Genetic diversity of bitter gourd (Momordica charantia L.) genotypes revealed by RAPD markers and agronomic traits

Bitter gourd or bitter melon (Momordica charantia L.) is considered as minor cucurbitaceous vegetable in spite of having considerable nutritional and medicinal properties. Although some reports on genetic diversity based on morphological characterization are available, no work has been conducted to estimate genetic diversity using molecular markers in this crop. In the present study, 38 genotypes of M. charantia including few commercially cultivars collected from different parts of India based on agro-ecological zones were analysed for diversity study both at morphological and molecular levels. Genomic DNA was extracted from young healthy leaves following the procedure of Doyle and Doyle [Doyle, J.J., Doyle, J.L., 1990. A rapid DNA isolation procedure from small quantity of fresh leaf material. Phytochem. Bull. 119, 11–15]. Pair-wise comparison of genotypes was calculated as per the procedure of Jaccard [Jaccard, P., 1908. Nouvelles recherches sur la distribution florale. Bull. Soc. Vaud. Sci. Nat. 44, 223–270]. Dendrogram was constructed using the unweighted pair group method with arithmetic averages (UPGMA) and the computation for multivariate analysis was done using the computer programme NTSYS-pc Version 2.0 [Rohlf, F.J., 1998. NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, Version 2.01. Exeter Software, Setauket, NY, USA]. Diversity based on yield related traits and molecular analysis was not in consonance with ecological distribution. Among 116 random decamer primers screened 29 were polymorphic and informative enough to analyse these genotypes. A total of 208 markers generated of which 76 (36.50%) were polymorphic and the number of bands per primer was 7.17 out of them 2.62 were polymorphic. Pair-wise genetic distance (GD) based on molecular analysis ranged from 0.07 to 0.50 suggesting a wide genetic base for the genotypes. The clustering pattern based on yield related traits and molecular variation was different.     

Effect of fasting duration on clinical pathology results in Wistar rats

Background: Fasting is an important preanalytical factor that may affect the interpretation of hematology and clinical biochemistry data in toxicology or pharmacology studies. Limited information is available on how the results may be affected by different durations of fasting. Objective: The purpose of this study was to assess the influence of fasting duration on clinical pathology results in male and female rats and to determine an optimum fasting time for preclinical studies. Methods: Male and female Wistar rats (10 each per group) were fasted for 0, 4, 8, 16, 24, and 48 hours. Changes in body weight and in the results of routine CBC and clinical chemistry analysis were evaluated by 1‐way ANOVA. Results: Body weight was significantly decreased by 4 hours of fasting in all rats, and hemoglobin concentration was significantly increased at 16 hours in male rats. Serum glucose and triglyceride concentrations in both sexes and cholesterol and high‐density lipoprotein‐C concentrations in female rats were also significantly decreased beginning at 16 hours. The creatinine concentration was increased in females after 16 hours of fasting. Serum alkaline phosphatase and alanine aminotransferase activities were significantly decreased after 8 hours in males and 16 hours in females. Conclusions: Fasting‐induced changes in clinical pathology results were consistent with hemoconcentration and altered nutrition and metabolic function. Most changes occurred at 16 hours, with minimal subsequent changes. Hence, a 16‐hour fasting duration may be recommended for preclinical studies involving clinical pathology measurements.     

Genetic analysis and pyramiding of two gall midge resistance genes (Gm-2 and Gm-6t) in rice ( Oryza sativa L.)

