Biofabrication of Ag nanoparticles using Moringa oleifera leaf extract and their antimicrobial activity

Objective:To formulate a simple rapid procedure for bioreduction of silver nanoparticles using aqueous leaves extract of Moringa oleifera(M.oleifera).Methods:10 mL of leaf extract was mixed to 90 mL of 1 mM aqueous of AgNO_3 and was heated at 60-80 ℃ for 20 min.A change from brown to reddish color was observed.Characterization using UV-Vis spectrophotometry, Transmission Electron Microscopy(TEM) was performed.Results:TEM showed the formation of silver nanoparticles with an average size of 57 nm.Conclusions:M.oleifera demonstrates strong potential for synthesis of silver nanoparticles by rapid reduction of silver ions(Ag^+ to Ag^0). Biological methods are good competents for the chemical procedures,which are eco-friendly and convenient.

Antioxidant Protection Mechanisms Reveal Significant Response in Drought-Induced Oxidative Stress in Some Traditional Rice of Assam, India

Drought tolerance levels and antioxidant protection mechanisms were evaluated for 21 traditional rice varieties of Assam, India, along with Sahbhagi Dhan （drought tolerant） and IR64 （drought sensitive） as controls. Drought was imposed in hydroponic culture with polyethylene glycol 6000 （PEG6000） that was initially standardized with different concentrations. All the rice varieties showed apparent decreases in growth characteristics under drought stress （initially at 15% for 7 d followed by 20% PEG6000 for 7 d in Yoshida medium）. On the basis of standard evaluation score （SES）, eight rice varieties showed high drought tolerance which were carried forward for further biochemical analyses. Based on different morpho- physiological parameters, SN03 （Bora）, SN04 （Prosad Bhog）, SN05 （Kola Joha）, SN06 （Helash Bora）, SN08 （Salihoi Bao）, SN12 （Kola Amona）, SN20 （Ronga Bora） and SN21 （Sok-Bonglong） were identified as promising drought tolerant varieties. The non-enzymatic antioxidants activities viz., glutathione, ascorbate and enzymatic antioxidant activities such as superoxide dismutase （SOD）, catalase （CAT）, guaiacol peroxidase （GPX）, ascorbate peroxidase （APX）, glutathione reductase （GR） in shoots and roots of all the selected varieties revealed significant level of protection mechanisms as compared with controls. Enhancement in activities of the overall antioxidant enzymes including SOD, GPX, CAT, GR and APX under drought stress reflects their role in the adaptation process under water stress.

Marker-Assisted Pyramiding of Genes Conferring ResistanceAgainst Bacterial Blight and Blast Diseases into Indian RiceVariety MTU1010

Two major bacterial blight （BB） resistance genes （Xa21 and xa13） and a major gene for blastresistance （Pi54） were introgressed into an Indian rice variety MTU1010 through marker-assistedbackcross breeding. Improved Samba Mahsuri （possessing Xa21 and xa13） and NLR145 （possessingPi54） were used as donor parents. Marker-assisted backcrossing was continued till BC2 generationwherein PCR based functional markers specific for the resistance genes were used for foregroundselection and a set of parental polymorphic microsatellite markers were used for background selectionat each stage of backcrossing. Selected BC2F1 plants from both crosses, having the highest recoveriesof MTU1010 genome （90% and 92%, respectively）, were intercrossed to obtain intercross F1 （ICF1） plants,which were then selfed to generate 880 ICF2 plants possessing different combinations of the BB andblast resistance genes. Among the ICF2 plants, seven triple homozygous plants （xa13xa13Xa21Xa21Pi54Pi54）with recurrent parent genome recovery ranging from 82% to 92% were identified. All the seven ICF2plants showed high resistance against the bacterial blight disease with a lesion lengths of only 0.53–2.28 cm, 1%–5% disease leaf areas and disease scoring values of ‘1’ or ‘3’. The seven ICF2 plants wereselfed to generate ICF3, which were then screened for blast resistance, and all were observed to behighly resistant to the diseases. Several ICF3 lines possessing high level of resistance against BB andblast, coupled with yield, grain quality and plant type on par with MTU1010 were identified and advanced forfurther selection and evaluation.

