Associated bacterial diversity of insecticide-susceptible and -resistant brown planthopper, <Emphasis Type="Italic">Nilaparvata lugens</Emphasis> (Homoptera: Delphacidae) analyzed by culture-dependent and -independent methods

Nilaparvata lugens (Stål) is one of the major pests of rice throughout tropical and temperate Asia. Indiscriminate use of insecticides for suppressing N. lugens has resulted in the development of resistance to multiple insecticide classes, causing frequent control failures in the field. Analysis of gut bacterial diversity within an insect host is the initial step towards understanding the ecological roles of the symbionts. Present study aimed to survey the bacterial diversity associated with laboratory-reared (insecticide-susceptible) and field-collected (insecticide-resistant) populations of N. lugens by culture-dependent and PCR-Denaturing Gradient Gel Electrophoresis (DGGE) methods. Seventeen bacterial isolates were obtained by the culture-dependent method. Molecular characterization using the 16S rRNA gene and phylogenetic analysis revealed that the isolates belonged to Firmicutes and Proteobacteria. Taxonomic assignment placed these isolates into seven families representing 10 genera. Enterobacteriaceae was the most dominant family with its occurrence in four out of the five populations studied. The DGGE profiles indicated a low complex gut bacteria associated with N. lugens with limited number of bands. The Shannon-Wiener index ranged from 0.898 in insecticide-susceptible population to 0.946–1.035 in resistant populations. Sequencing and phylogenetic analysis revealed that the DGGE bands belonged to Firmicutes, Proteobacteria and Bacteriodetes. Results of this study illustrated that gut bacterial community associated with N. lugens is dominated by Proteobacteria and Firmicutes. Present findings could provide the basis for future work on the possible role of the bacterial symbionts in insecticide resistance and to formulate potential resistance management strategies.     

Identification of mungbean lines with tolerance or resistance to yellow mosaic in fields in India where different begomovirus species and different <Emphasis Type="Italic">Bemisia tabaci</Emphasis> cryptic species predominate

The segmentation of symptoms during image analysis of diseased plant leaves is an essential process for detection and classification of diseases. However, there are challenges involved in the task, many of them related to the variability of image and host/symptom characteristics and conditions. As a result of those challenges, the methods proposed in the literature so far focus on a specific problem and are usually bounded by tight constraints regarding image capture conditions. This research explores a new automatic method for segmenting disease symptoms on plant leaves that was designed to be applicable in a wide range of situations. The proposed technique employs only color channel manipulations and Boolean operations applied on binary masks, thus being simpler and more robust compared to many previously described automatic methods. Its effectiveness is demonstrated by tests performed over a large database containing images of 77 different diseases of 11 plant species. A comparison with manual segmentation is also presented, further reinforcing the advantages of the proposed approach.     

Biocontrol potential of Alternaria sp. PMK2, against a devastating weed: Parthenium hysterophorus L

Parthenium hysterophorus (Asteraceae) is known as one of the most aggressive invasive weeds, causing severe economic, environmental, human and animal health problems in India and around the world. During a series of extensive surveys for natural enemies of P. hysterophorus, a leaf spot pathogen was isolated from the affected parts of the parthenium following the standard isolation techniques using potato dextrose agar (PDA) and parthenium dextrose agar (PeDA) media. Koch's postulates were performed and found satisfactory for the isolate and proved to be pathogenic to this weed. On the basis of cultural, morphological and molecular characteristics, the pathogen was identified as Alternaria sp. PMK2. The growth of the pathogen was studied on eight selected media and it exhibited varying degrees of growth on different media. Phytotoxicity of fungal cultural filtrates was also confirmed on parthenium leaves in laboratory conditions. Due to the virulent nature of the isolated pathogen, it may be selected for further studies to develop mycoherbicide for control of this devastating weed.     