Asian rice gall midge (Orseolia oryzae) is a major pest across much of south and southeast Asia. This pest is genetically diverse and many gall midge biotypes are known to exist in each country. During the last three decades, host plant resistance has proved to be the most effective mechanism of controlling the Asian rice gall midge. Seven genes conditioning resistance to gall midge larvae have been identified in rice (Oryza sativa) and are being used in cultivar improvement programs. However, some of these genes are rendered ineffective by new gall midge biotypes. Increased understanding of genetics, inheritance, allelic relationships and linkage is necessary to maximise the durability of major gene resistance by the pyramiding of these genes. The two genes, Gm-2 and Gm-6(t), are known to confer resistance against a number of biotypes in India and China, respectively. An F₃ population derived from a cross between Duokang #1 (donor of Gm-6(t)) and Phalguna (donor of Gm-2) was screened against Chinese gall midge biotype 4 at Guangdong, China, and Indian gall midge biotype 1 at Raipur, India. At each location, separately,a single gene governed resistance. The parallel segregation of 417 F₃progenies for both biotypes at two locations revealed that recombination had occurred between the two genes, establishing that the two genes are not allelic. However, the two genes Gm-2 and Gm-6(t), were found to be linked with a distance of ∼16.3 cM. A number of lines homozygous at one locus and segregating for the other locus were identified and selected. These lines were selfed to obtain lines homozygous for the favourable alleles at both loci (two locus pyramids). This is the first report on use of conventional host-pest interaction method for pyramiding two closely located Gm-resistance loci of dissimilar effects. The implications of deployment of these pyramids within and across country borders, with reference to the prevailing gall midge populations are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification of a diverse mini‐core panel of Indian rice germplasm based on genotyping using microsatellite markers

Identification of a small core germplasm set representing the available genetic diversity is essential for its proper evaluation and subsequent utilization in rice improvement programmes. For constituting a small diverse mini‐core panel of Indian rice germplasm, a representative set of 6912 accessions drawn based on their geographic origin from the whole rice germplasm collection available in the National Gene Bank was genotyped using 36 microsatellite markers. Automated fragment analysis of amplicons yielded a total of 435 alleles, with an average 12.4 and range of 3–29 alleles per locus. Polymorphism information content (PIC) ranged from 0.08 (RGNMS190) to 0.86 (RM552) with an average of 0.528. Based on genotyping data, a mini‐core consisting of 98 genotypes was identified. Ninety‐four per cent of the alleles present in the core set were present in the mini‐core. The identified small but diverse panel will be useful for further intensive trait‐specific evaluation and utilization in allele mining.     

Permeability determination and pharmacokinetic study of nobiletin in rat plasma and brain by validated high-performance liquid chromatography method

In the present study, we are reporting permeability and pharmacokinetics of nobiletin in rat plasma and brain, using a validated reverse phase high performance liquid chromatographic method. Protein precipitation method was used for the extraction of nobiletin and coumarin (IS) from rat plasma and brain tissue. The system was run in isocratic mode with mobile phase consisting of potassium dihydrogen ortho-phosphate (pH 4.5; 0.04 mM) and acetonitrile in ratio of 50:50, v/v. The total chromatographic run time was 9.0 min. The method was proved to be accurate and precise at linearity range of 0.05–10 μg/mL with a correlation coefficient (r) of ≥ 0.994 in rat plasma and ≥ 0.995 in rat brain. The intra- and inter-day precision and accuracy values are found to be within the assay variability limits as per the FDA guidelines. Nobiletin was found stable in the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and long term storage in a freezer at − 70 ± 10 °C. Maximum concentrations of nobiletin in both plasma and brain were observed at 1 h after single oral dosing (50 mg/kg). The maximum concentration in plasma and brain were 1.78 and 4.20 μg/mL, respectively. The AUC_0–t in plasma and brain were 7.49 and 20.66 μg·h/mL, respectively. The mean elimination half life (t_1/2) in plasma and brain were 1.80 and 11.42 h, respectively. The Parallel Artificial Membrane Permeability Assay (PAMPA) permeability of nobiletin was found to be high at both pH 4.0 and 7.0.     

A bifurcating pathway directs abscisic acid effects on stomatal closure and opening in Arabidopsis

Terrestrial plants lose water primarily through stomata, pores on the leaves. The hormone abscisic acid (ABA) decreases water loss by regulating opening and closing of stomata. Here, we show that phospholipase Dalpha1 (PLDalpha1) mediates the ABA effects on stomata through interaction with a protein phosphatase 2C (PP2C) and a heterotrimeric GTP-binding protein (G protein) in Arabidopsis. PLDalpha1-produced phosphatidic acid (PA) binds to the ABI1 PP2C to signal ABA-promoted stomatal closure, whereas PLDalpha1 and PA interact with the Galpha subunit of heterotrimeric G protein to mediate ABA inhibition of stomatal opening. The results reveal a bifurcating signaling pathway that regulates plant water loss.     