Tagging of Brown Planthopper Resistance Genes in F_2s of IR50 × Ptb33 of Rice by Using Bulked Segregant Analysis

Brown planthopper （Nilaparvata lugens Stal） is one of the most damaging pests causing hopper burn in rice, and thereby reducing the productivity and also the quality of the product. The effective management strategy to control this pest is the identification and transfer of desirable genes to local rice cultivars. The most important approach for developing resistant cultivars is the identification of markers, which can help in marker-assisted selection of more durable resistant genotype. The susceptible parent IR50 and the resistant parent Ptb33, and their F2 populations were used in bulked segregant analysis for identification of resistant genes with random amplified polymorphic DNA marker （RAPD） primers. The primers OPC7 and OPAG14 showed both dominant and susceptible specific banding pattern so called co-dominant markers. Moreover, OPC7697 and OPAG14680 showed resistant specific bands and thus being in coupling phase, whereas OPC7846 and OPAG14650 showed susceptible specific genotypic bands in bulked segregant analysis. Therefore, the coupling phase markers, OPC7697 and OPAG14680, are considered to be more useful in marker-assisted selection of rice genotypes in crop improvement.

Nature of Gene Action and Maternal Effects for Pod Borer, <i>Helicoverpa armigera</i>Resistance and Grain Yield in Chickpea, <i>Cicer arietinum</i>

Information on mechanisms and inheritance of resistance is critical to plan an effective strategy to breed for resistance to insect pests. Therefore, we evaluated a diverse array of chickpea genotypes (eight desi and one kabuli) with varying levels of resistance to the pod borer, Helicoverpa armigera to gain an understanding of the nature of gene action and possible maternal effects. The test genotypes were crossed in all possible combinations for a full diallel. The 72 F1s (36 direct and 36 reciprocal crosses) along with the parents were evaluated for resistance to H. armigera under field conditions, and for antibiosis mechanism of resistance (larval survival and larval weight gain) by using detached leaf assay under laboratory conditions, and grain yield under un-protected conditions in the field. Additive gene action governed the inheritance of resistance to H. armigera, while non-additive type of gene action was predominant for inheritance of antibiosis component of resistance (larval survival and larval weight) and grain yield. Greater magnitude of σ2 A(17.39 and 1.42) than σ2 D (3.93 and 1.21) indicated the preponderance of σ2 Ain inheritance of resistance to pod borer, H. armigera under laboratory and field conditions, respectively. There were no maternal effects for inheritance of resistance to pod borer and grain yield. Lines with significant gca effects for pod borer damage and grain yield were identified for further use in the resistance breeding program. The implications of the inheritance pattern of pod borer resistance and grain yield are discussed in the context of strategies to enhance pod borer resistance and grain yield in chickpea.

Soil Test Based Fertilizer Prescriptions through Inductive cum Targeted Yield Model for Sesamum on Alfisol

Studies on Soil Test Crop Response Based Integrated Plant Nutrition System(STCR-IPNS)were conducted for three years adopting the Inductive cum Targeted Yield Model,on alfisols of Unified Andhra Pradesh,Southern India during summer 2016-2018 in order to develop fertilizer prescriptions through IPNS for the desired yield targets of Sesamum under field conditions.The bases for making the fertilizer prescriptions viz.nutrient requirement(NR),contribution of nutrients from soil(Cs),fertilizer(Cf)and vermicompost(CVC)were computed using the field experimental data.Making use of these basic parameters,the fertilizer prescription equations were developed under NPK alone and under IPNS for the desired yield targets of Sesamum for a range of soil test values.The quantity of fertilizers contributed by the application of vermicompost was assessed.Nutrient requirement to produce 100 kg of sesame seed was worked out to be 10.20 kg N,3.90 kg P2O5 and 5.22 kg K2O.In the present investigation,the requirement of N was higher which is followed by K2O and P2O5.The requirement of N was 2.62 times higher than P and 1.95 times higher than K.The percent contribution of N,P and K was 12.25,15.75 and 6.00 from soils,41.68,22.85 and 59.97 from fertilizer and 9.87,6.74 and 18.65 from organic manures,respectively.Thus the Inductive cum Targeted Yield Model used to develop fertilizer prescription equations provides a strong basis for soil fertility maintenance consistent with high productivity and efficient nutrient management in farming for sustainable and enduring agriculture.