SRG1, Encoding a Kinesin-4 Protein,Is an Important Factor for Determining Grain Shape in Rice

Grain shape is one of the important agronomic traits, which is closely related to the yield of rice. A new rice mutant, named small and round grain (srg1), was isolated from an indica cultivar Zhenong 34 by ethyl methane sulfonate (EMS) mutagenesis. The microscopic analysis showed that the cell length of spikelet in srg1 was decreased compared with that in wild type (WT), which caused the grain length short. Meanwhile, the grains of srg1 were wider than those of WT because of the increased cell layers in spikelet in the lateral direction. Therefore, the inhibition of cell expansion and increased cell proliferation collectively led to the small and round grain. By map-based cloning, the gene SRG1 was located on the short arm of chromosome 9, which encodes a kinesin-4 protein, represented by the gene LOC_Os09g02650. A single nucleotide polymorphism, occurred in the 16th exon of SRG1, led to premature translation stop in mutant. The cell cycle-related genes were up-regulated in srg1, which conferred that SRG1 controlled grain width through the cell proliferation. Since the role of SRG1 in regulating grain shape was not clarified well before, it is valuable to explore the mechanism of grain development. This study could, hence, provide a morphogenesis and molecular basis for elucidating the function of SRG1, as well as a new germplasm resource for the further study of grain development.     

Induction of DNA damage in the peripheral blood of zebrafish (Danio rerio) by an agricultural organophosphate pesticide,monocrotophos

The use of organophosphate pesticides (OPs) in agricultural practices in India has gained popularity due to their broad spectrum, low cost and high potency. Monocrotophos (an OP) which is a systemic and contact broad-spectrum cholinesterase inhibitor is being widely used in India. However, its use entails the damage to other non-target organisms such as fish. In the present study, zebrafish (Danio rerio) were exposed to various doses of monocrotophos (0.125, 0.625 and 1.25 μL/L) and genotoxicity studies were carried out employing the comet assay and micronucleus test in the peripheral blood of these fish after 24, 48 and 72 h of exposure. Significant DNA damage in the form of % tail DNA and micronuclei was observed in all the treated fish compared to the control. Both % tail DNA and micronuclei frequency were found to increase significantly as the concentration increased. DNA damage was also observed at all the time intervals for all treatments except in the 0.125 μL/L-treated group. A positive correlation was also observed between the comet assay and micronucleus test. The study thus suggests the use of zebrafish as an experimental model to study the genotoxic effects of agricultural pesticides using the comet assay and micronucleus test.

     

Land-use effects on phosphorus fractions in Indo-Gangetic alluvial soils

Phosphorus (P) in soil exists both in organic and inorganic forms and their relative abundance could determine P supplying capacity of soil. Differential input of exogenous and plant-mediated phosphorus and carbon in soil under different land-uses could influence P availability and fertilizer P management. While the effect of land-use on soil organic carbon (SOC) is fairly well-documented, its effect on soil P fractions is relatively less known. We investigated the effect of different land-uses including rice–wheat, maize–wheat, cotton–wheat cropping systems and poplar-based agroforestry systems on soil P fractions and organic carbon accrual in soils. Total P concentration was the highest under agroforestry (569 mg P kg^?1) and the lowest under maize–wheat (449 mg P kg^?1) cropping systems. On the contrary, soils under rice–wheat had significantly higher available P concentration than the agroforestry systems, probably because of higher fertilizer P application in rice–wheat and prevailing wetland conditions during rice growth. In soils under sole cropping systems viz. rice–wheat, maize–wheat and cotton–wheat, inorganic P was the dominant fraction and accounted for 92.2–94.6% of total P. However, the soils under agroforestry had smaller proportion (73%) of total P existing as inorganic P. Among soil P fractions, water soluble inorganic P (0.13–0.26%) represented the smallest proportion inorganic P in soils under different land-uses. Agroforestry showed significantly (p < 0.05) higher concentrations of SOC than the other land-uses. Soil organic C was significantly correlated with soil P fractions. It was concluded that poplar-based agroforestry systems besides leading to C accrual in soil result in build-up of organic P and the P supplying capacity of soil.     

