Molecular characterization and PCR detection of a nitrogen-fixing Pseudomonas strain promoting rice growth

Nitrogen-fixing plant growth-promoting rhizobacteria (PGPR) from the genus Pseudomonas have received little attention so far. In the present study, a nitrogen-fixing phytohormone-producing bacterial isolate from kallar grass (strain K1) was identified as Pseudomonas sp. by rrs (16S ribosomal RNA gene) sequence analysis. rrs identity level was high with an uncharacterized marine bacterium (99%), Pseudomonas sp. PCP2 (98%), uncultured bacteria (98%), and Pseudomonas alcaligenes (97%). Partial nifH gene amplified from strain K1 showed 93% and 91% sequence similarities to those of Azotobacter chroococcum and Pseudomonas stutzeri, respectively. The effect of Pseudomonas strain K1 on rice varieties Super Basmati and Basmati 385 was compared with those of three non-Pseudomonas nitrogen-fixing PGPR (Azospirillum brasilense strain Wb3, Azospirillum lipoferum strain N4 and Zoogloea strain Ky1) used as single-strain inoculants. Pseudomonas sp. K1 was detected in the rhizosphere of inoculated plants by enrichment culture in nitrogen-free growth medium, which was followed by observation under the microscope as well as by PCR using a rrs-specific primer. For both rice varieties, an increase in shoot biomass and/or grain yield over that of noninoculated control plants was recorded in each inoculated treatment. The effect of Pseudomonas strain K1 on grain yield was comparable to those of A. brasilense Wb3 and Zoogloea sp. Ky1 for both rice varieties. These results show that nitrogen-fixing pseudomonads deserve attention as potential PGPR inoculants for rice.     

Quantitative Estimation of Genetic Variability in Morphological Characteristics of Eastern Cottonwood (Populus deltoides Bartr.) Clones

The study was carried out to analyze the genetic variability for different growth parameters in poplar clones at the age of 2 and 3 yr in the nursery. Forty-nine exotic and indigenous clones of poplar were evaluated for eight morphological traits. Clones were planted in randomized block design (RBD) with three replications with four clones in a block in each treatment. Observations for different characters were recorded on six selected competitive clones per genotype. Results showed a high interclonal variability for most parameters. Statistically significant differences among clones indicated that the majority of study characters are controlled by genetic factors, specific to each clone. Highly significant genotypic difference supported by wide range of variation of mean and range values were observed for the characters under study. Significant and positive correlation was observed between diameter at breast height (DBH) and plant height. Leaf lamina length showed positive and significant correlation with petiole length, total leaf length, leaf width, and L/B ratio. Total leaf length showed positive and significant correlation with leaf width and L/B ratio. High estimates of heritability (in a broad sense) were observed for almost all characters in the study. High genetic advance expressed as percent of mean was recorded for petiole length (60.90), followed by plant height (60.78) and collar diameter (44.19) at 2 yr. At the age of 3 yr, genetic advance was found maximum for petiole length (60.05), followed by collar diameter (47.62) and plant height (45.29). The efforts for selecting new clones and their field-testing must continue under a long-term improvement plan so that the best clones can be recommended for plantations and hybridization programs.     

Effect of ammonia-N on growth and feeding of juvenile Macrobrachium rosenbergii (De-Man)

Survival rate, growth and feed intake were determined for late juveniles (4.31 ± 0.18 g) of river prawn Macrobrachium rosenbergii in freshwater with total ammonia‐N (NH₃‐N+NH₄‐N) concentrations of 0.015 (control), 0.5, 1.0 and 1.5 mg L⁻¹ for 60 days at pH 7.53 ± 0.04 and temperature 24.0 ± 2.5°C. Survival rate was significantly (P<0.05) lower (54 ± 4.2–70 ± 5.4%) for total ammonia concentrations from 0.5 to 1.5 mg L⁻¹ [0.0139–0.0419 mg L⁻¹ of unionized ammonia (NH₃)]. Growth (0.026–0.030 g day⁻¹ range) of the prawns did not differ for the different NH₃ levels but were significantly (P<0.05) lower compared with control (0.056 g day⁻¹). Feed intake rates also diminished significantly (P<0.05) from 77.60 ± 2.45% at control (0.015 mg L⁻¹ NH₃‐N) to 48.69 ± 2.13% at 1.5 mg L⁻¹ NH₃‐N (0.0419 mg L⁻¹ of unionized NH₃).     

