakistan, and the Punjab Province is showing the highest significant vegetation trend at 88.51%. Our findings reveal that the response of vegetation to climate change varies by region and is influenced by local climatic conditions. However, the relationship between rainfall and annual NDVI is stronger than the temperature in the study area-Pakistan. The Hurst exponent value is above 0.5 in all four provinces, that is, the indication of consistent vegetation trends in the future. The highest values are observed in Punjab and Khyber Pakhtunkhwa (KPK). In the Punjab Province, 88.41% of the area showed positive development, with forests in particular showing a significant positive effect on land use classes. On the other hand, the Sindh Province has the highest negative result at 2.87%, with urban areas showing the highest negative development. To sum up, the NDVI pattern and change attribute suggest vegetation restoration in Pakistan.

Transcriptional Programs and Regulators Underlying Age-Dependent and Dark-Induced Senescence in Medicago truncatula.

In forage crops, age-dependent and stress-induced senescence reduces forage yield and quality. Therefore, delaying leaf senescence may be a way to improve forage yield and quality as well as plant resilience to stresses. Here, we used RNA-sequencing to determine the molecular bases of age-dependent and dark-induced leaf senescence in Medicago truncatula. We identified 6845 differentially expressed genes (DEGs) in M3 leaves associated with age-dependent leaf senescence. An even larger number (14219) of DEGs were associated with dark-induced senescence. Upregulated genes identified during age-dependent and dark-induced senescence were over-represented in oxidation-reduction processes and amino acid, carboxylic acid and chlorophyll catabolic processes. Dark-specific upregulated genes also over-represented autophagy, senescence and cell death. Mitochondrial functions were strongly inhibited by dark-treatment while these remained active during age-dependent senescence. Additionally, 391 DE transcription factors (TFs) belonging to various TF families were identified, including a core set of 74 TFs during age-dependent senescence while 759 DE TFs including a core set of 338 TFs were identified during dark-induced senescence. The heterologous expression of several senescence-induced TFs belonging to NAC, WKRY, bZIP, MYB and HD-zip TF families promoted senescence in tobacco leaves. This study revealed the dynamics of transcriptomic responses to age- and dark-induced senescence in M. truncatula and identified senescence-associated TFs that are attractive targets for future work to control senescence in forage legumes.

Promoting Healthcare Sustainability in Developing Countries: Analysis of Knowledge Management Drivers in Public and Private Hospitals of Pakistan.

Investing in a sustainable future has no alternative; the healthcare sector in developing countries has failed to achieve sustainability objectives. Knowledge management (KM) is a concrete application of sustainability in healthcare, as organizations (hospitals) that manage their knowledge assets will gain sustainable competitive advantage. Several organizations in developed countries are moving towards the adoption of knowledge management so that they can manage their knowledge well and improve their performance. Due to the effective implementation of KM in developed countries, developing countries are also considering adopting KM in their healthcare. In this study, an attempt has been made to identify the drivers of KM adoption in public and private hospitals of Pakistan. With the help of an extensive literature review and expert opinion, the drivers were identified and a hierarchical structure was developed. Nineteen drivers were identified and screened out by experts. The experts identified the contextual relationships between the drivers during a brainstorming session. The hierarchical model of the drivers for KM in the healthcare of Pakistan was eventually developed using interpretive structural modeling (ISM). The structure has 10 levels, in which "developed competitive advantage" formed the foundation of the structure and "job creation" and "improvement in the reputation of healthcare" formed the topmost level. The "Matrices d'Impacts Croises Multiplication Appliqué a un Classement" (MICMAC) analysis classified the drivers by categorizing them according to their driving and dependence powers. One driver is identified as autonomous, six drivers as dependent, seven drivers as linkage, and five drivers as independent. The analysis of KM drivers will provide a good understanding of the interdependence and interactions between them and support the effect adoption of KM in developing countries especially in Pakistan.

Enablers Supporting the Implementation of Knowledge Management in the Healthcare of Pakistan.

Knowledge is considered to be an important resource; it is the source of competitive advantage. However, if knowledge is managed well with Knowledge Management (KM), then it becomes a source for sustainable competitive advantage for organizations. If KM is implemented in an organization, it would improve the organizational competitiveness, performance, and productivity, and facilitate the efficient use of resources. Due to intense competition in the global market, many organizations are moving towards the adoption of KM. The healthcare sectors of many developed countries have moved towards the implementation of KM because it can improve the procuring of knowledge from ongoing activities by the effective use of data repositories. Developing countries have now realized the potential and benefits of KM adoption. Pakistan is one of the developing countries that have recently shown an inclination towards the adoption of KM in its healthcare sector to improve performance of its healthcare. This study was composed of two main research phases. Firstly, the enablers of KM were reviewed from earlier studies. Secondly, interpretive structural modeling (ISM) and MICMAC (Cross-Impact Matrix Multiplication Applied to Classification) techniques were used to show the interrelationships between KM enablers and driving and dependence power of each enabler. The application of ISM and MICMAC technique shows that policy incentive, long-term strategic planning, Information Technology (IT), and alignment of KM efforts with business strategy are the main enablers of KM adoption in the healthcare of Pakistan. Focusing on the identified enablers will help in the implementation of KM. Policy incentives can work as a catalyst to promote KM adoption in the healthcare of Pakistan.