Development of a <Emphasis Type="Italic">Cajanus cajanifolius</Emphasis>-based CMS Hybrid Technology in Pigeonpea

Pigeonpea is an important food legume of the semi-arid tropics grown mainly under subsistence agriculture. As a first step to address the five decades-long yield stagnation, a proof-of-concept for a commercially viable CMS-based hybrid technology with high standard heterosis and fertility restoration has been demonstrated. Six generations of backcrossing and selection for male sterility, and agronomic superiority using a Cajanus cajanifolius- (A₄ cytoplasm) based CMS donor (ICPA 2039) and a medium-maturity group recurrent parent ICPL 20176 (ICPB 2043) which resulted in a stable male sterile line (ICPA 2043) with near-perfect male sterility and superior agronomic traits. Following backcrossing and selections, the A-line (ICPA 2043) appeared to be morphologically similar in terms of various qualitative and quantitative traits compared to the B-line (ICPB 2043). However, differences between ICPA 2043 and ICPB 2043 were observed for days to maturity, 100-seed weight, pods plant^-1, and seed weight plant^-1. These differences may not be genetic but physiological. The best heterotic restorer line selected in a cross (ICPH 2671) combination restored a mean of 96.49% male fertility in 36 environments (12 locations, 3 years). Present investigation did not reveal significant G×E interaction for fertility restoration, indicating the possibility of obtaining high and stable grain yields in the pigeonpea growing areas of South and Central India.     

Vacuum press drying studies on two fast-growing Indian wood species

Two fast-growing Indian species, Melia composita Benth. and Eucalyptus tereticornis Sm., which have different sets of physical properties, were dried together in a vacuum press dryer(VPD) under two drying conditions,i.e., above boiling point(ABP) and below boiling point(BBP). The ABP and BBP conditions were maintained by keeping the temperature constant at 75 ℃ and maintaining two pressure levels: 300 mm of Hg(ABP) and 450 mm of Hg(BBP). In order to understand pressure conditions at the core during vacuum drying, a cylindrical brass pipe was inserted in both wood cores and attached with pressure gauges placed outside of the VPD. The results indicate that the Melia wood core attained equilibrium pressure immediately with the pressure of VPD, while Eucalyptus attained it very slowly, reaching equilibrium at later stages of drying when cracks and checks advanced to the core.The drying rate was higher for Melia than Eucalyptus under both drying conditions. The drying rate of Melia(ABP) was higher than Melia(BBP), however, the drying rate for Eucalyptus(ABP) was not significantly different from the BBP drying rate.     

Variability in physical properties of plantation-grown progenies of Melia composita and determination of a kiln-drying schedule

Plantation-grown progenies of Melia composita Willd.were studied for variability in several physical properties: density,radial and tangential shrinkage,longitudinal permeability of heartwood and sapwood.Furthermore,flat-sawn planks from each of the progeny were subjected to a quick-drying test for determination of kilndrying schedule.The mean density of the species was 0.39 g cm~(-3) and the wood may be categorized as light wood.Mean radial shrinkage(%) for the species was 2.8% with progeny-wise variation in radial shrinkage from 1.56 to 4.11%.Mean tangential shrinkage(%) for the species was 5.54% with progeny-wise variation in tangential shrinkage from 3.69 to 7.71%.The resultant tangentialradial shrinkage ratio was 1.98(less than two),which suggests that the wood is relatively stable with respect to drying behavior.Mean sapwood and heartwood longitudinal permeability of the species were 3.38 and 2.02 Darcy,respectively.Higher longitudinal permeability of sapwood and heartwood indicate better drying and preservative properties of the species.Terazawa quick-drying test method suggests that the species is less susceptible to drying defects.During the test,only moderate checks and cracks were observed.A tentative kiln-drying schedule was recommended based on these results.     

Genetics of Fe,Zn, β-carotene,GPC and yield traits in bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) using multi-locus and multi-traits GWAS

The present study was conducted to study the genetic architecture of grain micronutrients (Zn, Fe and β-carotene contents), grain protein content and four yield traits in a spring wheat reference set comprising 246 genotypes. Phenotypic data on these traits recorded at two locations and the genotyping data for 17,937 SNP markers (obtained through outsourcing) were used for genome wide association study, which gave following results after Bonferroni correction using four methods: (1) single locus single trait analysis gave 136 marker-trait associations; (2) multi-locus mixed model gave 587 MTAs; (3) multi-trait mixed model gave 28 MTAs and (4) matrix-variate linear mixed model gave 33 MTAs. As many as 73 epistatic interactions were also detected. Keeping all the results in mind, nine most important MTAs were selected for biofortification. These markers were associated with three traits (GPC, GFeC and GYPP). These MTAs can be used in wheat improvement programs either using marker-assisted recurrent selection or pseudo-backcrossing method.