Synthetic clonal reproduction through seeds

Cloning through seeds has potential revolutionary applications in agriculture, because it would allow vigorous hybrids to be propagated indefinitely. However, asexual seed formation or apomixis, avoiding meiosis and fertilization, is not found in the major food crops. To develop de novo synthesis of apomixis, we crossed Arabidopsis MiMe and dyad mutants that produce diploid clonal gametes to a strain whose chromosomes are engineered to be eliminated after fertilization. Up to 34% of the progeny were clones of their parent, demonstrating the conversion of clonal female or male gametes into seeds. We also show that first-generation cloned plants can be cloned again. Clonal reproduction through seeds can therefore be achieved in a sexual plant by manipulating two to four conserved genes.     

Volatile constituents from Cinnamomum zeylanicum fruit stalks and their antioxidant activities

Cinnamomum zeylanicum Blume is an important spice and aromatic crop having wide applications in flavoring, perfumery, beverages, and medicines. The steam-distilled volatile oil from cinnamon fruit stalks was analyzed with GC and GC-MS. It showed the presence of hydrocarbons (44.7%) and oxygenated compounds (52.6%). Twenty-seven compounds constituting ca. 95.98% of the volatile oil were characterized. (E)-Cinnamyl acetate (36.59%) and (E)-caryophyllene (22.36%) are found to be major compounds. The volatile oil was screened for its potential as an antioxidant by using in vitro models, such as the beta-carotene-linoleate and phosphomolybdenum complex method. The volatile oil showed 55.94% and 66.9% antioxidant activity at 100 and 200 ppm concentration, respectively. Also, the volatile oil showed good antioxidant capacity, using the formation of the phosphomolybdenum complex. A comparison of the chemical composition of the volatile oil was made with that of buds, flowers, and fruits. This is the first report on the chemical composition of volatile oil of the fruit stalks of this species and its antioxidant activity.     

Acylated and non-acylated flavonol monoglycosides from the Indian minor spice Nagkesar (Mammea longifolia)

A methanol extract of nagkesar (buds of Mammea longifolia), which showed strong radical scavenging activity, yielded 13 compounds by separations using column chromatography and HPLC. Structure elucidation of these compounds was achieved by (1)H and (13)C NMR, including DQF-COSY, TOCSY, DEPT, HMQC, HSQC, and HMBC. They include two new compounds, quercetin 3-O-(2' ',4' 'di-E-p-coumaroyl)-alpha-L-rhamno-pyranoside and quercetin 3-O-(3' ',4' '-di-E-p-coumaroyl)-alpha-L-rhamnopyranoside, along with known compounds kaempferol, quercetin, the isopropylidenedioxy derivative of shikimic acid, kaempferol 3-O-(2' ',4' '-di-E-p-coumaroyl)-alpha-L-rhamnopyranoside, kaempferol 3-O-(3' ',4' '-di-E-p-coumaroyl)-alpha-L-rhamnopyranoside, kaempferol 3-O-alpha-L-rhamnopyranoside, quercetin 3-O-alpha-L-rhamnopyranoside, shikimic acid, kaempferol 3-O-beta-D-glucopyranoside, quercetin 3-O-beta-D-glucopyranoside, and beta-sitosterol 3-O-beta-D-glucopyranoside.     

‘Soya Milk Tris‐based Phytoextender Reduces Apoptosis in Cryopreserved Buffalo (Bubalus bubalis) Spermatozoa’

The objective of this study was to investigate the effect of newly developed soya milk Tris (SMT)‐based phytoextender as an alternative to egg yolk Tris (EYT) extender used for cryopreservation of buffalo (Bubalus bubalis) spermatozoa on apoptosis. Fresh buffalo semen (control without dilution) was cryopreserved in conventional EYT (20% egg yolk v/v in Tris) and SMT (25% soya milk v/v in Tris) extender and used for the assessment of expression of apoptotic proteins. Proteins extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random were separated using SDS–PAGE followed by immunoblotting against caspase‐8, caspase‐9, caspase‐3, poly(ADP‐ribose)polymerase (PARP), cytochrome c and apoptosis inducing factor (AIF). In addition, fluorescence microscopy was used for the detection of mitochondrial membrane potential (JC‐1 assay) and apoptotic cells (annexin V‐FITC/PI assay). The results obtained clearly indicate the significant (p < 0.05) reduction in the expression of caspase‐3 (27 kDa), caspase‐8 (53 kDa), caspase‐9 (50 kDa) precursor and cytochrome c (17 kDa) in semen cryopreserved in SMT extender in comparison with EYT extender. A non‐significant (p > 0.05) reduction in expression of PARP‐DNA‐binding subunit (24 kDa) was observed in SMT extender. No expression of AIF was found in cryopreserved semen samples. A significant (p < 0.05) increase in the mean percentage of cells having high mitochondrial membrane potential and a non‐significant (p > 0.05) decrease in late apoptotic cells (AN+/PI+) was observed in SMT extender when compared to EYT extender. The results demonstrated that cryopreservation of buffalo semen in SMT‐based phytoextender can replace the traditional egg yolk extenders as it reduces the expression of apoptotic proteins maintaining high mitochondrial membrane potential and gives better protection to sperms in terms of its non‐animal origin.     