Tree age influences nutritional,pectin, and anatomical changes in developing ‘Kinnow’ mandarin (Citrus nobilis Lour × Citrus deliciosa Tenora) fruit

An understanding of variation in fruit quality in relation to tree age can help understand the issue of poor fruit quality in young orchards; however, limited information is available on the citrus fruit. In this study, the endogenous nutritional status in rind, rag, and leaves; pectin status in rind and rag; and anatomical fruit growth parameter in rind tissues were studied in ‘Kinnow’ mandarins fruit during their development on trees from three age (6-, 18-, and 35-year-old) groups. In older (35-year-old) trees, rind, rag, and leaf nitrogen (N), phosphorus (P), and potassium (K) concentrations were superior. In fruit from all tree age groups, total pectin and protopectin reduced; however water-soluble pectin (WSP) improved. In rind tissues harvested from young (6-year-old) trees, cell density was more while cell size was less. In all tree age groups, cell density in rind tissues correlated negatively with rind WSP.     

Molecular and biologic characterization of a phytoplasma associated with <Emphasis Type="Italic">Brassica campestris</Emphasis> phyllody disease in Punjab province,Pakistan

A phytoplasma-associated disease was identified in Brassica campestris (sarson) plants during a survey conducted in Punjab province of Pakistan in 2014–2016. The symptomatic plants showed characteristic symptoms of phyllody and witches’ broom. Phytoplasma presence was detected by polymerase chain reaction on 16S ribosomal and tuf DNAs, followed by RFLP analysis and sequence comparison of the 16S rRNA and tuf genes. The phytoplasma detected was classified in a new ribosomal subgroup designed 16SrIX-H. The phytoplasma presence in phloem tissues of symptomatic sarson samples was also confirmed through light microscopy and transmission studies to healthy plants through dodder and the leafhopper Orosius albicinctus. This is the first report of identification of 16SrIX-H subgroup phytoplasma associated with sarson and its natural vector in Pakistan.     

Suitability of Immobilized Pseudomonas fluorescens SM1 Strain for Remediation of Phenols, Heavy Metals, and Pesticides from Water

Immobilized microbial cells for the biological treatment have the potential to degrade toxic chemicals faster than conventional wastewater treatment systems. In the present study, suitability of immobilized Pseudomonas fluorescens SM1 strain in calcium alginate beads for remediation of the major toxicants in Indian water bodies was tested by means of GC/HPLC and AAS techniques. Roughly 80% reduction in the concentration of phenols was observed by immobilized SM1 cells compared with 60% by the free cells. Also, in the case of the bioremediation of heavy metals, immobilized SM1 cells were found to be more efficient compared with the free cells. Suspension of P. fluorescens SM1 cells in the test model water for 24 h brought down the concentrations of Cu⁺⁺, Cd⁺⁺, Ni⁺⁺, and Pb⁺⁺ by more than 75% under free cell state and 7?C9% better efficiency under the immobilized conditions. However, Cr(VI) could show only 44% removal by the cell immobilized system, whereas a mere 35% reduction in the Cr(VI) levels was shown in the test model water by the free SM1 cells under the same conditions. Moreover, a model water containing 2,000 ppb of BHC, 1,248 ppb mancozeb, and 312 ppb 2,4-D passed through the cell immobilized column resulted in the decline in their concentrations up to 362 ppb, 750 ppb, and 126 ppb, respectively. Generally, AAS, HPLC, and GC analyses of treated test model waters with the free and immobilized SM1 cells exhibited high potential of immobilized SM1 in detoxification of test water. From the results, we conclude that immobilized cells of P. fluorescens SM1 strain were quite effective in bioremediation of major toxicants present in Indian water bodies, and we also recommend the use of immobilized bacterial cells rather than the free cells for the bioremediation/detoxification process.     

In vitro activity of isothiocyanates against Fusarium graminearum

Isothiocyanates are biotoxic degradation products formed as a result of enzymatic hydrolysis of glucosinolates present in Brassica species. The application of biofumigant Brassica crops, as an alternative crop protection method for soilborne pathogens and pests is increasingly gaining interest. However, little is known of the potential of biofumigation to reduce the inoculum of Fusarium species affecting cereals. The aim of this study was to evaluate the antifungal activity of five isothiocyanates, namely allyl, benzyl, ethyl, 2-phenylethyl and methyl isothiocyanates, against germination and growth of Fusarium graminearum under in vitro conditions. Aromatic isothiocyanates were more inhibitory than the aliphatic isothiocyanates against mycelial growth, whereas the reverse was observed for conidial germination. Among the tested isothiocyanates, allyl and methyl isothiocyanates were more efficient overall, showing lower ED₅₀ values (35–150 mg/L) for conidial germination and mycelial radial growth. The findings suggest that Brassica plants containing allyl and methyl glucosinolates could have a suppressive effect, reducing the inoculum of F. graminearum in soil prior to cereal production.     