Toxicity of Bacillus thuringiensis Crystal Toxins to Field Populations of Rice Leaf Folder, Cnaphalocrocis medinalis （Guenee） and Establishment of Baseline Susceptibility to Cry1Ab

Eight insecticidal crystal proteins of Bacillus thuringiensis, CrylAa, CrylAb, CrylAc, CrylB, Cry2Aa, CrylC, CrylDa and Cry 1Ea were assessed for toxicity against 1 st instar larvae of rice leaf folder, Cnaphalocrocis medinalis （Guenee） at 48 HAT and 72 HAT. Bioassay results depicted CrylAa was the most toxic （LCso 2.35 ppm） followed by CrylBa （LCso 8,50 ppm） and CrylAb （LCso 8.73 ppm） at 48 HAT, whereas, at 72 HAT CrylAb proved to be highly toxic （LC50 0.50 ppm） followed by CrylAa （LCso 4.07 ppm）, CrylAc （LCso 4,84 ppm） and CrylBa （LCso 6.42 ppm）. Toxins Cry2Aa, CrylCa, CrylDa and CrylEa did not resulted in any mortality at 48 HAT and 72 HAT, respectively. Baseline estimates for CrylAb against 1st instar larvae of C. medinalis sampled from seven geographical locations revealed variation in LC50＇s from 0.37 ppm to LC50 16.25 ppm at 48 HAT and LC50 0.50 ppm to LC50 6.49 ppm 72 HAT, respectively with relative resistance ratios of 44-fold and 13-fold at 48 HAT and 72 HAT over the susceptible population.

Qualitative and Quantitative Changes in Biochemical Constituents in the Rice Plant Treated with Different Xenobiotics

Experiments were conducted at Entomology division of Directorate of Rice research to study the qualitative and quantitative changes in biochemical constituents in the rice plant treated with different xenobiotics, viz. insecticides like deltamethrin, imidacloprid, herbicide like 2,4-Dichlorophenoxyacetic acid, fungicide like carbendazim, green manure Calotropis gigantea leaves, botanicals like Neem Seed Powder and fertilizer urea along with control plants of TN1, PTB33 and Swama varieties comprising of total 15 treatments. Application of butachlor, carbendazim, deltamethrin and excessive nitrogen fertilization resulted in increased population build up of brown plant hopper （BPH）. Significantly, lower honeydew excretion （13 mme area） in the BPH nymphs was observed on resistant PTB33 variety compared to susceptible TN1 and Swarna varieties （116 and 110 mm2 respectively）. In susceptible TN1 variety, the sucrose content was significantly higher （9.35 mg/g tissue） compared to BPH resistant PTB33 （5.32 mg/g tissue）. The BPH resistant PTB33 variety exhibited significantly lower free amino acid content （1.23 mg/g tissue） as compared to TN1 （2.51 mg/g）. The C/N ratio was comparatively high in the resistant PTB33 alone （4.32）, but was low in PTB33 applied with 2,4-D ＋ carbendazim （2.73）, 2,4-D ＋ imidacloprid ~ carbendazim （2.3）, excess urea fertilizer （2.18）. The total phenol contents measured in resistant PTB33 （7.5 mg/g dry wt）, susceptible TN1 （5.865 mg/g dry wt） and the popular variety Swarna （5.57 mg/g dry wt） were almost same and no statistical difference was observed. Further, treatment of these varieties with different xenobiotics and botanicals also did not significantly alter their total phenol content. However, the phenol content of the resistant PTB 33 alone and treated with xenobiotics was higher compared to the susceptible TN 1 and Swarna varieties alone and treated with xenobiotics. The plant biochemical constituents such as free amino acids, sucrose were lower in untreated resistant PTB33 compared to susceptible TN1, and PTB33 treated with urea and deltamethrin. The high feeding rate and population build up on the susceptible variety TN1 is positively correlated with high quantities of sucrose, free amino acids and lower quantities of phenols and low C/N ratio.