Effect of Indigenous Plant Growth-Promoting Rhizobacteria on Wheat (Triticum Aestivum L.) Productivity and Soil Nutrients

The present study aimed at selection of efficient bacterial isolates with multiple plant growth-promoting (PGP) traits at variable doses of chemical fertilizers for enhanced wheat productivity and sustenance of soil health. Ten bacterial isolates from wheat (rhizosphere soil and root endosphere) were screened for PGP traits (indole acetic acid, phosphate solubilization, siderophore production, and ammonia production). Only three isolates (B2, SIR1, and BIS2) possessed all PGP traits. Net house evaluation of these isolates at graded doses of chemical fertilizers revealed that the potential of B2 isolate is significantly superior for enhancing wheat yield and soil properties. On the basis of 16S rDNA analysis, the potential isolate (B2) was identified as Serratia marcescens. Conjoint use of the B2 isolate at 80% recommended doses of fertilizers (RDF) significantly increased wheat growth and saved 18 kg nitrogen and 10 kg phosphorous on per hectare basis. The developed module not only increases profitability but also protects the environment and sustains soil health.     

Suppression of Reproduction in Poekilocerus pictus (F.) using Penfluron and Diflubenzuron

Abstract

Penfluron and diflubenzuron, both chitin synthesis inhibitors known to be highly effective against nymphs of Poekilocerus pictus (F.), were tested against the adult for their possible effects upon reproduction. These compounds were found to have the maximum effect, when no eggs were laid, if both sexes were treated. The eggs of treated males × untreated females did not hatch and those of treated females × untreated males gave rise to nymphs which died before reaching maturity.     

Resistance to blast (Magnaporthe grisea) in a mini-core collection of finger millet germplasm

Blast caused by Pyricularia grisea [teleomorph: Magnaporthe grisea] is an economically important and widespread disease of finger millet in the world. Host resistance is the most economical and effective means of combating this disease as finger millet is predominantly grown by resource-poor and marginal farmers. At the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), we evaluated a finger millet mini-core collection of 80 germplasm accessions (about 1 % of the total germplasm collection representing major trait variability) for blast resistance both in the field and greenhouse. Field evaluation was done using a refined screening technique that included new improved rating scales for leaf, neck and finger infection. Sixty six of the 80 accessions showed combined resistance to leaf, neck and finger blast in two seasons (2009 and 2010) of field screening. A highly significant and positive correlation was found between neck and finger blast ratings (r?=?0.92), whereas small but significant correlations were found between leaf blast and neck blast (r?=?0.25) and between leaf blast and finger blast (r?=?0.30). These accessions were also screened for leaf blast resistance in the greenhouse by artificial inoculation of seedlings to confirm field observations. Fifty-eight of the 80 accessions were resistant to leaf blast in the greenhouse screen as well. These resistant accessions represented one wild (africana) and four cultivated races (vulgaris, plana, elongate and compacta) of finger millet that originated from 13 countries in Asia and Africa and exhibited considerable diversity for agronomic traits, such as maturity period, plant height and panicle type. These blast resistant accessions from the mini-core collection would be useful in finger millet disease resistance breeding programs.     

Balanced and integrated nutrient management for enhancedand economic food production: case study from rainfed semi-arid tropics in India

Soil degradation in the semi-arid tropics (SAT) is mainly responsible for low crop and water productivity. In Madhya Pradesh and Rajasthan states in India, the soil analyses of farmers’ fields revealed widespread deficiencies of S (9–96%), B (17–100%) and Zn (22–97%) along with that of P (25–92%). Soil organic C was deficient in 7–84% fields indicating specifically N deficiencies and poor soil health in general. During on-farm evaluations in rainy seasons 2010 and 2011, the soil test based addition of deficient nutrient fertilizers as balanced nutrition (BN) increased crop yields by 6–40% (benefit to cost ratios of 0.81–4.28) through enhanced rainwater use efficiency. The integrated nutrient management (INM), however, decreased the use of chemical fertilizers in BN by up to 50% through on-farm produced vermicompost and recorded yields at par or more than BN with far better benefit to cost ratios (2.26–10.2). Soybean grain S and Zn contents improved with INM. Applied S, B, Zn and vermicompost showed residual benefits as increased crop yields for succeeding three seasons. Hence, results showed INM/BN was economically beneficial for producing more food, while leading to resilience building of SAT production systems.