Synthesis and characterization of chitin/curcumin blended polyurethane elastomers.

In this work, chitin-curcumin based polyurethane elastomers (PUEs) were prepared by step growth polymerization technique using hydroxy terminated polybutadiene (HTPB), hexamethylene diisocyanate (HDI), chitin and curcumin. The molecular characterization was done by using FTIR and SS 1HNMR techniques. The surface morphology and thermal stability was studied by scanning electron microscopy (SEM) and thermal gravimetric analysis (TGA), respectively. Degree of absorption and swelling characters were also determined in water as well as in DMSO. The crystalline behavior of prepared elastomers was checked by using X-ray diffraction (XRD) and differential scanning calorimeter (DSC). Results presented that crystallinity of elastomers increased by increasing the content of chitin due to formation of more ordered structure.

Wernicke's encephalopathy with atypical findings on MR imaging.

Wernicke's encephalopathy (WE) is a disease usually related to chronic alcoholism. The clinical diagnosis is often difficult to establish. MR imaging is a highly sensitive method for detecting lesions associated with Wernicke's encephalopathy. We report a case of atypical manifestation of hyperintense lesions on magnetic resonance imaging (MRI) in a 60-year-old male patient with Wernicke's encephalopathy.

Altered Expression of OsNLA1 Modulates Pi Accumulation in Rice (Oryza sativa L.) Plants.

Current agricultural practices rely on heavy use of fertilizers for increased crop productivity. However, the problems associated with heavy fertilizer use, such as high cost and environmental pollution, require the development of crop species with increased nutrient use efficiency. In this study, by using transgenic approaches, we have revealed the critical role of OsNLA1 in phosphate (Pi) accumulation of rice plants. When grown under sufficient Pi and nitrate levels, OsNLA1 knockdown (Osnla1-1, Osnla1-2, and Osnla1-3) lines accumulated higher Pi content in their shoot tissues compared to wild-type, whereas, over-expression lines (OsNLA1-OE1, OsNLA1-OE2, and OsNLA1-OE3) accumulated the least levels of Pi. However, under high Pi levels, knockdown lines accumulated much higher Pi content compared to wild-type and exhibited Pi toxicity symptoms in the leaves. In contrast, the over-expression lines had 50-60% of the Pi content of wild-type and did not show such symptoms. When grown under limiting nitrate levels, OsNLA1 transgenic lines also displayed a similar pattern in Pi accumulation and Pi toxicity symptoms compared to wild-type suggesting an existence of cross-talk between nitrogen (N) and phosphorous (P), which is regulated by OsNLA1. The greater Pi accumulation in knockdown lines was a result of enhanced Pi uptake/permeability of roots compared to the wild-type. The cross-talk between N and P was found to be nitrate specific since the knockdown lines failed to over-accumulate Pi under low (sub-optimal) ammonium level. Moreover, OsNLA1 was also found to interact with OsPHO2, a known regulator of Pi homeostasis, in a Yeast Two-Hybrid (Y2H) assay. Taken together, these results show that OsNLA1 is involved in Pi homeostasis regulating Pi uptake and accumulation in rice plants and may provide an opportunity to enhance P use efficiency by manipulating nitrate supply in the soil.

Morphological and thermal studies of chitin-curcumin blends derived polyurethanes.

The present study describes a novel ecofriendly series of chitin/curcumin/1,4-butane diol (BDO) blend derived polyurethanes (PUs), using hydroxy terminated polybutadiene (HTPB) and hexamethylene diisocyanate (HDI) along with different mole ratio of chitin, curcumin and BDO. The structural and morphological elucidation of the prepared films was done by FTIR and SEM techniques. The swelling behavior of the films was analyzed in both water and DMSO, which showed that incorporation of chitin increases the hydrophobicity and decreases the rate of swelling. Thermal analysis of synthesized PU blends revealed better thermal stability with following mole ratio 1:0.5:0.5 of chitin: curcumin: BDO as determined by TGA and DSC techniques.

The Arabidopsis Transcription Factor ANAC032 Represses Anthocyanin Biosynthesis in Response to High Sucrose and Oxidative and Abiotic Stresses.