Carboxylesterase and Glutathione-S-Transferase （GST＇s） Induced Resistance to Bacillus thuringiensis Toxin CrylAb in Rice Leaf Folder, Cnaphalocrocis medinalis （Guenee） Populations

The rice leaf folder （RLF）, Cnaphalocrocis medinalis （Guenee） （lnsecta： Lepidoptera： Pyralidae）, is an important pest, widely distributed in many rice growing areas of Asia. The over-use of broad-spectrum chemical insecticides has been cited as a major cause of outbreaks of C. medinalis as excessive spraying of insecticide disrupts natural biological control insecticides still remain the major control tactics against leaf folder. Carbofuran and fenthion, bendiocarb, acephate, carbosulfan, quinolphos, monocrotophos, phosphamidon and fenvalerate are the common ones used against rice leaf folder. Genetically, modified rice lines expressing B. thuringiensis insecticidal crystal proteins produced are highly tolerant to leidopteran pests. Though economic and environmental benefits of GM crops is well established, the matter of concern is the possibility of target insect pest developing resistance to this B. thuringiensis insecticidal toxins, evident from many laboratory and field experiments against many insect pests. The involvement of GSH S-transferase, carboxylesterase, and microsomal monooxygenase in insecticide resistance has been reported in insecticide-resistant strains of many insect species. Hence, the present study was taken up to monitor for cross resistance between B. thuringiensis cry toxins and synthetic insecticides in larvae of leaf folder as it is mediated by carboxylesterase titre and other enzymes by bioassay for two selected rice leaf folder field populations at the Entomology division of Directorate of Rice Research which showed 2-fold resistance ratio. Qualitative and quantitative changes of carboxylesterase （CarE） and glutathione-s-transferase （GST＇s） were worked out with midguts extracts of the two C. medinalis populations in the presence of a-napthyl acetate and chlorodi-nitro benzene substrates.

Efficacy of Acaricides alone and in Combination with Propiconozole against Rice Panicle/Sheath Mite, Steneotarsonemus spinki

Experiments were conducted for chemical management against panicle mite, Steneotarsonemus spinki using seven treatments. Based on the overall performance of chemicals against this pest, spiromesifen ~ propiconazole, diafenthuron ＋ propiconazole and fenpyroxymate alone were found effective against panicle mite and associated grain discolouration. Spiromesifen （1 mL） ＋ propiconazole （l mL） per litre of water was the best treatment which recorded 24.77% increase in grain yield over control.

Functional Genes in Relation to Residual Feed Intake in Murrah Buffalo Heifers

High Feed efficiency (FE) in growing heifers has economic importance in dairy, but remains less understood in buffaloes. Feed conversion efficiency is defined as dry matter intake (DMI) per unit body weight gain and is determined as residual feed intake (RFI), i.e., the difference between actual and predicted feed intake to gain unit body weight during a feed trial run for 78 days under control feeding. A large variation was identified ranging between -0.42 to 0.35 in growing buffalo heifers (n = 40) of age between 11 to 15 months. An average daily weight gain (ADG) varied between 382.0 and 807.6 g/day when compared with the control-fed heifers at an organized buffalo farm. The whole blood transcriptome data obtained from the selected growing heifers from extremes of estimated high and low RFI efficiency were compared with the reference assembly generated from the transcriptome of multiparous buffaloes (n = 16) of diverse age of maturity, period of regaining post partum cyclicity and level of milk production. Differentially expressed genes (DEGs) were identified using the reference genome of Mediterranean water buffalo. GO: terms (Padj 0.05, FDR 0.05) enriched by annotated DEGs and biological pathways in gene network for RFI efficiency trait were identified. GO: terms specific to pre-transcriptional regulation of nucleus and Chromatin organization under Nucleoplasm, Energy balancing, Immunity, Cell signaling, ROS optimization, ATP generation through the Electron Transport chain and cell proliferation were determined. The study reveals the indicators targeting the actual metabolic changes and molecular functions underlying the feed utilization capacity of buffaloes. Estimated RFI efficiency revealed a large variation over heifers which may lower the DMI even up to 13.6% thus, enabling an increase in ADG up to 16% by involving efficient heifers in breeding plan. The study revealed a scope of high gain by selective breeding for FE in heifers. FE variants catalogued in the study are useful breed-specific RFI markers for future reference. The study contributes to the understanding of feed efficiency in buffaloes and its association with key interactive traits such as reproduction and growth. This knowledge can be utilized to develop more effective breeding programs.