Diversity among melon (Cucumis melo L.) landraces from the Indo-Gangetic plains of India and their genetic relationship with USA melon cultivars

We report here the first broad genetic characterization of farmer-developed landraces of melon (Cucumis melo L.) from the Indo-Gangetic plains of India, an area overlooked in previous melon genetic diversity analyses of Indian melon germplasm. Eighty-eight landraces from three melon Groups in two subspecies (C. melo subsp. agrestis Momordica Group, and C. melo subsp. melo Cantalupensis Group and Reticulatus Group) were collected from the four agro-ecological regions (six sub-regions) of two northern states of the Indo-Gangetic plains of India, Uttar Pradesh and Uttarakhand. Significant differences were found among the landraces and eight USA Reticulatus Group reference cultivars for 18 plant and fruit traits: no. of primary branches per plant, days to marketable maturity, sex expression, fruit shape, flesh colour, netting, no. of fruit per plant, fruit weight, shelf life, total soluble solids (°Bx), ascorbic acid (mg/100 g), titratable acidity (%), fruit length and diameter, seed cavity length and diameter, flesh thickness, and resistance to Cucumber mosaic virus. The three melon groups differed significantly for 10 of the plant and fruit traits. Cantalupensis Group and Reticulatus Group accessions were andromonoecious, and the Momordica Group was monoecious. Neighbour-joining (NJ) tree and factorial correspondence analysis (FCA) of simple sequence repeat loci also revealed a high level of genetic variability in this germplasm. The 96 melon genotypes clustered into five groups in the NJ tree analysis: the 16 Indian Reticulatus Group accessions and eight USA reference cultivars formed a distinct group; and the 60 Cantalupensis Group accessions clustered in four other groups with the 12 Momordica Group accessions in a distinct subgroup of one of the Cantalupensis groups. The FCA plot largely confirmed the NJ tree with three distinct groups, one for each melon group. The close affinity of the Indian and USA Reticulatus melons was not unexpected, but it is not clear whether it was inherent in the group and maintained as Reticulatus melons moved from India through Central Asia and Europe to North America, or the result of recent intercrossing of Indian landraces with the USA-derived cultivars and selection for a broad range of Reticulatus type melons.     

Characterization and identification of leaf morphology of <Emphasis Type="Italic">Populus deltoides</Emphasis> Bartr. clones

Leaves are of fundamental importance to plants, representing their facility to generate power and are the sensing units of plants towards the environment. An attempt was made to characterize and compare the variations of leaf morphology of various Populus deltoides Bartr. clones by studying the winter buds and other leaf parameters of fully developed leaves. To achieve these objectives, forty-three exotic and indigenous clones of P. deltoides Bartr. were evaluated for different parameters. On the basis of various morphological characteristics the results reveal that each clone has a distinct color pattern of leaves. Different colors observed in these clones varied from light green through green to dark green. Two distinct lengths of the leaf apex were found, i.e., short and long; as well both acuminate and acute apex types were found. Erratic distribution of serration of leaves was also found. In this study, the morphological traits of leaves provided discriminatory grounds for separating various populations of P. deltoides Bartr. clones. Winter bud studies indicate that different clones vary considerably with regard to shape, color, shape of leaf scars and exudation.     

Potassium isotherm partitioning based on modified quantity‐intensity relation and potassium buffering characterization of soils of North India

Background : Potassium (K) availability in soil and plant uptake is restrained by the dynamic interactions among the different pools of K. Aims : To understand these interactions, a study was undertaken to assess the quantity–intensity (Q/I) and buffering characteristics of rainfed maize (Zea mays L.) growing soils. Ten contrasting soils were evaluated for K partitioning changes in exchangeable K (ΔEK) and non‐exchangeable K (ΔNEK) pools in the soil‐solution phase and buffering characteristics using a modified version of Q/I approach. Results : The partitioned Q/I isotherms showed strong adsorption with the increase in K concentration ratio (CR^K) and the changes due to ΔEK were higher than changes due to ΔNEK. Total buffering capacity (PBC^K) significantly correlated (r = 0.92, p <0.01) with clay content with a major share contributed by buffering capacity owing to non‐exchangeable K ( ${\mathrm{PBC}}_{\Delta \mathrm{NEK}}^K$) rather than exchangeable K ( ${\mathrm{PBC}}_{\Delta \mathrm{EK}}^K$). The fixation capacity (β) factor, the magnitude of added K converted into a non‐exchangeable pool, ranged from 41 to 63%, whereas release (α) factor, the magnitude of added K converted to the exchangeable pool, ranged from 19 to 36%. Both threshold solution K (CK_r) and threshold exchangeable K (EK_r) values were found to be high in Satran clay loam (S2) and lower in Doon silty clay loam (S3) soils. The equilibrium exchangeable K (EK_o) was found close to minimum exchangeable K (E_min) in Doon silty clay loam (S3) and Babaweyl sandy clay loam (S1) soils and overall E_min constituted about 8.94 to 0.57% of the EK_o. Conclusion : It may be concluded that K Q/I isotherm partitioning provides a valuable insight to assess the dynamic relations. The ratio of α/β (K recharge index) could be used to evaluate the K enrichment capacity of soil to K additions while EK_r and E_min can be potentially useful in the elucidation of exchangeable K as K fertility index especially in soils with poor K fertilizer management.