Production of anthocyanins is one of the adaptive responses employed by plants during stress conditions. During stress, anthocyanin biosynthesis is mainly regulated at the transcriptional level via a complex interplay between activators and repressors of anthocyanin biosynthesis genes. In this study, we investigated the role of a NAC transcription factor, ANAC032, in the regulation of anthocyanin biosynthesis during stress conditions. ANAC032 expression was found to be induced by exogenous sucrose as well as high light (HL) stress. Using biochemical, molecular and transgenic approaches, we show that ANAC032 represses anthocyanin biosynthesis in response to sucrose treatment, HL and oxidative stress. ANAC032 was found to negatively affect anthocyanin accumulation and the expression of anthocyanin biosynthesis (DFR, ANS/LDOX) and positive regulatory (TT8) genes as demonstrated in overexpression line (35S:ANAC032) compared to wild-type under HL stress. The chimeric repressor line (35S:ANAC032-SRDX) exhibited the opposite expression patterns for these genes. The negative impact of ANAC032 on the expression of DFR, ANS/LDOX and TT8 was found to be correlated with the altered expression of negative regulators of anthocyanin biosynthesis, AtMYBL2 and SPL9. In addition to this, ANAC032 also repressed the MeJA- and ABA-induced anthocyanin biosynthesis. As a result, transgenic lines overexpressing ANAC032 (35S:ANAC032) produced drastically reduced levels of anthocyanin pigment compared to wild-type when challenged with salinity stress. However, transgenic chimeric repressor lines (35S:ANAC032-SRDX) exhibited the opposite phenotype. Our results suggest that ANAC032 functions as a negative regulator of anthocyanin biosynthesis in Arabidopsis thaliana during stress conditions.

Recent developments in curcumin and curcumin based polymeric materials for biomedical applications: A review.

Turmeric (Curcuma longa) is a popular Indian spice that has been used for centuries in herbal medicines for the treatment of a variety of ailments such as rheumatism, diabetic ulcers, anorexia, cough and sinusitis. Curcumin (diferuloylmethane) is the main curcuminoid present in turmeric and responsible for its yellow color. Curcumin has been shown to possess significant anti-inflammatory, anti-oxidant, anti-carcinogenic, anti-mutagenic, anticoagulant and anti-infective effects. This review summarizes and discusses recently published papers on the key biomedical applications of curcumin based materials. The highlighted studies in the review provide evidence of the ability of curcumin to show the significant vitro antioxidant, diabetic complication, antimicrobial, neuroprotective, anti-cancer activities and detection of hypochlorous acid, wound healing, treatment of major depression, healing of paracentesis, and treatment of carcinoma and optical detection of pyrrole properties. Hydrophobic nature of this polyphenolic compound along with its rapid metabolism, physicochemical and biological instability contribute to its poor bioavailability. To redress these problems several approaches have been proposed like encapsulation of curcumin in liposomes and polymeric micelles, inclusion complex formation with cyclodextrin, formation of polymer-curcumin conjugates, etc.

Overexpression of the CC-type glutaredoxin, OsGRX6 affects hormone and nitrogen status in rice plants.

Glutaredoxins (GRXs) are small glutathione dependent oxidoreductases that belong to the Thioredoxin (TRX) superfamily and catalyze the reduction of disulfide bonds of their substrate proteins. Plant GRXs include three different groups based on the motif sequence, namely CPYC, CGFS, and CC-type proteins. The rice CC-type proteins, OsGRX6 was identified during the screening for genes whose expression changes depending on the level of available nitrate. Overexpression of OsGRX6 in rice displayed a semi-dwarf phenotype. The OsGRX6 overexpressors contain a higher nitrogen content than the wild type, indicating that OsGRX6 plays a role in homeostatic regulation of nitrogen use. Consistent with this, OsGRX6 overexpressors displayed delayed chlorophyll degradation and senescence compared to the wild type plants. To examine if the growth defect of these transgenic lines attribute to disturbed plant hormone actions, plant hormone levels were measured. The levels of two cytokinins (CKs), 2-isopentenyladenine and trans-zeatin, and gibberellin A1 (GA1) were increased in these lines. We also found that these transgenic lines were less sensitive to exogenously applied GA, suggesting that the increase in GA1 is a result of the feedback regulation. These data suggest that OsGRX6 affects hormone signaling and nitrogen status in rice plants.

Chitin based polyurethanes using hydroxyl terminated polybutadiene, Part II: morphological studies.

Chitin-hydroxyl terminated polybutadiene (HTPB) based polyurethane (PU) was prepared and structure of the pre-designed PU was confirmed using FT-IR spectrometer. The FT-IR analysis confirmed that the crosslinking density increased with increasing the chitin contents in the final PU. During the detailed FT-IR study, it was observed that tri-functional character of chitin is responsible for the formation of network structure. The scanning electron microscope (SEM) analysis also confirmed the cross-linked structure of the material. The amount of carbon, nitrogen and oxygen elements obtained from EDX-SEM microanalysis also supported the results. The resistance in solubility against the solvent also confirmed the crosslinking behavior of the prepared polyurethane.