Morpho-Physiological Responses of Grape Rootstock ‘Dogridge’ to Arbuscular Mycorrhizal Fungi Inoculation Under Salinity Stress

Effects of arbuscular mycorrhizal fungi (AMF) alone or in combination with bacterial consortium (AMF+BC) inoculation prior to induced salinity (NaCl @ 150 or 250 mM) were studied on root growth; plant biomass; leaf area; Na⁺ and K⁺ contents; leaf water potential (Ψ_w); osmotic potential (Ψ_π); photosynthesis rate (P_n); and contents of chlorophyll, phytohormones, and polyamines in the grape rootstock ‘Dogridge’, popular among Indian vine growers. AMF inoculation in the NaCl untreated rootstocks plants increased root growth, root and shoot biomass, and leaf area and improved leaf Ψ_w, Ψ_π, P_n, and chlorophyll content, and also countered the stress-induced decline in the NaCl treated plants. The abscisic acid (ABA), cytokinins, and polyamine-spermidine and spermine contents in the leaves of NaCl untreated or treated were significantly increased by the AMF inoculation. Among the treatments, AMF with BC was relatively more effective than AMF alone with respect to changes in above morpho-physiological characters. The results depicted that AMF (AMF alone or AMF+BC) inoculation significantly improved salinity tolerance of grape rootstock and tolerance is induced by improvements in plant water balance, K⁺:Na⁺ ratio, and P_n, besides distinct accumulations in ABA and polyamines-spermine and spermidine. The above findings have potential in suggesting the AMF usefulness in improving the efficacy of ‘Dogridge’ rootstock in grape cultivation under salt affected soils.     

Effect of probiotic supplementation on serum/yolk cholesterol and on egg shell thickness in layers

1. The effect of probiotic supplementation on egg production, on serum and yolk cholesterol and on egg shell thickness in 24 White Leghorn layers was studied from 28–38 weeks of age.

2. In 3 treatments the diet was supplemented with 0, 100 and 150 mg probiotic/kg food.

3. In the 100 mg probiotic group, egg production improved by 5%, and shell thickness improved slightly, with fewer thin‐shelled eggs than in the control (8.6% compared to 18.6%).

4. The initial serum cholesterol concentration of 170.2 mg/dl in control birds remained similar throughout the 10‐week experimental period, whereas in the 150 mg group the initial value of 176.5 mg/dl decreased to 114.3 mg by week 10.

5. Yolk cholesterol concentration was 14.69 mg in the control group and 11.28 and 11.37 mg/g in the 100 and 150 mg probiotic groups respectively. Overall mean total egg cholesterol was thus reduced by probiotic supplementation.     

Recombinant LipL32 antigen-based single serum dilution ELISA for detection of canine leptospirosis

A recombinant antigen-based single serum dilution enzyme-linked immunosorbent assay (ELISA) was developed to measure the specific antibody activity in sera of dogs with leptospirosis. The recombinant antigen developed and used in the assay was specific for the pathogenic serovars of Leptospira. A linear relationship was found to exist between the predicted antibody titres at a single working dilution of 1:1000 and the corresponding observed serum titres as determined by the standard serial-dilution method. Regression analysis was used to determine a standard curve from which an equation can be derived that allows demonstration of the mentioned correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay was proved to be sensitive, specific and accurate as compared to the standard microscopic agglutination test (MAT).     

矿山废弃地土壤基质改良研究

矿山废弃地对生态环境有着极大的破坏作用,危及人类的生存与健康,并影响到区域生态经济的可持续发展。矿山废弃地的生态修复和重建,已被看作是矿区土地修复的主要组成部分,而在生态恢复的植被重建过程中,基质改良是生态恢复的核心问题。研究借鉴目前国内外土壤基质改良的方法,较系统地介绍了矿山废弃地土壤基质物理、化学、生物改良技术措施。最后提出我国矿山废弃地土壤改良方法的选用原则：遵循因地制宜、就地取材,结合以往的研究成果,借鉴国际矿区土地复垦和生态修复的成功经验,研究废弃物土壤化演化的自然规律和机理,最终实现人工辅助的土壤化演化。     

Characterization of Spray-Dried Hydrolyzed Proteins from Pink Perch Meat Added with Maltodextrin and Gum Arabic

In the present study, spray-dried pink perch meat protein hydrolysate (PPH) and hydrolysate with maltodextrin and gum arabic (PPHMG) were characterized in relation to their physical, rheological, functional, antioxidant, thermal, and sensory properties. The Herschel–Bulkley model was suitable to explain the flow behavior of PPH and PPHMG emulsions. Both the samples exhibited functional and antioxidant properties. Antioxidant activities were significantly higher for PPH compared with PPHMG (p<0.05). PPH and PPHMG alone did not exhibit antimicrobial activity but enhanced the activity of chitosan against pathogens. Sensory acceptability of the samples in soup revealed that PPHMG of up to 4% was highly scored without any traces of bitterness, whereas for PPH, a concentration of up to 3% was acceptable.     

N-Dimethylamino Succinamic Acid (B-Nine) as a Potential Aid in Improvement of Medicago sativa L.

The influence of N-dimethylamino succinamic acid (B-nine) on lucerne plants cv. IGFRI-244 was investigated. B-nine (a growth regulant) was sprayed as a foliar sprays, employing 10, 100, 250, 500, 1000 and 5000 ppm alongwith water spray as control, and the effects were observed on plant growth, flowering, nutritive constituents and seed yield of the crop. Plant height was reduced from 6.19 to 26.85 % in various concentrations of B-nine being highest retardation in 5000 ppm, whereas 250 ppm produced more branches and leaves with dense foliage in comparison to control and other treatments. Delayed flowering by 2–15 days with significant increase in seed yield and 1000 seed weight B-nine bein maximum seed yield in 250 ppm. However, total seed yield was increased by 4.16 to 24.76 per cent over unsprayed control plants. Among the treated levels of B-nine, 250 ppm also gave higher values for Carbohydrate, IVTDMD in shoot of lucerne plants with considerable decreased in 5000 ppm. NDF condent decreased progressively with increased concentration of B-nine but always lesser than the value of IVTDMD.